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YALE UNIVERSITY SCHOOL OF MEDICINE

Vascular Biology and Therapeutics Program

Annual Report 2011-2012

VASCULAR BIOLOGY AND THERAPEUTICS ANNUAL REPORT 2011– 2012

TABLE OF CONTENTS

Message from the Director .................................................................................................... 1 Program Operations ............................................................................................................... 1

VBT Steering Committee ................................................................................................ 1 Administrative Operations .............................................................................................. 1 Program Faculty Membership ........................................................................................ 2

Program Activities .................................................................................................................. 3

Seminar Series ................................................................................................................. 3 Retreat .............................................................................................................................. 4

Yale-Cambridge Program in Cardiovascular Disease ......................................................... 4 Tissue Engineering Group ..................................................................................................... 4 VBT Research in Progress (RIP) Talks ................................................................................. 4 Research Accomplishments .................................................................................................. 5 Appendices

The Eleventh Annual VBT Retreat............................................................................... 1-1 The Tenth Annual Meeting of the Joint Yale-Cambridge Program in ...................... 2-1

Cardiovascular Research

On the Cover: Aortic cross section from a lineage tracing mouse expressing tomato red fluorescing protein in non-smooth muscle tissues and green fluorescent protein in smooth muscle cells. Green autofluorescence is also detected in adventitial adipocytes. Light blue (cyan) immunostaining reveals that both adventitial adipocytes and medial vascular smooth muscle cells express the cardioprotective hormone adiponectin. Dark blue staining marks cell nuclei. Orange autofluorescence indicates elastin fibers of the vessel media. Image credit: Catarina Carrao, Ph.D. and Kathleen Martin, Ph.D.

Vascular biology & therapeutics


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Vascular Biology and Therapeutics Program (VBT)

Annual Report 2011 – 2012 Message from the Director The past academic year has been filled with exciting scientific developments with many VBT faculty publishing in top notch journals in 2012. Clearly this is a testimony to the excellence and high quality of work here at Yale within the VBT program. VBT membership is growing and now includes new members: Barbara Ehrlich, Karen Hirschi, Yasuko Iwakiri, Jaime Grutzendler, Stefania Nicoli, and Jun Yu. In addition, Martin Schwartz and Themis Kyriakides have agreed to serve as members of our steering committee. The interactions of VBT with the Cardiovascular Research Center (CVRC) growing with continued collaborations. One example is the new PPG led by Michael Simons, with Bill Sessa, Themis Kyriakides, Laura Niklason, Martin Schwartz and Al Sinusas as key contributors to study the molecular mechanisms of arteriogenesis. Finally, I would like to applaud Themis Kyriakides and Kerry Russell who have done a wonderful job coordinating our joint seminar series and we appreciate their efforts. PROGRAM OPERATIONS VBT Steering Committee The Steering Committee serves as the principal advisory and leadership group for the program for the program. The current membership of the Steering Committee is listed in Table 1. Administrative Operations Ms. Carol Muzzey serves as the Program Manager and is assisted by Ms. Diane Strumpf. The program is served by the Central Administration Business Office.

Table I. VBT Steering Committee Jeffrey R. Bender, M.D., Professor of Internal Medicine (Cardiovascular Medicine) and Immunobiology Alfred L.M. Bothwell, Ph.D., Professor of Immunobiology Anne Eichmann, Ph.D., Professor Internal Medicine - Cardiology Jack A. Elias, M.D., Waldmar von Zedwitz Professor of Medicine, Professor of Immunobiology, Chairman of Internal Medicine Frank Giordano, M.D., Associate Professor of Internal Medicine (Cardiology) Themis Kyriakides, Ph.D. Associate Professor of Pathology & Biomedical Engineering Joseph A. Madri, M.D., Ph.D., Professor of Pathology and Molecular, Cellular and Developmental Biology Laura Niklason, M.D., Ph.D., Professor and Vice Chair of Anesthesia and Professor Biomedical Engineering Jordan S. Pober, M.D., Ph.D., Vice Chair, Immunobiology for Section of Human and Translational Immunology, Professor of Pathology, Immunobiology and Dermatology Nancy H. Ruddle, Ph.D., John Rodman Paul Professor, Epidemiology and Public Health, Professor of Immunobiology W. Mark Saltzman, Ph.D., Professor of Chemical and Biomedical Engineering, Chair of Biomedical Engineering William C. Sessa, Ph.D., Director Vascular Biology & Therapeutics, Professor and Vice Chair of Pharmacology Martin Schwartz, Ph.D., Professor Internal Medicine - Cardiology Michael Simons, M.D., RW Berliner Professor of Medicine & Cell Biology, Chief Section of Cardiovascular Medicine

George Tellides, M.D., Ph.D., Professor of Surgery (Cardiothoracic) and Chief, Cardiothoracic Surgery

VASCULAR BIOLOGY AND THERAPEUTICS ANNUAL REPORT 2011 – 2012

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Program Faculty Membership All faculties at Yale with a significant interest in vascular biology and/or therapeutics are eligible to join VBT. VBT members in academic year 2011-2012 are:

Table 2 VBT Membership Jeffrey R. Bender, M.D., Professor of Internal Medicine (Cardiovascular Medicine) and Immunobiology Anton Bennett, Ph.D., Associate Professor of PharmacologyAlfred L.M. Bothwell, Ph.D., Professor of Immunobiology Christopher Breuer, M.D., Associate Professor of Surgery (Pediatrics) David Calderwood, Ph.D., Associate Professor of Pharmacology Hyung J Chun, M.D., Assistant Professor of Medicine, Cardiovascular Medicine Alan Dardik, Ph.D., M.D., Associate Professor of Surgery (Vascular Surgery) Barbara E. Ehrlich, Ph.D., Professor, Departments of Pharmacology & Cellular & Molecular Physiology Anne Eichmann, Ph.D., Professor, Internal Medicine – Cardiology Jack A. Elias, M.D., Waldmar von Zedwitz Professor of Medicine, Professor of Immunobiology, Chairman of Internal Medicine Tarek Fahmy, Ph.D., Associate Professor of Biomedical Engineering and Chemical Engineering Richard Flavell, Ph.D., FRS, Sterling Professor and Chairman of Immunobiology, Investigator of Howard Hughes Medical Institute Arnar Geirsson, M.D., Assistant Professor of Surgery Frank J. Giordano, M.D., Associate Professor Internal Medicine (Cardiovascular Medicine) Daniel R. Goldstein, M.D., Associate Professor Internal Medicine (Cardiovascular Medicine) Daniel Greif, M.D., Assistant Professor, Internal Medicine - Cardiology Murat Gunel, M.D., Associate Professor of Neurosurgery Karen Hirschi, Ph.D., Professor Internal Medicine, Cardiology Jay Humphrey, Ph.D., Professor, Biomedical Engineering John Hwa, M.D., Ph.D., Associate Professor of Medicine, Cardiovascular Medicine Yasuko Iwakiri, Ph.D., Assistant Professor Internal Medicine – Digestive Disease Suk-Won Jin, Ph.D., Assistant Professor Internal Medicine - Cardiology Martin Kluger, Ph.D., Research Scientist, Immunobiology Diane Krause, M.D., Ph.D., Professor of Laboratory Medicine Sanjay Kulkarni, M.D., Associate Professor of Surgery (Transplantation & Immunology) Themis Kyriakides, Ph.D., Associate Professor of Pathology and Biomedical Engineering Jaime Grutzendler, M.D., Associate Professor of Neurology Patty J. Lee, M.D., Associate Professor of Internal Medicine, Pulmonary & Critical Care Joseph A. Madri, M.D., Ph.D., Professor of Pathology and Molecular, Cellular and Developmental Biology Kathleen A. Martin, Ph.D., Associate Professor of Medicine (Cardiovascular Medicine) and Pharmacology Laura R. Ment, M.D., Professor of Pediatrics (Neurology) Wang Min, Ph.D., Associate Professor with Tenure, Pathology Stefania Nicoli, Ph.D., Assistant Professor, Internal Medicine, Cardiovascular Medicine Laura Niklason, M.D., Ph.D., Professor & Vice Chair of Anesthesia; Professor Biomedical Engineering Jordan S. Pober, M.D., Ph.D., Vice Chair, Section of Human and Translational Immunology, Department of Immunobiology, Professor of Immunobiology, Pathology and Dermatology Nancy H. Ruddle, Ph.D., John Rodman Paul Professor Emerita & Senior Research Scientist, Departments of Epidemiology and Public Health and Immunobiology Kerry S. Russell, M.D., Ph.D., Associate Professor of Medicine, Cardiovascular Medicine Mehran M. Sadeghi, M.D., Associate Research Scientist of Internal Medicine (Cardiovascular Medicine) W. Mark Saltzman, Ph.D., Goizueta Foundation Professor of Chemical and Biomedical Engineering Martin Schwartz, Ph.D., Professor Internal Medicine - Cardiology William C. Sessa, Ph.D., Director Vascular Biology & Therapeutics and Professor and Vice Chair of Pharmacology Michael Simons, M.D., RW Berliner Professor of Medicine & Cell Biology, Chief Section of Cardiovascular Medicine Albert J. Sinusas, M.D., F.A.C.C., Professor of Internal Medicine (Cardiovascular Medicine) & Diagnostic Radiology Jeffrey Sklar, M.D., Ph.D., Professor of Pathology and Laboratory Medicine Edward Snyder, M.D., Professor Laboratory Medicine, Director, Apheresis/Cell Processing VBT Core Facility Bing Su, Ph.D., Associate Professor of Immunobiology George Tellides, M.D., Ph.D., Professor of Surgery (Cardiothoracic) Agnes Vignery, DDS, Ph.D., Associate Professor of Orthopaedics and Rehabilitation Dianqing (Dan) Wu, Ph.D., Professor of Pharmacology Jun Yu, M.D., Instructor, Internal Medicine - Cardiology


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Table 3 – VBT 2011-2012 Seminar Series

SEPTEMBER 2011 David T Shima, PhD , Department of Ocular Biology and Therapeutics, Institute of Ophthalmology, University College London, UK – “Cyto-architectural basis of endothelial barrier function: a tale of two extremes” October 2011 Alan Tall, MD,Tilden Weger Bieler Professor of Medicine, Columbia - “ABC transporters and ApoE regulate hematopoietic stem cell proliferation, leukocytosis and atherosclerosis” Christopher V. Carman, PhD, Assistant Professor of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School – “The Dynamic Endothelium: Insight to Barrier Regulation & Immune Functions” Elazer Edelman, M.D., PhD, Thomas D. And Virginia W. Cabot, Professor of Health Sciences & Technology, MIT - “Endothelial Angiocrine Regulation Convergence of Vascular & Cancer Biology” November 2011 Robert P. Mecham, PhD, Alumni Endowed Professor of Cell Biology and Physiology, Professor of Medicine, Pediatrics, and Bioengineering, Washington University School of Medicine – “Vascular extracellular matrix & vessel homeostasis” Salvador Moncada, M.D., Department of Cardiovascular Research, Director, Wolfson Institute for Biomedical Research, University College of London – “Finding the mechanism that coordinates metabolic supply with cell proliferation” UCL SEMINAR Jan Kitajewski, PhD, Professor of Clinical Pathology, Institute of Cancer Genetics and HICC, Columbia University – “Ligand-specific Notch signaling in developmental and tumor angiogenesis” December 2011 Douglas L. Mann, MD, Lewin Professor and Chief, Cardiovascular Division, Washington University School of Medicine, Cardiologist-in-Chief, Barnes Jewish Hospital, St. Louis – “Determinants of Myocardial Remodeling & Reverse Remodeling in the Failing Heart” Jaime Grutzendler, MD, Associate Professor Neurology, Yale University – “Cellular Imaging of the Neurovascular Unit in Health & disease” January 2012 Alvin C. Powers, MD, Joe C. Davis Chair in Biomedical Science, Professor of Medicine, Molecular Physiology and Biophysics, Director, Vanderbilt Diabetes Center, Vanderbilt University School of Medicine – “Islet and Endothelial Cells: Context and Connections” February 2012 Charles Cha, MD, Associate Professor of Surgical Oncology and Gastrointestinal Surgery, Yale University - “Role of intermedin in tumor angiogenesis: bystander effect or therapeutic target” Alpha Yap, PhD, FRACP, Professor, University of Queensland, NHMRC Senior Research Fellow, Head, Division of Molecular Cell Biology, Institute of Molecular Bioscience, University of Queenslandm – “Integration at junctions: cadherin adhesion and the actin cytoskeleton” March 2012 Karen Hirschi, PhD, Professor, Section of Cardiovascular Medicine, Yale University – “Parallel Development of Blood and Blood Vessels” Howard A. Rockman, MD, Professor of Medicine, Professor of Cell Biology, Professor of Molecular Genetics & Microbiology, Department of Medicine, Cardiology, Duke University School of Medicine - “Drosophila Screens to Identify Cardiomyopathy Genes” Colleen Mac Namara, MD, Professor, Cardiovascular Medicine, University of Virginia School of Medicine –“B Lymphocytes and Atherosclerosis” April 2012 Anthony Rosenzweig, MD, Cardiovascular Division Research, Department of Medicine, Beth Israel Deaconess Medical Center – Charting the Fate of the “Good Cholesterol” – Characterization of the HDL receptor SR-BI and its influence on Coronary Heart Disease George Tellides, MD, PhD, Professor of Surgery (Section of Cardiac Surgery) and of Investigative Medicine; Chief of Cardiothoracic Surgery, Veterans Affairs Medical Center – “Protective Role of TGF-beta against Arterial Medial Injury”

PROGRAM ACTIVITIES Seminar Series The VBT Monday afternoon seminars continue to serve as an intellectual focus of the vascular biology community at Yale. The series also serves as venue for assistance in the recruitment of faculty with research in vascular biology to various departments at Yale. The seminars are run by Dr. Themis Kyriakides. A list of seminar speakers and their titles are shown in Table 3.


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Hong Chen, Ph.D., Assistant Member, Cardiovascular Biology Research Program, Adjunct Assistant Professor, Department of Biochemistry and Molecular, Biology, University of Oklahoma Health Sciences Center, Oklahoma Medical Research Foundation - “Epsin-Mediated Endocytosis Regulates VEGF Signaling in Vascular Development and Tumor Angiogenesis” Jeffery D. Molkentin, PhD, Professor | Howard Hughes Medical Institute Investigator, Professor, UC Department of Pediatrics, Cincinnati Children’s Hospital - “TRPC6-dependent pathway for cardiac fibrosis and myofibroblast formation” May 2012 Brian H. Annex, MD, Professor of Medicine and Biomedical Engineering, Division Chief, Cardiovascular Medicine, Division of Cardiovascular Medicine - “Genetic Modifiers of Peripheral Arterial Disease” Raghu Kaluri, MD, Professor of Medicine, Harvard Medical School, Beth Israel Deaconess Medical Center - “Cancer cell autonomous defects must cooperate with stromal alterations to determine the rate of cancer progression and metastasis” June 2012 Monty Krieger, PhD, Professor of Biology, MIT - “Charting the Fate of the ‘Good Cholesterol’ Characterization of the HDL receptor SR-Bl and its influence on Coronary Heart Disease” Jonathan Chernoff, Professor, SVP Chief Scientific Officer Stanley P. Reimann Chair in Oncology Research, Co-Leader, Cancer Biology Co-Leader, Epigenetics and Progenitor Cells Keystone, Fox Chase Cancer Center - “p21 – activated kinases: Sculptor of Hearts and Vessels”

Retreat The annual retreat continues to be an extremely popular activity, bringing together over one hundred sixty-five scientists from the laboratories of VBT faculty members. This past year, the retreat was held on October 22, 2011 at the Grace Murray Hopper Auditorium, West Campus. The retreat continued the poster session competition with prizes for the best posters by a graduate student and by a post-doctoral fellow.

The Keynote Address at this years’ retreat was David Cheresh, Ph.D., Moores University California San Diego Cancer Center. The retreat Program is listed in Appendix 1.

Yale-Cambridge Program in Cardiovascular Disease

The research alliance with Cambridge has continued as an important activity, with 13 faculty from Cambridge visiting Yale in September 2011 for a two day scientific meeting. The program for this retreat is listed in Appendix 2. A visit by Yale members to Cambridge is scheduled for September 2012. Tissue Engineering Group This biweekly forum, sponsored by VBT and organized by Dr. Themis Kyriakides, brings together investigators from Yale Medical School and Yale’s central campus to exchange updates in research in progress and to foster new research collaborations. VBT Research in Progress (RiP) Talks A monthly series organized by Marty Kluger and Jun Yu featuring presentations by graduate students and postdoctoral fellows working in VBT laboratories.


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Anton M. Bennett, Ph.D.

Associate Professor of Pharmacology

1. Overall Goal(s) of the Research Program of the Laboratory: The broad research interests of this laboratory are to define the molecular mechanisms, physiological and pathophysiological roles of the protein tyrosine phosphatase (PTP) family of enzymes. Using mouse genetic approaches we are studying how PTPs are involved in regulating metabolism. This work suggests new modes of control through which PTPs participate in metabolic signaling and potentially the progression of obesity and diabetes. We are also investigating the pathophysiological mechanisms of the PTPN11 gene, which encodes for the PTP, SHP-2. PTPN11/SHP-2 mutations are found in ~50% of Noonan syndrome cases, an autosomal dominant disorder that causes congenital heart disease. SHP-2-associated Noonan syndrome mutations constitute the largest representation of non-chromosomal mutations in congenital heart disease. The broad goal of this project is to identify the signaling mechanisms induced by the mutant form of PTPN11SHP-2 that leads to congenital heart disease. 2. Specific Research Accomplishments in the last 12 months: We have found using phospho-proteomic approaches PTPN11/SHP-2-interacting proteins that are aberrantly tyrosyl phosphorylated in the hearts of knock-in mice containing a PTPN11 mutation. Our data suggest that these proteins represent targets through which PTPN11/SHP-2 signal in order to control cytoskeletal function in the developing heart. 3. Significance of Key Findings Relevant for the Mission of VBT: The identification of SHP-2-interacting proteins that are aberrantly tyrosyl phosphorylated in the hearts of knock-in mice containing a PTPN11 mutation may provide new insight into the mechanisms that cause congenital heart disease and other abnormalities observed in Noonan syndrome. 4. Publications: (Publications July 1, 2011– June 30, 2012) Roth Flach, R.J., Zhang, and Bennett, A.M. (2011) Loss of MAP Kinase Phosphatase-1

protects from hepatic steatosis by repression of CIDEC/Fat-specific protein 27, J. Biol. Chem., 286: 22195-22202. PMID: 21521693.

Diano, S., Liu, Z., Jeong, J. K., Dietrich, M. O., Ruan, H., Kim, E., Suyama, S., Kelly, K., Gyengesi, E., Arbiser, J. L., Belsham, D., Sarruf, D., Schwartz, M., Bennett, A. M., Shanabrough, M., Mobbs, C. V., Yang, X., Gao, X., and Horvath, T. L. (2011) Peroxisome proliferation-related hypothalamic control of ROS sets melanocortin tone and feeding in diet-induced obesity. Nature Medicine, 11: 1121-1129. PMID: 21873987.

Ruan, Hai-Bin, Han, X., Li, Mindian, Signh, J. P., Bennett, A.M., Yates III J. and Yang, X., (2012) O-GlcNAc Transferase/Host Cell Factor C1 Complex Regulates Gluconeogenesis by Modulating PGC-1 Stability. Cell Metabolism, 16: 226-237.

Lawan A, Shi H, Gatzke F and Bennett AM: Diversity and specificity of the mitogen-activated protein kinase phosphatase-1 functions. Cell Mol Life Sci. 2012 Jun 14; Epub 2012 Jun 14. PMID: 22695679


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David A. Calderwood, Ph.D.

Associate Professor, Department of Pharmacology and Cell Biology

1. Overall Goal(s) of the Research Program of the Laboratory: The overall goals of my lab are to understand signaling to and from integrin adhesion receptors and characterize their links to the actin cytoskeleton. Control of cell adhesion, spreading and migration requires the integrin cytoplasmic tails. These short tails bind intracellular ligands and connect integrins to signaling and cytoskeletal networks. Thus, integrins provide a link for the bidirectional transmission of mechanical force and biochemical signals across the plasma membrane. 2. Specific Research Accomplishments in the last 12 months: Over the past year we have continued our major interests in control of integrin activation and the actin cross-linking protein filamin. In collaboration with the group of Titus Boggon (Pharmacology, Yale) we have also extended our studies of the integrin-linked kinase (ILK) signaling complex. Integrin activation: we have extended our studies of the molecular mechanisms by which talin binding to the cytoplasmic tails of integrins regulates integrin activation. It is now clear that talin cooperates with kindlin proteins during activation of platelet, leukocyte and endothelial integrins and this may be a very general activation mechanism. Building on data generated over the past several years we are characterizing the roles of specific kindlin domains and their interactions, and have compared activities of different kindlins. Notably, we have also implicated a new protein, Zasp, in control of talin-mediated integrin activation. Mutations in Zasp lead to cardiomyopathies although if or how effects on integrin activation are connected to this remain unclear. Filamin: in recent years we have established that filamin levels are acutely regulated during leukocyte differentiation and myogenesis through the action of an E3 ubiquitin ligase ASB2. In the past year we characterized the interaction of filamin with ASB2,and established that the actin-binding domain of filamin is required for ASB2-mediated proteosomal degradation of filamin. Most recently have mapped putative ubiquitin acceptor sites in filamin and generated filamin mutants resistant to ASB2 mediated degradation - we are currently studying the functional effects of ASB2-resistant filamins. In separate studies we also revealed that filamin controls matrix degradation and cell invasion through effects on levels of the secreted metaloproteinase inhibitor TIMP2. ILK: with Titus Boggon we have continued to pursue structural studies of the ILK, PINCH, parvin complex and the role of parvin-paxillin interactions in focal adhesion targeting. 3. Significance of Key Findings Relevant for the Mission of VBT: Integrin activation and signaling is critical for platelet aggregation, angiogenesis and leukocyte trafficking and the links between integrins and actin play key mechano-signaling roles during cell- adhesion and tissue formation with clear relevance to the vascular system. Our recent studies also establish roles for filamins in leukocyte differentiation and matrix remodeling.


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4. Publications: (Publications July 1, 2011– June 30, 2012) Razinia Z., Baldassarre M., Bouaouina M., Lamsoul I., Lutz P.G., and Calderwood D.A.

(2011) The E3 ubiquitin ligase specificity subunit ASB2α targets filamins to proteasomal degradation by interacting with the filamin actin-binding domain. J. Cell Sci. 124, 2631-41 PMCID: PMC3138704

Lamsoul I, Burande C.F., Razinia Z., Houles T.C., Menoret D., Baldassarre M., Erard M., Moog-Lutz C., Calderwood D.A. and Lutz P.G. (2011) Functional and Structural Insights into ASB2α, a novel regulator of integrin-dependent adhesion of hematopoietic cells. J. Biol. Chem. 286, 30571-30581 PMCID: PMC3162417

Lawson C., Lim S-T., Uryu S., Chen X.L., Calderwood D.A., and Schlaepfer D.D. (2012) FAK promotes recruitment of talin to nascent adhesions to control cell motility. J. Cell Biol. 196:223-232 PMCID: PMC3265949

Bouaouina M., Goult B.T., Huet-Calderwood C., Bate N., Brahme N.N., Barsukov I.L., Critchley, D.R., and Calderwood D.A. (2012) A conserved lipid-binding loop in the kindlin FERM F1 domains is required for kindlin-mediated αIIbβ3 integrin co-activation. J. Biol. Chem. 287, 6979-6990 PMCID: PMC3293583

Guiet R., Verollet C., Lamsoul I., Cougoule C., Poincloux R., Labrousse A., Calderwood D.A., Glogauer M., Lutz P.G., and Maridonneau-Parini I. (2012) Macrophage mesenchymal migration requires podosome stabilization by Filamin A. J. Biol. Chem. 287(16):13051-62 PMCID: PMC3339984

Bandyopadhyay A., Rothschild G., Kim S., Calderwood D.A., and Raghavan S. (2012) Functional differences between kindlin-1 and kindlin-2 in keratinocytes. J. Cell Sci. 125:2172-2184 PMCID: PMC3367939

Li X., Zhang R., Draheim K.M., Calderwood D.A., and Boggon T.J. (2012) Structural basis for the small G-protein-effector interaction of Ras-related protein 1 (Rap1) and the adaptor protein Krev interaction trapped 1 (KRIT1). J. Biol. Chem. 287:22317-22327. PMCID: PMC3381192

Bouaouina M., Harburger D.S., and Calderwood D.A. (2012) Talin and signaling through integrins. Methods in Mol. Biol. 757:325-47. Integrin and Cell Adhesion Molecules: Methods and Protocols, Edited by Motomu Shimaoka.

Razinia Z, Mäkelä T., Ylänne J., and Calderwood D.A. (2012) Filamins in mechanosensing and signaling. Annu. Rev. Biophys. 41:227-246

Baldassarre M., Razinia Z., Brahme N.N., Buccione R., and Calderwood D.A. (2012) Filamin A controls matrix metalloprotease activity and regulates cell invasion in human fibrosarcoma cells. In Press J. Cell Sci

Coyer S.R., Singh A., Dumbauld D.W., Calderwood D.A., Craig S.W., Delamarche E., and García A.J. (2012) Balance between Integrin Cluster Adhesive Force and Cytoskeletal Tension Regulates Nanoscale Integrin Cluster Assembly. In press J. Cell Sci.

Stiegler A.L., Draheim K.M., Li X., Chayen N.E., Calderwood D.A., and Boggon T.J. Structural basis for paxillin binding and focal adhesion targeting of β-parvin. In press J. Biol. Chem.

Brahme N.N., and Calderwood D.A. (2012) Cell Adhesion: A FERM Grasp of the Tail Sorts Out Integrins. In press Curr. Biol.


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Hyung J. Chun, M.D.

Assistant Professor of Medicine (Cardiovascular Medicine)

1. Overall Goal(s) of the Research Program of the Laboratory: The main interest of my laboratory is to characterize the signalling pathways that are important in vascular homeostasis and in disease processes such as atherosclerosis and pulmonary arterial hypertension. Our ongoing characterization of the apelin-APJ GPCR signalling pathway has shed new light on the crosstalk and downstream mechanisms that are important in animal disease models, and we will continue to elucidate how this and other signaling mechanisms are relevant both in animal models and in the clinical setting. 2. Specific Research Accomplishments in the last 12 months: Our recent studies have identified two endothelially-expressed microRNAs that are significantly downregulated in pulmonary arterial hypertension. We identified that these microRNAs are critical regulators of the FGF2/FGFR1 signaling pathway, which is aberrantly activated in PAH. We found in experimental PAH models that restoration of these microRNAs can lead to significant amelioration of PAH, along with restoration of normal FGF signalling levels in the lungs. Moreover, we have found that apelin-APJ signalling pathway is a critical mediator of the statin effects in endothelial cells. Statins are widely known to induce an array of genes in endothelial cells that are thought to be atheroprotective. We found that apelin-APJ signalling is required for statins to induce the known target genes, including KLF2, eNOS, and thrombomodulin. 3. Significance of Key Findings Relevant for the Mission of VBT: We continue to explore vascular signaling pathways that are important in both vascular disease states as well as maintenance of vascular homeostasis. Better understanding of these signaling mechanisms will provide greater insights into the mechanisms of disease pathogenesis, as well as identify novel venues for potential therapies aimed at treating vascular diseases such as atherosclerosis and pulmonary arterial hypertension. 4. Publications: (Publications July 1, 2011– June 30, 2012) Won, C.H.*, Chun, H.J.* (*co-primary authors), Chandra, S.M., Sarinas, P.S., Chitkara,

R.K., Heidenreich, P.A.., Severe obstructive sleep apnea increases mortality in patients with ischemic heart disease and myocardial injury, Sleep and Breathing, 2012 Feb 1. [Epub ahead of print]

Jane-Wit, D., Chun, H.J., Mechanisms of dysfunction in senescent pulmonary endothelium, J Gerontol A Biol Sci Med Sci. 236-241, 2012


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Alan Dardik, M.D., Ph.D.

Associate Professor of Surgery (Vascular Surgery)

1. Overall Goal(s) of the Research Program of the Laboratory: The Dardik laboratory continues to study the healing and function of blood vessels as used in patients having vascular bypass surgery. We are currently trying to understand the fundamental molecular mechanisms by which vein graft adaptation results in positive remodeling and successful adaptation to the arterial environment, yet often proceeds, in the long-term, to neointimal hyperplasia and graft failure.

2. Specific Research Accomplishments in the last 12 months: The laboratory continues to study the mechanisms by which Eph-B4 controls wall thickness during vein graft adaptation to the arterial circulation. We continue to collaborate closely with several other labs in the VBT program, including examination of mechanisms of tissue engineered graft-adaptation to the circulation. Dr Rob Brenes, a postdoctoral fellow in the laboratory, won the Society for Vascular Surgery (SVS) Foundation Resident Research Prize, and presented his research at the opening plenary session of the 2012 SVS Vascular annual meeting, Washington DC.

3. Significance of Key Findings Relevant for the Mission of VBT: I am particularly pleased to report the publication of the book “Outpatient Surgery” with my coauthor Michael Gaunt, of Cambridge, UK. The collaboration that led to this surgical text was a direct result of the Yale-Cambridge collaboration of the VBT program.

3. Publications: (Publications July 1, 2011– June 30, 2012) Hibino N, Yi T, Duncan DR, Rathore A, Dean E, Naito Y, Dardik A, Kyriakides T, Madri J,

Pober JS, Shinoka T, Breuer CK. A Critical Role for Macrophages in Neovessel Formation and the Development of Stenosis in Tissue Engineered Vascular Grafts. Faseb Journal 25(12):4253-4263 (2011).

Eghbalieh SDD, Chowdhary P, Muto A, Ziegler KR, Kudo FA, Pimiento JM, Mirmehdi I, Model LS, Kondo Y, Nishibe T, Dardik A. Age-related neointimal hyperplasia is associated with monocyte infiltration after balloon angioplasty. Journal of Gerontology Series A: Biological Sciences 67(2):109-117 (2012).

Muto A, Panitch A, Kim N, Park K, Komalavilas P, Brophy CM, Dardik A. Inhibition of Mitogen Activated Protein Kinase Activated Protein Kinase II with MMI-0100 reduces intimal hyperplasia ex vivo and in vivo. Vascular Pharmacology 56:47-55 (2012).

Gallo A, Saad A, Ali R, Dardik A, Tellides G, Geirsson A. Circulating interferon-γ-inducible Cys-X-Cys chemokine receptor 3 ligands are elevated in humans with aortic aneurysms and Cys-X-Cys chemokine receptor 3 is necessary for aneurysm formation in mice. Journal of Thoracic and Cardiovascular Surgery 143(3):704-710 (2012).

Quint C, Arief M, Muto A, Dardik A, Niklason LE. Allogeneic human tissue-engineered blood vessel. Journal of Vascular Surgery 55(3):790-798 (2012).

Li X, Jadlowiec C, Guo Y, Protack C, Ziegler K, Lv W, Yang C, Shu C, Dardik A. Pericardial patch angioplasty heals via an Ephrin-B2 and CD34 positive cell mediated mechanism. PLoS ONE 7(6):e38844 (2012).

Brenes RA, Jadlowiec CC, Bear M, Hashim P, Protack CD, Li X, Lv W, Collins MJ, Dardik A. Toward A Mouse Model of Hind Limb Ischemia to Test Therapeutic Angiogenesis. Journal of Vascular Surgery in press (2012).

Dardik A and Gaunt M. Outpatient Surgery. Clinical decision making and board review. Radcliffe Publishing, London (2012).


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Anne Eichmann, Ph.D. Professor, Cardiovascular Research Department

1. Overall Goal(s) of the Research Program of the Laboratory: My laboratory studies vascular and lymphatic development, with particular interests in mechanisms that direct patterning and guidance. Specialized endothelial cells (EC) called tip cells located at the extremities of growing capillary sprouts mediate guided vascular patterning. Tip cells exhibit characteristic features, including extension of filopodia that explore the tip cell environment, lack of a lumen and a slow proliferation rate. Following behind tip cells, other EC termed stalk cells form the capillary lumen and proliferate. Our research is focused on: (1) Tip cell formation (2) Guidance of angiogenesis and lymphangiogenesis 2. Specific Research Accomplishments in the last 12 months: Tip cell formation Tip cell formation is coordinated by the interplay between VEGF and Notch signaling. VEGF promotes tip cell selection, while activation of Notch inhibits tip cells formation and promotes the stalk cell phenotype. Notch regulates expression levels of all three VEGFRs: VEGFR2 and 3 levels are decreased by Notch activation, while VEGFR1 levels are increased by Notch. VEGFR1, notably the soluble sflt1 form, negatively regulates tip cell formation. The VEGF-C receptor VEGFR3, which is critical for lymphangiogenesis, also contributes to coordinated tip cell sprouting. Inducible deletion of the Vegfr3 gene in postnatal retinal vessels leads to hypersprouting, while Vegfc haploinsufficiency reduces angiogenesis, suggesting that VEGFR3 activation occurs both in a ligand-dependent and -independent manner. Mechanistically, VEGFR-3 reinforces Notch signaling by activating expression of Notch target genes via the transcription factor FoxC2, thereby restricting formation of new tip cells (Tammela et al., Nat. Cell Biol. 2011). We have also shown that Activin-receptor like kinase 1 (Alk1) signaling is activated in stalk cells by circulating Bone Morphogenetic Protein 9 (BMP9). BMP9 activation of Alk1 leads to downstream activation of ERK and Smads. Smad signaling downstream of BMP9-Alk1 activation cooperates with Notch signaling to induce expression of Notch target genes as well as of VEGFR1 and the guidance receptor UNC5B, thereby dampening the VEGF response and preventing tip cell formation. Consequently, sequestering BMP9 binding to by adenoviral overexpression of an extracellular Alk1 trap leads to significant retinal hypervascularization and formation of arterio-venous shunts (Larrivée et al., Dev Cell 2012). Taken together, these observations show that capillary sprout formation is guided by tip cells in response to high VEGF levels, and that a regulatory feedback loop involving Notch and Alk1 signaling is initiated downstream of VEGF to prevent tip cell formation in stalks, thus ensuring proper capillary morphogenesis. Guidance of angiogenesis and lymphangiogenesis

Capillary sprouting shows morphological similarities to axon guidance. Like endothelial tip cells, axonal growth cones extend filopodia that sense and respond to guidance cues provided by soluble, cell- or matrix-bound ligands. Several key molecules regulate capillary and axon guidance, including Neuropilin receptors 1 and 2 (Nrp1, Nrp2). Nrp1 and Nrp2 regulate guidance of distinct sets of axons to their targets. In the vascular system, Nrp1 and 2 are expressed in distinct compartments, with Nrp1 mainly labeling arteries and Nrp2 veins and lymphatic vessels. Deletion of the genes encoding Nrp1 or Nrp2 specifically affects


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arterial or lymphatic vessel sprouting, respectively. Despite prominent expression of Nrp2 in lymphatic vessels, Nrp2 expression is absent from valve forming lymphatic EC (LECs). Valves prevent backflow of lymph, and abnormal valve formation leads to lymphedema formation. Lymphatic valves express the Nrp1 receptor and one of the four signal-transducing type A Plexins, PlexinA1. We have shown that Sema3A preferentially binds to lymphatic valves and that valve formation is deficient in mouse mutants for Sema3a, Plexna1 and the Sema3A binding site in Nrp1 (Bouvrée et al., Circ. Res. 2012). These experiments reveal essential, non-redundant function for Nrp signaling in lymphatic development: VEGF-C signaling through Nrp2-VEGFR3 directs lymphatic sprouting, while Sema3A-Nrp1 signaling directs valve morphogenesis. 3. Significance of Key Findings Relevant for the Mission of VBT: Our findings provide new insight into fundamental mechanisms directing angiogenesis and blood and lymphatic vessel assembly. 4. Publications: (Publications July 1, 2011– June 30, 2012) Tammela T, Zarkada G, Nurmi H, J acobsson L, Heinolainen K, Tvogorov D, Mutomäki A,

Franco C, Aranda E, Yla-Herttuala S, Fruttiger M, Mäkinen T, Eichmann A, Pollard J, Gerhardt H, Alitalo K. VEGFR-3 reinforces N otch signaling through FoxC2 to control angiogenesis. Nat Cell Biol 2011; 11: 1202-13.

Eichmann A, Thomas JL. Molecular Parallels between Ne ural and Vascular Development. Cold Spring Harb Perspect Med. 2011; doi: 10.1101/cshperspect.a006551, 159-173.

Larrivée B, Prahst C, Gordon E, del Toro R, Mathivet T, Duarte A, Simons M, Eichmann A. Alk1 signaling inhibits angiogenesis by cooperating with the Notch p athway. Dev. Cell 2012; 22: 489-500.

Bouvrée K, Brunet I, del Toro R, Gordon E, Prahst C, Cristofaro B, Mathivet T, Xu Y, Soueid J, Fortuna V, Miura, N, Aigrot MS, Maden CH, Ruhrberg C, Thomas JL, Eichmann A. Semaphorin3A, Neuropilin-1 and PlexinA1 are required for lymphatic valve formation. Circ. Res. 2012; 111: 437-445.

Kim, J, Ka ng, H, Larri vée B, Lee, M, Mettlen, M, Schmid, S, Roman BL, Qyang, Y, Eichmann, A, Jin, S. Context dependent pro-angiogenic function of bone morphogenetic protein signaling is mediated by Disabled homolog 2. Dev. Cell 2012; 23, 441-448.

Eichmann A, Simons M. VEGF signaling inside vascular endothelial cells and beyond. Curr. Op. Cell Biol. 2012; 24: 188-193.

Simons M, Eichmann A. “On target” cardiac effects of anti-cancer drugs: lessons from new biology. JACC 2012; in press.

HUMAN AND TRANSLATIONAL IMMUNOLOGY ANNUAL REPORT 2011 – 2012

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Jack A. Elias, M.D.

Waldemar Von Zedtwitz Professor of Medicine Professor of Immunobiology and Chair, Department of Internal Medicine

1. Overall Goal(s) of the Research Program of the Laboratory:

Our laboratory is actively engaged in murine, in vitro and human studies of lung injury, repair and remodeling. A particular focus of the laboratory revolves around the biology of the chitinases and chitinase-like proteins and mechanisms of COPD.

2. Specific Research Accomplishments in the last 12 months: During this past year we have: A. Demonstrated that the major role of the chitinase-like protein, chitinase 3-like-1, relates to its

roles in anti-pathogen responses. These studies demonstrated that chitinase 3-like-1 contributes to anti-bacterial responses where it controls macrophage pyroptosis and contributes to host disease tolerance. The latter was mediated by the ability of chitinase 3-like-1 to inhibit inflammasome activation and purinergse activation.

B. Demonstrated that cigarette smoke decreases the expression of NRX1 allowing for unchecked RIG-like helicase activation which leads to inflammation and emphysema.

3. Significance of Key Findings Relevant for the Mission of VBT:

These findings provide entirely new concepts of the roles that chitinases and chitinase-like proteins play in the pathogenesis of injury, inflammation and repair all of which involve blood vessels in an intimate way.

4. Recent Publications: (up to 5 publications July 1, 2011– June 30, 2012) Ma, B., Dela Cruz, C., Takyar, S., Lee, C.G. and Elias, J.A. RIG-like helicase innate immunity

inhibits VEGF tissue responses via a Type I interferon-dependent mechanism. Am. J. Resp. Crit. Care Med. 183:1322-1335, 2011.

Matsuura, H., Hartl, D., Kang, M-J, Dela Cruz, C.S., Koller, B., Chupp, G.L., Homer, R.J., Zhou, Y., Cho, W-K, Elias, J.A., Lee, C.G. Role of breast regression protein (BRP)-39 in the pathogenesis of cigarette smoke-induced inflammation and emphysema. Am. J. Respir. Cell Mol. Biol. 44:777-786, 2011.

Kang, M-J, Choi, J-M, Kim, B.H., Lee, C-M, Cho, W-K, Choe, G., Kim, D-H, Lee, C.G. and Elias, J.A. IL-18 induces emphysema, and airway and vascular remodeling via IFN-, IL-17A and IL-13. Am. J. Resp. Crit. Care Med. 185:1205-1217, 2012.

Dela Cruz, C.S., Liu, W., He, C.H., Jacoby, A., Gornitzky, A., Ma, B., Flavell, R.F., Lee, C.G. and Elias, J.A. Chitinase 3-like-1 (Chi311) Regulation of Streptococcus pneumoniae Lung Infection. Cell Host and Microb. (in press).

Lee, C.G., Herzog, E., Ahangari, F., Zhu, Y., Gulati, M., Lee, C-M, Peng, X., Feghali-Bostwick, C., Jimenez, S.A., Varga, J. and Elias, J.A. Chitinase1 is a biomarker for and therapeutic target in scleroderma-interstitial lung disease that augments TGF-β1 signaling. J. Immunol. (in press).


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Richard Flavell, Ph.D., FRS Sterling Professor and Chairman of Immunobiology, Investigator of Howard Hughes Medical Institute

1. Overall Goal(s) of the Research Program of the Laboratory: We have used knockout mice to investigate the role of innate immune sensors known as inflammasomes in chronic inflammatory disease. Specifically we have found a new pathway that is expressed in the colonic epithelium that controls the homeostasis of the microbiota of the intestine. In the last year, we have examined the effect of the deficiency in the inflammasome pathway which controls the microbiota on systemic chronic diseases. The human population in the developed world suffers from a variety of afflictions including obesity, fatty liver disease, type II diabetes and cardiovascular disease, which is the ultimate cause of death in most cases. We found that mice which exhibit dysbiosis as a consequence of inflammasome deficiency leads to the development of metabolic syndrome, including fatty liver disease, obesity and type II diabetes. This work is highly relevant to the Vascular Biology community and has significant implications. The most striking of these is that we found that the metabolic syndrome developed by these mice is transmissible to genetically normal animals. This is as a consequence of the infectious microbiota that develops in the absence of the defense mechanisms provided by the inflammasome pathway. The implications of this for human health are significant. 2. Specific Research Accomplishments in the last 12 months: As described over, we have found that dysbiosis downstream of inflammasome deficiency can cause metabolic syndrome including diabetes, obesity and fatty liver disease. Second, we have found that these deficiencies are infectious and can be transmitted to genetically normal mice as a consequence of the dysbiosis was this that is transmitted to them from the inflammasome deficient mouse. 3. Significance of Key Findings Relevant for the Mission of VBT: The above findings that lead to the development of metabolic syndrome, the leading cause of death in the developed world, including fatty liver disease, obesity and type II diabetes are highly relevant to the mission of VBT. 4. Publications: (Publications July 1, 2011– June 30, 2012) Henao-Mejia J, Elinav E, Jin C, Hao L, Mehal WZ, Strowig T, Thaiss CA, Kau AL,

Eisenbarth SC, Jurczak MJ, Camporez J-P, Shulman GI, Gordon JI, Hoffman HM, Flavell RA. Inflammasome-mediated dysbiosis regulates progression of NAFLD and obesity. Nature 482:179-185 (2012). PMC3276682

Strowig T, Henao-Mejia J, Elinav E, Flavell RA. Inflammasomes in health and disease. Nature 481:278-286 (2012) PMC3276682

Huber S, Gagliani N, Zenewicz LA, Huber FJ, Hedl W, O’Connor W, Murphy AJ, Valenzuela DM, Yancopoulos GD, Booth CJ, Cho JH, Ouyang W, Abraham C, Flavell RA. IL-22BP axis is regulated by the inflammasome and modulates tumorogenesis in the intestine. Nature (in press) (2012)

VASCULAR BIOLOGY AND THERAPEUTICS ANNUAL REPORT 2011-2012

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Arnar Geirsson, M.D. Assistant Professor of Surgery Overall Goals of the Research Program of the Laboratory: The laboratory pursues two main research projects. The first project involves assessment of the role of miRNAs in both cardiac remodeling following ischemic injury and in maintenance of normal cardiac function. Second project involves analyzing TGF- signaling in mitral regurgitation and degenerative mitral valve disease. Specific Research Accomplishments in the Last Year (7/1/11-6/30/12): Significant progress has been made in the miRNA project over the last year. Inducible cardiac specific Dicer knock-out mouse model demonstrated a dramatic rapid decline in cardiac function coinciding with a reduction in miR-1 levels. Potential biological target has been identified as Sorcin a ryanodine receptor 2 modulator that provides a mechanistic explanation of our phenotype. Significant dysregulation of calcium signaling in cultured cardiomyocytes was confirmed as well as similar finding were noted in wild type mouse model treated with antagomir-1. Reciprocal findings are observed in clinical speciment of heart failure. The result have now been published and further studies planed. Dissection of the role of TGF- in myxomatous mitral valve disease has progressed significantly. The TGF- pathway is upregulated corresponding to extracellular matrix deposition in myxomatous mitral valve tissue. Immunohistochemistry and immunofluorescence have further delineated specific cell types and activation of valvular interstitial cells. Isolation and culturing of valvular interstitial cells was successful and allow dissection of the signaling pathways involved. Angiotensin receptor blockers effectively modulate TGF- signaling that involves SMAD2/3, PI3K and p38 but is independent of ERK. A surprising findings is the total absence of the angiotensin II receptor type 1 in valvular interstitial cells. Angiotensin receptor blockers appear to modulate both TGF- signaling and downstream extracellular matrix targets in clinical tissue of patients on losartan. The results of these studies have been published and further extensive confirmatory studies are in progress. Significance of Key Findings Relevant for the Mission of VBT: Both projects remain with significant interdepartmental collaboration in all aspects of the research projects. Publications and Abstracts: Amy Gallo, Ahmad Saad, Rahmat Ali, Alan Dardik, George Tellides, Arnar Geirsson.

Circulating interferon--inducible CXCR3 ligands are elevated in humans with aortic aneurysm and CXCR3 is necessary for aneurysm formation in mice. Journal of Thoracic and Cardiovascular Surgery. 143;704-710, 2012.

Sabet W. Hashim, Samuel J. Youssef, Bassem Ayyash, Anthony J Rousou, Sigurdur Ragnarsson, Susan Collazo, Arnar Geirsson. Pseudo-Prolapse of the Anterior Leaflet in Chronic Ischemic Mitral Regurgitation: Identification and Repair. Journal of Thoracic and Cardiovascular Surgery. 143;S33-37, 2012.

Rahmat Ali, Yan Huang, Stephen E. Maher, Richard W. Kim, Frank J. Giordano, George Tellides, Arnar Geirsson. miR-1 mediated suppression of Sorcin regulates

myocardial contractility through modulation of Ca2+ signaling. Journal of Molecular and Cellular Cardiology. 52;1027-1037, 2012.

VASCULAR BIOLOGY AND THERAPEUTICS ANNUAL REPORT 2011-2012

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Arnar Geirsson, Mansher Singh, Rahmat Ali, Hussain Abbas, Wei Li, Juan A. Sanchez, Sabet Hashim, George Tellides. Modulation of TGF-β Signaling and Extracellular Matrix Production in Myxomatous Mitral Valves by Angiotensin II Receptor Blockers. Circulation. 126;S189-S197, 2012.

Arnar Geirsson, Mansher Singh, Rahmat Ali, Hussain Abbas, Wei Li, Juan A. Sanchez, Sabet Hashim, George Tellides. Modulation of Transforming Growth Factor-β Signaling and Extracellular Matrix in Myxomatous Mitral Valve Degeneration by Angiotensin II Receptor Blocker Losartan. American Heart Association Scientific Sessions 2011. Orlando, FL. Nov 15, 2011.

Rahmat Ali, Frank J. Giordano, George Tellides, Arnar Geirsson. Regulation of myocardial function by miR-1 occurs through modulation of Sorcin and Ca2+ signaling. Poster. American Heart Association Scientific Sessions 2011. Orlando, FL. Nov 14, 2011.


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Daniel R. Goldstein, M.D.

Associate Professor of Internal Medicine and Immunobiology

1. Overall Goal(s) of the Research Program of the Laboratory: Understand how aging modifies inflammation and how inflammation alters organ transplantation 2. Specific Research Accomplishments in the last 12 months: Discovered that haptoglobin is a novel innate immune ligand that activates inflammation

after organ transplantation. Determined that naïve CD8+ T cells may be cellular mediators of transplant tolerance

resistance with aging. Discovered that with aging vascular smooth muscle cells exhibit pro atherogenic

features. Evaluated the efficacy of a vaccine that links TLR5 activation with influenza peptides to

induce protection from influenza viral infection with aging.

3. Significance of Key Findings Relevant for the Mission of VBT: Our findings provide fundamental insights as to how aging alters inflammation and how modifying innate immunity may improve organ transplantation. 4. Publications: (Publications July 1, 2011– June 30, 2012) Du W, Shen H, Galan H and Goldstein DR. An age-specific CD8+ T cell pathway that

impairs the effectiveness of strategies to prolong allograft survival. The Journal of Immunology 2011 (187): 3631-40

Shirali AC, Look, M, Du W, Fahmy T and Goldstein DR. Nanoparticle delivery of mycophenolic acid upregulates PD-L1 on dendritic cells to prolong murine allograft survival. The American Journal of Transplantation 2011 (12): 2582-92.

Leng J, Stout-Delgado HW, Kavita U, Jacobs A, Tang J, Tussey L and Goldstein DR. Efficacy of a vaccine that links viral epitopes to flagellin in protecting aged mice from influenza viral infection Vaccine 2011 (29): 8147-8155

Song Y, Shen H, Schenten D, Lee PJ and Goldstein DR. Aging enhances the basal production of IL-6 and CCL2 in vascular smooth muscle cells. Arteriosclerosis, Thrombosis and Vascular Biology, 2012 Jan;32(1):103-9

Shen H, Song Y, Colangelo C, Wu T, Bruce C, Scabia G, Maffei M and Goldstein DR. Haptoglobin activates innate immunity to enhance acute transplant rejection in mice The Journal of Clinical Investigation 2012, (122):383-387


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Daniel M. Greif, M.D. Assistant Professor of Medicine, Cardiovascular Section

1. Overall Goal(s) of the Research Program of the Laboratory: My laboratory investigates blood vessel morphogenesis, the maintenance of the adult blood vessel and diseases of the vasculature. We initially determined the patterns of smooth muscle and adventitial cell differentiation, proliferation and migration in the developing pulmonary arterial wall in the mouse and the role of the platelet derived growth factor pathway in these processes. Currently, we are studying the morphogenesis of the walls of other vessels, such as the aorta, and comparing and contrasting their morphogenesis with that of the pulmonary artery. In addition, little is known about the maintenance of blood vessels, and we aim to determine the patterns and underlying mechanisms of cell turnover, proliferation and migration in the adult vessel wall. Moreover, diseases of the vasculature are thought to largely involve a recapitulation of developmental programs, and we are investigating animal models of vascular diseases that involve ectopic and aberrant smooth muscle cells, such as atherosclerosis, arterial stenosis and pulmonary artery hypertension. Finally, we will study clinical samples obtained from patients with vascular diseases and relate them to our findings in animal models. 2. Specific Research Accomplishments in the last 12 months: We found that the developing pulmonary arterial wall in the mouse is radially patterned, from the inside out, by two distinct but coordinated processes: i) sequential induction of successive cell layers from surrounding lung mesenchyme, and ii) controlled invasion of outer layers by inner layer cells through developmentally-regulated cell reorientation and radial migration. In addition, we have determined the patterns of smooth muscle progenitor migration and differentiation in the aorta during development and compared this process to that in the pulmonary artery. We have also discerned the origins of the excess smooth muscle cell burden in mouse models of pulmonary hypertension. 3. Publications: (Publications July 1, 2011– June 30, 2012) Greif, DM*, Kumar, M., Lighthouse, JK, Hum, J., An, A., Ding, L., Red-horse, K., Espinoza,

FH, Olson, L., Offermanns, S., Krasnow, MA*. Radial construction of an arterial wall. Developmental Cell - in press. (*Corresponding authors.)

Greif, DM. Invited chapter: Vascular embryology and angiogenesis. In Vascular Medicine, A Companion to Braunwald’s Heart Disease, 2nd edition, ed. M.A. Creager, J.A. Beckman, and J. Loscalzo, Elsevier Inc., Philadelphia, PA. In press.


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Jay D. Humphrey, Ph.D.

Professor, Department of Biomedical Engineering

1. Overall Goal(s) of the Research Program of the Laboratory: The primary goal of my laboratory is to understand better and mathematically model the roles of mechanobiological mechanisms in tissue-level vascular growth and remodelling. Specific interests include arterial changes in response to hypertension, altered flow, and aging, the design and assessment of tissue engineered arteries, venous changes when used as arterial grafts, and the progression of large artery diseases such as aortic and cerebral aneurysms. We are also interested in exploiting genetically modified mice to elucidate individual contributions of extracellular matrix constituents (e.g., fibrillin-1, fibulin-5, elastin, collagens I, III, and IV, and proteoglycans) to tissue-level arterial structure and function. 2. Specific Research Accomplishments in the last 12 months The primary accomplishments included the elucidation, via mechanical testing and modeling, of an important coupling between elastic fibers and collagen fibers that significantly influences the evolving stiffening of large arteries in aging, finding a closed-form mathematical solution to a longstanding tissue engineering experiment that clarifies smooth muscle and fibroblast behavior in collagen-based tissue equivalents, and development of large-scale computational models of the aortic and cerebral circulations, which will allow quantification of hemodynamic-induced loads on vascular cells. Finally, we secured additional new NIH and NSF support for the laboratory. 3. Significance of Key Findings Relevant for the Mission of VBT: Our ability to assess biomechanically the multiaxial mechanical properties of mouse arteries and veins supported two different projects by other investigators in the VBT and has enabled the initiation of two additional collaborative projects that bring together bioengineers and vascular biologists within the VBT. 4. Publications: (Publications July 1, 2011– June 30, 2012) Ferruzzi J, Collins MJ, Yeh AT, Humphrey JD (2011) Mechanical assessment of elastin integrity in

fibrillin-1 deficient carotid arteries: Implications for Marfan syndrome. Cardiovasc Res 92: 287-295.

Collins MJ, Bersi M, Wilson E, Humphrey JD (2011) Mechanical properties of suprarenal and infrarenal abdominal aorta: Implications for mouse models of aneurysms. Med Phys Engr 33: 1262-1269.

Masson I, Beaussier H, Boutouyrie P, Laurent S, Humphrey JD, Zidi M (2011) Carotid artery mechanical properties and stresses quantified using in vivo data from normotensive and hypertensive human subjects. Biomech Model Mechanobiol 10: 867-882.

Karsaj I, Humphrey JD (2011) A multilayered wall model of arterial growth and remodeling. Mech Material 44: 110-119.

Naito Y, Williams-Fritze M, Duncan DR, Church SN, Hibino N, Madri JA, Humphrey JD, Shinoka T, Breuer CK (2012) Characterization of the natural history of extracellular matrix production in tissue-engineered vascular grafts during neo-vessel formation. Cells, Tissues, Organs 195: 60-72.

Collins MJ, Eberth JF, Wilson E, Humphrey JD (2012) Acute mechanical effects of elastase in a mouse model of abdominal aortic aneurysms. J Biomech 45: 660-665.

Humphrey JD, Holzapfel GA (2012) Mechanics, mechanobiology, and modeling of human abdominal aorta and aneurysms. J Biomech 45: 805-814.

Simon DD, Humphrey JD (2012) On a class of admissible constitutive behaviors in free-floating engineered tissues. Int J Nonlin Mech 47: 173-178.


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Zhang P, Huang A, Ferruzzi J, Mecham RP, Starcher BC, Tellides G, Humphrey JD, Giordano FJ,

Niklason LE, Sessa WC (2012) Inhibition of microRNA 29 enhances elastin levels in cells haploinsufficient for elastin and in bioengineered vessels. Arterioscl Thromb Vasc Biol 32: 756-759.

Genovese K, Collins MJ, Lee YU, Humphrey JD (2012) Regional finite strains in an angiotensin-II infusion model of dissecting abdominal aortic aneurysms. J Cardiovasc Engr Tech 3: 194-202.

Di Achille P, Humphrey JD (2012) Towards large scale computational fluid-solid-growth models of intracranial aneurysms Yale J Biol Med 85: 217-228.


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John Hwa, M.D., Ph.D. Associate Professor of Internal Medicine - Cardiology 1. Overall Goal (s) of the Research Program of the Laboratory: The major focus of our laboratory is the pharmacology and genetics (pharmacogenetics) of the human prostacyclin and thromboxane receptors. The central hypothesis for these studies are; “suppression of PGI2 activity by either COX-2 inhibition, or through genetically defective human prostacyclin receptor (hIP), results in accelerated atherothrombosis. In contrast, suppression of thromboxane activity by either COX-1 inhibition, or through genetically defective human thromboxane receptor (hTP), results in reduced atherothrombosis”. A second series of studies focuses on the relationship between diabetes mellitus, thromboxane, and platelet hyperactivity. For these studies our hypothesis is; “aldose reductase is a major transducer of the hyperglycemic response, resulting in increased thromboxane release and platelet hyperactivity”. To test all our hypotheses we study human patients and human tissues at multiple levels (clinical, pathophysiology, pharmacology, cell biology, molecular biology, and bioinformatics). These are also supported by animal models and cell line studies. Our ultimate goals are to identify patients at increased risk for atherothrombosis, and to develop novel therapies targeting atherothrombosis. 2. Specific Research Accomplishments in the last 12 months: We have now demonstrated that multiple hIP variants are associated with increased coronary artery disease (atherosclerosis) burden on coronary angiography and increased cardiovascular events. In search of the mechanisms, the dysfunctional hIP variants are able to heterodimerize with wild type receptor, exerting a dominant negative effect. We also demonstrated that reduced hIP function can lead to VSMC proliferation and dedifferentiation, in part through reduced prostacyclin-induced prostacyclin release from human VSMC. We are currently assessing cAMP/PKA-dependent and novel PKA-dependent and independent pathways for prostacyclin signaling. Such studies in addition to identifying patients at increased risk of developing atherothrombosis, will provide important insights into critical components in the human prostacyclin receptor structure that are required for binding ligand and activation. We have also identified and characterized naturally occurring mutations in the human thromboxane receptor from sequencing approximately 1,000 cardiovascular patients and extensive database searches. At least 4 of the approximately 30 mutations identified demonstrate distinct functional defects. We are currently assessing protection from or promotion of atherothrombosis arising from these mutations. We anticipate that these studies may also provide structural insights in the development of prostacyclin and thromboxane based drugs in combating atherothrombosis. For our second series of studies we recently demonstrated in human platelets that aldose reductase (AR) modulates platelet response synergistically, to both hyperglycemia and collagen exposure, through a pathway involving ROS/PLC2/PKC/p38 MAPK. These results were supported with clinical studies in patients with platelet activation (deep vein thrombosis, and patients with saphenous vein graft occlusion after coronary bypass surgery) where significant increases in urinary levels of a major enzymatic metabolite of TX (i.e. 11-dehydro-TXB2, TX-M) were observed, particularly in diabetic patients (even in the presence of low dose aspirin). This suggests that many patients with persistently raised TX-M despite the use of low dose aspirin may have underlying collagen exposure with thrombo-vascular disease (reflecting endothelial damage). We are currently exploring the use of AR inhibitors to combat cardiovascular disease in diabetes mellitus. Our studies additionally


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provide multiple signaling targets for combination chemotherapy to inhibit the synergistic platelet activation observed with hyperglycemia and collagen exposure. 3. Significance of Key Findings Relevant for the Mission of VBT

Identification of novel mechanisms that modulate atherothrombosis Identification of patients that are at increased risk (hIP variants), or protected from

(hTP variants), atherothrombosis Identification of the role played by aldose reductase in platelets Identification of potential novel therapeutic targets

4. Publications in the last 12 months:

Menon, I., Huber T., Sanyal S., Banerjee S., Barre P., Warren D., Hwa J., Sakmar T.P. Menon A.K. Opsin is a phopholipid flippase. Curr Biol 2011 21(2):149-153

Stitham J., Arehart E., Elderon L., Gleim S.R., Douville K., Kasza Z., Fetalvero K., MacKenzie T., Robb J., Martin K.A., Hwa J. Comprehensive biochemical analysis of rare prostacyclin receptor variants: Study of association of signaling with coronary artery obstruction. J. Biol. Chem 2011 286(9):7060-7069

Garcia-Rodriguez L.A., Gonzalez-Perez A., Bueno H., Hwa J. NSAID use selectively increases the risk of non-fatal myocardial infarction: A systematic review of randomized trials and observational studies. PLoS One 2011 6(2):e16780

Ding M., Xie Y., Wagner R.J., Jin Y., Carrao, A.C., Liu L., Guzman A.K., Powell R.J., Hwa J., Rzucidlo E.M., Martin K.A. Adiponectin Induces Vascular Smooth Muscle Cell Differentiation via mTORC1 and FoxO4. Arterioscler Thromb Vasc Biol 2011 1;31(6):1403-1410

Stitham J., Midgett C., Martin K.A., Hwa J. Prostacyclin pathophysiology: An inflammatory paradox. Frontiers in Pharmacology 2011 2:24

Midgett C., Stitham J., Martin K.A., Hwa.J. Prostacyclin receptor regulation---from transcription to trafficking. Current Molecular Medicine 2011 11(7):517-28

Tang WH, Stitham J, Gleim S, Febbo C.D., Porreca E., Fava C, Tacconelli S, Capone M, Evangelista V, Levantesi G, Wen L., Martin K.A., Minuz P, Rade J, Patrignani P, Hwa J. Glucose and collagen regulate platelet activity through aldose reductase induction of thromboxane; implications for diabetes mellitus. J Clin Inves 2011 121(11):4462-76

Tang W.H., Martin K.A., Hwa J Aldose reductase, oxidative stress and diabetes mellitus Frontiers in Pharmacology 2012 3:87

Ding M., Carrao A.C., Wagner R.J., Xie Y., Jin Y., Rzucidlo E.M., Li W., Tellides G., Hwa J., Aprahamian T.R., Martin KA. Vascular smooth muscle cell-derived adiponectin: A paracrine regulator of contractile phenotype. J Mol Cell Card 2012 52:474-484

Chakraborty R., Pydi SP., Gleim S., Dakshinamurti D., Hwa J., Chelikani P. Site-directed mutations and the polymorphic variant Ala160Thr in the human thromboxane receptor uncover a structural role for transmembrane helix 4. PLoS One 2012 7(1): e29996

Gleim S., Stitham J., Tang W., Martin K.A., Hwa J. An eicosanoid-centric view of atherothrombotic risk factors. Cell Mol Life Sci 2012 (in Press)


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Suk-Won Jin, Ph.D.

Assistant Professor, Dept. of Internal Medicine, Section of Cardiovascular Medicine

1. Overall Goal(s) of the Research Program of the Laboratory: My laboratory is interested in understanding molecular and cellular mechanisms that regulate developmental and pathological angiogenesis, using zebrafish as a model system. 2. Specific Research Accomplishments in the last 12 months: We have identified the novel role of Disabled homolog2 (DAB2) in mediating endocytosis of Bone Morphogenetic Protein 2 (BMP2) signaling complex (Kim et al, in press). In addition, we have found that BMP2 signaling can function as an anti-lymphangiogenic cue during vertebrate development (Dunworth et al, submitted). 3. Significance of Key Findings Relevant for the Mission of VBT: Considering the essential role of BMP2 signaling on venous morphogenesis (Wiley et al, 2011), delineating how BMP2 signaling is transduced within endothelial cells will help us to better understand the molecular and cellular mechanisms underlying developmental and pathological angiogenesis. In addition, since there is no effective therapeutic agent to manipulate lymphatic development and maintenance, identifying BMP2 as a novel inhibitory signal for lymphangiogensis will provide theoretical background to develop clinical intervention for lymphatic dysfunctions. 4. Publications: (Publications July 1, 2011– June 30, 2012) Kim JD, Kang H, Larrivée B, Lee MY, Mettlen M, Schmid SL, Roman BL, Qyang Y, Eichmann A,

and Jin SW. Context Dependent Pro-Angiogenic Function of Bone Morphogenetic Protein Signaling is Mediated by Disabled Homolog 2. Dev. Cell. In press.

Pi X, Schmitt CE, Xie L, Portbury AL, Wu Y, Lockyer P, Dyer LA, Moser M, Bu G, Flynn EJ, Jin SW, and Patterson C. An LRP1-Dependent Endocytic Mechanism Governs the Signaling Output of the BMP System in Endothelial Cells and in Angiogenesis. Circ Res. In press.

Schmitt CE, Holland MB, and Jin SW. Visualizing vascular networks in zebrafish: An introduction to microangiography. Methods Mol Biol. 2012;843:59-67.

Wiley DM and Jin SW. Bone Morphogenetic Protein functions as a context-dependent angiogenic cue in vertebrates. Semin Cell Dev Biol. 2011;22(9):1012-8

Tao Y, Neppl RL, Huang ZP, Chen J, Tang RH, Cao R, Zhang Y, Jin SW, and Wang DZ. The histone methyltransferase Set7/9 promotes myoblast differentiation and myofibril assembly. J Cell Biol. 2011;194(4):551-65.


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Martin S. Kluger, Ph.D. Research Scientist, Department of Immunobiology 1. Overall Goals of the Research Program of the Laboratory: a. To understand how tumor necrosis factor (TNF) targets endothelial cell (EC) intercellular junctions to produce capillary leak. We aim to identify TNF pathways that affect EC junctional molecules, especially claudin-5, and lead to capillary leak, the uncontrolled paracellular transit of fluid and proteins in capillary beds, which is critical to the pathogenesis of blood sepsis. b. To understand how EC interaction with accessory cells (pericytes) can stabilize nascent microvessels. We seek to characterize the maturing effects of pericytes on vascular networks formed in 3-D matrices as a step toward therapeutic re-vascularization of engineered replacement tissue. 2. Specific Research Accomplishments in the Last 12 Months: This year Dr. Kluger obtained RO1 funding as a NHLBI co-Principal Investigator in a project entitled, “Proteins of the Endothelial Cell Surface” with Dr. Jordan Pober. This grant funded a collaborative study characterizing a new tight junction dependent model of barrier responses to TNF involving cultured human microvascular EC that was presented at the VBT-sponsored Yale-Cambridge Program and at the national meeting of the North American Vascular Biology Association (NAVBO). Conclusions from our study are (1) in human blood vessels outside the central nervous system, claudin-5 expression correlates with known TJ frequencies and barrier strengths; (2) claudin-5 expression is required but alone insufficient to establish paracellular barriers in cultured EC; (3) claudin-5 maintains paracellular barriers of cultured EC monolayers independently from the major adherens junction molecule VE-cadherin. Research progress toward our second overall goal relating to optimizing therapeutic revascularization was documented in two separate manuscripts in progress. Recognizing a need for an in-house forum dedicated to communicating the diverse approaches to scientific discovery within our VBT program, Dr. Kluger organized and now coordinates with Dr. Jun Yu a new VBT Research-In-Progress (RIP) seminar series in the Amistad building presented by graduate students and post-docs that is attended by VBT faculty. This year he served as the local Yale representative for the nationally based NAVBO. 3. Significance of Key Findings Relevant for the Mission of VBT: Our findings that claudin-5 expression patterns differ among different microvascular segments (e.g., capillaries compared to post-capillary venules) implicate claudin-5 and claudin-5 interactive proteins as potential segment-specific targets for vascular therapy. We are now investigating this possibility with in vivo models and are using our in vitro model to study mechanisms of how TNF modulates EC barriers in a TJ-dependent fashion. Because capillary leak syndrome in sepsis affects 750,000 Americans per year resulting in death in up to 70% of cases, our work is relevant to the translational mission of VBT. 4. Publications: Kluger, M.S., Clark, P. R., Gerke, V. and Pober, J.S. Claudin-5, not VE-Cadherin, Limits

Paracellular Permeability of Human Dermal Microvascular Endothelium. In revision, Arteriosclerosis, Thrombosis, and Vascular Biology.

Chang, W.G., Andrejecsk, J.W., Kluger, M.S., Saltzman, W.M., and Pober, J.S. Platelet-Derived Growth Factor Elicits Pro- and Anti-Angiogenic Responses from Pericytes. In revision, Arteriosclerosis, Thrombosis, and Vascular Biology.

Waters, J.P., Kluger, M.S., Graham, M. Chang, W.G., and Pober, J.S. In vitro assembly of human pericyte-supported endothelial microvessels in three-dimensional co-culture: a simple model for interrogating endothelial:pericyte interactions. Submitted.


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Diane Krause MD, Ph.D.

Professor of Laboratory Medicine

1. Overall Goal(s) of the Research Program of the Laboratory: The overall goals of my research are to characterize bone marrow derived stem and progenitor cells, and to define the molecular mechanisms (signal transduction, biomechanical, and epigenetic) that regulate the self-renewal and differentiation of these cells. Our recent emphasis has been on megakaryocyte development, megakaryoblastic leukemia, as well as platelet formation. We have a renewed interest now on platelet function. We are studying the roles of G-proteins, the SRF signal transduction pathway, and RNA binding proteins in order to better understand and treat hematopoietic diseases including myelodysplasia, myeloproliferative disease and leukemia, and on vascular diseases related to thrombus formation and acute infarction. Projects include work with embryonic stem cells as well as hematopoietic stem cells from mice and humans. Our work provides insights not only into normal blood cell development, but also to the pathogenesis of myeloid leukemia. Studies on epithelial engraftment of bone marrow derived cells, which is also referred to ‘adult stem cell plasticity,’ are focused on the mechanisms of repair in response to tissue injury and disease in the lung. 2. Specific Research Accomplishments in the last 12 months Over the past year, we have published 7 papers. The highest impact study, published in Developmental Cell, is focused on guanine exchange factor regulation of polyploidization of megakaryocytes. We also published that the effects of SRF are likely to be mediated by MKL1 and MKL2 because simultaneous knockdown of both MKL1 and MKL2 in the megakaryocytic lineage causes severe abnormalities in platelet formation and function analogous to those observed in SRF KO mice. Lastly, we have discovered how components of the cytoskeleton regulate the complex cellular responses to activation of megakaryocytic differentiation. MKL1, an activator of serum response factor (SRF) transcriptional activity, plays critical roles in muscle, neuron, and megakaryocyte differentiation. Regulation of MKL1 subcellular localization is one mechanism by which a cell can control SRF activity with MKL1 localization to the nucleus being critical for its function as a transcriptional regulator. MKL1 localization is cell type specific; MKL1 is predominantly cytoplasmic in some muscle cell types. In contrast, neuronal cells have constitutively nuclear localization. We have found that subcellular localization and regulation of MKL1 in megakaryocytes is dependent on RhoA activity and actin organization. Induction of megakaryocytic differentiation of human erythroleukemia (HEL) cells and primary megakaryocytes by TPA and thrombopoietin, respectively, promotes MKL1 nuclear localization. This MKL1 localization is blocked by drugs inhibiting either actin polymerization or RhoA activity. Once in the nucleus, MKL1 then activates transcription of SRF target genes, which are needed for megakaryocyte maturation. This work has been submitted for publication. 3. Significance of Key Findings Relevant for the Mission of VBT Because platelets play a key role in atherosclerosis and inflammation and perhaps in development of lymphatic endothelial lining cells as well, the means by which they are produced, and the transcriptional regulation of the genes expressed by platelets are highly relevant to vascular biology. In addition, the SRF/MKL1 pathways on which we focus is highly relevant to the dysfunction of smooth muscle cells in vascular diseases.


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4. Publications: Qian L, Krause DS, Saltzman WM. Enhanced growth and hepatic differentiation of fetal liver

epithelial cells through combinational and temporal adjustment of soluble factors. Biotechnology Journal 7:440-8, 2012. PMID: 21922669.

Rogers FA, Lin SS, Hegan DC, Krause DS, Glazer PM. Targeted gene modification of hematopoietic progenitor cells in mice following systemic administration of a PNA-peptide conjugate. Molecular Therapy. 20, 109-118, 2011. PMID: 21829173.

Kassmer SH, Bruscia EM, Zhang P, Krause DS. Non-hematopoietic cells are the primary source of bone marrow derived lung epithelial cells. Stem Cells 30: 491-499, 2012. PMID: 22162244.

Guo JK, Marlier A, Shi H, Shan A, Ardito TA, Du ZP, Kashgarian M, Krause DS, Biemesderfer D, Cantley LG. Increased tubular proliferation as an adaptive response to glomerular albuminuria. J Am Soc Nephrol. 23:429-37, 2012. PMID: 22193389.

Truman LA, Bentley KL, Smith EC, Massaro SA, Gonzalez DG, Haberman AM, Hill M, Jones D, Min W, Krause DS, Ruddle NH. ProxTom lymphatic vessel reporter mice reveal Prox1 expression in the adrenal medulla, megakaryocytes, and platelets. The American Journal of Pathology 180:1715-25, 2012. PMID: 22310467

Gao,Y, Smith E, Ker E, Campbell P, Cheng E, Zou S, Lin S, Wang L, Halene S, Krause DS. Role of RhoA specific guanine exchange factors in regulation of endomitosis in megakaryocytes. Developmental Cell 22:573-84, 2012. PMID: 22387001

Smith EC, Thon JN, Devine MT, Lin S, Schulz VP, Gao Y, Massaro SA, Halene S, Gallagher P, Italiano JE, Krause DS. MKL1 and MKL2 play redundant and crucial roles in megakaryocyte maturation and platelet formation. Blood, 2012. PMID: 22806889


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Themis R. Kyriakides Ph.D. Associate Professor, Pathology and Biomedical Engineering 1. Overall Goal (s) of the Research Program of the Laboratory The main area of my research is the elucidation of the molecular events that dictate the course of healing and especially inflammation and angiogenesis following ischemia, injury and the implantation of biomaterials and scaffolds for tissue engineering applications. Our primary research focus is on two molecules, MCP-1 and TSP-2 that we have shown to be critical to various aspects of these processes. In a new research effort we are exploring the inter-relationship between TSP2 and the Akt/eNOS signaling axis. In addition, through the process of molecular dissection of cell-matrix interactions, we aim to incorporate rational design in the development of bioengineering applications such as tissue-engineered vascular grafts. Finally, we aim to define role of TSP2 in the maintenance and repair of the blood brain barrier in the context of brain-biomaterial interactions. 2. Specific Research Accomplishments in the last 12 months (7/1/11-6/30/12) We have continued our investigation of the participation of TSP-2 in angiogenesis and arteriogenesis. Furthermore, we have explored the participation of TSP2/AKt/eNOS signaling axis in endothelial function. We have generated double eNOS/TSP2-null mice and Akt1/TSP2-null mice and discovered that the absence of TSP2 ameliorates the phenotypes of the eNOS-null and Akt1-null mice. More recently, we have shown in in vivo and vitro studies that nitric oxide and hypoxia suppress TSP2 expression at the transcriptional level. In our biomedical engineering-related research we have discovered that MCP-1 contributes to the disruption of the blood brain barrier in the foreign body response. More importantly, we have found that MCP-1-null mice display reduced leakage of the blood brain barrier and enhanced neuronal survival. Finally, we have discovered that cell-biomaterial interactions trigger activation of the inflammasome, which augments the inflammatory response. Within the VBT program we have continued our collaborations with the following investigators: Sessa, Giordano, Saltzman, Niklason, Simons, Bender and Tellides. 3. Significance of Key Findings Relevant for the Mission of VBT Studies in angiogenesis, arteriogenesis, and engineering of vascular grafts are central to the mission of the VBT. In addition, our studies investigating the link between TSP2, Akt1, and eNOS are of importance to many processes in vascular biology. 4. Publications Skokos E.A., Charokopos A, Khan K., Wanjala J., Kyriakides T.R. Lack of TNF--induced

MMP-9 production and abnormal E-cadherin redistribution associated with compromised fusion in MCP-1-null macrophages. American Journal of Pathology 178:2311-21, 2011.

Tian W., Sawyer A., Kocaoglu F.B., Kyriakides T.R. Astrocyte-Derived Thrombospondin-2. Is Critical for the Repair of the Blood-Brain Barrier. American Journal of Pathology 179: 860-8, 2011.

Liu J, Jiang Z, Zhang S, Liu C, Gross RA, Kyriakides TR, Saltzman WM. Biodegradation, biocompatibility, and drug delivery in poly(ω-pentadecalactone-co-p-dioxanone) copolyesters. Biomaterials. 32: 6646-54, 2011

Lynn AD, Blakney AK, Kyriakides TR, Bryant SJ. Temporal progression of the host response to implanted poly(ethylene glycol)-based hydrogels. J Biomed Mater Res A. 96: 621-31, 2011.


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MacLauchlan S., Yu J., Parrish M., Schleicher M., Krady M.M., Zeng J., Huang P.L., Sessa

W.C., Kyriakides T.R. eNOS controls the expression of the angiogenesis inhibitor thrombospondin 2. P.N.A.S. 108: E1137-45. 2011

Malik A, Hoque R, Ouyang X, Ghani A, Hong E, Khan K., Moore L.B., Ng G., Munro F., Flavell R.A., Shi Y., Kyriakides T.R., Mehal W. (2011) Inflammasome components ASC and Caspase-1 mediate biomaterial-induced inflammation and foreign body response. P.N.A.S. 108: 20095-100.

Zhang J., Modi Y., Yarovinsky T, Yu J., Collinge M., Kyriakides T.R., Zhu Y, Sessa W.C.,Pardi R., Bender J.R. (2012) Macrophage β2 Integrin–Mediated, HuR-Dependent Stabilization of Angiogenic Factor–Encoding mRNAs in Inflammatory Angiogenesis. Am. J.Pathol. 180:1751-60.

Hibino N, Villalona G, Pietris N, Duncan DR, Schoffner A, Roh JD, Yi T, Dobrucki LW, Mejias D, Sawh-Martinez R, Harrington JK, Sinusas A, Krause DS, Kyriakides TR, Saltzman WM, Pober JS, Shin'oka T, Breuer CK. Tissue-engineered vascular grafts form neovessels that arise from regeneration of the adjacent blood vessel. FASEB J. 25: 2731-9. 2011

Hibino N, Yi T, Duncan DR, Rathore A, Dean E, Naito Y, Dardik A, Kyriakides TR, Madri J, Pober JS, Shinoka T, Breuer CK. A critical role for macrophages in neovessel formation and the development of stenosis in tissue-engineered vascular grafts. FASEB J. 25: 4253-63. 2011


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Patty J. Lee, M.D.

Associate Professor of Medicine, Section of Pulmonary, Critical Care & Sleep Medicine

1. Overall Goal(s) of the Research Program of the Laboratory: My laboratory investigates mechanisms of lung and endothelial injury and repair responses. Our overall goal is to identify novel signal transduction pathways that can be modulated to protect the lungs and lung endothelium from oxidant damage. We identified the importance of the heat shock protein heme oxygenase-1 (HO-1) and its gaseous reaction product, carbon monoxide (CO), in resisting oxidant-induced endothelial cell death via mitochondrial pathways. We found that a family of signaling molecules, mitogen-activated protein kinases (MAPKs), mediates HO-1 and CO’s protective effects and, more recently, critical innate immune responses. The innate immune system consists of pattern-recognition receptors called toll-like receptors (TLRs), of which TLR4 is the LPS-responsive receptor. We discovered that TLR4 is required for lung endothelial cell survival in aging and oxidant-challenged conditions. These studies represent important paradigm shifts in our understanding of TLR and lung biology and are now the basis of translational studies in people with acute lung injury and age-related chronic lung disease. 2. Specific Research Accomplishments in the last 12 months: We successfully generated vascular endothelial-specific transgenic mice and lentiviral silencing RNA constructs targeted TLR4 and HO-1. We completed testing of the new mouse lines and constructs by showing endothelial specificity and functionality. We also conducted survival and lung injury experiments demonstrating the crucial role of endothelial TLR4 and HO-1 in protective responses against acute oxidant injury, cigarette smoke- and age-induced chronic lung disease. We identified that VEGF-induced endothelial cell survival during acute oxidant stress depends on TLR4. We also found that TLR4 regulates the pleiotropic cytokine macrophage migration inhibitory factor (MIF) in lung endothelial cells during age- and cigarette smoke-related chronic lung disease. 3. Significance of Key Findings Relevant for the Mission of VBT: We were the first to demonstrate the utility of intranasal, lung-targeted and endothelial-targeted silencing RNA (siRNA) constructs in vivo. These unique tools can be used to specifically dissect the role of lung endothelium in a variety of experimental models. In parallel, we have also generated endothelial-targeted transgenic and knockout mouse models to specifically interrogate the role of the endothelium using genetic approaches. Our coordinated use of siRNA technology and genetic technology in both endothelial cell and mouse models offer immense insight into vascular diseases and may identify novel therapeutic targets. 4. Publications: (Publications July 1, 2011– June 30, 2012) Volkova M, Zhang Y, Shaw AC, Lee PJ. The role of toll-like receptors in age-associated

lung diseases, J Gerontol A Biol Sci Med Sci 2012; 67A(3):247-253. Vaz Fragoso C, Lee PJ The Aging Lung, J Gerontol Biol Sci 2011, 2012 5:67A(3):233-

235. Song, Y., Shen H., Schenten, D., Shan, P., Lee, P.J. and Goldstein, D.R. Arteriosclerosis,

Aging enhances the basal production of IL-6 and CCL2 in vascular smooth muscle cells, Arterioscler Thromb Vasc Biol, 32: 103-109, 2012.

Mannam, P., Zhang, X., Shan, P., Zhang, Y., Shinn, A., Zhang, Y. and Lee PJ Endothelial MKK3 is a critical mediator of lethal endotoxemia and acute lung injury, Submitted


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Mannam, P., Shinn, A., Neaum, RF, Kang, MJ, Dela Cruz, CS., West, P, Shadel, GS., Shen,

J., Elias, JA and Lee PJ, MKK3 regulates endothelial mitophagy and susceptibility to lethal sepsis, Submitted

Zhang, Y., Zhang, X.P., Shan, P., Pandita, T. and Lee, P.J. Endothelial Hsp70 mediates the protective effects of TLR4 in lethal hyperoxic lung injury, Submitted


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Joseph A. Madri, M.D., Ph.D. Professor, Dept. of Pathology and Molecular, Cellular and Developmental Biology 1. Overall Goal(s) of the Research Program of the Laboratory: During the past year we have continued our investigations into extracellular matrix-endothelial cell interactions and their dynamic interactions during vessel formation and also initiated investigations in collaboration with investigators in the physics department evaluating novel approaches in brain imaging. Lastly, we have continued our investigations of T cell trafficking and endothelial permeability, focusing on the roles of CD44. 2. Significance of Key Findings Relevant for the Mission of VBT: Our findings impact both the basic and translational missions of the VBT in that we have advanced our understanding of the roles of the vasculature in the development, maintenance and responsiveness of neurovascular niches in the CNS. 3. Publications: (Publications July 1, 2011– June 30, 2012) Naito, Y., Williams-Fritz, M., Madri, J.A., Shinoka, T., Breuer, C., Characterization of the

natural history of extracellular matrix production in tissue engineered vascular grafts during neovessel formation, Cells Tissues Organs. 195(1-2):60-72, 2012 Oct 12

Frey M.A., Michaud, M., VanHuten, J.N., Insogna, K.L., Madri, J.A., Barrett, S.E., Phosphorus-31 MRI of hard and soft solids using quadratic echo line-narrowing, In Proc Natl Acad Sci, USA, 109(14):5190-5195, 2012.


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Kathleen A. Martin, Ph.D. Associate Professor of Medicine (Cardiovascular Medicine) and Pharmacology

1. Overall Goal(s) of the Research Program of the Laboratory: The primary goals of the Martin lab are to understand the signaling mechanisms that regulate VSMC phenotype in order to suggest therapeutic strategies for intimal hyperplasia, graft arteriosclerosis, atherosclerosis, and hypertension. 2. Specific Research Accomplishments in the last 12 months: A major interest in our lab has been signaling through the mTOR pathway in vascular smooth muscle cells, since this is the pathway targeted by the highly effective stent therapeutic rapamycin. We have been examining the role of the cardioprotective adipokine adiponectin in VSMC and have reported that adiponectin induces VSMC differentiation through inhibition of the mTORC1 pathway. Notably, we found that in contrast to rapamycin, adiponectin has no adverse effects on endothelial cells, but promotes endothelial cell function and survival. We additionally reported that adiponectin, traditionally thought to be derived mainly from adipocytes, is also synthesized by VSMC, and that this VSMC-derived adiponectin regulates VSMC phenotype in an autocrine/paracrine manner. We have reviewed our findings and the literature on adiponectin in the vasculature in a comprehensive book chapter. In our close collaboration with Dr. John Hwa in the Department of Cardiology, we have reported that glucose-induced upregulation of aldose reductase leads to increased thromboxane synthesis, thromboxane receptor expression, and platelet activation/aggregation in diabetes. We also assisted the laboratory of Dr. Yibing Qyang in Cardiology in their important studies generating and characterizing smooth muscle cells derived from induce pluripotent stem cells from SVAS and WBS patients. 3. Significance of Key Findings Relevant for the Mission of VBT: The work in our own laboratory has identified new roles for adipokines in the vasculature. Most importantly, we have found for the first time that smooth muscle cells are a source of the protective adipokine adiponectin, as this was previously thought to be produced only by adipocytes. As VBT is a multi-disciplinary and highly collaborative program, we are enthusiastic about our collaborations with the Hwa lab, in which our expertise in signal transduction has helped them in making exciting new discoveries about the effects of glucose on platelets in diabetics and normal subjects. Our expertise in smooth muscle biology has also been helpful in our collaboration with Dr. Qyang’s lab in their cutting edge studies which allowed them to generate an ongoing source of patient-dervied smooth muscle cells in order to study the molecular mechanisms of genetic vascular disease. 4. Publications: (Publications July 1, 2011– June 30, 2012) Ding M, Xie Y, Wagner RJ, Jin Y, Carrao AC, Liu LS, Guzman AK, Powell RJ, Hwa J,

Rzucidlo EM, Martin KA. Adiponectin induces vascular smooth muscle cell differentiation via repression of mTORC1 and FoxO4. Arterioscler Thromb Vasc Biol. 2011 1;31(6):1403-1410.

Tang WH, Stitham J, Gleim S, Febbo C.D., Porreca E., Fava C, Tacconelli S, Capone M, Evangelista V, Levantesi G, Wen L., Martin KA, Minuz P, Rade J, Patrignani P, Hwa J. Glucose and collagen regulate platelet activity through aldose reductase induction of thromboxane; implications for diabetes mellitus. J Clin Invest. 2011 1:1212(11):4462-4476.


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Ding M*, Carraro AC*, Wagner RJ, Xie Y, Rzucidlo EM, Li W, Tellides G, Hwa J,

Aprahamian TR, Martin KA. Vascular smooth muscle cell-derived adiponectin: a Paracrine regulator of contractile phenotype. J. Mol. Cell. Cardiol. 2012; 52(2):474-484. *Denotes equal contribution.

*Ge X, *Ren Y, *Bartulos O, Lee MY, Yue Z, Kim K-Y, Li W, Amos PJ, Bozkulak EC, Iyer A, Zheng W, Zhao H, Martin KA, Kotton DN, Tellides G, Park I-H, Yue L, Qyang Y. Modeling Supravalvular Aortic Stenosis Syndrome using human induced pluripotent stem cells. Circulation, in press.

Midgett C, Stitham J, Martin KA, Hwa J. Prostacyclin receptor regulation—from transcription to trafficking. Current Molecular Medicine. 2011, 11(7):517-528

Stitham J, Midgett C, Martin KA, Hwa J. Prostacyclin: An inflammatory paradox. Frontiers in Pharmacotherapy of Inflammation, 2011, 2:24.

Gleim S, Stitham J, Tang WH, Martin KA, Hwa J. An eicosanoid-centric view of atherothrombotic risk factors. Cellular and Molecular Life Sciences, published online April 11, 2012.

Tang, WH, Martin KA, Hwa J. Aldose reductase, oxidative stress and diabetes mellitus. Frontiers in Experimental Pharmacology and Drug Discovery, 2012, 3:87.

*Ding M, *Rzucidlo EM, Davey J, Xie Y, Liu R, Jin Y, Stavola L, and Martin KA.“Adiponectin in the heart and vascular system” Chapter 11 in Volume 90, Vitamins and Hormones, Academic Press/Elsevier, Gerry Litwack, editor, in press. *Denotes equal contribution.


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Wang Min, Ph.D.

Associate Professor of Pathology

1. Overall Goal(s) of the Research Program of the Laboratory: Concise description of research program(s): Understanding of the fundamental molecular mechanisms for vasculogenesis, arteriogenesis and angiogenesis may lead to improved therapeutic strategies for treatment of vascular diseases. The goal in my lab is to dissect the signaling pathways in vasculature involved in vascular development, remodeling and repair related to human diseases such as vascular malformation, atherosclerosis, stroke, graft transplant rejection and tumor metastasis. In the past ten years, my lab has extensively employed biochemical, cell biological and mouse genetic approaches to define the critical molecules mediating vascular development, remodeling and repair. 2. Specific Research Accomplishments in the last 12 months: 1. Angiogenesis, lymphangiogenesis and vascular diseases. One of the focuses in the lab is to understand how VEGFRs signaling is regulated in vitro and during pathogenesis. We have summarized recent progresses on the role of lymphangiogenesis in cardiovascular diseases (Jones and Min, 2011). We have helped Dr. Ruddle’s lab in their work on the ProxTom mouse model for visualizing lymphatics in vivo (Truman et al, 2012). We are the first one to report that VEGFR3 is regulated by a microRNA MiR-1236 (Jones et al, 2012). We have previously shown that Bmx, a bone marrow non-receptor tyrosine kinase, mediates ischemia-mediated arterioegenesis and angiogenesis by regulating VEGFR2-dependent signaling (He, Y. et al., 2006 J. Clin Invest), and mediates inflammation-dependent lymphangiogenesis by regulating VEGFR3 signaling (Jones 2010, ATVB). Our recent studies suggest that Bmx plays a role in tumor progression and metastasis (Holopainen et al, 2012). We have also explored a potential therapeutic approach to treat retinopathy by inhibiting angiogenesis (Liang et al., 2012). 2. AIP1 in Graft arteriosclerosis models: Graft arteriosclerosis (GA) is the major cause of late allograft failure. Pathologic features include arterial intimal hyperplasia due to recruitment and proliferation of VSMC, which eventually causes luminal obstruction and allograft ischemia. We have established and characterized two mouse artery transplant models - a single minor histocompatibility antigen (male to female)-dependent aorta transplantation model and IFN--induced syngeneic graft transplantation model. We have demonstrated, for the first time, IFN- is a critical mediator in mouse GA models. These two models will offer us great tools to define role of a specific gene involved in GA. As an example, we have defined the role of AIP1 in these two newly established models. AIP1 (also called DAB2IP), a novel member of the Ras GTPase-activating protein family, was identified as apoptosis signal-regulating kinase 1(ASK1)-interacting protein in our lab in 2003. We have extensively investigated the role of AIP1 in vascular system. Our recent study has demonstrated that AIP1 functions as a negative regulator in IFN--induced intimal formation, in part, by downregulating IFN--JAK2-STAT1/3–dependent migratory and proliferative signaling in vascular smooth muscle cells (Yu, L. et al., 2011). We have summarized our model and studies in an invited review article (Min and Pober, 2011). We have also demonstrated that AIP1 regulates TNFR2 function (Ji et al., 2012). Our studies have defined AIP1 as a potential therapeutic target for the prevention of GA and other vascular diseases. 4. Publications: (Publications July 1, 2011– June 30, 2012) Jones, D., and Min, W. (2011) An overview of lymphatic vessels and their merging role in

cardiovascular disease (Review) J Cardiovasc. Dis. Res. 2)3):141-52. PMCID: PMC3195192.


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Truman LA, Bentley KL, Smith EC, Massaro SA, Gonzalez DG, Haberman AM, Hill M,

Jones D, Min W, Krause DS, Ruddle NH. (2012) ProxTom lymphatic vessel reporter mice reveal prox1 expression in the adrenal medullar, megakaryocytes, and platelets. Am J Pathol. 180(4):1715-25. PMCID: PMC3349900.

Jones, D, Li, Y, He, Y, Xu, Z, Chen, H, Min W*. (2012) Mirtron microRNA-1236 inhibits VEGFR-3 signaling during inflammatory lymphangiogenesis. Arterioscler Thromb Vasc Biol. 32(3):633-42. PMCID: PMC32288963.

Holopainen, T., Alpuche, VL., Zheng, W., Heljasvaara, R., Jones, D., He, Y., Tvorogov, D., D’Amico, G., Wiener, Z., Andersson, LC., Pihlajaniemi, T., Min, W., and Alitalo, K. (2012) Deletion of the endothelial Bmx tyrosine kinase decreases tumor angiogenesis and growth. Cancer Res. 72(14):3512-3521.

Liang, XL., Zhou, H., Ding, Y., Yang, C., He, Y., Xu, Z., Luo, Y., Li, S., Sun, G., Liao, X., Min, W. (2012) TMP prevents retinal neovascularization and imparts neuroprotection in an oxygen-induced retinopathy model. Invest Ophthalmol Vis Sci. 53(4):2157-69.

Yu, L., Qin, L., Zhang, H., He, Y., Chen, H., Pober, J.S., Tellides, G., Min, W*. (2011) AIP1 prevents arteriosclerosis by inhibiting IFN-γ-dependent smooth muscle cell proliferation and intimal expansion. Cir Res. 109(4):418-27. PMCID: PMC3227522

Min, W*., and Pober, JS (2011) AIP1 in Graft Arteriosclerosis (Review). Trends Cardiovascular Med. 21(8):229-33.

Ji, W., Li, Y., Wan, T., Wang, J., Zhang, H., Chen, H., and Min, W*. (2012) Both internalization and AIP1 association are required for TNFR2-mediated JNK Signaling. Arterioscler Thromb Vasc Biol. 32(9):2271-9


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Laura E Niklason MD, PhD. Professor & Vice-Chair, Departments of Anesthesia & Biomedical Engineering

1. Overall Goal(s) of the Research Program of the Laboratory: Dr. Niklason’s research program focuses on cardiovascular and lung tissue engineering, utilization of stem cells for tissue regeneration, and on mechanical characteristics of native and engineered vascular structures. 2. Specific Research Accomplishments in the last 12 months: Niklason’s group has continued to work on paradigms for allogeneic vascular engineering. Specifically, we have identified alternative synthetic polymer scaffolds that permit robust vascular cell proliferation and matrix deposition, while exhibiting rapid degradation so as to minimize the impact of polymer fragments on final tissue mechanics. In addition, we have worked to further refine conditions by which mesenchymal stem cells – both adult and human iPS-derived – are stimulated to differentiate into vascular smooth muscle cells. Using human iPS cells, we have begun to translate differentiation strategies to this adult-derived stem cell type to produce human iPS-derived vessels in vitro. Lastly, we are working to delineate the impact of discrete bi-axial physical forces on collagen and elastin deposition and on tissue mechanics. With respect to lung regeneration, we have advanced our approaches for cellular repopulation of acellular lung scaffolds, and are beginning to study the impact of scaffold components on epithelial differentiation. We have completed proteomics studies of lung matrix components, in collaboration with an investigator at the University of Colorado Boulder. Working with human iPS cells, we have identified means of producing highly pure populations of lung epithelium expressing multiple markers of type II alveolar cells, which are a resident progenitor cell in adult and neonatal lung. Seeding of these cells onto acellular lung scaffolds has resulted in maintenance of epithelial phenotype and some anatomic preference for airway vs. alveolar repopulation, depending upon the mileur of soluble factors used in culture. In addition, we have developed strategies to isolate and purify type II alveolar epithelial cells from rodents, and have used these cells to trace differentiation events of cells seeded inside lung matrices. 3. Significance of Key Findings Relevant for the Mission of VBT: Related to the mission of VBT, our work in vascular regeneration is continuing to define the impact of physical forces on vessel growth, remodeling and mechanics. This work particularly pertains to arterial development and healing responses after injury or surgery. Our recent success in modulating elastin synthesis in vitro, using either micro-RNA antagonism or using bi-axial physical forces, will allow us to tease apart the contributions of elastin and collagen, as distinct from smooth muscle cells, in vascular mechanical behavior. Members of VBT with whom we have published in the last calendar year include Hirschi, Dardik, Giordano, Tellides, Humphrey and Sessa. 4. Publications: (Publications July 1, 2011– June 30, 2012) Correia, C., Grayson, W.L., Park,M., Hutton, D., Zhou, B., Guo, X.E., Niklason, L., Sousa,

R.A., Reis, R.L., Vunjak-Novakovic, G., “In vitro model of vascularized bone: syntergizing vascular development and osteogenesis”, PLoS One 2011; 6(12):e28352. Epub 2011 Dec 2.

Badylak, S.F., Hirschi, K.K., Niklason, L.E., “Cardiovascular regenerative biology”, Cells Tissues Organs 2012; 195(1-2): 4.


36

Petersen, T.H., Calle, E.A., Colehur, M.B. and Niklason L.E. Matrix composition and

Mechanics of Decellularized lung scaffold. Cells Tissues Organs 2012; 195(3): 222-231. Zhang, P., Huang, A., Ferruzzi, J., Mecham, R.P., Starcher, B.C., Tellides, G., Humphrey,

J.D., Giordano, F.J., Niklason, L.E., Sessa, W.C. “Inhibition of microRNA-29 enhances elastin levels in cells haploinsufficient for elastin and in bioengineered vessels –brief report”, Arterioscler Thromb Vasc Biol 2012; 32(3): 756-759.

Quint, C., Arief, M., Muto, A., Dardik, A., Niklason, L.E., “Allogeneic human tissue-engineered blood vessel”, J Vasc Surg 2012; 55(3): 790-798.

Dahl, S.L., Blum, J.L., Niklason L.E., “Bioengineered vascular grafts: can we make them off-the-shelf?”, Trends Cardiovasc Med 2012 21(3): 83-89.

Sundaram, S., Niklason, L.E., “Smooth muscle and other cell sources for human blood vessel engineering”, Cells Tissues Organs 2012; 195(1-2): 15-25.

Schriefl, A.J., Collins, M.J., Pierce, D.M., Holzapfel, G.A., Niklason, L.E., Humphrey, J.D. “Remodeling of intramural thrombus and collagen in an Ang-II infusion ApoE-/- model of dissecting aortic aneurysms”, Thromb Res 2012; May 3 In press


37

Jordan S. Pober M.D., Ph.D.

Professor and Vice-Chair, Immunobiology

1. Overall Goal(s) of the Research Program of the Laboratory: The Pober laboratory studies interactions between the human immune and vascular systems. We seek to understand how vascular cells are involved in normal innate and adaptive immune reactions, how the immune system can alter and/or injure blood vessels, and how this knowledge can be applied to human diseases with a particular focus on organ replacement by allografts or engineered tissues. 2. Specific Research Accomplishments in the last 12 months: We have developed new humanized mouse models to explore how human blood vessels can be altered by injury or by therapeutics in a manner that changes the nature of the adaptive immune response to that vessel. 3. Significance of Key Findings Relevant for the Mission of VBT: Research conducted by the Pober lab is focused on developing therapeutic approaches that alter the vasculature in a manner that will prevent or reduce immune-mediated pathologies. 4. Publications: (Publications July 1, 2011– June 30, 2012) Pober JS. Just the FACS or stalking the elusive circulating endothelial progenitor cell.

Arterioscler Thromb Vasc Biol. 2012 Apr;32(4):837-8. Fogal B, Yi T, Wang C, Rao DA, Lebastchi A, Kulkarni S, Tellides G, Pober JS.

Neutralizing IL-6 reduces human arterial allograft rejection by allowing emergence of CD161+ CD4+ regulatory T cells. J Immunol. 2011 Dec 15;187(12):6268-80.

Yi T, Fogal B, Hao Z, Tobiasova Z, Wang C, Rao DA, Al-Lamki RS, Kirkiles-Smith NC, Kulkarni S, Bradley JR, Bothwell AL, Sessa WC, Tellides G, Pober JS. Reperfusion injury intensifies the adaptive human T cell alloresponse in a human-mouse chimeric artery model. Arterioscler Thromb Vasc Biol. 2012 Feb;32(2):353-60.

Pober JS, Tellides G. Participation of blood vessel cells in human adaptive immune responses. Trends Immunol. 2012 Jan;33(1):49-57.

Hibino N, Yi T, Duncan DR, Rathore A, Dean E, Naito Y, Dardik A, Kyriakides T, Madri J, Pober JS, Shinoka T, Breuer CK. A critical role for macrophages in neovessel formation and the development of stenosis in tissue-engineered vascular grafts. FASEB J. 2011 Dec;25(12):4253-63.

Steenblock ER, Fadel T, Labowsky M, Pober JS, Fahmy TM. An artificial antigen-presenting cell with paracrine delivery of IL-2 impacts the magnitude and direction of the T cell response. J Biol Chem. 2011 Oct 7;286(40):34883-92.

Lebastchi AH, Khan SF, Qin L, Li W, Zhou J, Hibino N, Yi T, Rao DA, Pober JS, Tellides G. Transforming growth factor beta expression by human vascular cells inhibits interferon gamma production and arterial media injury by alloreactive memory T cells. Am J Transplant. 2011 Nov;11(11):2332-41.

Yu L, Qin L, Zhang H, He Y, Chen H, Pober JS, Tellides G, Min W. AIP1 prevents graft arteriosclerosis by inhibiting interferon-γ-dependent smooth muscle cell proliferation and intimal expansion. Circ Res. 2011 Aug 5;109(4):418-27.

Tobiasova Z, Zhang L, Yi T, Qin L, Manes TD, Kulkarni S, Lorber MI, Rodriguez FC, Choi JM, Tellides G, Pober JS, Kawikova I, Bothwell AL. Peroxisome proliferator-activated receptor-γ agonists prevent in vivo remodeling of human artery induced by alloreactive T cells. Circulation. 2011 Jul 12;124(2):196-205.

Pober JS. Interleukin-17 and atherosclerotic vascular disease. Arterioscler Thromb Vasc Biol. 2011 Jul;31(7):1465-6.


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Nancy H. Ruddle, Ph.D. Senior Research Scientist and Professor Emerita

1. Overall Goal(s) of the Research Program of the Laboratory: The overall goals of the laboratory are to understand cell trafficking in inflammation and lymphoid organ development and to relate these two seemingly disparate phenomena. Our long-term goal is to understand how the vasculature of lymphoid organs, lymphatic vessels and high endothelial venules (HEVs), controls the immune response. We study HEV regulation in canonical secondary lymphoid organs and “tertiary lymphoid organs” (TLOs), ectopic lymphoid accumulations arising in situations of chronic inflammation in autoimmunity, graft rejection, and microbial infection. We are particularly interested in Type 1 diabetes, multiple sclerosis, and Sjogren’s syndrome. Visualization of HEVs and lymphatic vessels in living mice is providing a better understanding of the dynamics of their interaction in lymph nodes and TLOs with therapeutic potential in autoimmunity and malignancy. 2. Specific Research Accomplishments in the last 12 months: a) Characterization of mice transgenic for human insulin – crossing to MISTERG mice of

Richard Flavell b) Characterization of the TLOs of a new model of experimental autoimmune

encephalomyelitis c) Development of a mouse model of Sjögren’s syndrome d) Characterization of requirements for self antigen presentation in a mouse model of

multiple sclerosis e) Development of a mouse with fluorescent HEVs and lymphatic vessels to study cell

trafficking in real time in the immune system 4. Publications: Motallebsadeh RM, Rehakova S, Conlon T, Win TS, Callaghan CJ, Goddard M, Bolton EM,

Ruddle, NH, Bradley JA, Pettigrew GJ. Blocking lymphotoxin signaling abrogates the development of ectopic lymphoid tissue within cardiac allografts and inhibits effector antibody responses. FASEB J. 26:51-62, 2012. PMID: 21926237.

Liao S, Cheng G, Connor DA, Huang Y, Kucherlapati RS, Munn LL, Ruddle NH, Jain RK, Fukumura, D, Padera TP. Impaired lymphatic contraction associated withimmunosuppression. PNAS108:18784-9, 2011. PMID:22065738.

Truman LA, Bentley K, Smith E, Massaro S, Gonzalez DG, Haberman A, Hill M, Jones D, Min W, Krause D, Ruddle NH. ProxTom lymphatic vessel reporter mice reveal Prox1 expression in the adrenal medulla, megakaryocytes, and platelets. Am. J. Pathol. 180:1715-1725, 2012. PMID: 22310467.

Bergman, CM, Marta, CB, Maric, M, Steven E. Pfeiffer, SE, Cresswell, PE, Ruddle, NH. A switch in pathogenic mechanism in myelin oligodendrocyte glycoprotein-induced EAE in GILT-free mice. J. Immunol. 188:6001-6009, 2012.


39

Mehran M. Sadeghi, M.D.

Associate Professor of Medicine (Cardiology)

1. Overall Goal(s) of the Research Program of the Laboratory: The main goal of our research is to develop novel molecular imaging approaches for cardiovascular diseases, with an emphasis on the vascular system. Vascular remodeling, changes in the vessel geometry and/or composition, is a common feature of a broad spectrum of vasculopathies, from atherosclerosis to graft arteriosclerosis and aneurysm. We have focused on four examples of vascular remodeling, namely injury-induced vascular remodeling, graft arteriosclerosis, aneurysm, and atherosclerosis. For each process studied, we identify specific imaging targets based on pathophysiology or genomic and proteomic screening, develop novel ligands for imaging or use existing radiotracers, establish relevant animal models, and use a dedicated microSPECT/CT small animal imaging system to detect the process in vivo. Studies of the pathophysiology of vascular remodeling, to identify novel targets for imaging and to understand the biology of the targets identified, is an integral part of our research. 2. Specific Research Accomplishments in the last 12 months: Over the past several years, we have demonstrated that MMP-targeted imaging can detect and monitor the effect of therapeutic interventions, including dietary modification and lipid lowering therapies, in a murine model of injury-induced vascular remodeling and atherosclerosis. In a murine model of aneurysm, we demonstrated that MMP-targeted imaging may be used to track the remodeling process in vivo. Using this approach we can predict a murine aneurysm’s propensity to expansion in vivo. Other work focused on imaging protease activation in atherosclerosis, demonstrating the heterogeneity of protease activation along the mouse aorta. Using molecular imaging in conjunction with histology we demonstrated the presence of discordance between anatomy and biology in atherosclerosis. In parallel, we demonstrated established a role for VEGF imaging in tracking vascular remodeling in transplant vasculopathy. Together, these studies established VEGF receptor-, integrin- and protease-targeted molecular imaging for detection of remodeling and inflammation in the vessel wall and set the stage for future clinical studies of molecular imaging to identify patients at high risk for cardiovascular events. 4. Publications: (Publications July 1, 2011 – June 30, 2012) Tavakoli S, Razavian M, Zhang J, Nie L, Marfatia R, Dobrucki DS, Sinusas AJ, Edwards DS,

Robinson S, Sadeghi MM. “Monitoring the Progression of Vascular Remodeling and Response to Dietary Modification by Molecular Imaging of Matrix Metalloproteinase Activation”, Atherosclerosis, Thrombosis and Vascular Biology, 2011, 31:102-109.

Sadeghi MM, et al “Cardiovascular Nuclear Imaging: Balancing Proven Clinical Value and Potential Radiation Risk”. Journal of Nuclear Medicine, 2011, 52:1162-4

Razavian M, Tavakoli S, Zhang J, Nie L, Dobrucki LW, Sinusas AJ, Azure M, Robinson S, Sadeghi MM. “Atherosclerosis Plaque Heterogeneity and Response to Therapy Detected by in vivo Molecular Imaging of Matrix Metalloproteinase Activation”. Journal of Nuclear Medicine, 2011, 52:1795-802.

Razavian M, Marfatia R, Mongue-Din H, Tavakoli S, Sinusas AJ, Zhang J, Nie L, Sadeghi MM. “Integrin-targeted imaging of inflammation in vascular remodeling”, Arterioscler Thromb Vasc Biol. 2011, 31:2820-6.

Fernandez AB, Wong TY, Klein R, Collins D, Burke G, Cotch MF, Klein B, Sadeghi MM, Chen J. “Age-related macular degeneration and incident cardiovascular disease: The multi-ethnic study of atherosclerosis”, Ophthalmology, 2012, 119:765-70.

Zhang J, Razavian M, Tavakoli S, Nie L, Tellides G, Backer JM, Backer MV, Bender JR, Sadeghi MM. “Molecular imaging of VEGF receptors in graft arteriosclerosis”, Arterioscler Thromb Vasc Biol. 2012, 32:1489-55.


40

Martin Schwartz, Ph.D.

Professor of Medicine and Cell Biology

1. Overall Goal(s) of the Research Program of the Laboratory: Our laboratory studies signaling by integrins and mechanotransduction in the vascular system. We are especially interested in how endothelial cells respond to forces from flowing blood. The goals are to understand basic mechanisms of signal transduction, and to apply this information to both atherosclerosis and flow-dependent artery remodeling. 2. Specific Research Accomplishments in the last 12 months: We have made significant advances in understanding the mechanisms of endothelial alignment in fluid flow and identified a major anti-inflammatory mechanism in the flow response. We have also made substantial progress toward elucidating the mechanism by which the junctional complex (PECAM-1, VE-cadherin and VEGFR2) mediates flow signalling. 3. Significance of Key Findings Relevant for the Mission of VBT: These results advance our understanding of basic mechanisms by which endothelial cells respond to fluid shear stress and the downstream pathways by which flow triggers vascular remodeling and atherosclerosis. 4. Publications: (Publications July 1, 2011– June 30, 2012) Hoffman B.D., Grashoff C. and Schwartz MA. (2011) Dynamic molecular processes

mediate cellular mechanotransduction. Nature, 475:316-23. Norambuena A. and Schwartz, MA (2011) Effects of integrin-mediated cell adhesion on

plasma membrane lipid raft components and signaling. Mol. Biol. Cell, 22:3456-64. Hahn C., Chong, W., Orr, A.W., Coon, B.G, Schwartz, M.A. (2011) JNK2 promotes

endothelial cell alignment under flow. Plos One, 6:e24338. Schwartz MA, Simons M., (2012) Lymphatics thrive on stress: mechanical force in

lymphatic development. EMBO J. 31(4):781-2. Batra N, Burra S, Siller-Jackson AJ, Gu S, Xia X, Weber GF, Desimone D, Bonewald LF,

Lafer EM, Sprague E., (2012) Mechanical stress-activated integrin α5β1 induces opening of connexin 43 hemichannels Proc Natl Acad Sci U S A. 109(9):3359-64.

Conway D, Schwartz MA. (2012) Lessons from the endothelial junctional mechanosensory complex. F1000 Biol Rep. 4:1.

Wang C., Schwartz M.A., A novel in vitro flow system for changing flow direction on endothelial cells. J. Biomechanics, 45(7):1212-8.

Yaqoob U., Cao S., Shergill U., Jagavelu K., Geng Z., Yin M., Szabolcs A., Thorgeirsson S., Schwartz M.A., Yang J.D., Ehman R., Roberts L., Mukhopadhay D., Shah V.H. (2012) Neuropilin-1 enhances tumor growth through stimulatory effects on fibronectin fibril assembly within the tumor microenvironment, Cancer Res, in press.

Jhaveri KA., Debnath P., Chernoff J., Sanders J., Schwartz MA., (2012) The role of p21-activated kinase in the initiation of atherosclerosis. BMC Cardiovascular Disorders, in press.

Park MH., Lee HS., Lee CS., You ST., Kim DJ, Jeong YJ., Kang MJ, Lee JH, Do HW., Park BH., Shin EY., Schwartz MA., and Kim EG. (2012) p21-activated kinase 4 promotes prostate cancer progression through CREB. Oncogene, in press.


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William C. Sessa, Ph.D. Alfred Gilman Professor of Pharmacology, Director, VBT Program 1. Overall Goal (s) of the Research Program of the Laboratory: Our laboratory is very interested in endothelial cell biology, signaling and regulation of post-natal angiogenesis/ arteriogenesis and atherosclerosis. 2. Specific accomplishments in the last year: In the past year, we have made successful inroads into several areas of nitric oxide signaling, caveolin function and miRNAs. 3. Publications: Bernatchez, P., Sharma, A., Bauer, P.M., Marin, E. and Sessa, W.C. A noninhibitory mutant

of the caveolin-1 scaffolding domain enhances eNOS-derived NO synthesis and vasodilation in mice. J Clin Invest. 121(9):3747-55 (2011).

Karuppiah, K., Druhan, L.J., Chen, C.A., Smith, T., Zweier, J.L., Sessa, WC and Cardounel, A.J. Suppression of eNOS-derived superoxide by caveolin-1: a biopterin-dependent mechanism. Am J Physiol Heart Circ Physiol. 301(3):H903-11 (2011).

Maclauchlan, S., Yu, J., Parrish, M., Asoulin, T.A., Schleicher, M., Krady, .M.M., Zeng, J., Huang, P.L., Sessa. W.C. and Kyriakides, T.R. Endothelial nitric oxide synthase controls the expression of the angiogenesis inhibitor thrombospondin 2. Proc Natl Acad Sci USA , 108(46):E1137-45 (2011).

Zhang, P., Huang, A., Ferruzzi, J., Mecham, R.P., Starcher, B.C., Tellides, G.,Humphrey, J.D., Giordano, F.J., Niklason, L.E. and Sessa, W.C. Inhibition of MicroRNA-29 enhances elastin levels in cells haploinsufficient for elastin and in bioengineered vessels. Arterioscler Thromb Vasc Biol. 32(3):756-9 (2012).

Yu. J., Zhang, Y., Zhang, X., Rudic, R.D., Bauer, P.M., Altieri, D.C. and Sessa, W.C. Endothelium Derived Nitric Oxide Synthase Negatively Regulates the PDGF-Survivin Pathway during Flow-Dependent Vascular Remodeling. PLoS One. 7(2):e31495 (2012).

Sangwung, P., Greco, T.M., Wang, Y., Ischiropoulos, H., Sessa, W.C. and Iwakiri Y. Proteomic Identification of S-Nitrosylated Golgi Proteins: New Insights into Endothelial Cell Regulation by eNOS-Derived NO. PLoS One. 7(2):e31564 (2012).

Marin, E.P., Derakhshan, B., Lam, T.T., Davalos, A. and Sessa, W.C. Endothelial cell palmitoylproteomics identify novel lipid-modified targets and potential substrates for protein acyl transferases. Circulation Research,110(10):1336-44 (2012).

VASCULAR BIOLOGY AND TRANSPLANTATION ANNUAL REPORT 2011 – 2012

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Michael Simons, M.D. RW Berliner Professor of Medicine & Cell Biology, Chief, Section of Cardiovascular Medicine 1. Overall Goals of the Research Program of the Laboratory Our laboratory is interested in regulation of arterial and lymphatic morphogenesis and angiogenic growth factor signaling. These process are investigated at all levels, including in vitro signaling studies, in vivo mouse transgenic and knock-out models and translational studies in larger animal models and early phase clinical trials. 2. Specific Research Accomplishments in the last 12 months We have made significant advances in unraveling the impact of FGF signaling on VE-cadherin-dependent regulation of vascular integrity and in the role of receptor trafficking in control of FGFR1 signaling. 3. Significance of Key Findings Relevant for the Mission of VBT These findings advance our knowledge of molecular details of regulation of vascular development and signaling and should eventually enable the development of new therapeutic paradigms. 4. Key Publications Larrivee B, Prahst C, Gordon E, Del Toro R, Mathivet T, Simons M, Eichmann, A. Alk1

signaling inhibits angiogenesis by cooperating with the Notch pathway. Dev Cell 2012; 22:489-500.

Elfenbein A, Lanahan AA, Zhou TX, Yamasaki A, Tkachenko E, Matsuda M, Simons M. Syndecan 4 regulates endothelial FGFR1 signaling by directing macropinocytosis. Science Signaling 2012; 5:ra36.

Christoffersson G, Zang G, Zhuang ZW, Vågesjö E, Simons M, Phillipson M, Welsh M. Vascular adaptation to a dysfunctional endothelium as a consequence of Shb deficiency. Angiogenesis 2012; 15:469-80.

Hatanaka K, Lanahan AA, Murakami M, Simons M. Fibroblast Growth Factor signaling potentiates VE-cadherin stability at adherens junctions by regulating SHP2. PLOS One 2012;7:337600,


43

Albert J. Sinusas, M.D.

Professor of Medicine (Cardiology) and Diagnostic Radiology; Director, Cardiovascular Nuclear Imaging and Stress Laboratories; Director, Animal Research Laboratories-Section of Cardiovascular Medicine; Director, Yale Translational Research Imaging Center (Y-TRIC)

1. Overall Goal(s) of the Research Program of the Laboratory: Research in the Sinusas laboratory is directed at development of noninvasive imaging approaches for the assessment of myocardial viability, angiogenesis, arteriogenesis, and post-infarction remodeling. The laboratory has been employing the 3-D modalities of single photon emission computed tomography (SPECT), positron emission tomography (PET), echocardiography, X-ray tomography, and magnetic resonance (MR) imaging for assessment of a wide range of physiological and molecular processes primarily focused in the cardiovascular system. The laboratory is currently focused on targeted molecular imaging, developing non-invasive nuclear imaging strategies for identifying the hypoxic stimulus for angiogenesis, and targeted imaging of selected integrins previously established to modulate the angiogenic process, and the interrelationship of angiogenesis and arteriogenesis. These studies involve the use of rodent and large animal models of myocardial ischemia as well as hindlimb ischemia. 2. Specific Research Accomplishments in the last 12 months: Application of quantitative tools for automated physiologically based segmentation of the coronary and peripheral vasculature. Development of CT and MR approached for evaluation of tissue oxygenation and arteriogenesis. Development of novel multimodality nanoparticle-based contrast agents. 3. Significance of Key Findings Relevant for the Mission of VBT: Demonstrated the phenomenon of augmented CT attenuation due to nano-confinement with the development of novel CT containing multimodality imaging agents. This development has significant implications for targeted molecular CT-based imaging of the vascular tree. 3. Publications: (Publications July 1, 2011– June 30, 2012) Sinusas AJ, Thomas JD, Mills G. The future of molecular imaging. JACC Cardiovasc Imaging.

2011 Jul:4(7): 799-806. PMID:21757172 Pearlman, P.C., Tagare, H.D., Lin, B.A., Sinusas, A.J. & Duncan, J.S. (2011). Segmentation of 3D

RF echocardiography using a multiframe spatio-temporal predictor. Proceedings of Information Processing in Medical Imaging 2011, Lecture Notes in Computer Science 6801:37-48.

Compas, C.B., Lin, B.A., Sampath, S., Jia, C., Wei, Q., Sinusas, A.J. & Duncan, J.S. (2011). Multi-frame radial basis functions to combine shape and speckle tracking for cardiac deformation analysis in echocardiography. Proceedings of Functional Imaging and Modeling of the Heart 2011, Lecture Notes in Computer Science 6666:113-120.

Criscione JM, Sinusas AJ, Fahmy TM. Multimodaltiy/fusion imaging toward imaging of structure and function. Chapter In Stem Cell Labeling for Delivery and Tracking Using Noninvasive Imaging. Edited by Kraitchman DL, and Wu J. CRC Press, London, UK, September 2011 (ISBN: 978-1-4398075-1-4)

Jiang Y, Zhuang ZW, Sinusas AJ, Staib LH and Papademetris X. Vessel connectivity using Murray's hypothesis. Medical image computing and computer-assisted intervention : MICCAI International Conference on Medical Image Computing and Computer-Assisted Intervention. 2011;14(Pt 3):528-36. PMID: 22003740.


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Razavian M, Marfatia R, Mongue-Din H, Tavakoli S, Sinusas AJ, Zhang J, Nie L, Sadeghi MM. Integrin-targeted imaging of inflammation in vascular remodeling. Arterioscler Thromb Vasc Biol. 2011 (12):2820-6. PMID:21940943

Razavian M, Tavakoli S, Zhang J, Nie L, Dobrucki LW, Sinusas AJ, Azure M, Robinson S, Sadeghi MM. Atherosclerosis plaque heterogeneity and response to therapy detected by in vivo molecular imaging of matrix metalloproteinase activation. J Nucl Med. 2011 52(11):1795-802. PMID:21969358

C. B. Compas, B. A. Lin, S. Sampath, C. X. Jia, Q. Wei, A. J. Sinusas, and J. S. Duncan, Combining shape and speckle tracking for deformation analysis in echocardiography using radial basis functions, IEEE International Symposium on Biomedical Imaging: From Nano to Macro (ISBI), 1322–1325, 2011.

C. B. Compas, B. A. Lin, S. Sampath, L. Huang, Q.Wei, A. J. Sinusas, and J. S. Duncan. Comparing Shape Tracking, Speckle Tracking, and a Combined Method for Deformation Analysis in Echocardiography, Healthcare Informatics, Imaging, and Systems Biology (HISB), San Jose, pp. 120-125, 2011.

Criscione JM, Dobrucki LW, Zhuang ZW, Papademetris X, Simons M, Sinusas AJ, Fahmy TM. Development and application of a multimodal contrast agent for SPECT/CT hybrid imaging. Bioconjug Chem. 2011 21:22(9):1784-92. PMID:21851119

Pearlman PC, Tagare HD, Bin BA, Sinusas AJ, Duncan JS. Segmentation of 3D RF echocardiography using a multiframe spatio-temporal predictor. Inf Process Med Imaging. 2011:22:37-48. PMID:21761644

P.C. Pearlman, H.D. Tagare, B.A. Lin, A. J. Sinusas, and J. S. Duncan. Segmentation of 3D radio frequency echocardiography using a spatio-temporal predictor. Medical Image Analysis 16, no. 2, pp. 351-360, 2012.

X. Huang, B. A. Lin, C. B. Compas, A. J. Sinusas, L. H. Staib, and J. S. Duncan. Segmentation of left ventricles from echocardiographic sequences via sparse appearance representation. IEEE Workshop on Mathematical Methods in Biomedical Image Analysis (MMBIA), pages 305-312, 2012.

X. Huang, D. P. Dione, C. B. Compas, X. Papademetris, B. Lin, A. J. Sinusas, and J. S. Duncan. A Dynamical Appearance Model Based on Multiscale Sparse Representation: Segmentation of the Left Ventricle from 4D Echocardiography. In Medical Image Computing and Computer-Assisted Intervention (MICCAI), 2012. (in press).

C. B. Compas, E. Y.Wong, X. Huang, S. Sampath, B. A. Lin, X. Papademetris, K. Thiele, D. P. Dione, A. J. Sinusas, M. O’Donnell, and J. S. Duncan. A Combined Shape Tracking and Speckle Tracking Approach for 4D Deformation Analysis in Echocardiography. In IEEE International Symposium on Biomedical Imaging (ISBI), pages 458-461, 2012.

Liu H, Hu H, Sinusas AJ, Shi P. An H() approach for elasticity properties reconstruction. Med Phys. 2012 39(1):475-81. PMID:22225318.

Stacy MR, Maxfield MW, Sinusas AJ. Targeted molecular imaging of angiogenesis in PET and SPECT: a review. Yale J Biol Med. 2012 85(1):75-86. PMID:22461745.

Suh JW, Kwon OK, Scheinost D, Sinusas AJ, Cline GW and Papademetris X. CT-PET weighted image fusion for separately scanned whole body rat. Medical physics. 2012;39(1):533-42. PMID: 22225323; PMCID: PMC3266828

Hedhli N, Dobrucki LW, Kalinowski A. Zhuang ZW, Wu X, Russell RR 3rd, Sinusas AJ, Russell KS. Endothelial-deried neuregulin is an important mediator of ischaemia-induced angiogenesis and arteriogenesis. Cardiovasc Res. 2012 1:93(3):516-24. PMID:22003740.


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Edward L. Snyder, M.D. Professor Laboratory Medicine, Director, Apheresis/Cell Processing VBT Core Facility (Core D) 1. Overall Goal(s) of the Research Program of the Laboratory: The Apheresis/Cell Processing Core Facility played a critical role in the Vascular Biology and Transplantation Program. The Cell Processing Core Laboratory is designed to support the needs of the VBT Program users who are performing basic science and clinical research involving mononuclear and other cell types, by providing five specific functions. First, the Apheresis section of the Cell Processing Core Laboratory procured and provided normal donor specimens in support of research projects. These samples, obtained under IRB approved protocols from fresh specimens, were made available to VBT membership. Second the VBT Core D Cell Collection and Processing Laboratory provided, as requested, cell purification services. Third, the Cell Processing Core provided large-scale processing capabilities in support of specific research studies involving human MNCs as well as CD34 positive and other cell types. Included within this section was the development of cell selection and culturing techniques to support the novel cell therapy protocols, as well as the pre-clinical validation of research procedures. The VBT Core resource provided the critical instrumentation and technical expertise in cell processing and cryopreservation, needed for the in vitro use of cells, or infusion of cells into animals. Fourth, the Core provided collections of MNC and could as, and if, needed, provide CD34+ cells from G-CSF stimulated donors. Fifth, the Apheresis/ Cell Processing VBT Core Facility maintained compliance with institutional, NIH, FDA, AABB and FACT guidelines, and ensured that the protocols were safely and effectively applied. Included with this objective was training of new investigators in Compliance and Quality Control issues. Thus, this resource provided access to cell collection, selection, processing and culturing technologies, as well as services and scientific consultation to enhance the productivity of the VBT members. This technically sophisticated resource was critical to the Vascular Biology and Transplantation Section’s research progress. 2. Specific Accomplishments in the last 12 months: In 2011 - 2012, Core D performed 32 MNC apheresis collections for Program Leaders’ research. 4. Publications: Cooper DL, Pratt K, Medoff E, Conkling-Walsh A, Foss F, Snyder E, Yen W. Seropian SE.

Late afternoon dosing of plerixafor for stem cell mobilization: a practical solution. Clinical lymphoma, myeloma & leukemia 2011;11:267-72

Choate J, Snyder EL. The rise of cellular therapy. Transfusion & Apheresis Science 2011;45:91-7

Hibino, N, Nalbandian A, Devine L, Martinez RS, McGillicuddy E, Yi T, Karandish S, Ortolano GA, Shinoka T, Snyder E, Breuer CK. Comparision of human bone marrow mononuclear cell isolation methods for creating tissue-engineered vascular grafts: novel filter system versus traditional density centrifugation method. Tissue Engineering – Part C: Methods 2011;17:993-8

Majhail NS, Murphy EA, Denzen EM, Ferguson SS, Anasetti C, Bracey A. Burns L, Champlin R, Hubbard N, Markowitz M, Maziarz RT, Medoff E, Neumann J, Schmit-Pokorny K, Weisdorf DJ, Yolin Raley DS, Chell J, Snyder EL. The National Marrow Donor Program’s Symposium on Hematopoietic Cell Transplantation in 2020: a health care resource and infrastructure assessment. Biology of Blood & Marrow Transplantation 2012;18:172-82


46

Bing Su, Ph.D.

Associate Professor of Immunobiology

1. Overall Goal(s) of the Research Program of the Laboratory: The overall goals of the research program of this laboratory are to understand the biology of signal transduction mediated by the mitogen-activated protein kinase (MAPK) pathways, and the mammalian target of rapamycin (mTOR) pathway. We study these two signal transduction pathways using combined approaches of mouse genetics, biochemistry and molecular biology. We specifically investigate the function and regulation of the MEKK2/3-MAPK signals, and the mTOR complex 2 pathway in the immune system and vascular system. 2. Specific Research Accomplishments in the last 12 months: In the past 12 months, we have studied the roles of MEKK2 and MEKK3 in T cell development, survival and effector differentiation. We have revealed critical functions of MEKK2 and MEKK3 in T cell homeostasis, cytokine production, and development. We have also studied the role of MEKK3 in endothelial cells in regulating the brain and retinal angiogenesis during development. We demonstrate that MEKK3 plays an essential role in these processes. In addition, we also investigate the role of Sin1-mTORC2 in embryonic angiogenesis and development. We also identified a critical role of Sin1-mTORC2 in B cell development and survival. Finally, we show that by inhibiting both the molecular chaperone pathway and the mTORC2 signal, we may be able to more efficiently inhibit tumor cell growth and survival. Both the MAPK pathways and the mTOR pathway control numerous physiological and pathologic processes ranging from cell growth, stress-responses, aging, survival, to diabetes, autoimmunity and cancer, it is important to understand their roles in these processes, especially by focusing on their roles in the vascular system and in immune responses. Our findings from studying both the MEKK3 and Sin1-mTORC2-Akt pathway are relevant for the mission of VBT. 4. Publications Xing Chang, Fang Liu, Xiaofang Wang, and Su B. MEKK2 and MEKK3 regulate TGF-b-

mediated helper T cell differentiation. Immunity. 34: 201-212, 2011. You-Tong Wu, Weiming Ouyang, Adam S Lazorchak, Dou Liu, Han-Ming Shen, and Su B.

mTOR Complex 2 Targets Akt for Proteasomal Degradation via Phosphorylation at the Hydrophobic Motif. J Biol Chem. 286: 14190-14198, 2011.

Xiaofang Wang, Fan Zhang, Fanping Chen, Dou Liu, Yi Zheng, Yongliang Zhang, Chen Dong, and Su B. MEKK3 regulates IFNg production in T cells through the Rac1/2 dependent MAPK cascades. J. Immunol. 186: 5791-5800, 2011.

Xiaoqing Gan, Jiyong Wang, Su B, Dianqing Wu. Evidence for direct activation of mTORC2 kinase activity by phosphatidylinositol 3,4,5-trisphosphate. J Biol Chem. 286: 10998-11002, 2011.

Adam S. Lazorchak, Su B. Perspectives on the role of mTORC2 in B lymphocyte development, immunity and tumorigenesis. Protein & Cell 2011, 2(7): 523–530.

Su B and Estela Jacinto. Mammalian TOR signaling to the AGC kinases. Crit Rev Biochem Mol Biol. 2011 Dec;46(6):527-47.

Ajibade AA, Wang Q, Cui J, Zou J, Xia X, Wang M, Tong Y, Hui W, Liu D, Su B, Wang HY, Wang RF. TAK1 negatively regulates NF-κB and p38 MAP kinase activation in Gr-1+CD11b+ neutrophils. Immunity. 2012 36(1):43-54. PMID:22226633.


47

Xing Chang, Adam S. Lazorchak, Dou Liu and Su B. Sin1 regulates Treg-cell development

but is not required for T-cell growth and proliferation. Eur J Immunol. 2012 42(6):1639-47. PMID: 22678916.

Fan Zhang, Adam Lazorchak, Dou Liu, Fanping Chen, Su B. Inhibition of the mTORC2 and chaperone pathways to treat leukemia.. Blood. 2012 119(25):6080-8. PMID:22566604

Al-Tawashi A, Jung SY, Liu D, Su B, Qin J. Protein implicated in nonsyndromic mental retardation regulates protein kinase A (PKA) activity. J Biol Chem. 2012, 287(18):14644-58. PMID:22375002.


48

George Tellides, M.D., Ph.D.

Professor of Surgery (Cardiothoracic)

1. Overall Goal(s) of the Research Program of the Laboratory: Our research program encompasses smooth muscle cell biology as related to vascular disease. Our primary research interest is immune-mediated vascular remodeling focusing on the effects of T cells and their products on vascular smooth muscle cell proliferation, apoptosis, and inflammatory responses and conversely on the regulation of artery-infiltrating T cell immune responses by vascular smooth muscle cells. 2. Specific Research Accomplishments in the last 12 months: We have further dissected the cytokine network between vascular cells and leukocytes contributing to arteriosclerosis. 3. Significance of Key Findings Relevant for the Mission of VBT: Participation in multiple collaborative research projects. 4. Publications: (Publications July 1, 2011– June 30, 2012) Tobiasova Z, Zhang L, Yi T, Qin L, Manes TD, Kulkarni S, Lorber MI, Rodriguez FC, Choi

JM, Tellides G, Pober JS, Kawikova I, Bothwell AL. Peroxisome Proliferator-Activated Receptor-gamma Agonists Prevent In Vivo Remodeling of Human Artery Induced by Alloreactive T Cells. Circulation. 2011;124:196-205.

Yu L, Qin L, Zhang H, He Y, Chen H, Pober JS, Tellides G, Min W. AIP1 Prevents Graft Arteriosclerosis by Inhibiting Interferon-y-Dependent Smooth Muscle Cell Proliferation and Intimal Expansion. Circ Res. 2011;109:418-427.

Lebastchi AH, Khan SF, Qin L, Li W, Zhou J, Hibino N, Yi T, Rao DA, Pober JS, Tellides G. Transforming growth factor Beta expression by human vascular cells inhibits interferon gamma production and arterial media injury by alloreactive memory T cells. Am J Transplant. 2011;11:2332-2341.

Lebastchi AH, Qin L, Khan SF, Zhou J, Geirsson A, Kim RW, Li W, Tellides G. Activation of human vascular cells decreases their expression of transforming growth factor-beta. Atherosclerosis. 2011;219:417-424.

Fogal B, Yi T, Wang C, Rao DA, Lebastchi A, Kulkarni S, Tellides G, Pober JS. Neutralizing IL-6 Reduces Human Arterial Allograft Rejection by Allowing Emergence of CD161+ CD4+ Regulatory T Cells. J Immunol. 2011;187:6268-6280.

Ding M, Carrão AC, Wagner RJ, Xie Y, Jin Y, Rzucidlo EM, Yu J, Li W, Tellides G, Hwa J, Aprahamian TR, Martin KA. Vascular smooth muscle cell-derived adiponectin: A paracrine regulator of contractile phenotype. J Mol Cell Cardiol. 2012;52:474-484.

Yi T, Fogal B, Hao Z, Tobiasova Z, Wang C, Rao DA, Al-Lamki RS, Kirkiles-Smith NC, Kulkarni S, Bradley JR, Bothwell AL, Sessa WC, Tellides G, Pober JS. Reperfusion Injury Intensifies the Adaptive Human T Cell Alloresponse in a Human-Mouse Chimeric Artery Model. Arterioscler Thromb Vasc Biol. 2012;32:353-360.

Gallo A, Saad A, Ali R, Dardik A, Tellides G, Geirsson A. Circulating interferon-γ-inducible Cys-X-Cys chemokine receptor 3 ligands are elevated in humans with aortic aneurysms and Cys-X-Cys chemokine receptor 3 is necessary for aneurysm formation in mice. J Thorac Cardiovasc Surg. 2012;143:704-10.

Zhang P, Huang A, Ferruzzi J, Mecham RP, Starcher BC, Tellides G, Humphrey JD, Giordano FJ, Niklason LE, Sessa WC. Inhibition of MicroRNA-29 Enhances Elastin Levels in Cells Haploinsufficient for Elastin and in Bioengineered Vessels--Brief Report. Arterioscler Thromb Vasc Biol. 2012;32:756-9.


49

Sucher R, Fischler K, Oberhuber R, Kronberger I, Margreiter C, Ollinger R, Schneeberger S,

Fuchs D, Werner ER, Watschinger K, Zelger B, Tellides G, Pilat N, Pratschke J, Margreiter R, Wekerle T, Brandacher G. IDO and Regulatory T Cell Support Are Critical for Cytotoxic T Lymphocyte-Associated Ag-4 Ig-Mediated Long-Term Solid Organ Allograft Survival. J Immunol. 2012;188:37-46.

Ali R, Huang Y, Maher SE, Kim RW, Giordano FJ, Tellides G, Geirsson A. miR-1 mediated suppression of Sorcin regulates myocardial contractility through modulation of Ca(2+) signaling. J Mol Cell Cardiol. 2012;52:1027-37.

Zhang J, Razavian M, Tavakoli S, Nie L, Tellides G, Backer JM, Backer MV, Bender JR, Sadeghi MM. Molecular imaging of vascular endothelial growth factor receptors in graft arteriosclerosis. Molecular imaging of vascular endothelial growth factor receptors in graft arteriosclerosis. Arterioscler Thromb Vasc Biol. 2012;32:1849-55.

Pober JS, Tellides G. Participation of blood vessel cells in human adaptive immune responses. Trends Immunol. 2012;33:49-57.


50

Agnès Vignery, D.D.S., Ph.D. Associate Professor of Orthopaedics and Cell Biology 1. Overall Goals of our Research Program Research in our laboratory focuses on two lines of investigation, the commonality of which is osteoporosis, a disease that is at the cross road of the immune, vascular and nervous systems. The first line of investigation regards the differentiation of osteoclasts, which resorb bone, and giant cells, which resorb foreign bodies, with particular emphasis on the molecular mechanisms of fusion of their mononucleate precursor cells that belong to the monocyte-macrophage lineage. Macrophages are mononucleate cells that seed all tissues and can fuse with themselves to differentiate into multinucleate osteoclasts, in bone, or giant cells, in chronic inflammatory reactions and cancer. Although osteoclasts and giant cells play a central role in these diseases, the molecular mechanisms that are responsible for the fusion of macrophages remain poorly understood. The second research focus is the targeted induction of new bone to specific sites of the skeleton. 2. Specific Research Accomplishments in the last 12 months 2011 Board member, The Journal of Immunology 2011 Session Leader and Speaker at the third Gordon Research Conference on Cell-Cell Fusion 3. Significance of Key Findings Relevant for the Mission of VBT To initiate the formation of intramembranous bone, in contrast with endochondral bone, requires extensive vascularization. Our finding that new bone forms in targeted sites of the skeleton as a result of marrow ablation and daily treatment with parathyroid hormone has revealed an essential link between the formation of new bone and vascularization, which we are actively exploring. In addition, our long-term collaboration with Dr Richard Flavell has led to the publication of an article in PNAS. 4. Publications Zhang Q, Carlson J, Gilligan J, Mehta N, Vignery A. Aging potentiates the site-specific

formation of new bone after marrow ablation and treatment with PTH in rats. Submitted Jin C, Frayssinet P,Pelker R, Cwirka D, Hu B, Vignery A, Eisenbarth SC, Flavell RA. The

NLRP3 inflammasome plays a critical role in the pathogenesis of hydroxyapatite- associated arthropathy" 2011 Proc Natl Acad Sci U S A. 2011 108(36):14867-72.


51

Dianqing (Dan) Wu, Ph.D.

Professor of Pharmacology

1. Overall Goal(s) of the Research Program of the Laboratory: The overall objective of our research activities is to understand the mechanisms and functions of signal transduction activated by chemoattractants and Wnts. 2. Specific Research Accomplishments in the last 12 months: The key research accomplishment includes: 1) Characterization of a signaling pathway that regulate actin reorganization in neutrophils and elucidation of a key role of the interactions of different chemoattractant signaling pathways in the regulation of neutrophil polarization and chemotaxis. 2) Characterization of a signaling pathway that underlying persistent PKC activation by mTORC2 and demonstration of the important role of this pathway in fibroblast migration and pulmonary fibrosis. 3) Identification of small molecule inhibitors for Wnt antagonist DKK, which help to uncover an important of role of DKK2 in regulation of glucose metabolism. 4. Publications (Publications July 1, 2011– June 30, 2012): Bao, J., Zheng, J.J., and Wu, D. (2012). The structural basis of DKK-mediated inhibition of

Wnt/LRP signaling. Sci Signal 5, pe22. Gan, X., Wang, J., Wang, C., Sommer, E., Kozasa, T., Srinivasula, S., Alessi, D.,

Offermanns, S., Simon, M.I., and Wu, D. (2012). PRR5L degradation promotes mTORC2-mediated PKC-delta phosphorylation and cell migration downstream of Galpha(12). Nat Cell Biol 14, 686-696. PMID: 3389271

Li, X., Shan, J., Chang, W., Kim, I., Bao, J., Lee, H.J., Zhang, X., Samuel, V.T., Shulman, G.I., Liu, D., Zheng, J.J., and Wu, D. (2012). Chemical and genetic evidence for the involvement of Wnt antagonist Dickkopf2 in regulation of glucose metabolism. Proc Natl Acad Sci U S A 109, 11402-11407. PMID: 3396493

Qian, F., Le Breton, G.C., Chen, J., Deng, J., Christman, J.W., Wu, D., and Ye, R.D. (2012). Role for the guanine nucleotide exchange factor phosphatidylinositol-3,4,5-trisphosphate-dependent rac exchanger 1 in platelet secretion and aggregation. Arterioscler Thromb Vasc Biol 32, 768-777. PMID: 3288658

Rajasekaran, D., Keeler, C., Syed, M.A., Jones, M.C., Harrison, J.K., Wu, D., Bhandari, V., Hodsdon, M.E., and Lolis, E.J. (2012). A Model of GAG/MIP-2/CXCR2 Interfaces and Its Functional Effects. Biochem. In Press

Shi, T., Bao, J., Wang, N.X., Zheng, J., and Wu, D. (2012). Identification Of Small Molecule TRABID Deubiquitinase Inhibitors By Computation-Based Virtual Screen. BMC Chemical Biology 12, 4.

Tang, W., Zhang, Y., Xu, W., Harden, T.K., Sondek, J., Sun, L., Li, L., and Wu, D. (2011). A PLCbeta/PI3Kgamma-GSK3 signaling pathway regulates cofilin phosphatase slingshot2 and neutrophil polarization and chemotaxis. Dev Cell 21, 1038-1050. PMID: 3241930

Xiao, W., Kashiwakura, J., Hong, H., Yasudo, H., Ando, T., Maeda-Yamamoto, M., Wu, D., Kawakami, Y., and Kawakami, T. (2011). Phospholipase C-beta3 regulates FcvarepsilonRI-mediated mast cell activation by recruiting the protein phosphatase SHP-1. Immunity 34, 893-904. PMID: 3124618


52

Jun Yu, M.D.

Instructor of Internal Medicine - Cardiology

1. Overall Goal(s) of the Research Program of the Laboratory: The primary research goals in my laboratory are to understand the molecular control of vascular remodelling and angiogenesis/arteriogenesis in response to arterial injury, atherosclerosis and ischemia. We have intensively used mouse genetic, cell biological and biochemical approaches to achieve these goals. Among the laboratory's current major active projects are: a. To study the role of Reticulon-4B (Nogo-B) in macrophage biology and atherosclerotic

plaque progression. b. To identify the function and molecular mechanism of endothelial Nogo-B in

arteriogenesis. c. To understand the role of primary ciliary localized protein, polycystins -1 and -2, in shear

stress induced inflammatory responses. 2. Specific Research Accomplishments in the last 12 months: In the past year, we have made significant progresses in several research areas. (a) In the study of examine the role of Nogo-B in macrophage function and atherogenesis, we have found that loss of Nogo-B accelerates atherosclerotic plaque formation and leads to formation of morphologically more complex lesion in vivo. Macrophages that lack of Nogo-B are more susceptible to free cholesterol induced apoptosis. We have found that Nogo-B is necessary for regulating ER-mitochondria complex and intracellular calcium transfer. Furthermore, we have generated a new mouse model carrying a floxed allele of Nogo, which provides a novel tool to study cell specific function of Nogo in vivo. (b) We have discovered that Nogo-B is a key regulator of endothelium function. Endothelium specific over-expression of Nogo-B negatively regulates flow dependent vascular remodeling in vivo and ex vivo. Mechanistically, Nogo-B modulates NO bioavailability by influencing eNOS level and its cellular localization. (c) In a collaboration with Drs. Martin Schwartz and Stefan Somlo, we are uncovering a new potential functions of polycystins (PC) -1 and -2 in regulating shear stress induced inflammatory responses in endothelial cells in vitro. We are now in the process of investigating the contribution of PC -1 and -2 in atherogenesis in vivo. (d) We have been collaborating continuously with other research groups in the School of Medicine and have contributed to several publications during the past year as reflected below. 4. Publications: (Publications July 1, 2011– June 30, 2012) Zhang Y, Liu J, Kou J, Yu J, Yu B. DT-13 suppresses MDA-MB-435 cell adhesion and

invasion by inhibiting MMP-2/9 via the p38 MAPK pathway. Mol Med Report. 2012; Doi: 10.3892/mmr.2012.1047

Yu J*, Zhang Y, Zhang X, Rudic RD, Bauer PM, Altieri DC, Sessa WC. Endothelium derived nitric oxide synthase negatively regulates the PDGF-survivin pathway during flow-dependent vascular remodeling. PLoS ONE 2012; 7(2):e31495. PMCID: PMC3280303 (*corresponding author)

Zhang J, Modi Y, Yarovinsky T, Yu J, Collinge M, Kyriakides T, Zhu YH, Sessa WC, Pardi R, Bender J. Macrophage ß2 intergrin-mediated, HuR-dependent stabilization of angiogenic factor-encoding mRNAs in inflammatory angiogenesis. Am J Pathol. 2012; 180(4):1751-60. PMCID: PMC3349897


53

Ding M, Carrao AC, Wagner RJ, Xie Y, Jin Y, Rzucidlo EM, Yu J, Li W, Tellides G, Hwa J,

Aprahamian TR, Martin KA. Vascular smooth muscle cell-derived adiponectin: a paracrine regulator of contractile phenotype. J Mol Cell Cardiol. 2012; 52(2):474-84. PMCID: PMC3264700

MacLauchlan S, Yu J, Parrish M, Asoulin T, Schleicher M, Krady M, Zeng J, Huang P, Sessa WC, Kyriakides T. eNOS controls the expression of the angiogenesis inhibitor thrombospondin 2. Proc Natl Acad Sci U S A. 2011; 108(46):E1137-45. PMCID: PMC3219156


APPENDIX 1

The Eleventh Annual VBT Retreat

1‐1

The Eleventh Annual Retreat of the Vascular Biology and Therapeutics Program held in conjunction with

Cardiovascular Medicine

Saturday, October 22, 2011

The Grace Murray Hopper Auditorium, Building W-B25, West Campus

8:00-8:30 Registration -Continental Breakfast

Session I 8:30-10:00

Tissue Engineering Session Chair – Laura Niklason, M.D., Ph.D.

8:30-9:00 Themis Kyriakides, Ph.D. “Tunable cell-derived extracellular matrix in vascular engineering”

Questions

9:00-9:30 Toshi Shinoka, M.D. “The Translation of the tissue engineered vascular graft”

Questions

9:30-10:00

Tarek Fahmy, Ph.D. "Paracrine Cytokine Delivery from Bioparticles: Implications for Immunotherapy"

Questions

10:00-10:20 Break

SESSION II 10:20-11:50

Inflammation - Session Chair – Jordan Pober, M.D., Ph.D.

10:20:10:50 Jeffrey Bender, M.D. "Endothelial transmembrane steroid hormone receptors"

Questions

10:50-11:20 Nancy Ruddle, Ph.D. “In Vivo Imaging of the Immune Response”

Questions

11:20-11:50 Arnar Geirsson, M.D. “Angiotensin II receptor blocker-mediated modulation of TGF-β signaling in myxomatous mitral valve disease”

Questions

11:50-1:30 Lunch & Poster Session

1‐2

The Eleventh Annual Retreat of the Vascular Biology and Therapeutics Program held in conjunction with

Cardiovascular Medicine

SESSION III 1:30-3:20

Signaling - Session Chair – Anne Eichmann, Ph.D.

1:30-1:50 Martin Schwartz, Ph.D. “How endothelial cells sense fluid shear stress”

1:50-2:00 Questions

2:00-2:30 Suk-Won Jin, Ph.D. “BMP signaling in vascular development”

Questions 2:30-3:00 William Sessa, Ph.D.

“miRNAs and vessel function” Questions 3:00-3:20

Break

3:20-3:30 William Sessa, Ph.D. Introduction of Keynote Speaker

3:30-4:20 Keynote Address David Cheresh, Ph.D., Professor & Vice-Chair for Research and Development, Associate Director for Translational Research, Moores UCSD Cancer Center "Regulation of the Angiogenic Switch"

4:20-4:30 Announcement of Poster Contest Winners

Vascular Biology & Therapeutics

VASCULAR BIOLOGY AND `THERAPEUTICS ANNUAL REPORT 2011 – 2012

APPENDIX 2

The 10th Annual Yale – Cambridge Collaboration Meeting

2‐1

Saturday, September 24 1:25 – Will be picked up at the JFK by Premier Limo: Meeting the bus/van: Step 1 - After picking up your luggage and everyone is together and accounted for, call Premier dispatch center and inform the dispatcher of the airport terminal that you are at. (860-828-9111 Option 3) Step 2 - The dispatcher will contact the driver and let him know what terminal you are at. You will be given a description of the vehicle and a license plate number, so you can easily identify your chauffeur. 4:00 – Arrive and Check-in Omni 5:30 – Bus will pick-up Cambridge Attendees from Omni Hotel 6:00 - Dinner at Sage American Grill & Oyster Bar, 100 South Water Street, New Haven, CT

203-787-3466 8:30 – Bus will return Attendees to Omni Hotel Sunday, September 25 11:00 – 12:00 Brunch - Amistad

12:00 – 5:00 Session 1 – Genetics and Immunology - Chair – Robert Sherwin

12:00 – 12:15 Judy Cho: Genetics and genomics of autoimmune susceptibility loci

12:15 – 12:20 Q & A

12:20 – 12:35 Linda Wicker: Autoimmune phenotypes in type 1 diabetes

12:35 – 12:40 Q & A

12:40 – 12:55 PART 1: David Hafler : Yale-Cambridge phenogenetic project: from genotype to

phenotype

12:55 – 1:00 Q & A

1:00 – 1:15 PART 2: John Todd: Yale-Cambridge phenogenetic project: from genotype to phenotype

1:15 – 1:20 Q & A

1:20 – 1:35 Clara Abraham: NOD2 regulation of human macrophages

1:35 – 1:40 Q & A

1:40 – 1:55 Arthur Kaser: Endoplasmic reticulum stress at the epithelial surface

1:55 – 2:00 Q & A

2:00 – 2:15 Frank Waldon-Lynch: Development of a humanized mouse skin transplant model

2:15 – 2:20 Q&A

2‐2

2:20 – 2:35 Andres Floto: Autophagy in mycobacterial infection

2:35 – 2:40 Q & A

2:40 – 3:00 BREAK

3:00 – 3:15 Augusto Rendon: Genetic loci uncovering new gene functions in megakaryopoiesis and platelet formation

3:15 – 3:20 Q & A

3:20 – 3:35 Diane Krause: Rho GEFs and mitotic control of megakaryocytopoiesis

3:35 – 3:40 Q & A

3:40 – 3:55 Willem Ouwehand: Identification of NBEAL2 as the causative gene for gray platelet syndrome

3:55 – 4:00 Q & A

4:00 – 4:15 Insoo Kang: Inflammasome and autoimmunity

4:15 – 4:20 Q & A

4:20 – 4:35 John Trowsdale: Functions of genes in the leucocyte receptor complex

4:35 – 4:40 Q & A

4:40 – 4:55 Chris Breuer: Tissue engineered vascular graft development

4:55 – 5:00 Q & A

6:00 Dinner at L’Orcio, 806 State Street, New Haven, CT 06511-3922 (203) 777-6670 (Yale attendees will drive Cambridge attendees to and from restaurant)

Monday, September 26 8:30-9:00 Continental Breakfast – Amistad Building

9:00 – 5:00 Session 2 –Transplantation and Immunology - Chair – John Wallwork

9:00 – 9:15 Jordan Pober: Tolerizing T cells by endothelial cells

9:15 - 9:20 Q & A

9:20 - 9:35 Gavin Pettigrew: Humoral immunity in transplantation

9:35 – 9:40 Q & A

9:45 – 10:00 Joao Pereira: Oxysterols direct B cell migration during T-dependent Ab responses

10:00 – 10:05 Q & A

10:05 – 10:30 Break

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