agenda book - novonordisk.com.cnin biopharmaceuticals will be held in tianjin institute of...

Agenda Book

Tianjin, November 7, 2014

Invitation

Novo Nordisk – Chinese Academy of Sciences Research Fund was established in

2007 by a donation of 2 million USD from Novo Nordisk, a biopharmaceutical

company headquartered in Denmark and with a major R&D Centre in Beijing.

Novo Nordisk has since then contributed with additional donations allowing the Fund

to support more than 50 projects engaging scientists from both 10 different CAS

Institutes and from Novo Nordisk scientists in China, Denmark, and the US.

To learn more about NN-CAS Research Fund and the projects it has funded, visit:

http://www.novonordisk.com.cn/documents/article_page/document/04_english_03_01.asp

To celebrate the achievements of the Fund so far, a NN-CAS Symposium on

Translational Research in Biopharmaceuticals will be held at CAS Tianjin Institute of

Industrial Biotechnology on Friday 7 November 2014. The purpose of the

Symposium is also to give selected CAS and Novo Nordisk scientists the opportunity

to report on progress and plans in relation to their NN-CAS projects as well as to

present competencies and interests of CAS and Novo Nordisk research departments

to facilitate the formation of joint research projects in the future.

As co-chairmen of the Symposium and on behalf of Professor Yaping Zhang, VP of

CAS and Prof. Peter Kurtzhals, SVP of Novo Nordisk R&D (chairman and

vice-chairman of the steering committee of the Fund) and Professor Ruiming Xu,

CAS, and Dr. Per Falk, SVP of Novo Nordisk R&D (chairman and vice-chairman of the

board of the Fund), we are happy to invite you to attend the Symposium.

Please let us know at your earliest convenience whether you are able to accept our

invitation.

Kind regards

Professor Børge Diderichsen, Vice President, Novo Nordisk R&D

Professor Tao Xu, Director-General, Institute of Biophysics, CAS

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http://www.novonordisk.com.cn/documents/article_page/document/04_english_03_01.asp

Introduction

Novo Nordisk – Chinese Academy of Sciences Research Fund was established in

March 2007. Since then, Novo Nordisk has donated more than 4 mio USD to the

Fund.

In celebration of 7 years of partnership between Chinese Academy of Sciences (CAS)

and Novo Nordisk based on this Fund, a Symposium titled Translational Research

in Biopharmaceuticals will be held in Tianjin Institute of Industrial Biotechnology

(TIB), Chinese Academy of Sciences, on 7th November, 2014.

The objectives of the Novo Nordisk – Chinese Academy of Sciences Research Fund

(NN-CAS Fund) are to fund or co-fund activities of common interest within the fields

of diabetes and biopharmaceuticals. The Fund was established because “Chinese

and Danish scientists share a history of excellent collaborations which the new Fund

will further strengthen”, commented by Professor Chen Zhu, then CAS vice

president and the founding Chairman of the NN-CAS Fund Board and now Vice

Chairman of the Standing Committee of the National People’s Congress of China,

when announcing the establishment of the Fund, “Novo Nordisk is well recognized in

China for its major role in preventing and treating diabetes. We are very pleased to

collaborate with Novo Nordisk and believe the collaboration will be of great benefit

to both parties.”

The Fund started when Chinese bioscience research was about to take off amongst

the tremendous success of the Chinese economic reform and development. This

attracted overseas Chinese scientists to return to China as well as investments from

multinational pharmaceuticals companies to establish R&D in China. Novo Nordisk

was a frontrunner in exploring the potential of innovative bioscience research in

China, and the first multinational pharmaceutical company to establish an R&D

center in China back in 1997. For Novo Nordisk, “This cooperation illustrates the

great attention Novo Nordisk pays to China as well as our long-term commitment to

be a partner in developing the Chinese healthcare system,” commented by Professor.

Mads Krogsgaard Thomsen, Executive Vice President and Chief Science Officer of

Novo Nordisk.

With direct attention of CAS leadership and Novo Nordisk R&D management, the

Fund has so far supported 55 joint projects, including 39 research projects, 8

2

Workshops, 4 PhD fellowships, and 4 Postdoc Fellowships. These projects cover

research in diabetes and biopharmaceuticals with a variety of technologies and

disciplines including protein chemistry, immunology, toxicology, endocrinology and

drug delivery. In addition, Great Wall Professorships was set up to promote the

recruitment of top level scientists in China. Eight professors have received this

honorable support. The collaborations between CAS and Novo Nordisk have greatly

promoted the understanding and networking between Chinese and Novo Nordisk

scientists in China, Denmark and the US. This fruitful collaboration has also resulted

in over 100 research publications and a few patent applications acknowledged the

support from the Fund. Ten PIs, who received the support from the Fund, will

present their work at the Symposium.

As the work presented by these PIs represent only a fraction of the projects funded

by the Fund, selected information from projects funded so far is listed in this booklet

to provide snap shots of the Fund’s footprint in the past 7 years. The booklet is

intended for the Symposium participants and interested scientists in CAS and Novo

Nordisk. To learn more about NN-CAS Research Fund and the projects it has funded,

visit: www.novonordisk.com.cn (English>Cooperation>NN-CAS Fund).

Novo Nordisk and CAS are both committed to continue their support to the NN-CAS

Research Fund. Accordingly, a new agreement to extend the program was signed on

6th November 2014.

In the Symposium, Novo Nordisk will introduce their current technology platforms,

and CAS presenters will introduce major drug discovery platforms and programs

being undertaken in China. Such information exchange aims to inspire new

collaboration opportunities.

In summary, it has been a fruitful journey of cooperation in past 7 years. Both CAS

and Novo Nordisk are pleased with the achievements and committed to continue

this unique and long-term collaboration. We hope this Symposium will be a bridge to

connect the success of the past and the bright future that will be created through

closer collaboration between CAS and Novo Nordisk scientists, who have established

friendship and trust through the Fund.

Sincerely,

Ruiming Xu, Ph.D.

Chairman, NN-CAS Fund Board,

Director-General

Bureau of Frontier Sciences and

Education, CAS

Per Falk, Ph.D.

Vice Chairman, NN-CAS Fund Board,

Senior Vice President

Novo Nordisk

3

Information on the Novo Nordisk and

the Chinese Academy of Sciences

Novo Nordisk

Novo Nordisk is a global healthcare company with 90 years of innovation and

leadership in diabetes care. The company also has leading positions within

haemophilia care, growth hormone therapy and hormone replacement therapy.

Headquartered in Denmark, Novo Nordisk employs approximately 40,700

employees, of which 18% of the employees are within Research & Development, in

75 countries, and markets its products in more than 180 countries.

In 1997, Novo Nordisk established the first biotechnology R&D center among

multinational pharmaceutical companies in Beijing, China. Novo Nordisk Research

Center China is currently also one of the biggest R&D centers set up by multinational

pharmaceutical companies in China.

For more information please refer to our web page: http://www.novonordisk.com.

Chinese Academy of Sciences

Being China’s top academic institution in science and technology and the national

integrated R&D center in natural sciences and high technology, the Chinese

Academy of Sciences has made important contributions to China’s scientific

advancement and social development since its establishment in 1949. It has about

100 research institutes, two universities, and about 60,000 professional staff and

45,000 graduate students.

The Chinese Academy of Sciences engages above all in the basic research, strategic

high technology research and research relevant to the sustainable development of

economy and society, focuses on the solutions to science and technology issues of

fundamental, strategic and forward looking importance in China’s modernization

process, and provides scientific basis and sources of technological means for

promoting the overall sustainable development of China’s economy and society.

For more information please refer to our web page: http://english.cas.cn or contact

us at [email protected].

4

Meeting Information

Organizers: Chinese Academy of Sciences and Novo Nordisk R&D

Co-Organizer: Tianjin Institute of Industrial Biotechnology, CAS

Sponsor: Novo Nordisk-Chinese Academy of Sciences Research Fund

Co-Chairpersons: Prof. Børge Diderichsen and Prof. Tao Xu

Symposium Organizing Committee: Prof. Børge Diderichsen, Prof. Tao Xu,

Prof. Xuerong Xing, Dr. Baoping Wang, Prof. Mingwei Wang

Secretariat: Ms. Dongyao Wang, Ms. Qianqian Chai, Ms. Hao Wang

Time: November 7, 2014, 9:10 – 19:30

Venue: B301, Tianjin Institute of Industrial Biotechnology (TIB), Chinese Academy

of Sciences, 32 West 7th Avenue, Tianjin Airport Economic Area, Tianjin

300308, China.

Notes: indicates TIB, CAS

5

Program

09:10 Registration / coffee

Opening Session:

Chairs: Prof. Børge Diderichsen, Novo Nordisk R&D and Prof. Tao Xu,

CAS

09:30 - 9:50 Welcoming addresses

Prof. Ruiming Xu, DG, Bureau of Frontier Sciences and

Education, CAS

Per Falk, SVP, Biopharmaceuticals Research Unit, Novo

Nordisk R&D

9:50-10:00 Introduction of Tianjin Institute of Industrial

Biotechnology, CAS

Prof. Jibin Sun, DD, Tianjin Institute of Industrial

Biotechnology, CAS

10:00 - 10:15 NN-CAS history & achievements

Prof. Børge Diderichsen, VP, Novo Nordisk R&D

Plenary Lectures:

Chairs: Prof. Ruiming Xu, CAS and Dr. Baoping Wang, President,

Novo Nordisk Research Center China

10:15 - 10:35 The Chinese National Compound Library - a long

cherished dream shared with Novo Nordisk

Prof. Ming-Wei Wang, Director, the National Center for Drug

Screening, SIMM, CAS

10:35 - 10:55 Sustainable biotechnology – an industrial perspective

on next generation cell culture processes

Dr. Jakob Helding Rasmussen, CVP, Biopharmaceuticals

Research Unit, Novo Nordisk R&D

6

10:55 - 11:10 Group Photo and Coffee Break

Session 2: Diabetes and Beyond

Chairs: Prof. Tao Xu, CAS IBP, and Dr. Rasmus Jørgensen, Novo

Nordisk RCC

11:10 - 11:30 Rasmus Jørgensen, Novo Nordisk RCC: Novo Nordisk

diabetes research in China

11:30 - 11:50 Dehua Yang, CAS SIMM: Functional hot spots in the human

GLP-1 receptor structure

11:50 - 12:10 Birgitte Andersen, Novo Nordisk DRU R&D: Endocrine

fibroblast growth factors and treatment of the metabolic

syndrome

12:10 - 12:30 John Speakman, CAS IGDB: Impacts of gastric bypass

surgery on non-obese patients

12:30 - 13:30 Lunch

13:30 - 13:50 Fang Zhang, Novo Nordisk RCC: Diabetes biology,

pharmacology and China focus group at Novo Nordisk

Diabetes Research China

13:50 - 14:10 Xu Lin, CAS SIBS: The productive role of omics-based

biomarkers in the development of type 2 diabetes

14:10 - 14:30 Wanzhu Jin, CAS IZ: Isolation and characterization of new

brown adipokine

Session 3: Protein Engineering for Biopharmaceuticals

Chairs: Prof. Ruiming Xu, CAS HQ, CAS, and Xujia Zhang, Novo

Nordisk RCC

14:30 - 14:50 Ruiming Xu, CAS HQ and CAS-Novo Nordisk Great Wall

Professor: Structural basis for Sirt1 activation

14:50 - 15:10 Xujia Zhang, Novo Nordisk RCC: Protein production in E.coli

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15:10 - 15:30 Caihong Zhou, CAS SIMM: Polymorphism study and assay

development for potential drug targets

15:30 - 15:50 Liu Yun, Novo Nordisk RCC: Discovery of the improved

antagonistic prolactin variants by library screening

15:50 - 16:05 Coffee Break

Session 4: Advanced Technology

Chairs: Prof. Ming-Wei Wang, CAS SIMM and Dr. Zhiru Yang, Novo

Nordisk RCC

16:05 - 16:25 Wei Yang, Novo Nordisk RCC: NN6K: an unparalleled

platform for protein candidate identification

16:25 - 16:45 Rong Zeng, CAS SIBS: Systemswide evaluation of treatment

outcome in diabetes patients

16:45 - 17:05 Jianhe Chen, Novo Nordisk RCC: State-of-the-art fully

human Fab phage display library

17:05 - 17:30 Dengning Xia (representing Yong Gan), CAS SIMM:

Functional nanocarriers for overcoming biological barriers of

insulin absorption

Closing: NN-CAS Future Perspectives

17:30 - 17:45 Per Falk, SVP, Biopharmaceuticals Research Unit (BRU), Novo

Nordisk R&D

17:45 - 18:00 Prof. Ruiming Xu, DG, Bureau of Frontier Sciences and

Education, CAS

18:00-19:30 Dinner

8

Summary and Publication

2007

CAS PI NN PI Grant Number of Project

Name Affiliation/Job title Name Affiliation/Job title

Zhi-Jie Liu Prof. ,IBP Carsten Behrens Dr. NNCAS-2007-2

Title Engineered Glycosyltransferases Transfering Non-Natural Sugar Substrates

Executive

Summary

A set of clones were constructed includes wild type bovine 1,4-galT, and the single point mutant M1(R349H), M2(N353A) and the

multi point mutation M3(K352G,N353A,P355V) and M4(R349H, K352G,N353A,P355V). Both E.Coli expression system and

Baculovirus expression system were tried but failed to get soluble enzymes. Protein denaturing and refolding method was tried

to get soluble protein, unfortunately, the recovery rate was too low to get enough enzyme for function assay.



Xiyun Yan Prof. ,IBP Birgitte Ursø Dr. NNCAS-2007-3

Title Cancer Stem Cell markers and tumour specific antibodies

Executive

Summary

KG-1a-immunized phage-displayed antibody scFv library was constructed and subjected to difference-screening against KG-1a

and the other leukemic lymphoid cells, such as HL60, U937 and CEM, respectively. Two clones with high specific binding affinity

to KG-1a but not the other leukemic lymphoid cells were selected as the candidate CSC-specific scFvs. Further characterization

assays with WB, FACS and IF showed that the two scFvs bound to the surface molecules on CD34+CD38- subpopulation cells in

KG-1a, with a MW of 131kD and 91kD, respectively. To identify the two proteins, IP and MS were performed. The very low portion

of the positive subpopulation delayed the project to some degree.

Some other efforts were attempted to explore CSC markers. In our study on a cancer-specific mAb and its antigen, we found that

this mAb stained some stem-like cells in the crypt bottom on intestine tumor tissues specifically in IHC assay. Although stem cells

in intestine crypt had been assumed for many years, there was no specific marker for these stem cells, yet. Our findings may

inspire some new thoughts to explore CSC markers and the roles of CSC in tumorigenesis.

13



Ling Zhou Prof., SIMM Tobias Cornvik NNCAS-2007-8

Title DNA array to Protein array

Executive

Summary

DAPA is a novel concept in protein array technology that creates protein arrays directly from DNA array templates. In earlier

published work it has been shown that DNA in solution can be used to synthesize tagged proteins by cell-free systems, and that

the proteins could be simultaneously immobilized on the surface of a microtiter well (protein in situ array or PISA). Using a similar

approach it is possible to generate protein arrays on glass slides from immobilized DNA arrays.DAPA promises considerable

advantages for protein array generation, in terms of increased stability (protein arrays are produced on demand from stable DNA

templates, minimizing the requirement for storage) and ease of production (fewer steps, no requirement for cloning, in vivo

expression and purification). Plasmodium falciparum subtilisin-like protease-1 (PfSUB1) is a protein belonging to the

subtilisin-like superfamily of serine proteases (subtilases). It is synthesized during maturation of the intraerythrocytic parasite

and accumulates in a set of merozoite secretory organelles, suggesting that it may play a role in host cell invasion or

post-invasion events and could be a potential target for malaria therapy. In this project we aim to establish a prototype process

of DAPA including: 1. Identify an appropriate expression cassette for in vitro translation; 2. Establish the in vitro translation

system for producing active target protein; 3. Optimize parameters like spot density and quantity of DNA immobilized for DNA

array preparation; 4. Optimize conditions like the amount of protein immobilized for protein array preparation. By establishing

the process in the context of PfSUB1, we also aim at exploring the possibility of screening small molecule inhibitors of PfSUB-1.



George Fu Gao Prof. ,Institute of Microbiology Baoping Wang Dr. NNCAS-2007-9

Title Workshop: International Symposium on Immune Regulation and Its Clinical Application in Human Diseases

Executive Summary

The symposium was held in Oct 25-27, 2008 in Beijing. It was a world level symposium on T regulatory cells, with more than 30 world renowned experts in the field gathered in Beijing to present the most recent findings as well as the advancement in recent years in the field. Based on the success of that inauguration meeting, the symposium have been held once in every two years in Shanghai or Beijing, and has become one of best meetings in regulatory T cells worldwide.

14

2008

CAS PI NN PI

Name Affiliation/Job title Name Affiliation/Job title Grant Number of Project

JianFeng Chen Prof., Institute of Biochemistry and Cell Biology,

SIBS Wei Yang

Dr., Head of Molecular Biology

Department NNCAS-2008-1

Title To develop antibodies specifically block active forms of integrin α4β7 and α4β1 for the treatment of autoimmune diseases

List of

Papers

1. Qi JP, Zhang K, Zhang Q, Sun Y, Fu T, Li GH, Chen JF*. (2012) Identification, characterization and epitope mapping of a human

monoclonal antibody J19 that specifically recognizes the activated integrin a4b7. J Biol Chem. 287(19): 15749-59.

Other

Outcomes 1. Patent (pending): A humanized monoclonal antibody against the activated integrin

CAS PI NN PI


Zeng Rong Prof., SIBS Flemming Pociot NNCAS-2007-12

Title Proteome analysis for detecting early stage diabetic nephropathy biomarkers

List of Papers

1.Gao X, Chen W, Li R, Wang M, Chen C, Zeng R, Deng Y. Systematic variations associated with renal disease uncovered by

parallel metabolomics of urine and serum. BMC Syst Biol. 2012; 6:S14.2. Chen H, Zheng Z, Li R, Lu J, Bao Y, Ying X, Zeng R, Jia

W. Urinary pigment epithelium-derived factor as a marker of diabetic nephropathy. Am J Nephrol. 2010;32:47-56.3. Li QR, Fan

KX, Li RX, Dai J, Wu CC, Zhao SL, Wu JR, Shieh CH, Zeng R. A comprehensive and non-prefractionation on the protein level

approach for the human urinary proteome: touching phosphorylation in urine. Rapid Commun Mass Spectrom. 2010;

2:823-32.4. Li SJ, Peng M, Li H, Liu BS, Wang C, Wu JR, Li YX, Zeng R. Sys-BodyFluid: a systematical database for human body

fluid proteome research. Nucleic Acids Res. 2009；37:D907-12.

15

CAS PI NN PI


Pingyong Xu Prof., IBP Søren Tullin Prof. NNCAS-2008-3

Title Establish and utilise a novel GLUT4 translocation assay for HTS of compound library to identify drug leads for treating insulin resistance

Executive

Summary

During the project, we have developed a novel probe based on a pH-sensitive red fluorescent protein mOrange2-labeled IRAP that can

monitor the translocation process of GLUT4 to the plasma membrane and established stable cell lines (3T3-L1 and L6) that expressing the

functional probe that responds efficiently upon insulin stimulation. Based on the probe, the project has developed a high throughput

method for detecting GLUT4 translocation using Total Internal Reflection Fluorescence (TIRF) Microscopy. The project has screened two

compound libraries of 56000 low-molecular-weight compounds and identified successfully one precursor drug that facilitate GLUT4

translocation to the plasma membrane. We confirmed the biological function of the screened compound A2 that A2 can promote the probe

translocation and glucose uptake in L6 cell line. Last the mechanism study shows that A2 can promote GLUT4 translocation by activating

AMPK pathway through decreasing ATP synthesis and membrane potential of mitochondrial.

Other

Outcomes 1. Patent “A probe for drug screen anti-insulin resistance”, 200710064381.X, China

CAS PI NN PI


Qian Li

Zhiyun Zhang Prof., SIMM

Cecilia Nilsson

Kirsten Raun Dr. NNCAS-2008-4

Title Impact of fetal diet program on fetal and adult metabolic and endocrine phenotype and window of intervention to reverse the fetal program

Executive

Summary

Using chocolate and fructose beverage as supplement of normal diet, we have established a high-energy diet intake rat model to

investigate the effects of high fat, high carbohydrate and high energy on foetal programming. Our study suggests that dams, living in a high

fat (mainly from chocolate) and high-energy environment during gestation, tend to significantly consume more energy and gain more

weight. Over nutrition with chocolate reduced chow intake in dams leading to lower protein consumption. As a result, pups delivered by

these dams exhibited low birth weights that lasted until adulthood. It appears that the dietary impact on fat metabolism in the pup is more

pronounced when taking place during lactation than during gestation Obviously, over supply of carbohydrate such as chocolate and

fructose either in gestation or in lactation has negative impact on the well being of pups. Pregnant woman should properly control their

craving for sweet foods.

16

CAS PI NN PI


Sheng Jiang Prof., GIBH Wei Yi Dr. NNCAS-2008-8

Title Novel bicyclic peptide IMSF-8 and analogs as potential anti-cancer and anti-inflammation drug leads by selectively inhibit type II

transmembrane serine proteases matriptase

List of

Papers

1. J. Zheng, B. Yin, W. Huang, X. Li, H. Yao, S. Jiang*(corresponding author) Tetrahedron Letters, 2009，50，5094-5097 2. X. Zeng, B. Yin, Z. Hu, C. Liao, J. Liu, S. Li, Z. Li, M. C. Nicklaus, G. Zhou, and S. Jiang* (corresponding author) Organic Letters, 2010，12，1368-1371. 3. Q. Xiao, Y. Liu, Y. Qiu, G. Zhou, C. Mao, Zheng Li, Z.-J. Yao, and S. Jiang*, (corresponding author) J. Med. Chem. 2011, 54, 525-533 4. Z. Yao, X. Zeng, W. Yi and S. Jiang*, (corresponding author) Letters in Organic Chemistry. 2011, 8, 66-69. 5. S. Jiang*, C. Liao, L. Bindu, B. Yin, K. W. Worthy, R. J. Fisher, T. R. Burke, Jr., M. C. Nicklaus, P. P. Roller. (corresponding author) Bioorg. Med. Chem. Lett. 2009, 19, 2693-2698.

Other

Outcomes

1.“Method for synthesis of largazole and its analogs as antitumor agents.”

Inventors: S. Jiang, G. Zhou, B. Yin, X. Zeng, and Z. Hu.

Patent No. CN 101781321

2. S. Jiang, Z-J. Yao, G. Zhou, Q. Xiao, Y. Liu

Synthesis and application of simplified chiral annonaceous acetogenins compound.

Faming Zhuanli Shenqing Gongkai Shuomingshu. 2010, (CN2010102884066).

CAS PI NN PI


Feifan Guo Prof., Institute for Nutritional Sciences, SIBS Kirsten Raun Dr. NNCAS-2008-10

Title Identification of key protein/hormones secreted in the serum which critically regulate the rapid loss of abdominal fact mass under leucine

deprivation

17

List of

Papers 1. Cheng Y*, Meng Q* (co-author), Wang C, Li H, Huang Z, Chen S, Xiao F, Guo F*. Leucine deprivation decreases fat mass by stimulation

of lipolysis in WAT and upregulation of UCP1 in BAT. Diabetes. 2010, 59:17-25

2. Wang C，Huang Z, Du Y, Cheng Y, Chen S and Guo F*. ATF4 regulates lipid metabolism and thermogenesis. Cell Res. 2010, 20:174–184.



Ruiming Xu Prof., IBP NNCASGWP-2008-11

Title

List of

Papers

1. C.P. Liu, C. Xiong, M. Wang, Z. Yu, N. Yang, P. Chen, Z. Zhang, G. Li, and R.M. Xu (2012) Structure of the variant histone H3.3-H4

heterodimer in complex with its chaperone DAXX, Nat. Struct. Mol. Biol., 2012 Dec;19(12):1287-92.

2. L. Wang, F. Wang, D. Zhang, Z. Chen, R.M. Xu, K.H. Nierhaus, W. Gong, and Y. Qin (2012) A conserved proline switch on the ribosome

facilitates the recruitment and binding of trGTPases. Nat. Struct. Mol. Biol. 19, 403-410.

3. N. Yang and R.M. Xu (2012) Structure and function of the BAH domain in chromatin biology. Crit. Rev. Biochem. Mol. Biol. 2013

May-Jun;48(3):211-21. 4. H.C. Hsu, C.L. Wang, M. Wang, N. Yang, Z. Chen, R. Sternglanz, and R.M. Xu (2013.1.10) Structural basis for

allosteric stimulation of Sir2 activity by Sir4 binding. Genes & Dev., 27: 64-73.

5. K. Liu, C. Bian, H. Liu, Y. Guo, R. Lam, F. MacKenzie, L.A. Rojas, D. Reinberg, M. Bedford, R.M. Xu*, and J. Min* (2012) Crystal structure

of TDRD3 and methyl-arginine binding characterizations of TDRD3, SMN and SPF30. PLoS ONE ,7(2): e30375, .

6. P.C. Lin, and R.M. Xu (2012) Structure and assembly of the SF3a splicing factor complex of U2 snRNP. EMBO J. 31, 1579-1590.

7. N. Yang, W. Wang, Y. Wang, M. Wang, Q. Zhao, Z. Rao, B. Zhu, and R.M. Xu (2012) Distinct mode of methyl-H3K4 recognition by tandem

tudor-like domains of Spindlin1, Proc. Natl. Acad. Sci .USA, Vol. 109, 2012 Oct 30;109(44):17954-9.

8. L. Sun, M. Wang, Z. Lv, N. Yang, Y. Liu, S. Bao, W. Gong, and R.M. Xu (2011) Structural insights into protein arginine symmetric

dimethylation by PRMT5. Proc. Natl. Acad. Sci. USA 108, 20538-20543.

9. H. Hu, Y. Liu, M. Wang, J. Fang, H. Huang, N. Yang, Y. Li, J. Wang, X. Yao, Y. Shi, G. Li, and R.M. Xu (2011) Structure of a CENP-A-Histone

H4 heterodimer in complex with chaperone HJURP. Genes & Dev. 25, 901-6.

10. Q. Qiao, Y. Li, Z. Chen, M. Wang, D. Reinberg, and R.M. Xu (2011) The structure of NSD1 reveals an autoregulatory mechanism

underlying histone H3K36 methylation. J. Biol. Chem. 286, 8361-8.

11. H. Liu, J.Y. Wang, Y. Huang, Z. Li, W. Gong, R. Lehmann, and R.M. Xu (2010) Structural basis for methylarginine-dependent recognition

of Aubergine by Tudor. Genes & Dev .2010 Sep 1;24(17):1876-81.

18

2009

CAS PI NN PI Grant Number of

Project Name Affiliation/Job title Name Affiliation/Job title

Huarong

Zhou

Prof., Key lab of systems biology,

SIBS

Clausen Trine

Ryberg Senior Scientist

NNCAS-2009-1,

NNCAS-PostDoc-2011-1

Title The role of islets improvement in the diabetes control after bariatric surgery

List of

Papers

1. Rui Zhang*, Caifeng Yan*, Xinrong Zhou*, Bangguo Qian, Fuqiang Li, Yidan Sun, Chen Shi, Bing Li, Shigeru Saito, Katsuhisa Horimoto and

Zhou H. Association of Rev-erba in Adipose Tissues with Type 2 Diabetes Mellitus Amelioration after Gastric Bypass Surgery in Goto-Kakizaki

Rats. American Journal of Physiology- Regulatory, Integrative and Comparative Physiology. 2013;305:R134-146.

2. Li B， Zhou X, Wu J, Zhou H. From gut changes to type 2 diabetes remission after Gastric Bypass Surgeries. Frontiers of Medicine,

2013;7:191-200.

3. Wang J, Sun Y, Zheng S, Zhang XS*, Zhou H*, and Chen L*. APG: an Active Protein-Gene Network Model to Quantify Regulatory Signals

in Complex Biological Systems. Science Report, 2013; 3:1097. (*:co-corresponding authors)

4. Zhou X, Saito S, Chen L, Horimoto K, Zhou H. Tissue Relationship along Diabetes Progression in Goto-Kakizaki Rats by Network Inference

and Clustering. BMC Systems Biology, 2013 In press.

5. Sun Y, Saito S, Horimoto K, Zhou H. Functional Networks in Diabetes-Progression by Comparison of Gene Expression in Three Tissues of

Goto-Kakizaki Rats. Current Bioinformatics. 2013; 8(1):63-71.

6. Yu T, Toshimori A, Xia J, Saito S, Horimoto K, Zhou H. Cellular relationships of testicular germ cell tumors determined by

7. partial canonical correlation analysis of gene expression signatures. Current Bioinformatics. 2013; 8(1): 72-79.

Other

Outcomes

1.The grants have supported discovery of 47 compound candidates whose activation potentially able to treating diabetes and its

complications.

2. The grants have supported researches for 8 publications.

3. The grants have supported 5 international invited oral presentations.



Caihong

Zhou Prof. SIMM Minghuang Zhang Dr. NNCAS-2009-2

19

Title Pharmacological characterization of cyclosporins on a panel of human formyl peptide receptors with different single nucleotide

polymorphisms

List of

Papers

1. Caihong Zhou, Yan Zhou, Jia Wang, Yang Feng, Haonan Wang, Jinglun Xue, Yani Chen, Richard D. Ye and Ming-Wei Wang (2013). V101L

of human formyl peptide receptor 1 (FPR1) increases the receptor affinity and augments the antagonism mediated by cyclosporins.

Biochemical Journal, 451 (part 2), 245-255.

Other

Outcomes

1. The haplotype pattern and frequency of human FPR1 haplotypes in 209 healthy Han Chinese subjects was characterized using five single

nucleotide polymorphisms (SNPs).

2. Variants of FPR1 carrying a single amino acid substitution of leucine for valine at position 101 were found to improve the receptor affinity

of CsA and CsH significantly.



Pingsheng Liu Prof., IBP Xujia Zhang Senior Scientist NNCAS-2009-3

Title The Function of Lipid Droplet-associated Apolipoprotein A1 in Skeletal Muscle

List of

Papers

1.Pu J, Liu P: Fatty Acids Stimulate Glucose Uptake by the PI3K/AMPK/Akt and PI3K/ERK1/2 Pathways. In: Protein Phosphorylation in Human

Health. Edited by Huang C: InTech; 2012: 129-148.

2.Peng G, Li L, Liu Y, Pu J, Zhang S, Yu J, Zhao J, Liu P: Oleate blocks palmitate-induced abnormal lipid distribution, endoplasmic reticulum

expansion and stress, and insulin resistance in skeletal muscle. Endocrinology 2011, 152(6):2206-2218.

3. Pu J, Peng G, Li L, Na H, Liu Y, Liu P: Palmitic acid acutely stimulates glucose uptake via activation of Akt and ERK1/2 in skeletal muscle

cells. J Lipid Res 2011, 52(7):1319-1327.

4.Zhang H, Wang Y, Li J, Yu J, Pu J, Li L, Zhang S, Peng G, Yang F, Liu P: Proteome of skeletal muscle lipid droplet reveals association with

mitochondria and apolipoprotein a-I. J Proteome Res 2011, 10(10):4757-4768.

5. Li ZY, Na HM, Peng G, Pu J, Liu P: Alteration of microRNA expression correlates to fatty acid-mediated insulin resistance in mouse

myoblasts. Mol Biosyst 2011, 7(3):871-877.



Bin Li Prof., IPS Lingyun Wang

Yi Luo

Scientist Protein Chemistry,Beijing Novo Nordisk

Pharmaceuticals Sci & Tech Co. Ltd NNCAS-2009-5

20

Title Evaluation of GITR-GITR-L interaction in immune system as a therapeutic target for autoimmune diseases

List of

Papers

1.Zhang J*, Chen C*, Hou X*, Gao Y, Lin F, Tsun A, Shi G and Li B, Identification of the E3 deubiquitinase USP21 as a positive regulator of

GATA3. J Biol Chem.2013. 288(13):P.9373-82

2. Tsun A*, Chen Z and Li B*. Romance of the three kingdoms: RORgammat allies with HIF1alpha against Foxp3 in regulating T cell

metabolism and differentiation. Protein & Cell. (2011), DOI 10.1007/s13238-011-1114-2.

3. Tsun A, Li YY and Li B*. You, Me, and Foxp3: Immune Regulation for Two. Immunotherapy. 2011 Oct; 3(10): 1139-42.

4. Gao Y, Lin F, Su J, Gao Z, Li Y, Yang J, Deng Z, Tsun A* and Li B*. Molecular Mechanisms Underlying the Regulation and Functional

Plasticity of FOXP3+ Regulatory T Cells. GENES & IMMUNITY, (2011) .DOI: 10.1038/gene.2011.77.

5. Chen Z, Lin F, Gao Y, Li Z, Zhang J, Xing Y, Deng Z, Yao Z, Tsun A, Li B*. FOXP3 and RORγt: Transcriptional regulation of Treg and Th17.

International Immunopharmacol. (2011)11(5):536-42. Epub 2010 Nov 23.

6．Chen, Z., et al., The ubiquitin ligase Stub1 negatively modulates regulatory T cell suppressive activity by promoting degradation of the

transcription factor Foxp3. Immunity, 2013. 39(2): p. 272-85.

7. Shan, Z., et al., Negative regulation of interferon-induced transmembrane protein 3 by SET7-mediated lysine monomethylation. J Biol

Chem, 2013. 288(49): p. 35093-103.

8. Li, Y., et al., 60-kDa Tat-interactive protein (TIP60) positively regulates Th-inducing POK (ThPOK)-mediated repression of eomesodermin

in human CD4+ T cells. J Biol Chem, 2013. 288(22): p. 15537-46.

9. Gao, Z.M., et al., Synergy between IL-6 and TGF-beta signaling promotes FOXP3 degradation. International Journal of Clinical and

Experimental Pathology, 2012. 5(7): p. 626-633.

CAS PI NN PI


Grant Number of

Project

Zhihai QIN Prof., IBP Jianhe CHEN NNCAS-2009-6

Title Induction and Functional Analysis of Regulatory B cells

21

Executive

Summary

We have constructed several expression plasmids of the fusion protein GMCSF-IL15 with human or mouse DNA in a retrovirus vector (HyTK),

and transfected 293T cells with them. We have demonstrated the expression of GMCSF-IL15 by the Western Blot. The novel proteins were

about 37KD in size. The concentration of GMCSF-IL15 in cell culture supernatant was also determined by ELISA or CBA analysis.

Furthermore, we have constructed several new variants of the fusion gene GMCSF-IL15 by shortening the linker between GMCSF and IL15.

The protein structures of these fusokines will be further analyzed and this may help the design of novel peptides with therapeutic effects in

autoimmune diseases.

At the same time, we have established an autoimmune disease model, namely, the experimental autoimmune encephalomyelitis (EAE) in

mice. This animal model is very important for our further study of the fusion protein functions in vivo. Currently, the immune regulatory cells

were induced with the novel fusokines and tested for their efficacy in treating EAE by adoptive transfer. Furthermore, we have found that the

IFNγR expression on endothelial cells could prevent brain inflammation by keeping blood-brain barrier intact during EAE. The construction

of expression plasmids, induction of immune regulatory cells as well as the establishment of animal models will help us in developing novel

strategies to treat autoimmune diseases.



He Min Prof., SIMM Kristian

Strømgaard Dr. NNCAS-2009-7

Title Targeting protein-protein interaction involving PDZ domains

Executive

Summary

This report describes the results of a high-throughput screening (HTS) campaign for small-molecule inhibitors of the protein-protein

interaction between PSD-95 and the C-terminal tail of NMDAR. The C-terminal peptide (AB1) has been labeled with three different dyes, and

thus the property measured in the assay is the change in fluorescence polarization (FP) upon binding/unbinding of the peptide to the protein.

The differently labeled peptides were used for both the primary and secondary screening. A total of 316,000 compounds from the Novo

Nordisk donated compound library were screened in X-Y plates with the FP assay, however, no hits (high-affinity inhibitors of the

PSD-95/NMDAR interaction) were found.



Yong Gan Prof., SIMM Lars Hovgaard Dr. NNCAS-2009-10

Title Design of functional lipid nanoparticles for oral delivery of protein/peptide drugs and study of absorption mechanism

List of

Papers

1) Xiuying Li, Dan Chen, Chaoyi Le, Chunliu Zhu, Yong Gan*, Lars Hovgaard, Mingshi Yang. Novel mucus-penetrating liposomes as potential

oral drug delivery system: preparation, in vitro characterization, and enhanced cellular uptake. Int J Nanomed. 2011,6:3151-62.

22

2010



Guangxun Meng Prof., IPS Lishan Kang Dr., Associate Scientist NN-CAS-2010-2

Title Targeting Pannexin-1 to regulate NLRP3 inflammasome associated Autoimmune and Autoinflammatory diseases

List of Papers

1. Wang H, Xing Y, Mao L, Luo Y, Kang L, Meng G*. Pannexin-1 influences peritoneal cavity cell population but is not involved in NLRP3

inflammasome activation. Protein Cell. 2013 Apr;4(4):259-65. (IF=3.22).PMID: 23549611.

2. Lei G#, Chen M#, Li H#, Niu JL, Wu S, Mao L, Lu A, Wang H, Chen W, Xu B, Leng Q, Xu C, Yang G, An L, Zhu LP*, Meng G*. Biofilm

from a clinical strain of Cryptococcus neoformans activates the NLRP3 inflammasome. Cell Res. 2013 Jul;23(7):965-8.

(IF=10.216).PMID: 23567555.

3. Wang Y#, Yang C#, Mao K, Chen S, Meng G*, Sun B*. Cellular localization of NLRP3 inflammasome. Protein Cell. 2013

Jun;4(6):425-31. (IF=3.22).PMID: 23609011.

4. Xu Y#, Li H#, Chen W#, Yao X, Xing Y,Wang X, Zhong J*, Meng G*. Mycoplasma hyorhinis Activates the NLRP3 Inflammasome and

Promotes Migration and Invasion of Gastric Cancer Cells. PLOS ONE. 2013 Nov 6;8(11):e77955. (IF=4.244). PMID: 24223129.

5.Chen W#, Xu Y#, Li H, Tao W, Xiang Y, Huang B, Niu J, Zhong J*, Meng G*. HCV Genomic RNA Activates the NLRP3 Inflammasome

in Human Myeloid Cells. PLOS ONE. 2013 (IF=4.244).PMID: 21100125.

6. Lei G, Mao L, Li H, An L, Yang G, Meng G*. Function of Inflammasomes in Anti-Microbial Infections. Chinese Journal of Cell biology.

2011 Dec, 33(12). Invited Review. Cover Page.



Qin Yan Prof., IBP Zhang Xujia Senior Scientist NNCAS-2010-3

Title The function of the mitochondrial translational factor GUF1 in the Diabetes pathogenesis.

List of Papers

1.Zhang D and Qin Y* (2013) The paradox of elongation factor 4: highly conserved, yet of no physiological significance? Biochem. J.,

452 (2), 173-181

2. Zhang D, Liu G, Xue J, Lou J, Nierhaus K H, Gong W* and Qin Y* (2012) Common chaperone activity in the G-domain of trGTPase

protects L11–L12 interaction on the ribosome, Nucleic Acids Res., 40 (21) 10851-10865.

3. Wang L, Yang F, Zhang D, Chen Z, Xu R, Nierhaus K H, Gong W and Qin Y* (2012) (A conserved proline switch on the ribosome

facilitates the recruitment and binding of trGTPases, Nat. Struct. Mol. Biol., 19(4)403-410

23



Jiang Xingyu Prof., NCNST Yang Li Dr., Beijing Novo Nordisk NNCAS-2010-5

Title Comprehensive Biomarker Analysis in Arthritis and Build-up of Bio-repository

List of Papers

1. Tuning the Composition of Aupt Bimetallic Nanoparticles for Antibacterial Application, Zhao Y, Ye C, Liu W, Chen R, Jiang X, Angew

Chem Int Ed, 53(31): 8127-8131. (2014).

2. A Peptide-Based Nanofibrous Hydrogel as a Promising DNA Nanovector for Optimizing the Efficacy of HIV Vaccine, Tian Y, Wang HM,

Liu Y, Mao LN, Chen WW, Zhu ZN, Liu WW, Zheng WF, Zhao YY, Kong DL, Yang ZM, Zhang W, Shao YM, Jiang XY, Nano Lett, 14(3):

1439-1445. (2014).

3. Facile, One-Pot Synthesis, and Antibacterial Activity of Mesoporous Silica Nanoparticles Decorated with Well-Dispersed Silver

Nanoparticles, Tian Y, Qi JJ, Zhang W, Cai Q, Jiang XY, ACS Appl Mater Inter, 6(15): 12038-12045. (2014).

4. Nanomaterials for Ultrasensitive Protein Detection, Zhang Y, Guo YM, Xianyu YL, Chen WW, Zhao YY, Jiang XY, Adv Mater, 25(28):

3802-3819. (2013).

5. Matrix-Localization for Fast Analysis of Arrayed Microfluidic Immunoassays, Zhang Y, Wang XW, Song LS, Xu CL, Ma LY, Li ZH, Xi JH,

Jiang XY, Anal Methods-Uk, 4(10): 3466-3470. (2012).

6. Towards a High-Throughput Label-Free Detection System Combining Localized-Surface Plasmon Resonance and Microfluidics, Zhang

Y, Tang YF, Hsieh YH, Hsu CY, Xi JZ, Lin KJ, Jiang XY, Lab Chip, 12(17): 3012-3015. (2012).

7. Tissue-specific Mechanical and Geometrical Control of Cell Viability and Actin Cytoskeleton Alignment, Sci Rep-Uk,

doi:10.1038/srep06160 (2014).



Andy Tsun Dr., IPS Luo Yi Scientist in Protein Chemistry

Department NNCAS-PostDoc-2010-1

Title The effects of soluble TNF/TNFR-family ligands on human FOXP3+ Treg cells: a potential therapeutic tool against rheumatoid arthritis

24

List of Papers 1. Tsun A*, Chen Z and Li B*. Romance of the three kingdoms: RORgammat allies with HIF1alpha against Foxp3 in regulating T cell metabolism and differentiation. Protein & Cell. (2011), DOI 10.1007/s13238-011-1114-2. 2. Tsun A, Li YY and Li B*. You, Me, and Foxp3: Immune Regulation for Two. Immunotherapy. 2011 Oct; 3(10): 1139-42. 3. Gao Y, Lin F, Su J, Gao Z, Li Y, Yang J, Deng Z, Tsun A* and Li B*. Molecular Mechanisms Underlying the Regulation and Functional Plasticity of FOXP3+ Regulatory T Cells. GENES & IMMUNITY, (2011) .DOI: 10.1038/gene.2011.77. 4. Shan, Z., et al., Negative regulation of interferon-induced transmembrane protein 3 by SET7-mediated lysine monomethylation. J Biol Chem, 2013. 288(49): p. 35093-103. 5. Li, Y., et al., 60-kDa Tat-interactive protein (TIP60) positively regulates Th-inducing POK (ThPOK)-mediated repression of eomesodermin in human CD4+ T cells. J Biol Chem, 2013. 288(22): p. 15537-46.

CAS PI NN PI


Wang Ming-weiNi Guan Jiejie Deng

Prof., SIMM Zhang Jun Dr. NNCAS-PhD-2010-1

Title Development and validation of label-free T Cell activation and CRAC channel assays

List of Papers Ni Guan, Jiejie Deng, Ting Li, Xiao Xu, Jeffrey T. Irelan and Ming-Wei Wang (2013). Label-free monitoring of T cell activation by the impedance-based xCELLigence system. Molecular BioSystems, 9, 1035-1043.

CAS PI NN PI


Wang Ming-wei/

Li Ting Prof., SIMM

Michael Christian Gerstenberg/

Mats Wikström

NNCAS-PhD-2010-2

Title Interaction matrix of IGFBPs and its IGF-signalling modulating mechanism

Executive

Summary

1) Protein expression and functional characterization were carried out at the Novo Nordisk Foundation Center for Protein Research in

Copenhagen where IGFBP2, IGFBP3 and IGFBP5 were produced using the E. coli expression system. These proteins were subsequently

validated for their biological relevance by showing that they have high affinity binding to IGF-1, using surface plasma resonance.

2) It follows that a prototype protease enzyme assay was preliminarily developed at Copenhagen and transferred to NCDS for

optimization to the high-throughput screening (HTS) format. This method was validated to be suitable to large-scale compound

screening of non-peptidic inhibitors. HTS campaigns will be launched upon receipt of a large batch of the enzyme from Denmark.

25

2011



Fuquan Yang Prof. IBP Zhiguo Li Dr. NNCAS-2011-1

Title Study on the pathogenesis of type 2 diabetes mellitus (T2DM) using quantitative proteomics approach

List of Papers

1. Chen X, Wei S, Yang F*., Mitochondria in the pathogenesis of diabetes: a proteomic view., Protein Cell. 2012,3(9):648-660.

2. Xiulan Chen, Ziyou Cui, Shasha Wei, Junjie Hou, Zhensheng Xie,Xue Peng, Jing Li, Tanxi Cai, Haiying Hang, Fuquan Yang

Chronic high glucose-induced INS-1β cell mitochondrial dysfunction: a comparative mitochondrial proteome with SILAC Proteomics

2013,13, 3030-3039



Lei Pan Dr., IBP Mark Applebby Principal Scientist NNCAS-2011-2

Title Novel genes and their essential functions in innate immunity

Executive

Summary

During the two years of this project, we have achieved the following goals: firstly, a bioinformatics algorithm for better prediction

of candidate genes involved in the regulatory network of innate immunity has been developed. Based on this algorithm, the score

of each gene in the whole Drosophila genome for the likelihood of being an innate immune regulator has been accessed. Secondly,

different infectious models (such as septic injury infectious model, natural oral feeding infectious model and nano-inject viral

infectious model) have been set up for screening the biology function of these predictive candidates in vivo. Thirdly, we have

finished genetic screening of around 100 candidates according to their order in predictive list and validated their immune functional

readout. Additionally, we have focused on several interesting genes to explore their roles on innate immune regulation post

infection.

Other Outcomes

1. Establish a Drosophila immune-genetic lab in IBP, CAS.

2. Develop a bioinformatics algorithm to predict innate immune regulator in flies.

3. Genetic screen hundreds of mutant or RNAi fly strains post bacterial or viral infection.



Bai Li Dr., USTC John Linu, Trine Ryberg Clausen Dr. NNCAS-2011-5

26

Title NKT Cell as a Modulator in Obesity Associated Tissue Inflammation

List of Papers 1.Bai L*, Deng SL*, Reboulet R*, Mathew R, Teyton L, Savage PB, Bendelac A. Natural killer T (NKT)-B-cell interactions promote prolonged antibody responses and long-term memory to pneumococcal capsular polysaccharides. Proc Natl Acad Sci U S A. 2013 Oct 1; 110(40):16097-102.

CAS PI NN PI


Liu Qing Prof., SIMM Lau Jesper Dr. NNCAS-2011-7

Title GLP-1/glucagon receptor chimeras, ligand binding characterization and crystal structure eetermination

List of Papers 1.Fai Yiu Siu, Min He, Chris De Graaf, Dehua Yang, Zhiyun Zhang, Caihong Zhou, Gye Won Han, Qingpin Xu, Daniel Wacker, Jeremiah S. Joseph, Wei Liu, Vadim Cherezov, Jesper Lau, Vsevolod Katritch, Ming-Wei Wang and Raymond C. Stevens (2013). Structure of the human glucagon class B G-protein-coupled receptor. Nature, 499, 444–449

CAS PI NN PI


Zhang Chenghai/Xu Eric Prof., SIMM Steffen Reedtz-Runge Dr. NNCAS-2011-8

Title Structure-Based Design of Fibroblast Growth Factor 19 that specifically Retains its Metabolic but not Mitogenic Activities

Executive

Summary

Fibroblast growth factor 19 (FGF 19) and 21 (FGF21) are atypical members of the FGF family that function as endocrine hormones. They are powerful regulators of glucose and lipid metabolism. However, FGF19 has been implicated as an associated factor with hepatocellular carcinoma. A closely related member of the FGF19 subfamily, FGF21, has also been shown to have similar effects with an improvement of insulin sensitivity in diabetic animal models. Unlike FGF19, FGF21 does not show significant mitogenic effects in vivo. The reason is that both FGF19 and FGF21 are able to activate FGF receptors (FGFRs) 1c, 2c, and 3c in the presence of β-Klotho co-receptor. Only FGF19 activates FGFR4, the predominant receptor in the liver. According to recent research, FGF21 is a potential drug for the treatment of obesity and diabetes that is currently in clinical trials. However, FGF21 also is a critical rheostat for bone turnover and a key integrator of bone and energy metabolism probably via the actions of FGF21 in CNS. Thus, our objectives of this project are; 1) to design specific FGF 19 variants that are defective in their interactions with FGFR4 but retain their activities with other FGFRs and β-Klotho; 2) to design Fc-FGF 21 fusion proteins, which are expected not to enter the CNS system due to its enlarged molecular weight. We will then test the functional activities of these FGF19 variants or Fc-FGF 21 to determine their activities on energy metabolism and bone using cell-based assays and animal models of diabetes.

27



Wang Ming-wei, Zhiyun

Zhang, Yue Zhu

Prof., SIMM Arne Staby Dr. NNCAS-PhD-2011-1

Title Improving recombinant protein production: addressing the frequent issue of insoluble proteins

Executive Summary

The primary aim of this project is improving recombinant protein production, however, due to bio-safety regulations in our new

laboratory (within limit to human residence), we are prohibited to conduct bacterium-related experiments. The project was this

changed to measure expression of membrane receptors, mainly, GPCRs. Whole cell and membrane binding are two classical

receptor binding assays widely used in G-protein coupled receptor (GPCR) studies. In this research, we successfully developed two

experimental systems, i.e., FACS and Western blotting, as basic methods to measure membrane expression levels of GPCRs such

as glucagon and glucagon-like peptide-1 receptors.



Huarong Zhou

Li Fuqiang, postdoc

Prof., SIBS Trine Ryberg Clausen Dr. NNCAS-PostDoc-2011-1

Title MECHANISMS OF BETA-CELL RECOVERY IN THE DIABETIC GK RAT AFTER BARIATRIC SURGERY

Executive

Summary

As we all know, diabetic patients cured by bariatric surgery usually rebuild glucose homeostasis not only instantly by gut hormones

in generally thinking, but also sustainably by beta-cell regeneration and islet recovery. The latter contributes to advantage of this

therapy compared to general pharmaceutical therapy usually targeting to instant decrease blood glucose levels. Thus, research of

mechanism of islet recovery in the diabetic rat after bariatric surgery will supply much information or support for developing a new

type of drug targeting to rebuild glucose homeostasis sustainably. To reach this target, we firstly clarify whether existing beta cell

proliferation and/or stem cell differentiation contribute(s) mainly to beta-cell regeneration in the diabetic rats after bariatric

surgery. Secondly, screen the possible key targets which control the proliferation or differentiation process by gene array and

related calculation. Thirdly, study the detailed roles of some of those key targets on proliferation or differentiation process for

beta-cell regeneration in vivo and in vitro.



Zheng Zhou Prof., IBP NNCASGWP-2011-5

Title

28

List of Papers 1.Hong JJ, Feng HQ, Zhou Z, Ghirlando R, Bai YW. Identification of Functionally Conserved Regions in the Structure of the Chaperone/CenH3/H4 Complex. J. Mol. Biol. 2013, 425(3): 536-545 2.Mao Z, et al. Anp32e, a higher eukaryotic histone chaperone directs preferential recognition for H2A.Z. Cell Research. 2014

2012

CAS PI NN PI


Chang Chen Prof., IBP Dan Han Dr. NNCAS-2012-2

Title The function of PI4KIIα in type 2 diabetes mellitus (T2DM)

Executive

Summary

Based on the original objective, which is to investigate whether PI4kIIα could be a new target for T2DM therapy, we carried out this

project from two aspects. On one hand, we found that the expression of PI4KIIα is upregulated in T2DM mice models, including KK mice

and db/db mice. Also we investigated the roles of PI4KIIαin blood glucose control and insulin secretion in PI4KIIα transgenic mice (TG

mice) and INS-1 cells. We found that the blood glucose of PI4KIIαoverexpressed mice are higher than the control ones, and the glucose

tolerance and glucose-stimulated insulin secretion of TG mice are both impaired, while the Insulin tolerance has no difference between

TG mice and control mice. So we suspected that PI4KIIα overexpression could impaired the function of pancreas islet, and then result

in glucose tolerance impaired. We further confirmed this conclusion in cell lines, the results indicated that PI4KIIα overexpression can

markedly reduce insulin content both in cell lysate and cultured medium. And we generated PI4KIIα knockout mice in order to make

it clear that whether PI4KIIα inhibition can resist T2DM progression and could be a way for T2DM therapy. On the other hand, based

on the structure of PI4KIIα, which was resolved by our lab recently, we are screening its specific inhibitors now.

CAS PI NN PI


Wanzhu Jin Prof., IOZ Xujia Zhang Dr. NNCAS-2012-8(A)

Title The role of Nectin-2 in brown adipose tissue activation

29

Executive

Summary

Recently we successfully isolated brown adipose tissue (BAT) secreted protein (BATOkine)-nectin2. To investigate the physiological

role of Nectin2, pCDH-Nectin2 expression cDNA plasmid were constructed and over-expressed in the white adipose tissue progenitor

cell line 3T3-L1. We found that ectopic expression of Nectin2 in the WAT progenitor cell line 3T3-L1 could dramatically induce fatty acid

oxidation related gene expression. Plasma nectin2 levels were significantly increased upon cold acclinimation and post high fat diet. We

asked if nectin2 have any effect in high fat diet (HFD) induced body weight gain. When nectin2 injected to the HFD induced obesity

mice, body weight gain was significantly inhibited. These results strongly suggest that nectin2 might influence both blood glucose and

BAT activity.



Youcun Qian Prof., Institute of Health Sciences, SIBS Jacques Peschon Novo Nordisk 530 Fairview Ave

North Seattle NNCAS-2012-9(A)

Title Identify IL-17C as a potential drug target for autoimmune diseases

List of Papers

1.Song X, Gao H, Lin Y, Yao Y, Zhu S, Wang J, Liu Y, Yao X, Meng G, Shen N, Shi Y, Iwakura Y, Qian Y*. Alterations in the Microbiota

Drive Interleukin-17C Production from Intestinal Epithelial Cells to Promote Tumorigenesis. Immunity. 2014 Jan 16;40(1):140-52.



Rong Zeng Prof., SIBS Mads Grønborg Dr. NNCAS-2012-11

Title

Quantitative analysis of acute and long-term effects of RouX-en-Y gastric bypass on plasma proteome in subjects with T2D and normal

glucose tolerance

30

Executive

Summary

Totally, we obtained plasma samples from 19 individuals. Of them, 10 subjects were with normal glucose tolerance (NGT), and 9 ones

were with T2D. For each subject, we harvested plasma samples at 4 time points, i.e. before RYGB (Pre), 1 week (1w), 3 months (3m)

and 1 year (1y) after RYGB. At each time point, an oral glucose tolerance test (OGTT) was conducted, and plasma samples were

harvested at fasting status, 30 min and 45 min after OGTT respectively. That is to say, we obtained 12 plasma samples for each subject

at different statuses. From clinical characteristics we knew that plasma glucose was dramatically decreased from 8.8±2.3 to 7.0±1.2

mmol/l 1 week after RYGB in T2D individuals, but there was no significant decrease of BMI (from 43.1±5.1 to 42.4±5.3 kg/m2).

The existence of high-abundance plasma proteins would severely influence identification and quantification of other proteins.

Therefore, we depleted top 7 high-abundance plasma proteins from each sample via immunodepletion column. The depletion efficiency

and variation were well controlled (Rsd less than 0.08). With the aid of high-abundance plasma-protein depletion, the number of

identified and quantified protein could increase by approximately 50% (from 238 to 341) in one assay.

Here, we used TMT-labelling strategy to relatively quantify dynamic protein expressions before and after RYGB, which was more precise

than prevalent shotgun label-free strategy. Based on their temporal profiles, we clustered proteins with significant changes into

different dynamic models. Furthermore, we compared proteins’ expression profiles between NGT individuals and T2D ones. Eventually,

we mapped quantified plasma proteins into key functional items so as to reveal the potential mechanisms responsible for early glucose

metabolism improvement by RYGB.

CAS PI NN PI


Xiangdong Fang

Hongzhu Qu

Prof., BIG Wenfeng Xu Dr. NNCAS-2012-12(A)

Title Identify Novel SLE Targets Using GWAS and ENCODE Non-coding Variants.

31

Executive

Summary

Genome-wide association (GWAS) studies have identified many variants associated with common diseases including systemic lupus

erythematosus (SLE). However, more than 90% of disease- and trait- associated variants emerging from these studies lie within

non-coding sequence, complicating their functional evaluation. Previous studies suggest the involvement of a proportion of such

variants in transcriptional regulatory mechanisms, including modulation of promoter and enhancer elements. We will study the

genome-wide DHSs profiles as well as other accessible high-throughput sequencing data under the ENCODE Project and the Roadmap

Epigenomics Program with systematic biological strategies and bioinformatical pipelines to delineate the functions of SLE-association

variation in the non-coding regulatory DNA regions and identify their target genes which will be selected for bio-functional analysis and

novel therapeutic targets for SLE patients. We modified the previous pipeline reported in the Science paper [2012; 337: 1190-5] to

identify the target of SNPs residing in non-coding regulatory DNA regions, including the expansion of SNP pools and the addition of the

EQTL data. We derived the GWAS SNP set associated with SLE from the NHGRI GWAS Catalog, GWAS Integrator, and

Phenotype-Genotype Integrator (as of 4/15/2013). We obtained 136 new SNPs residing in the non-coding regulatory DNA regions

compared with the published set in the Science paper. Using the special strategy of correlating the non-coding DHS harbouring a SNP

associated with SLE with its target DHS in a promoter region, 10 additional SLE association SNPs were assigned a target gene for them.

We will use these 10 new target genes and 11 target genes identified previously to do bio-functional analysis and novel therapeutic

targets for SLE patients.

CAS PI NN PI


Dehua Yang Prof., SIMM Jesper Lau Dr. NNCAS-2012-15

Title The roles of GLP-1 receptor in mesencephalon dopamine neuron development and glia cells mediated neuroprotection as well as

neuroimmunoregulation

Executive

Summary

T2DM has been identified as a risk factor for PD. GLP-1R, an important drug target for T2DM, has also been implicated as a potential target for the treatment of PD. Previous studies have demonstrated the neuroprotection effect of GLP-1 and exendin-4 on dopamine neurons in vitro and in vivo. However, the effect and molecular mechanism of GLP-1R in regulation of dopamine neuron development, glia cell mediated neuroprotection and neuroimmunoregulation remain largely unknown. In this study, we aim at evaluating the roles of ligands of GLP-1R in neurogenesis, neuroprotection and neuroimmunoregulation, including GLP-1, exendin-4, Boc5 (a non-peptidic GLP-1R agonist) etc. We will also attempt to elucidate the molecular mechanism involved. In 2013, we established a technology platform for culturing neuroblastoma and microglia cells, as well as primary culture of astrocytes, dopamine neuron and microglia. We also set up a method to study dopamine neuron differentiation of mouse embryonic stem cells with the GFP reporter system. Meanwhile, we detected the expression of GLP-1R neurons, astrocytes and microglia. In addition, we found that the ligands of GLP-1R exerted significant neuroprotective effect on neurons and neuroinflammation in microglia. Finally, we explored the possible regulation of microRNAs on neuroprotection and GLP-1R expression.

32

CAS PI NN PI


Mingdong HuangProf. Fujian Institute of Research on the Structure of Matter

Peter Andreasen; Egon Persson

Aarhus Univ/Professor;

Investigator/NN NNCAS-2012-17

Title Chinese Academy of Science (CAS) meeting on Serine Protease Enzyme Systems in Health and Disease

Executive

Summary

We held an international meeting “Serine Protease Enzyme Systems in Health and Disease” successfully in Guilin, Guangxi during the

period of October 26th – 30th, 2013. This important meeting is possible as the result of this grant (Novo Nordisk (NN) - Chinese

Academy of Science (CAS) meeting grant).

We evaluated the outcome of this meeting by an anonymous survey of participants, and found that its scientific value was highly

acknowledged.

This meeting strengthens the current ongoing scientific collaborative relationships between meeting participants. We highly appreciate

the support from NN-CAS foundation!

For news coverage of this international meeting in English, see http://english.fjirsm.cas.cn/ns/icn/201311/t20131105_111997.html

Other

Outcomes

We host the Novo Nordisk - Chinese Academy of Science (CAS) workshop on Serine Protease Enzyme Systems in Health and Disease

in Guilin, Guangxi in October 26th – 30th, 2013. About 50 scientists and student from Denmark, Australia, Belgium, Italy and China

participate the meeting. The meeting was evaluated by an anonymous survey of participants, which showed that the participants are

highly satisfied with the academic standards of this meeting. See:

http://www.fjirsm.ac.cn/xwzx/dtbd/201311/t20131106_3968375.html ;

http://english.fjirsm.cas.cn/ns/icn/201311/t20131105_111997.html

CAS PI NN PI


Zhihua Liu Prof., IBP NNCASGWP-2012-2

Title

List of Papers 1.Liu Z, Lenardo MJ. The role of LRRK2 in inflammatory bowel disease. Cell Research. 2012 Jul;22(7):1092-4.

CAS PI NN PI


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Yong Gan Prof. SIMM Lars Hovgarrd Oral Formulation Development, Principal Scientist

NNCAS-2009-10 and 2012-04

Title Design of functional lipid nanoparticles for oral delivery of protein/peptide drugs and study of absorption mechanism

List of Papers

1.Xiuying Li, Miaorong Yu, Weiwei Fan, Yong Gan, Lars Hovgaard, Mingshi Yang. Orally active targeted drug delivery systems for proteins and peptides. Expert opinion on drug delivery. Accepted. May, 2014.2.Xiuying Li, Shiyan Guo, Chunliu Zhu, Quanlei Zhu, Yong Gan, Jukka Rantanen, Ulrik Lytt Rahbek, Lars Hovgaard, Mingshi Yang. Intestinal mucosa permeability following oral insulin delivery using core shell corona nanolipoparticles. Biomaterials. 2013, 34(37):9678-87. 3. Xiuying LI, Quanlei Zhu, Lars Hovgaard, Mingshi Yang, Yong Gan. Enhanced oral absorption of insulin by the proton-coupled folate transporter (PCFT) targeting chitosan nanocomplexes. 2013 AAPS Annual Meeting & Exposition (Nov. 2013, USA) 4.Xiuying Li, Yong Gan, Lars Hovgaard, Mingshi Yang. Design of lipid modified polymeric nanoparticles for improvement of oral absorption of insulin. In proceeding of 2nd Electronic Conference on Pharmaceutical Sciences 2012.5.Xiuying Li, Dan Chen, Chaoyi Le, Chunliu Zhu, Yong Gan, Lars Hovgaard, Mingshi Yang. Novel mucus-penetrating liposomes as potential oral drug delivery system: preparation, in vitro characterization, and enhanced cellular uptake, International Journal of Nanomedcine. 2011, 6: 3151–3162.

CAS PI NN PI


Ming-Wei Wang Prof., SIMM Wisniewsk

Magdalena, Mats Wikström

NNCAS-PostDoc-2012-1

Title Biophysical and Structural Studies of the interaction matrix of IGFBPs

Executive Summary

We have developed a mammalian (HEK293) expression system that enables the expression and secretion of full-length IGFBPs. Using this approach we have been able to successfully express all 6 human IGFBPs in the folded and active states that will allows us to perform further detailed biophysical and structural studies of this family of proteins.

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