anx5 therapy - ashley davis
TRANSCRIPT
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Recombinant Human Annexin A5 Therapy – Inhibition of Proinflammatory Response with Improved Cardiac Function and Survival Rate
in Mice with Endotoxemia
Ashley DavisUniversity of North Florida
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Collaborative Research• Paul Arnold & Fatemeh Armirahmadi – Department of
Physiology and Pharmacology at the University of Western Ontario, London, Canada
• Xiangru Lu MD - Center of Critical Illness Research at the Lawson Health Research Institute ON, Canada
• Katharina Brandl PhD – Department of Genetics at the The Scripps Research Institute La Jolla, California
• Qingpin Feng MD, PhD – Department of Medicine at the University of Western Ontario, London, Canada
• Dr. Feng is a career investigator at Heart and Stroke Foundation of Canada
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Endotoxemia✧ Bacteria enter bloodstream and release endotoxins
✧ WBCs react to the presence of bacteria and release inflammatory substances – excessive cytokine production
✧ 18 million cases annually
✧ Increased endothelial permeability• Stage (II) - Severe Sepsis
• Organ dysfunction
• Mortality rate: 30-40%
• Stage (III) - Septic shock
• Myocardial depression
• Mortality rate: 40-80%
NIAID/RML, Science 2002
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Host’s Immunity✧ Host’s innate immunity activated
✧ TLR2 = Gram-positive
✧ TLR4 = Gram-negative
✧ Activation of TLR4 via LPS binding
✧ Initiates signaling pathway that leads to
✧ activation of MAPKs (mitogen-activated protein kinases)
✧ production of inflammatory cytokines - tumor necrosis factor (TNF-α) and Interleukin (IL-β)
Celeste Thorpe MD 2004
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Roll of Toll-like Receptors
Modlin, Nature 2000
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LPS• Lipopolysaccharide
• From Gram-negative bacteria
• LPS binds to TLR4 in CD14
• Binding of LPS activations signaling pathway that leads to activation of MAPKs and production of inflammatory cytokines
• Necrosis factor alpha (TNF-α)
• Leading cause of cardiac dysfunction
• Interleukin-1 beta (IL-β)
Feng et al. Crit Care Med, 2014
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Previous Treatment• Xigris = Activated Protein C
• Indications: for severe sepsis & septic shock
• Acute organ failure
• Properties: Anticoagulant, Anti-inflammatory, Profibrinolytic
• Side Effect: Serious Bleeding
• Was the only approved therapy by the U.S Food and Drug Administration
• No longer used due to severe bleeding risk
• Results from trial showed no survival benefit over placebo
• Xigris was voluntarily withdrawn from market in 2011
FDA Medwatch, 2011
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Annexin A5• A 35-kDa phospholipid binding protein
• High affinity binding to negatively charged phospholipids
• Has antiapoptotic and anticoagulant properties – by forming 2D crystallized shield over the surface of cells
• Shield decreases ability to initiate phagocytosis or thrombosis
• Anx5 interacts with cell receptors to inhibit their function in binding to leucine-rich repeats
• LPS binds to extracellular domain of TLR4 at leucine-rich repeats
• Anx5 inhibits LPS binding to TLR4 and thus decreases downstream MAPK signaling
HTI Inc, 2009
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Results• Anx5 improved cardiac function and survival rate during
endotoxemia by inhibiting LPS binding to TLR4/myeloid differentiation factor 2 (MD-2) receptor complex
• Anx5 decreased myocardial TNF-α and IL-βexpression
• Anx5 restored cardiac function to control levels
• via the inhibition of activation of NF-kB
Feng et al. Crit Care Med, 2014
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Anx5 decreases myocardial MAPKs and NK-κB
Feng et al. Crit Care Med, 2014
CardiacTissue
MAPK
Side Pathway
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Treatment with Anx5 significantly decreased myocardial TNF-αand IL-1β
Feng et al. Crit Care Med, 2014
mRNA
Protein
Plasma
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Anx5 Improves Cardiac Function
Feng et al. Crit Care Med, 2014
Measuring meanarterial pressure
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Improved Survival Rates5 day animal survival was significantly improved by either the immediate or 4-hour delayed
Anx5 treatment following LPS challenge.
Feng et al. Crit Care Med, 2014
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References1. Angus DC, Linde-Zwirble WT, et al. Epidemiology of severe sepsis in the United States:
Analysis of incidence, outcome, and associated costs of care. Critical Care Medicine 2001; 29:1303-1310.
2. Zhang T, Feng Q: Nitric oxide and calcium signaling regulate myocardial tumor necrosis factor- expression and cardiac function in sepsis. Can J Physiol Pharmacol 2010;88;92-104.
3. Parrillo JE, Parker MM, Natanson C, et al: Septic shock in humans. Advances in the understanding of pathogenesis, cardiovascular dysfunction, and therapy. Ann Intern Med 1990; 113:227-242.
4. Toussaint S, Gerlach H: Activated protein C for sepsis. N Engl J Med 2009;361:2646-2652.
5. Gentry CA, Gross KB, Sub B, et al: Adverse outcomes associated with the use of drotrecogin alfa (activated) in patients with severe sepsis and baseline bleeding precautions. Crit Care Med 2009; 37:19-25
6. Gerke V, Moss SE: Annexins: From structure to function. Physiol Rev 2002; 82:331-371
7. Reutelingsperger CP, van Heerde QL: Annexin V, the regulator of phosphatidylserine-catalyzed inflammation and coagulation during apoptosis. Cell Mol Life Sci 1997;53:527-532
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References8. Markoff A, Bogdanova N, Knop M, et al: Annexin A5 interacts with polycystin-1 and interferes with
polycystin-1 stimulated recruitment of E-cadherin into adherens junctions. J Mol Biol 2007;369:954-966
9. Bell JK, Mullen GE, Leifer CA, et al: Leucine-rich repeats and pathogen recognition in Toll-like receptors, Trends Immunol 2003; 24:528-533
10. Peng T, Lu X, Lei M, et al: Inhibition of p38 MAPK decreased myocardial TNF-alpha expression and improves myocardial function and survival during acute endotoxemia in mice. Cardiovasc Res 2003;59: 123-134
11. Ranieri VM, Thompson BT, Sarie PS, et al: PROWESS-SHOCK Study Group: Drotrecogin alfa (activated) in adults with septic shock. N Engl J Med 2012;366:2055-2064
12. US Food and Drug Administration: Voluntary market withdrawal of Xigris [drotrecogin alfa (activated)] due to failure to show a survival benefit. 2011.
13. Xiang FL, Lu X, Hammoud L, et al: Cardiomyocyte-specific overexpression of human stem cell factor improves cardiac function and survival post myocardial infarction in mice. Circulation 2009; 120: 1065-1074
14. Zhang T, Lu X, Beier F, et al: Rac1 activation induces tumor necrosis factor- expression and cardiac dysfunction in endotoxemia. J Cell Mol Med 2011; 15:1109-1121
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References15. Cour O, Kumar A, Parrillo JE, et al: Clinical review: Myocardial depression in sepsis and septic shock. Crit
Care 2002; 6:500-508
16. Peng T, Lu X, Lei M, et al: Endothelial nitric-oxide synthase enhances lipopolysaccharide-stimulated tumor necrosis factor-alpha expression via cAMP-mediated p38 MAPK pathway in cardiomyocytes. J Biol Chem 2003; 278:8099-8105
17. Grandel U, Fink L, Blum A, et al: Endotoxin-induced myocardial tumor necrosis factor-alpha synthesis depressed contractility of isolated rat hearts: Evidence for a role of sphingosine and cyclooxygenase-2-derived thromboxane production. Circulation 2000; 102:2758-2764
18. Damazo AS, Yona S, D’Acquisto F, et al: Critical protective role for annexin 1 gene expression in the endotoxemic murine microcirculation. Am J Pathol 2005; 166:1607-1617
19. Bouter A, Gounou C, Berat R, et al: Annexin-A5 assembled into two-dimensional arrays promotes cell membrane repair. Nat Commun 2011; 2:270
20. Lee WL, Liles WC: Endothelial activation, dysfunction, and permeability during severe infections. Curr Opin Hematol 2011; 18: 191-196
21. Romisch J, Seiffge D, Reiner G, et al: In-vivo antithrombotic potency of placenta protein 4 (annexin V). Thromb Res 1991; 61; 93-104