Abstract 3128: Cardio-respiratory Protection In Sepsis By Endothelial-targeted Recombinant Cd39.

Abstract

Introduction: Sepsis results in inflammatory cytokine storm and endothelial activationleading to ARDS and cardiac stress. CD39 is an endothelial cell(EC)-anchored enzyme working in concert with CD73 to hydrolyze extracellular ATP (pro-inflammatory) /ADP (pro-thrombotic) to adenosine which is anti-inflammatory, vasodilatory,and anti-thrombotic. 'Anti-VCAM1-CD39’ is novel therapeutic containing a scFv recognizingVCAM-1 (receptor expressed on activated ECs) fused to human soluble CD39. Aim: To demonstrate that Anti-VCAM-CD39 can reduce inflammation and cardiac stressafter lipopolysaccharide (LPS) challenge. Methods: Mice were challenged with LPS 5mg/kg and treated with either Anti-VCAM-CD390.8mg/Kg or buffer control after 1 hr and anlysed at 3, 6, 24 and 48 h. Results: 1: LPS challenge leads to upregulation of VCAM-1 in the lungs (Q-PCR) and in the heart (imaged with MRI after administration of magnetic particles of iron oxidecoated with VCAM1 antibody or IgG control) (data not shown).2: LPS leads to reduction in plasma levels of protective adenosine (P<05).3: Treatment with anti-VCAM-CD39 reduces hypothermia, decreases pulmonary cytokinestorm and hypoxic drive (HIF1a) (Fig 1) and alleviatesmyocardial stress expressed (transcription of atrial natriuretic peptide (Nppa) and troponinat 24h) (Fig 2), reduces features of ARDS on lung histology (not shown) and suppressesinflammasome (NLRP3 by Western blot, not shown) Conclusion: Anti-VCAM-CD39 suppresses inflammation and offers cardiorespiratoryprotection in sepsis.Fig 1: Treatment with anti-VCAM-CD39 0.8mg/Kg reduces pulmonary cytokine storm (IL-1B,IL-6, TNFalpha, MCP1/CCL2) and hypoxic drive (HIF1a)(Statistics: One-way ANOVA; *=0.034, **=0.009)Fig 2: Treatment with anti-VCAM-CD39 0.8mg/Kg alleviates myocardial stress expressed(transcription of atrial natriuretic peptide (Nppa) and troponin at 24h)1 Additional Data referenced in the Abstract but could not be included due to space: LPS leads to reduction in plasma levels of protective adenosine(One-way Anova, P values shown)Treatment with anti-VCAM-CD39 0.8mg/Kg reduces hypothermia(Statistics: One-way ANOVA)Treatment with anti-VCAM-CD39 0.8mg/Kg suppresses inflammation (NLRP3 by Westernblot)