Tonix Pharmaceuticals Holding Corp. 8-K

Exhibit 99.02

 

Pilot Evaluation Of A Clinical Xeno Heart Transplant Regimen In A Preclinical Model I. Ileka 1 , R. Chaban 1 , K. Kinoshita 1 , G. McGrath 1 , Z. Habibabady 1 , A. Calhoun 1 , A. Maenaka 1 , M. Ma 1 , V. Diaz 1 , S. Sanatkar 1 , S. Low 2 , W. Qin 2 , K. Hall 2 , K. Whitworth 3 , D.K.C. Cooper 1 R.N. Pierson III 1 1 Center for Transplantation Sciences, Massachusetts General Hospital and Harvard Medical School, Boston, MA 2 eGenesis, Inc., Cambridge, MA 3 National Swine Resource and Research Center (NSRRC), Columbia, MO

  

 

Relevant Disclosures • R. Chaban was supported by the Benjamin Research Fellowship from the German Research Foundation (DFG) • I. Ileka is supported by T32 5T32 AI007529 - 24 • Gene edited pigs were provided by eGenesis and NSRRC (NIH grant U42OD011140) • Tonix Pharmaceuticals provided TNX - 1500*, a humanized, Fc - modified, dimeric anti - CD154 mAb • This work was supported by NIH grants UO1 AI153612 (Pierson), U19 AI090959 (Cooper), and sponsored research agreements with Tonix Pharmaceuticals and eGenesis • D.K.C. Cooper is a consultant to eGenesis * TNX - 1500 is an investigational new biologics and is not approved for any indication

  

 

Background • Gene - edited (GE) pigs for Xenotransplantation. – Remove CHO antigen targets of preformed Ab • TKO (Gal - 1,3 - a Gal, Neu5Gc, Sd ) – Add human regulatory molecules • Complement: CD46, CD55, CD59 • Coagulation: TBM, EPCR, TFPI – Add anti - inflammatory ‘transgenes’ • CD47, HLA - E/ b 2, HO - 1, A20, CD39

  

 

Background • We evaluated hearts from multi - GE pigs in baboon transplants treated with a novel costimulation - based immunosuppressive regimen and cold - perfused ‘ischemia minimization’.

  

 

Methods • Pig Donors – 3 - , 11 - , or 15 - GE pig hearts – novel costimulation - based immunosuppressive regimen – cold - perfused storage technique designed to minimize graft ischemia . • Five baboons' recipients received heterotopic heart transplants using Steen’s cold - perfusion ischemia minimization – 3 Reference pig hearts ( Ntl . Swine Resource & Research Ctr: NSRRC) • 3 - GE pigs: GTKO. β4 GALNT2KO.CD55 – 2 Multi - GE pig hearts ( eGenesis ) (TKO = GTKO.CMAHKO. β4 GALNT2KO) • TKO.hCD46.CD55.TBM.TFPI.HLAE/ β2 M.CD47. CD59.CD39.A20.HO1.PD - 1 ( EGEN - 4737 [15 - GE]) • TKO.hCD46.CD55.TBM.TFPI.HLAE/ β2 M.CD47. EPCR ( EGEN - 4417 [11 - GE])

  

 

Methods: Heterotopic Heart Transplantation • Standard UW cold perfusate. • Ischemia minimization: Steen method – battery - powered portable perfusion circuit – 4 o C Steen Solution – Protocol 2 - hr perfusion • Heterotopic abdominal xenograft transplant

  

 

Methods: Steen Ischemia Minimization 7 • Steen: buffered extracellular solution (laboratory - made) • 240ml of washed human RBCs; 760 ml Steen solution • Aortic perfusion at 4 o C, 40 - 50 mmHg, immersed in reservoir; LV vent

  

 

Methods: Recipient Immunosuppresive Treatment Regimen • Induction Therapy – Antithymocyte Globulin (ATG) – α CD20 • Day of Surgery Therapy – Thromboxane inhibitor (BIA) – TNF Inhibitor (Etanercept) – Interleukin 6 inhibitor receptor blocker (Tocilizumab) • Maintenance Therapy – α CD154 (TNX - 1500), Mycophenolate Mofetil(MMF), Corticosteroids – Interleukin 6 inhibitor receptor blocker (Tocilizumab)

  

 

Results: Summary Final Biopsy Graft Survival (Days) Major complication Anticoagulati on GE Animal ID Antibody mediated rejection 3 Graft rupture Heparin IV 3G B3022 Antibody mediated rejection 5 Graft rupture Heparin IV 3G B2922 No evidence of rejection 0 Refractory Ventricular fibrillation Heparin IV 3G B8522 Antibody mediated rejection 20 Abdominal dehiscence None EGEN - 4737 (15G) B2521 Acute cellular rejection (3R) with severe antibody mediated rejection 300 None None EGEN - 4417 (11G) B2621

  

 

Results : Graft Survival and Function

  

 

Results: Cardiac Echo, 11 - GE Pig POD 21 POD 21 POD 240 POD 240 Relative to POD 21, interventricular septal thickness remained stable on weekly measurements until POD 240. Immunosuppression intensity was reduced on POD 200; hypertrophy progressed and contractility diminished after POD 240, until graft failure and euthanasia on POD 300.

  

 

Results: Anti - pig Antibody in 3 - GE pigs IgM on CD3+ POD Fold change relative donor Pig BL (0)1 hr 2 hr D1 D3 D7 D8 0 1 2 3 4 5 6 7 8 B3022 B2922 B8522 IgG on CD3+ POD Fold change relative donor Pig BL (0)1 hr 2 hr D1 D3 D7 D8 0 1 2 3 4 5 6 7 8 B3022 B2922 B8522

  

 

Results: Anti - pig Antibody in multi - GE pigs IgM on CD3+ POD F o l d c h a n g e r e l a t i v e d o n o r P i g BL (0) D7 D49 D119 D189 D300 0 1 2 3 4 5 6 7 8 B2521 B2621

  

 

Results: Histology H&E H&E C4d/C4d/H&E A B3022 (3GE) D#3 B B8522 (3GE) D#0 H B2621 (11GE) D# 110 G B2621 (11GE) D# 50 F B2521 (15GE) D# 20 E B2521 (15GE) D# 20 D B2521 (15GE) D# 20 C B8522 (3GE) D# 0 I B2621 (11GE) D# 300

  

 

Conclusion • This pilot experience with multi - GE porcine hearts in the context of a clinically applicable ischemia minimization and immunosuppressive regimen supports our working hypothesis that is effective to prolong heart xenograft survival. • We plan to further test this hypothesis in an orthotopic heart xenograft model.

  

 

Thank you! Key Contributors: R Chaban A Calhoun V Diaz I Ileka RN Pierson III