We Are Ready for Prime Time Invest Today! Joseph Tector MD PhD, University of Alabama Birmingham School of Medicine

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1 We Are Ready for Prime Time Invest Today! Joseph Tector MD PhD, University of Alabama Birmingham School of Medicine

2 Disclosure I have founded Xenobridge LLC and have applied for patents related to pig engineering and xenotransplantation Portions of this project were funded by United Therapeutics.

3 Where We Are With Regards to Clinical Xenotransplantation Greater than 1-year survival in preclinical life supporting renal pig-to-primate model Understanding of pathophysiology and graft failure Familiar paradigm for posttransplant success Greater than 2-year survival in heterotopic heart transplant model Donor pigs with a negative CDC and Flow crossmatch to patients on the waitlist Detailed understanding of histocompatibility testing for clinical xenotransplantation

4 Where We Are With Regards to Clinical Xenotransplantation Progress regarding Zoonotic viral concerns Viral testing for donor pigs Greater understanding of PERV risk Clinical grade pig facility Community ethics outreach program

5 Heterotopic survival exceeding 2 years

6 Greater than 1-Year Survival in Preclinical Life Supporting Renal Pigto-Primate Model

7 Experimental Design

8 Graft Survival

9 Experimental Design

10 Graft Survival and Function

11 Antibody Dominates the Pathology of Long-Term Survivors GTKO/hDAF Rhesus macaque: T cell depletion, anti- CD154, MMF/steroids Glomerulopathy and thrombotic microangiopathy Basement membrane thickening Immunoglobulin and C4d deposition

12 Paradigm for Initial Clinical Xenotransplantation Trials

13 Donor Pigs with a Negative CDC and Flow Crossmatch to Patients on the Waitlist Detailed Understanding of Histocompatibility Testing for Clinical Xenotransplantation

14 Histocompatibility Testing

15 Histocompatibility Testing in Humans Flow cytometric crossmatch >1,000 waitlisted patients screened 45-55% of patients have negative crossmatch

16 cpra > 80 HLA Class I SLA Cross Reactivity

17 Limitations of the Preclinical Model No Rhesus or Baboon has a negative crossmatch to a GGTA1/B4GALNT2 KO pig Crossmatch to GGTA1/CMAH/B4GALNT2 KO pig is worse than to GGTA1/B4GALNT2 KO pig Baboons and Rhesus express Neu5Gc because they have an active CMAH gene Significant AMR is expected and not reflective of a crossmatch negative transplant

18 Rhesus Antibody Binding Rhesus Macaque serum antibodies tested on: GGTA1 KO GGTA1/B4GalNT 2 KO IgG IgM No Rhesus has a negative crossmatch

19 NHP vs. Human Antigen Elimination Better Crossmatch In Humans

20 We Cannot Test the Pig We Intend to Use in Patients in Our Preclinical Model

21 Triple Knockout Pig GGTA1/CMAH/B4GALNT2 KO Has a negative crossmatch Genetic engineering relatively simple minimizing Risk to recipient Graft can participate in immune response (fight infection) Risk to pig Immune system intact Risk to public Minimize off target effects Healthy pigs

22 Crossmatch in Non-Human Primates is Not Reflective of Human Scenario

23 Progress Regarding Zoonotic Viral Concerns

24 Porcine Endogenous Retrovirus (PERV) Never caused disease in pig or human Never infected non-transformed human primary cell Multiple anti-viral drugs are effective against PERV

25 Infectious Disease Testing for Donor Pigs

26 Infectious Disease Testing for We engineered an assay system to evaluate serum specimens for 30 viruses that need to be excluded in donor Donor Pigs

27 Clinical Grade Pig Facility

28 Building A Gilt Acclimation and Gestation Unit Configuration

29 Procedural Areas Behind Barrier Operating Room Animal Prep Room

30

31 Local Outreach Program Preparing the community for first trial Religious and civic leaders State Board of health Hospital operations committee Tuskegee ethics committee

32 Patient Selection for Initial Trial Important to remember when talking about expectations from xenotransplant Many patients will not ever be transplanted Bar is not being transplanted not allotransplant Preclinical data is very reasonable > 6 months Cyclosporine Increased survival from 32 to 42 days v.s. Immuran as monotherapy Limitations of caring for primates Therapeutic drug monitoring Not treating rejection

33 Acknowledgements UAB: Matt Tector Jose Estrada Luz Reyes Zhang Yu Wang Greg Martens Joe Ladowski Devin Eckhoff Emory: Andrew Adams Steven Kim Mandy Ford Ken Newell

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