Core A: PI: David L. Ayares, PhD Abstract: Precise genetic manipulation of the pig genome through a combination of somatic cell nuclear transfer and sophisticated gene modification tools offers the opportunity to provide an unlimited source of human-compatible donor organs, cells, and tissues for transplantation. As Core A, Revivicor will apply their advanced transgenic pig production platform to provide the GE source pig organs (kidney, heart, lung, and liver) necessary to meet the goals of the specific aims in Project 1 and Project 2. Elimination of immunogenic Gal?1,3Gal (Gal) sugars through inactivation of the ?1,3-galactosyl transferase gene (GTKO) was a critical first step, resulting in the elimination of hyperacute rejection (HAR), and prolonged survival of xenografts in non- human primate studies, compared to wild-type controls. To provide further protection protection from non-gal-mediated humoral rejection, next-generation GTKO pigs were produced with constitutive high-level expression of the complement regulatory genes, hCD46 and hCD55, individually and as triple transgenic pigs. In addition, for inhibition of coagulation and thrombosis, human anti-coagulant genes (EPCR, CD39, TFPI, and thrombomodulin), under endothelial cell-specific promoter systems, were added alone or in combination to produce donor pigs with up to six genetic modifications. The use of multi-cistronic ?2A? vectors has facilitated the production of these multi- transgenic pigs in which the added genes are not only co-expressed, but co-integrated in the genome, thus allowing propagation of new lines with reduced transgene segregation. Core A will utilize this multi-transgenic 2A pig platform to produce multiple combinations of six-gene (6GE) pigs that have already shown improved efficacy, and will further modify the source pig to inhibit T cell activation (expression of a dominant negative inhibitor of SLA class II (CIITA), and immunomodulation, through overexpression of the ?cloaking gene?, human CD47, to prevent activation of monocytes and other immune cells. In addition, towards prevention of spontaneous aggregation and sequestration of recipient platelets (and resulting thrombocytopenia in NHP), specific exons of porcine vWF will be replaced by their human equivalents using gene editing nuclease technologies. Core A will identify the optimal multi- gene mix, with an efficacious balance of gene expression levels, bioactivity, and synergistic effects, while maintaining a healthy herd, as needed to support the aims of Project 1 & 2.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program--Cooperative Agreements (U19)
Project #
5U19AI090959-07
Application #
9115987
Study Section
Special Emphasis Panel (ZAI1)
Project Start
Project End
Budget Start
2016-08-01
Budget End
2017-07-31
Support Year
7
Fiscal Year
2016
Total Cost
Indirect Cost
Name
University of Pittsburgh
Department
Type
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Yamamoto, Takayuki; Li, Qi; Hara, Hidetaka et al. (2018) B cell phenotypes in baboons with pig artery patch grafts receiving conventional immunosuppressive therapy. Transpl Immunol 51:12-20
Jagdale, Abhijit; Iwase, Hayato; Klein, Edwin et al. (2018) Will donor-derived neoplasia be problematic after clinical pig organ or cell xenotransplantation? Xenotransplantation :e12469
Zhang, Zhongqiang; Hara, Hidetaka; Long, Cassandra et al. (2018) Immune Responses of HLA Highly Sensitized and Nonsensitized Patients to Genetically Engineered Pig Cells. Transplantation 102:e195-e204
French, Beth M; Sendil, Selin; Sepuru, Krishna Mohan et al. (2018) Interleukin-8 mediates neutrophil-endothelial interactions in pig-to-human xenogeneic models. Xenotransplantation 25:e12385
Yamamoto, Takayuki; Iwase, Hayato; King, Timothy W et al. (2018) Skin xenotransplantation: Historical review and clinical potential. Burns 44:1738-1749
Li, Qi; Hara, Hidetaka; Zhang, Zhongqiang et al. (2018) Is sensitization to pig antigens detrimental to subsequent allotransplantation? Xenotransplantation 25:e12393
Laird, Christopher T; Hassanein, Wessam; O'Neill, Natalie A et al. (2018) P- and E-selectin receptor antagonism prevents human leukocyte adhesion to activated porcine endothelial monolayers and attenuates porcine endothelial damage. Xenotransplantation 25:e12381
Zhang, Guoqiang; Hara, Hidetaka; Yamamoto, Takayuki et al. (2018) Serum amyloid a as an indicator of impending xenograft failure: Experimental studies. Int J Surg 60:283-290
Jagdale, Abhijit; Cooper, David K C; Iwase, Hayato et al. (2018) Chronic dialysis in patients with end-stage renal disease: Relevance to kidney xenotransplantation. Xenotransplantation :e12471
Iwase, Hayato; Yamamoto, Takayuki; Cooper, David K C (2018) Episodes of hypovolemia/dehydration in baboons with pig kidney transplants: A new syndrome of clinical importance? Xenotransplantation :e12472

Showing the most recent 10 out of 115 publications