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Reduced human platelet uptake by pig livers deficient in the asialoglycoprotein receptor 1 protein
Author(s) -
Paris Leela L.,
Estrada Jose L.,
Li Ping,
Blankenship Ross L.,
Sidner Richard A.,
Reyes Luz M.,
Montgomery Jessica B.,
Burlak Christopher,
Butler James R.,
Downey Susan M.,
Wang ZhengYu,
Tector Matthew,
Tector A. Joseph
Publication year - 2015
Publication title -
xenotransplantation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.052
H-Index - 61
eISSN - 1399-3089
pISSN - 0908-665X
DOI - 10.1111/xen.12164
Subject(s) - xenotransplantation , asialoglycoprotein receptor , somatic cell nuclear transfer , microbiology and biotechnology , biology , platelet , gene targeting , receptor , miniature swine , immunology , transplantation , andrology , hepatocyte , gene , medicine , biochemistry , embryo , in vitro , blastocyst , embryogenesis
Abstract Background The lethal thrombocytopenia that accompanies liver xenotransplantation is a barrier to clinical application. Human platelets are bound by the asialoglycoprotein receptor ( ASGR ) on pig sinusoidal endothelial cells and phagocytosed. Inactivation of the ASGR 1 gene in donor pigs may prevent xenotransplantation‐induced thrombocytopenia. Methods Transcription activator‐like effector nucleases ( TALEN s) were targeted to the ASGR 1 gene in pig liver‐derived cells. ASGR 1 deficient pig cells were used for somatic cell nuclear transfer ( SCNT ). ASGR 1 knock out ( ASGR 1 −/− ) fetal fibroblasts were used to produce healthy ASGR 1 knock out piglets. Human platelet uptake was measured in ASGR 1 +/+ and ASGR 1 −/− livers. Results Targeted disruption of the ASGR 1 gene with TALEN s eliminated expression of the receptor. ASGR 1‐/‐ livers phagocytosed fewer human platelets than domestic porcine livers during perfusion. Conclusions The use of TALEN s in liver‐derived cells followed by SCNT enabled the production of healthy homozygous ASGR 1 knock out pigs. Livers from ASGR 1‐/‐ pigs exhibit decreased human platelet uptake. Deletion of the ASGR 1 gene is a viable strategy to diminish platelet destruction in pig‐to‐human xenotransplantation.