Genetic engineering of porcine endothelial cell lines for evaluation of human-to-pig xenoreactive immune responses

Abstract Xenotransplantation (cross-species transplantation) using genetically-engineered pig organs offers a potential solution to address persistent organ shortage. Current evaluation of porcine genetic modifications is to monitor the nonhuman primate immune response and survival after pig organ x...

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Autores principales:	Ping Li, Julia R. Walsh, Kevin Lopez, Abdulkadir Isidan, Wenjun Zhang, Angela M. Chen, William C. Goggins, Nancy G. Higgins, Jianyun Liu, Randy R. Brutkiewicz, Lester J. Smith, Hidetaka Hara, David K. C. Cooper, Burcin Ekser
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3ae22fee438c45fa9103891970992214
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spelling	oai:doaj.org-article:3ae22fee438c45fa91038919709922142021-12-02T18:02:53ZGenetic engineering of porcine endothelial cell lines for evaluation of human-to-pig xenoreactive immune responses10.1038/s41598-021-92543-y2045-2322https://doaj.org/article/3ae22fee438c45fa91038919709922142021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-92543-yhttps://doaj.org/toc/2045-2322Abstract Xenotransplantation (cross-species transplantation) using genetically-engineered pig organs offers a potential solution to address persistent organ shortage. Current evaluation of porcine genetic modifications is to monitor the nonhuman primate immune response and survival after pig organ xenotransplantation. This measure is an essential step before clinical xenotransplantation trials, but it is time-consuming, costly, and inefficient with many variables. We developed an efficient approach to quickly examine human-to-pig xeno-immune responses in vitro. A porcine endothelial cell was characterized and immortalized for genetic modification. Five genes including GGTA1, CMAH, β4galNT2, SLA-I α chain, and β2-microglobulin that are responsible for the production of major xenoantigens (αGal, Neu5Gc, Sda, and SLA-I) were sequentially disrupted in immortalized porcine endothelial cells using CRISPR/Cas9 technology. The elimination of αGal, Neu5Gc, Sda, and SLA-I dramatically reduced the antigenicity of the porcine cells, though the cells still retained their ability to provoke human natural killer cell activation. In summary, evaluation of human immune responses to genetically modified porcine cells in vitro provides an efficient method to identify ideal combinations of genetic modifications for improving pig-to-human compatibility, which should accelerate the application of xenotransplantation to humans.Ping LiJulia R. WalshKevin LopezAbdulkadir IsidanWenjun ZhangAngela M. ChenWilliam C. GogginsNancy G. HigginsJianyun LiuRandy R. BrutkiewiczLester J. SmithHidetaka HaraDavid K. C. CooperBurcin EkserNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ping Li Julia R. Walsh Kevin Lopez Abdulkadir Isidan Wenjun Zhang Angela M. Chen William C. Goggins Nancy G. Higgins Jianyun Liu Randy R. Brutkiewicz Lester J. Smith Hidetaka Hara David K. C. Cooper Burcin Ekser Genetic engineering of porcine endothelial cell lines for evaluation of human-to-pig xenoreactive immune responses
description	Abstract Xenotransplantation (cross-species transplantation) using genetically-engineered pig organs offers a potential solution to address persistent organ shortage. Current evaluation of porcine genetic modifications is to monitor the nonhuman primate immune response and survival after pig organ xenotransplantation. This measure is an essential step before clinical xenotransplantation trials, but it is time-consuming, costly, and inefficient with many variables. We developed an efficient approach to quickly examine human-to-pig xeno-immune responses in vitro. A porcine endothelial cell was characterized and immortalized for genetic modification. Five genes including GGTA1, CMAH, β4galNT2, SLA-I α chain, and β2-microglobulin that are responsible for the production of major xenoantigens (αGal, Neu5Gc, Sda, and SLA-I) were sequentially disrupted in immortalized porcine endothelial cells using CRISPR/Cas9 technology. The elimination of αGal, Neu5Gc, Sda, and SLA-I dramatically reduced the antigenicity of the porcine cells, though the cells still retained their ability to provoke human natural killer cell activation. In summary, evaluation of human immune responses to genetically modified porcine cells in vitro provides an efficient method to identify ideal combinations of genetic modifications for improving pig-to-human compatibility, which should accelerate the application of xenotransplantation to humans.
format	article
author	Ping Li Julia R. Walsh Kevin Lopez Abdulkadir Isidan Wenjun Zhang Angela M. Chen William C. Goggins Nancy G. Higgins Jianyun Liu Randy R. Brutkiewicz Lester J. Smith Hidetaka Hara David K. C. Cooper Burcin Ekser
author_facet	Ping Li Julia R. Walsh Kevin Lopez Abdulkadir Isidan Wenjun Zhang Angela M. Chen William C. Goggins Nancy G. Higgins Jianyun Liu Randy R. Brutkiewicz Lester J. Smith Hidetaka Hara David K. C. Cooper Burcin Ekser
author_sort	Ping Li
title	Genetic engineering of porcine endothelial cell lines for evaluation of human-to-pig xenoreactive immune responses
title_short	Genetic engineering of porcine endothelial cell lines for evaluation of human-to-pig xenoreactive immune responses
title_full	Genetic engineering of porcine endothelial cell lines for evaluation of human-to-pig xenoreactive immune responses
title_fullStr	Genetic engineering of porcine endothelial cell lines for evaluation of human-to-pig xenoreactive immune responses
title_full_unstemmed	Genetic engineering of porcine endothelial cell lines for evaluation of human-to-pig xenoreactive immune responses
title_sort	genetic engineering of porcine endothelial cell lines for evaluation of human-to-pig xenoreactive immune responses
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/3ae22fee438c45fa9103891970992214
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Genetic engineering of porcine endothelial cell lines for evaluation of human-to-pig xenoreactive immune responses

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