Reduced human platelet uptake by pig livers deficient in the asialoglycoprotein receptor 1 protein

Leela L. Paris, Jose L. Estrada, Ping Li, Ross L. Blankenship, Richard A. Sidner, Luz M. Reyes, Jessica B. Montgomery, Christopher Burlak, James R. Butler, Susan M. Downey, Zheng Yu Wang, Matthew Tector, A. Joseph Tector

Research output: Contribution to journalArticle

24 Citations (Scopus)

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 ASGR1 gene in donor pigs may prevent xenotransplantation-induced thrombocytopenia. Methods Transcription activator-like effector nucleases (TALENs) were targeted to the ASGR1 gene in pig liver-derived cells. ASGR1 deficient pig cells were used for somatic cell nuclear transfer (SCNT). ASGR1 knock out (ASGR1-/-) fetal fibroblasts were used to produce healthy ASGR1 knock out piglets. Human platelet uptake was measured in ASGR1+/+ and ASGR1-/- livers. Results Targeted disruption of the ASGR1 gene with TALENs eliminated expression of the receptor. ASGR1-/- livers phagocytosed fewer human platelets than domestic porcine livers during perfusion. Conclusions The use of TALENs in liver-derived cells followed by SCNT enabled the production of healthy homozygous ASGR1 knock out pigs. Livers from ASGR1-/- pigs exhibit decreased human platelet uptake. Deletion of the ASGR1 gene is a viable strategy to diminish platelet destruction in pig-to-human xenotransplantation.

Original languageEnglish (US)
Pages (from-to)203-210
Number of pages8
JournalXenotransplantation
Volume22
Issue number3
DOIs
StatePublished - May 1 2015

Fingerprint

Asialoglycoprotein Receptor
Swine
Blood Platelets
Liver
Heterologous Transplantation
Proteins
Thrombocytopenia
Cytophagocytosis
Gene Deletion
Gene Silencing
Phagocytosis
Genes
Endothelial Cells
Perfusion
Fibroblasts

Keywords

  • ASGR1
  • asialoglycoprotein
  • genetically modified
  • liver xenotransplantation
  • pigs
  • platelet
  • thrombocytopenia
  • transcription activator-like effector nucleases

Cite this

Paris, L. L., Estrada, J. L., Li, P., Blankenship, R. L., Sidner, R. A., Reyes, L. M., ... Tector, A. J. (2015). Reduced human platelet uptake by pig livers deficient in the asialoglycoprotein receptor 1 protein. Xenotransplantation, 22(3), 203-210. https://doi.org/10.1111/xen.12164

Reduced human platelet uptake by pig livers deficient in the asialoglycoprotein receptor 1 protein. / 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, Zheng Yu; Tector, Matthew; Tector, A. Joseph.

In: Xenotransplantation, Vol. 22, No. 3, 01.05.2015, p. 203-210.

Research output: Contribution to journalArticle

Paris, LL, Estrada, JL, Li, P, Blankenship, RL, Sidner, RA, Reyes, LM, Montgomery, JB, Burlak, C, Butler, JR, Downey, SM, Wang, ZY, Tector, M & Tector, AJ 2015, 'Reduced human platelet uptake by pig livers deficient in the asialoglycoprotein receptor 1 protein', Xenotransplantation, vol. 22, no. 3, pp. 203-210. https://doi.org/10.1111/xen.12164
Paris LL, Estrada JL, Li P, Blankenship RL, Sidner RA, Reyes LM et al. Reduced human platelet uptake by pig livers deficient in the asialoglycoprotein receptor 1 protein. Xenotransplantation. 2015 May 1;22(3):203-210. https://doi.org/10.1111/xen.12164
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, Zheng Yu ; Tector, Matthew ; Tector, A. Joseph. / Reduced human platelet uptake by pig livers deficient in the asialoglycoprotein receptor 1 protein. In: Xenotransplantation. 2015 ; Vol. 22, No. 3. pp. 203-210.
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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 ASGR1 gene in donor pigs may prevent xenotransplantation-induced thrombocytopenia. Methods Transcription activator-like effector nucleases (TALENs) were targeted to the ASGR1 gene in pig liver-derived cells. ASGR1 deficient pig cells were used for somatic cell nuclear transfer (SCNT). ASGR1 knock out (ASGR1-/-) fetal fibroblasts were used to produce healthy ASGR1 knock out piglets. Human platelet uptake was measured in ASGR1+/+ and ASGR1-/- livers. Results Targeted disruption of the ASGR1 gene with TALENs eliminated expression of the receptor. ASGR1-/- livers phagocytosed fewer human platelets than domestic porcine livers during perfusion. Conclusions The use of TALENs in liver-derived cells followed by SCNT enabled the production of healthy homozygous ASGR1 knock out pigs. Livers from ASGR1-/- pigs exhibit decreased human platelet uptake. Deletion of the ASGR1 gene is a viable strategy to diminish platelet destruction in pig-to-human xenotransplantation.",
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AU - Li, Ping

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AU - Sidner, Richard A.

AU - Reyes, Luz M.

AU - Montgomery, Jessica B.

AU - Burlak, Christopher

AU - Butler, James R.

AU - Downey, Susan M.

AU - Wang, Zheng Yu

AU - Tector, Matthew

AU - Tector, A. Joseph

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N2 - 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 ASGR1 gene in donor pigs may prevent xenotransplantation-induced thrombocytopenia. Methods Transcription activator-like effector nucleases (TALENs) were targeted to the ASGR1 gene in pig liver-derived cells. ASGR1 deficient pig cells were used for somatic cell nuclear transfer (SCNT). ASGR1 knock out (ASGR1-/-) fetal fibroblasts were used to produce healthy ASGR1 knock out piglets. Human platelet uptake was measured in ASGR1+/+ and ASGR1-/- livers. Results Targeted disruption of the ASGR1 gene with TALENs eliminated expression of the receptor. ASGR1-/- livers phagocytosed fewer human platelets than domestic porcine livers during perfusion. Conclusions The use of TALENs in liver-derived cells followed by SCNT enabled the production of healthy homozygous ASGR1 knock out pigs. Livers from ASGR1-/- pigs exhibit decreased human platelet uptake. Deletion of the ASGR1 gene is a viable strategy to diminish platelet destruction in pig-to-human xenotransplantation.

AB - 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 ASGR1 gene in donor pigs may prevent xenotransplantation-induced thrombocytopenia. Methods Transcription activator-like effector nucleases (TALENs) were targeted to the ASGR1 gene in pig liver-derived cells. ASGR1 deficient pig cells were used for somatic cell nuclear transfer (SCNT). ASGR1 knock out (ASGR1-/-) fetal fibroblasts were used to produce healthy ASGR1 knock out piglets. Human platelet uptake was measured in ASGR1+/+ and ASGR1-/- livers. Results Targeted disruption of the ASGR1 gene with TALENs eliminated expression of the receptor. ASGR1-/- livers phagocytosed fewer human platelets than domestic porcine livers during perfusion. Conclusions The use of TALENs in liver-derived cells followed by SCNT enabled the production of healthy homozygous ASGR1 knock out pigs. Livers from ASGR1-/- pigs exhibit decreased human platelet uptake. Deletion of the ASGR1 gene is a viable strategy to diminish platelet destruction in pig-to-human xenotransplantation.

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