Effect of Genetic Modifications on Pig Heart and Kidney Graft Survival in Baboons

基因改造对狒狒猪心和肾移植物存活的影响

• 批准号: 8009656
• 负责人: DAVID KC COOPER
• 金额: $ 53.12万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8009656
• 关键词: Acute; Adrenal Cortex Hormones; Antibodies; Antithymoglobulin; B-Lymphocytes; Blood Coagulation Disorders; Blood Platelets; Coagulation Process; Complement; Data; Development; Endothelial Cells; Engineering; Euthanasia; Family suidae; Functional disorder; Future; Galactosyltransferases; Genetic; Graft Survival; Heart; Heart Transplantation; Heterogeneity; Human; Immune response; Immunity; Immunosuppressive Agents; Instruction; Investigation; Kidney; Kidney Transplantation; Life; Light; Maintenance; Modification; Molecular; Natural Immunity; Natural Killer Cells; Neoadjuvant Therapy; Organ; Organ Transplantation; Papio; Platelet Activation; Play; Prevention; Primates; Protocols documentation; Regimen; Role; Source; Thrombocytopenia; Thrombomodulin; Thrombosis; Transgenic Organisms; Vascular Endothelium; Vascular Graft; Xenograft procedure; activated protein C receptor; adaptive immunity; genetic regulatory protein; graft failure; immunosuppressed; knock-down; knockout gene; macrophage; mutant; mycophenolate mofetil; nonhuman primate; novel; prevent; response; success

The proposed studies are directed towards resolving two inter-related current problems in pig-tononhuman primate (NHP) organ xenotransplantation using GTKO.CD46.CD55 pigs (designated GE pigs) (i) to identify a clinically-applicable regimen that prevents an adaptive immune response, and (ii) to identify a means of preventing coagulation dysfunction - in the form of thrombocytopenia and/or consumptive coagulopathy (CC) with associated thrombotic microangiopathy. The immune response may influence the development of coagulation disorders, and coagulation dysregulation may influence the immune response. The exact causative factors of CC remain uncertain, but (i) heightened innate immunity (antibody, complement, platelets, macrophages, NK cells), (ii) the adaptive immune response (T and B cells), and (iii) molecular incompatibilities between pig and NHP, may all play roles. Graft vascular heterogeneity may also be evident as CC and thrombocytopenia occur more rapidly in NHPs with pig kidney rather than heart grafts. We propose that genetic modifications in pigs and novel therapies may protect against these factors. We shall use CIITA mutant pigs (with knock-down of SLA Class II) and other pigs transgenic for human thromboregulatory factors on the GTKO background. Aim 1: To investigate the efficacy in baboons of a clinically-applicable immunosuppressive regimen in preventing the innate and adaptive immune responses after heterotopic heart Tx from (A) GE pigs (n=6) and (B) GE.CIITA pigs (n=6), and to determine whether prevention of elicited xenogeneic immunity correlates with delay in" the onset or prevention of coagulation dysfunction. Aim 2: To investigate the coagulation disorders that develop after (A) heterotopic heart Tx (n=6) or (B) lifesupporting kidney Tx (n=6) from GE.CIITA pigs additionally transgenic for human TBM in baboons immunosuppressed with a clinically-applicable regimen, and to determine the causes of graft failure and coagulation dysfunction. Aim 3: To investigate the coagulation disorders that develop after (A) heterotopic heart Tx (n=6) or (B) lifesupporting kidney Tx (n=6) from GE.CIITA.TBM pigs additionally transgenic for human EPCR in baboons immunosuppressed with a clinically-applicable regimen, and to determine the causes of graft failure and coagulation dysfunction.

所提出的研究是针对解决两个相互关联的当前问题，在猪-人 使用GTKO.CD46.CD55猪（指定为GE猪）的灵长类动物（NHP）器官异种移植（i） 鉴定预防适应性免疫应答的临床适用方案，和（ii）鉴定预防适应性免疫应答的临床适用方案， 预防凝血功能障碍的方法-以血小板减少和/或消耗性血小板减少的形式 凝血病（CC）与相关的血栓性微血管病。免疫反应可能会影响 凝血障碍的发展和凝血失调可能影响免疫应答。 CC的确切致病因素仍不确定，但（i）先天免疫力增强（抗体， 补体、血小板、巨噬细胞、NK细胞），（ii）适应性免疫应答（T和B细胞），和（iii） 猪和NHP之间的分子不相容性可能都起作用。移植血管异质性也可能 因为CC和血小板减少症在猪肾而不是心脏移植物的NHP中发生得更快。 我们认为，猪的遗传修饰和新的疗法可能会保护这些因素。我们 应使用CIITA突变猪（SLA II类基因敲低）和其他人源转基因猪 GTKO背景下的血栓调节因子。 目的1：研究临床适用的免疫抑制方案在狒狒中的疗效， 预防来自（A）GE猪（n=6）的异位心脏Tx后的先天性和适应性免疫应答，以及 (B)CIITA猪（n=6），并确定预防引起的异种免疫是否与 延迟凝血功能障碍的发作或预防凝血功能障碍。 目的2：研究异位心脏Tx（n=6）或（B）生命支持后发生的凝血功能障碍 来自GE.CIITA猪的肾脏Tx（n=6），狒狒中的人TBM额外转基因 用临床适用的方案进行免疫抑制，并确定移植失败的原因， 凝血功能障碍 目的3：研究（A）异位心脏Tx（n=6）或（B）生命支持后发生的凝血功能障碍 来自GE.CIITA.TBM猪的肾脏Tx（n=6），狒狒中的人EPCR额外转基因 用临床适用的方案进行免疫抑制，并确定移植失败的原因， 凝血功能障碍