Achieving Xenograft Tolerance through Thymic Programming in Primates

通过灵长类动物的胸腺编程实现异种移植耐受

• 批准号: 10553284
• 负责人: KAZUHIKO YAMADA
• 金额: $ 79万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-15 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10553284
• 关键词: Achievement; Acids; Address; Affect; Animals; Anti-CD40; Antibodies; B-Cell Acute Lymphoblastic Leukemia; B-Lymphocytes; Biological Assay; Bone Marrow; Bone Marrow Transplantation; CD3 Antigens; CD47 gene; CD80 gene; CTLA4-Ig; Cells; Chimerism; Clinical; Clinical Trials; Collaborations; Complement; Data; Development; Donor person; Down-Regulation; Endothelial Cells; Euthanasia; FDA approved; Family suidae; Generations; Genetic; Goals; Graft Rejection; Graft Survival; Growth; Health; Human; Immunosuppression; In Vitro; Interleukin-3 Receptor; Ischemia; Kidney; Kidney Transplantation; Learning; Life; Macrophage; Methods; Modeling; Modification; Monoclonal Antibodies; Natural Killer Cells; Organ; Organ Harvestings; Organ Preservation; Outcome; PECAM1 gene; PTPNS1 gene; Papio; Phagocytosis; Pharmaceutical Preparations; Play; Primates; Proteinuria; Protocols documentation; Regimen; Regulatory T-Lymphocyte; Renal function; Risk; Role; Specificity; Sphingomyelinase; T-Lymphocyte; TNFSF5 gene; Therapeutic; Thymus Gland; Time; Transgenes; Transgenic Organisms; Transplantation; Transportation; Treatment Protocols; Up-Regulation; Vascularization; Xenograft Model; Xenograft procedure; antibody-mediated rejection; bone; clinical application; clinical trial protocol; effective therapy; genetic regulatory protein; humanized mouse; kidney xenograft; natural antibodies; organ growth; pathogen; peripheral blood; podocyte; post-transplant; preservation; prevent; rituximab; side effect; thrombotic; thymus xenograft; treatment strategy

Project 1 Summary: The overall goal of Project 1 is to induce tolerance of porcine kidneys in baboons by co-transplantation (Tx) of vascularized donor thymus. We have made significant progress during the current project period (2016-2020). First, we have identified the mechanism responsible for proteinuria, a significant obstacle in our GalT-KO pig-to-baboon kidney xenotransplant (XTx) model for the long-term survival of life-supporting kidney xenografts, and developed effective treatment strategies for overcoming it. Down-regulation of sphingomyelin phosphodiesterase acid-like 3b and up-regulation of CD80 on pig podocytes were found to play critical roles in proteinuria and species incompatibility between pig CD47 and baboon SIRPα, which caused phagocytosis of pig endothelial cells and podocytes by baboon macrophages, was observed. We learned that either the administration of CTLA4-Ig and rituximab mAb for GalT-KO grafts or the use hCD47 transgenic (Tg) GalT-KO kidney grafts inhibited the development of post-transplant proteinuria, and markedly prolonged baboon survival up to 193 days. While the exponential growth of pig grafts or drug-related side effects triggered the euthanasia of recipient baboons, no evidence of graft rejection was seen and both pig-specific unresponsiveness in vitro, and the development of new baboon T cells were observed in multiple recipients VT+K XTx. Second, in collaboration with Project 2, we have achieved prolonged peripheral blood macrochimerism lasting >60 days accompanied by gradual loss of anti-non-Gal antibodies (abs) in baboons following hCD47+ pig intra-bone bone marrow Tx. Such durable porcine chimerism has not been previously achieved in any primate. Our renewal is focused on achieving two aims. Aim 1: Optimize the protocol for pig kidney XTx with vascularized thymic grafts by determining: (1) acceptable non-Gal ab levels; (2) requirement for human complement regulatory protein Tg for recipients with high non-Gal ab levels; (3) a strategy to prevent rejection in recipients with high non-Gal abs; (4) acceptable preservation times of pig kidney and thymus grafts using an FDA-approved continuous organ preservation machine; and (5) the ability to completely terminate immunosuppression. Aim 2: Determine the ability of combined VT+KTx and IBBMTx using hCD47+ human IL3 receptor (hIL3r) transgenic (Tg) pigs as BM donors on (achieve robust, durable T cell tolerance by combined deletional and regulatory mechanisms. Additionally, we will also explore the ability of this combined approach to permit long-term kidney xenograft tolerance in baboons with high non-Gal natural antibody levels. We anticipate that the above studies will lead to achievement of the short- term goal of extending pig VTL+KTx grafts to clinical applications in the next five years, and the long- term goal of inducing robust tolerance of pig xenografts, respectively.

项目1摘要：项目1的总体目标是在 血管供体胸腺的共移植（TX）狒狒。我们取得了重大进展 在当前项目期（2016-2020）。首先，我们确定了负责的机制 蛋白尿症，我们的galt-ko-ko-ko-wig-baboon肾脏替身（XTX）模型的重要障碍 支持生命的肾脏Xen仪的长期生存，并制定了有效的治疗策略 为了克服它。鞘氨质磷酸二酯酶酸样3B的下调和上调 发现猪足细胞上的CD80在蛋白尿中起关键作用，物种不兼容 在猪CD47和狒狒SirPα之间，引起猪内皮细胞的吞噬作用和 观察到狒狒巨噬细胞的足细胞。我们了解到的是 用于Galt-KO移植物或使用HCD47转基因（TG）Galt-KO肾脏的CTLA4-IG和Rituximab mAb 移植物抑制了移植后蛋白尿症的发展，并显着延长了狒狒的生存 长达193天。而猪移植物或与药物相关的副作用的指数生长触发了 接受者狒狒的安乐死，没有看到移植排斥的迹象，并且两种猪特异性 在体外反应症，并在多个中观察到新的狒狒T细胞的发展 收件人VT+K XTX。其次，在与项目2合作中，我们实现了延长的外围 血液宏观主义持续> 60天，通过抗杀性抗体的年级损失（ABS）完成了60天 在HCD47+猪内骨髓骨髓TX之后的狒狒中。这种耐用的猪嵌合没有 我们以前在任何灵长类动物中都取得了成就。我们的续约专注于实现两个目标。目标1： 通过确定：（1），用血管化胸腺移植物优化猪肾XTX方案 可接受的非GAL AB水平； （2）对接受者的人体完成调节蛋白TG的要求 具有高的非GAL AB水平； （3）防止非GAL ABS的受体排斥的策略； （4） 使用FDA批准的继续持续的猪肾和胸腺移植的可接受的保存时间 器官保存机器； （5）完全终止免疫抑制的能力。目标2： 使用HCD47+人IL3受体（HIL3R）确定合并VT+ KTX和IBBMTX的合并能力 转基因（TG）猪作为BM供体（在合并中实现坚固，耐用的T细胞耐受性 缺失和监管机制。此外，我们还将探讨这一点的能力 合并的方法允许在非gal的狒狒中允许长期肾脏特征耐受性 天然抗体水平。我们预计上述研究将导致实现短期 在未来五年内将Pig VTL+KTX移植物扩展到临床应用的术语目标，而长期 分别诱导猪异种移植耐受性的术语目标。