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

基本信息

批准号：
10328001
负责人：
KAZUHIKO YAMADA
金额：
$ 81万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
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/10328001
关键词：
Achievement Acids Address Affect Animals Anti-CD40 Antibodies B-Cell Acute Lymphoblastic Leukemia B-Lymphocytes Biological Assay Blood Vessels Bone Marrow Bone Marrow Transplantation CD3 Antigens CD47 gene CD80 gene CTLA4-Ig Cells Chimerism Clinical Clinical Trials Clinical trial protocol document 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 Life Methods Modeling Modification Monoclonal Antibodies Natural Killer Cells Organ Organ Harvestings Organ Preservation Outcome PECAM1 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 Xenograft Model Xenograft procedure antibody-mediated rejection bone clinical application effective therapy genetic regulatory protein humanized mouse kidney xenograft macrophage natural antibodies organ growth pathogen peripheral blood podocyte post-transplant preservation prevent rituximab side effect thrombotic thymus xenograft treatment strategy

项目摘要

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概要：项目1的总体目标是诱导猪肾的耐受性通过血管化供体胸腺联合移植（Tx）来治疗狒狒，我们已经取得了重大进展。在当前项目期间（2016-2020年），我们确定了负责机制。蛋白尿，这是我们的 GalT-KO 猪狒狒肾异种移植 (XTx) 模型中的一个重大障碍维持生命的肾异种移植物的长期存活，并制定了有效的治疗策略鞘磷脂磷酸二酯酶酸样 3b 的下调和上调。研究发现猪足细胞上的 CD80 在蛋白尿和物种不相容性中发挥关键作用猪CD47和狒狒SIRPα之间的相互作用，引起猪内皮细胞的吞噬作用我们观察到狒狒巨噬细胞的足细胞。用于 GalT-KO 移植物的 CTLA4-Ig 和 rituximab mAb 或使用 hCD47 转基因 (Tg) GalT-KO 肾移植物抑制了移植后蛋白尿的发生，并显着延长了狒狒的存活时间长达193天，而猪移植物的指数增长或药物相关的副作用引发了这种情况。对受体狒狒实施安乐死，没有发现移植排斥的证据，并且都是猪特异性的体外无反应，并且在多个体内观察到新狒狒 T 细胞的发育接受者 VT+K XTx 其次，与项目 2 合作，我们实现了外周延长。血液大嵌合持续>60天，伴随抗非Gal抗体（abs）逐渐丧失在狒狒中，hCD47+猪骨内骨髓Tx没有这种持久的猪嵌合状态。此前已在任何灵长类动物中实现过，我们的更新重点是实现两个目标：通过确定以下因素来优化猪肾 XTx 与血管化胸腺移植物的方案：(1) 可接受的非 Gal ab 水平；(2) 受者对人补体调节蛋白 Tg 的要求；非 Gal ab 水平较高；(3) 预防非 Gal ab 水平较高的受者出现排斥反应的策略；使用 FDA 批准的连续保存法对猪肾和胸腺移植物进行可接受的保存时间器官保存机；以及（5）完全终止免疫抑制的能力。使用 hCD47+ 人 IL3 受体 (hIL3r) 确定 VT+KTx 和 IBBMTx 组合的能力转基因（Tg）猪作为 BM 供体（通过组合实现稳健、持久的 T 细胞耐受性）此外，我们还将探索其能力。联合方法允许狒狒具有高非 Gal 的长期肾异种移植耐受性我们预计上述研究将实现短期目标。未来五年将猪 VTL+KTx 移植物扩展到临床应用的长期目标，以及长期目标分别诱导猪异种移植物的稳健耐受性的长期目标。