Thromboregulatory Barriers to Xenotransplantation

异种移植的血栓调节障碍

• 批准号: 7609174
• 负责人: SIMON C. ROBSON
• 金额: $ 32.7万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 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 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7609174
• 关键词: Abbreviations; Acute; Address; Agonist; Anti-Inflammatory Agents; Antibodies; Anticoagulants; Antithrombin III; Antithrombins; Area; Blood Coagulation Factor; Blood Platelets; Blood Vessels; Bone Marrow; Bone Marrow Cells; Bone Marrow Transplantation; CD31 Antigens; Cells; Chimerism; Clinical; Coagulation Process; Consumption; Cyclophosphamide; Cyclosporine; Cyclosporins; Data; Development; Disease; Endothelial Cells; Engraftment; Excision; Family suidae; Fibroblasts; Genes; Genetic; Goals; Graft Survival; Heterophile Antigens; Human; Humoral Immunities; Inflammatory; Infusion procedures; Injury; Intervention; Kidney; Kinetics; Link; MALDI-TOF Mass Spectrometry; Mediating; Mediator of activation protein; Miniature Swine; Modality; Modeling; Molecular; Molecular Profiling; Mus; Nature; Organ; Outcome; Papio; Pathway interactions; Phenotype; Platelet Activating Factor; Platelet Activation; Platelet Glycoproteins; Prevention; Primates; Principal Investigator; Process; Proteins; Proteomics; Purinergic P1 Receptors; Reaction; Reagent; Research; Resistance; Role; SolCD39; Sus scrofa; TFPI; Testing; Therapeutic; Thrombocytopenia; Thrombomodulin; Thromboplastin; Thrombotic Thrombocytopenic Purpura; Thymic Tissue; Time; Toxic effect; Transgenic Mice; Transgenic Organisms; Umbilical vein; Up-Regulation; Viral Proteins; Xenograft procedure; antithrombin III-protease complex; clinically relevant; gene therapy; implantation; in vivo; kidney xenograft; novel; nuclear transfer; nucleoside triphosphate; protein expression; research study; response; therapeutic gene; vascular inflammation; vector; von Willebrand Factor

Xenotransplantation may be clinically feasible once the mechanisms of rejection are understood and graft survival can be achieved without compromising the recipient to the extent that systemic toxicity is encountered. Thrombotic and inflammatory reactions to porcine bone marrow (BM)-derived cells and vasculature are linked to difficulties in establishing mixed discordant chimerism in primates and the development of thrombotic microangiopathy in vascularized grafts. These responses may be associated with humoral immunity to xenogeneic grafts and intrinsic molecular barriers between the discordant species. The development of the GalT-KO pig and removal of the dominant xenoantigen has been a major advance in this area. However, there are still problems in inducing tolerance by generating mixed chimerism, either by vascularized thymic tissues or the bone marrow BM-derived cell approach; limited xenograft survival times and graft injury are still a concern. The goals of this project are directed at delineating mechanisms of the thrombotic sequelae associated with the GalT-KO pig-to-baboon xenotransplant model. We will identify and characterize porcine antigenic targets of both natural and elicited xenoreactive antibodies directed against Gal negative xenografts in vivo by MALDI-TOF Mass Spectrometry. Vascular markers of thrombotic injury will be also determined by porcine gene mini-arrays. Protein expression profiling will be undertaken to validate these vascular markers of xenograft rejection. The role of antithrombotic interventions will be determined addressing both pharmacological and genetic modalities. Antithrombin agents will be administered to baboon recipients of porcine cell infusions and renal grafts, as dual anti-thrombotic and anti- inflammatory agents, at the time of graft implantation and with rejection episodes. We will also study treatment with solCD39 (an ectonucleotidase) and ATL146e (an adenosine receptor A2a agonist), in combination with this antithrombin strategy. Gene therapeutic vectors will be employed to over-express CD39, thrombomodulin and tissue factor pathway inhibitor in GalT-KO BM-derived cells, prior to their infusion into baboons. Outcomes with respect to inflammatory or thrombotic sequelae and engraftment will be examined. We will finally evaluate transgenic approaches to over-express CD39 and the natural human anticoagulant factors in mice and pigs. Our studies will be judged successful if novel and clinically relevant antithrombotic strategies can be then developed and applied.

一旦排斥反应的机制被了解，异种移植在临床上是可行的， 可以在不损害接受者的情况下实现存活， 遇到的。对猪骨髓（BM）衍生细胞的血栓形成和炎症反应， 血管系统与在灵长类动物中建立混合不协调嵌合体的困难有关， 血管化移植物中血栓性微血管病的发展。这些反应可能与 对异种移植物的体液免疫和不一致物种之间的内在分子屏障。的 GalT-KO猪的开发和显性异种抗原的去除是这方面的主要进展。 区然而，通过产生混合嵌合体诱导耐受性仍然存在问题， 血管化胸腺组织或骨髓BM衍生细胞方法;有限的异种移植物存活时间， 移植物损伤仍然是一个问题。本项目的目标是阐明 与GalT-KO猪-狒狒异种移植模型相关的血栓后遗症。我们将查明和 表征天然和诱发的异种反应性抗体的猪抗原靶点， 通过MALDI-TOF质谱法测定体内Gal阴性异种移植物。血栓性损伤的血管标志物 也将通过猪基因微阵列来确定。将进行蛋白质表达谱分析， 验证这些异种移植排斥反应的血管标志物。抗血栓干预的作用将是 确定解决药理学和遗传学模式。抗凝血酶剂将是 给予狒狒接受猪细胞输注和肾移植，作为双重抗血栓形成和抗- 炎症因子，在移植物植入时和发生排斥反应时。我们还将研究 用solCD 39（外核苷酸酶）和ATL 146 e（腺苷受体A2 a激动剂）处理， 与这种抗凝血酶策略相结合。基因治疗载体将被用于过表达 CD 39、血栓调节蛋白和组织因子途径抑制剂在GalT-KO BM衍生细胞中的表达， 注入狒狒体内炎症或血栓性后遗症和植入的结局将 接受检查。最后，我们将评估过表达CD 39的转基因方法和天然的人类细胞。 小鼠和猪的抗凝因子。如果研究具有新颖性和临床相关性，则我们的研究将被判定为成功 然后可以开发和应用抗血栓形成策略。