Intragraft DepTOR and transplant rejection

移植内 DepTOR 和移植排斥

• 批准号: 9331928
• 负责人: David M. Briscoe
• 金额: $ 22.13万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-15 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9331928
• 关键词: Acute; Address; Adhesives; Adipose tissue; Allogenic; Allografting; Alpha Cell; Anatomy; Binding; Biological; Biology; CD4 Positive T Lymphocytes; Cardiac; Cell Adhesion Molecules; Cell Communication; Cells; Characteristics; Chronic; Clinical; Collaborations; Data; Development; Endothelial Cells; Ensure; Event; Evolution; Excision; Exploratory/Developmental Grant for Diagnostic Cancer Imaging; FRAP1 gene; Future; Graft Rejection; Graft Survival; Immune; Immune response; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Injury; Knock-in Mouse; Knock-out; Knockout Mice; Laboratories; Leukocytes; Life; Location; LoxP-flanked allele; MAP Kinase Gene; Modeling; Molecular; Mus; Muscle Cells; Organ Transplantation; Outcome; Pathway interactions; Pharmacology; Phenotype; Process; Proteins; Publishing; Reagent; Recruitment Activity; Reporting; Research; Research Proposals; Resources; Role; STAT1 gene; Savings; Signal Transduction; Small Interfering RNA; Solid; Suggestion; T-Lymphocyte; Testing; Time; Transgenic Mice; Transgenic Organisms; Translating; Transplantation; Ubiquitination; Vascular Endothelial Cell; allograft rejection; angiogenesis; base; cancer cell; cell type; chemokine; clinically relevant; cohesion; genetic regulatory protein; heart allograft; immunogenicity; immunoregulation; in vitro Assay; in vivo; in vivo Model; isoimmunity; knock-down; novel; response

Project Summary/Abstract An increasing body of evidence indicates that the state of activation, phenotype and functional response of the microvasculature to alloimmune injury creates an intragraft microenvironment that both initiates and sustains the progression of allograft rejection. The unique anatomic location of the graft endothelial cell (EC) ensures that it is a target of the alloimmune response, and characteristic phenotypes either augment or inhibit interactions with immune cells. We recently identified DepTOR as a cell intrinsic mTOR-binding partner that modulates mTOR, MAPK and STAT-induced signaling in microvascular EC. In addition, we found that it is potent to regulate activation responses, angiogenesis and EC-dependent mechanisms of inflammation in vitro. These observations allowed us to conclude that the removal of its negative/regulatory effects are a mechanistic component of the EC pro-inflammatory phenotype. Our findings also suggest that augmented levels of expression and/or function of DepTOR may inhibit intragraft EC activation and their immunogenicity to sustain immunoregulation in vivo. However, formal testing of all these possibilities requires models in which DepTOR expression may be regulated in vivo selectively within allografts post transplantation. In this R21 proposal, we plan to develop and use novel DepTOR transgenic mice as donors of cardiac allografts in well-established models in vivo. We will also use cultured EC from transgenic mice for mechanistic studies in vitro. In this manner, we will be able to evaluate whether DepTOR is functional within allografts to promote long-term graft survival, and whether it effects are dependent on its ability to regulate EC-dependent mechanisms of rejection. Our central hypothesis is that cell intrinsic expression of DepTOR within the graft is immunomodulatory and regulates EC activation responses, EC-dependent interactions with CD4+ T cells and acute and chronic rejection. We will test this hypothesis in two specific aims in which we will 1), determine the function of DepTOR within the graft, and its association with acute and chronic allograft rejection in vivo, and 2), determine the function of DepTOR in EC-dependent mechanisms of alloimmunity. Collectively, our proposed studies are novel and address clinically relevant questions and our approach provides for cohesiveness to translate in vitro findings into relevant transplant models in vivo. These exploratory studies will, for the first time, define a mechanism whereby the expression of a regulatory protein within the transplanted organ itself may be a determinant of the phenotype and outcome of the rejection process.

项目总结/摘要 越来越多的证据表明，细胞的激活状态、表型和功能反应与细胞的功能有关。 同种免疫损伤的微血管系统产生了一种移植物内微环境， 维持同种异体移植排斥反应的进展。移植内皮细胞（EC）的独特解剖位置 确保它是同种免疫应答的靶点，并且特征表型增加或 抑制与免疫细胞的相互作用。我们最近发现DepTOR是一种细胞内在的mTOR结合蛋白， 在微血管EC中调节mTOR、MAPK和STAT诱导的信号传导的伴侣。另外我们 发现它能有效调节激活反应、血管生成和内皮依赖性机制， 体外炎症。这些观察使我们得出结论，消除其负面/监管 作用是EC促炎表型的机制成分。我们的研究结果还表明， DepTOR的表达和/或功能水平的提高可以抑制移植物内EC活化及其 免疫原性以维持体内免疫调节。然而，对所有这些可能性的正式测试 需要模型，其中DepTOR表达可以在同种异体移植物后体内选择性地调节。 移植在这个R21提案中，我们计划开发和使用新型DepTOR转基因小鼠作为供体 心脏同种异体移植物在良好建立的体内模型。我们还将使用转基因小鼠培养的EC 用于体外机制研究。通过这种方式，我们将能够评估DepTOR是否有效 在同种异体移植物中促进长期移植物存活，以及它是否有效取决于其能力， 调节EC依赖的排斥机制。我们的中心假设是， 移植物内的DepTOR是免疫调节性的，并调节EC活化反应，EC依赖性 与CD 4 + T细胞的相互作用以及急性和慢性排斥反应。我们将在两个具体的实验中验证这一假设。 我们的目标是：1）确定移植物内DepTOR的功能，以及它与急性胰腺炎的关系。 和慢性同种异体移植物排斥反应，以及2）确定DepTOR在EC依赖性 同种免疫机制。总的来说，我们提出的研究是新颖的，并解决了临床相关性 问题和我们的方法提供了凝聚力，将体外发现转化为相关的移植 体内模型。这些探索性研究将首次确定一种机制， 移植器官本身内调节蛋白的表达可能是移植器官功能恢复的决定因素。 表型和排斥过程的结果。