Identification of the immunomodulatory mechanisms of nanocarrier-enhanced costimulation blockade in an allogeneic portal vein islet transplantation model

异体门静脉胰岛移植模型中纳米载体增强共刺激阻断的免疫调节机制的鉴定

• 批准号: 10494100
• 负责人: Evan A. Scott
• 金额: $ 18.98万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-24 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10494100
• 关键词: Address; Adolescent; Allogenic; American; Amputation; Anti-Inflammatory Agents; Antigen-Presenting Cells; Antigens; Benchmarking; Biodistribution; Biological; Biological Assay; Blindness; Blood Glucose; CD8-Positive T-Lymphocytes; CD80 gene; CD8B1 gene; CTLA4 gene; CTLA4-Ig; Cell Cycle; Cells; Cessation of life; Chimeric Proteins; Clinical; Collaborations; Data; Disease; Drug Carriers; Encapsulated; Endocrine System Diseases; Engineering; Erythrocytes; Flow Cytometry; G1 Phase; Graft Survival; Heart; Histocompatibility Antigens; Hydrophobicity; Hypoglycemia; Immune; Immunoglobulins; Immunologics; Immunosuppression; Immunosuppressive Agents; Immunotherapy; Inflammatory; Injury; Insulin; Insulin-Dependent Diabetes Mellitus; Interleukin-2; Intestines; Islets of Langerhans Transplantation; Kidney; Kidney Failure; Liver; Major Histocompatibility Complex; Mediating; Methods; Mixed Lymphocyte Culture Test; Neuropathy; Patients; Peripheral Blood Lymphocyte; Polymers; Population; Portal vein structure; Production; Protocols documentation; Regimen; Regulation; Regulatory T-Lymphocyte; Rodent Model; Self Administration; Sirolimus; Specificity; Splenocyte; Structure of beta Cell of islet; Sulfides; T-Cell Proliferation; T-Lymphocyte; TNFRSF5 gene; Testis; Therapeutic; Therapeutic immunosuppression; Toxic effect; Transplantation; Up-Regulation; Work; alternative treatment; anti-CTLA4; base; clinically relevant; cost; drug efficacy; ethylene glycol; experience; immunoregulation; improved; in vivo; islet; liver transplantation; mTOR Inhibitor; mTOR inhibition; monocyte; mouse model; nanocarrier; nanomaterials; nanoscale; novel; post-transplant; prevent; propylene; side effect; standard of care; subcutaneous; transplant model; uptake

PROJECT SUMMARY Dysregulation of blood glucose due to T1D leads to both short term consequences such as hypoglycemia induced injury and death, and long-term complications including amputation, blindness, kidney failure, and neuropathy. Islet transplantation has the potential to provide biologic glycemic regulation for all T1D patients, effectively curing the disease. However, this promising therapy is plagued by the requirement for ineffective immunosuppressive therapies. Currently, only 50 to 70% of grafts remain viable at 5 years post-transplantation, largely due to the immunosuppressive therapy causing toxicity and failing to fully protect the graft. Currently, a costly cocktail of immunosuppressive drugs is given in a rarely successful attempt to provide broad immune coverage with side effect mitigation. Thus, a need exists for simpler, lower cost, and more effective alternatives to these immunosuppressive regimens that can maintain islet transplant survival without off-target side-effects. The objective of this proposal is to mechanistically understand how repurposing a single component of these immunosuppressive cocktails, rapamycin, using engineered nanoscale drug carriers can provide sustained islet protection. Vesicular polymeric nanocarriers (i.e. polymersomes, PS) encapsulating rapamycin (rPS) were found to uniquely change the cellular biodistribution of rapamycin to avoid side-effects and significantly improve efficacy. Importantly, rapamycin normally has a wide cellular biodistribution and functions by directly inhibiting T cell proliferation, but rPS completely avoids T cells and instead switches the immunosuppressive mechanism to a selective and potent costimulation blockade of antigen presenting cells (APCs). This novel cell-selective nanocarrier-enhanced costimulation blockade was characterized by an upregulation of CD8+ regulatory T cells and double positive CD4+CD8+ T cells, achieving sustained normoglycemia in a rigorous fully major histocompatibility complex (MHC) mismatched allogenic intraportal (liver) islet transplantation mouse model. Here, this proposal will investigate and benchmark the immunological mechanism of rPS against the clinically relevant therapeutic belatacept, a CTLA4-IgG fusion protein that induces costimulation blockade via an alternative method: the blocking of CD80/86 coreceptors on APCs. The following aims will be achieved: Aim 1: Determine whether rPS induces general immunosuppression or antigen-specific tolerance. It is currently not known whether rPS induce antigen-specific or general systemic immunosuppression. A novel ex vivo mixed lymphocyte reaction assay and in vivo dual antigen-specific / non-specific islet transplantation will address this question. Aim 2: Compare the immunomodulatory mechanisms of costimulation blockade induced by rPS vs. CTLA4-IgG for enhanced allogenic islet graft survival. High parameter flow cytometry using T-distributed Stochastic Neighbor Embedding (tSNE) analysis and assessment of long-term graft survival following fully-MHC mismatch islet transplantation will be performed.

项目摘要 由于T1D引起的血糖失调导致两种短期后果，例如低血糖 诱发伤害和死亡以及长期并发症，包括截肢，失明，肾衰竭和 神经病。胰岛移植具有为所有T1D患者提供生物学血糖调节的潜力 有效治愈疾病。但是，这种有希望的疗法困扰着无效的要求 免疫抑制疗法。目前，在移植后5年，只有50％至70％的移植物保持生存 很大程度上是由于免疫抑制疗法引起的毒性，无法完全保护移植物。目前， 免疫抑制药物的昂贵鸡尾酒是在很少成功提供广泛免疫的尝试中提供的 覆盖范围，副作用缓解。因此，需要更简单，更低的成本和更有效的替代方案 对于可以维持胰岛移植生存的这些免疫抑制方案，而无需靶向副作用。 该提案的目的是机械地了解如何重新利用这些组成部分 雷帕霉素免疫抑制鸡尾酒，使用工程纳米级药物可以提供持续的胰岛 保护。发现囊泡纳米载体（即聚合物体，PS）封装了雷帕霉素（RPS） 独特地改变雷帕霉素的细胞生物分布以避免副作用并显着改善 功效。重要的是，雷帕霉素通常通过直接抑制，具有广泛的细胞生物分布和功能 T细胞增殖，但RPS完全避开T细胞，而是切换免疫抑制机制 对抗原呈递细胞（APC）的选择性和有效的共刺激阻碍。这种新颖的细胞选择性 纳米载体增强的共刺激阻滞的特征是CD8+调节T细胞的上调 和双阳性CD4+ CD8+ T细胞，在严格的完全专业中达到持续的正常血糖 组织相容性复合物（MHC）不匹配的同种异体内（肝）胰岛移植小鼠模型。 在这里，该提案将调查和基准基准RPS针对临床的免疫学机制 相关的治疗性Belatacept，一种CTLA4-IGG融合蛋白，可通过A 替代方法：在APC上阻止CD80/86共感受器。将实现以下目标： 目标1：确定RPS是诱导一般免疫抑制还是抗原特异性耐受性。目前是 不知道RP是诱导抗原特异性或一般全身免疫抑制。一个小说的离体混合 淋巴细胞反应测定和体内双重抗原特异性 /非特异性胰岛移植将解决此问题 问题。 AIM 2：比较RPS与CTLA4-IGG引起的共刺激阻滞的免疫调节机制 为了增强同种异体胰岛移植的存活。使用T分布的随机邻居高参数流式细胞仪 完全MHC不匹配胰岛后，嵌入（TSNE）分析和评估长期移植物存活率 将进行移植。