TARGETING HUMAN PLASMA CELLS TO OVERCOME HUMORAL RESPONSES IN TRANSPLANTATION

靶向人类浆细胞以克服移植中的体液反应

• 批准号: 10350564
• 负责人: James Joseph Driscoll
• 金额: $ 47.14万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-12-17 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10350564
• 关键词: 26S proteasome; Active Sites; Acute; Allografting; Antibodies; Antibody Formation; Autoantibodies; Autophagocytosis; B-Lymphocytes; Basic Science; Binding; Biochemical; Biological Assay; Blood Transfusion; Bone Marrow; Catabolic Process; Cell Survival; Cell Therapy; Cells; Clinical; Coculture Techniques; Complex; Degradation Pathway; Disease; Drug Combinations; Drug Design; Drug Screening; Drug Targeting; Drug resistance; Educational workshop; Evaluation; Event; Excision; Exhibits; Gene Proteins; Genetic Heterogeneity; Genomic approach; Genomics; Goals; Graft Rejection; Health; Heart Transplantation; Heterogeneity; Human; Immunosuppression; Individual; Intravenous Immunoglobulins; Isoantibodies; Kidney Transplantation; Knowledge; Longevity; Lysosomes; MS4A1 gene; Measures; Mediating; Medicine; Mission; Monitor; Nucleosome Core Particle; Organ; Outcome; Patients; Pharmaceutical Preparations; Pharmacology; Plasma Cells; Plasmapheresis; Population; Pre-Clinical Model; Pregnancy; Process; Proteasome Inhibitor; Proteomics; Refractory; Research; Resistance; Role; Science; Secretory Cell; Serum; Source; Steroids; Stromal Cells; System; Testing; Therapeutic; Toxic effect; Translating; Transplantation; Ubiquitin; Variant; Wait Time; Work; antibody-mediated rejection; arm; bioinformatics tool; cell stroma; clinically relevant; desensitization; donor-specific antibody; exhaust; functional adaptation; genomic signature; genomic tools; genomic variation; heart allograft; improved; in vivo; inhibitor; inhibitor therapy; kidney allograft; liver transplantation; lung allograft; multicatalytic endopeptidase complex; novel; novel therapeutic intervention; organ transplant rejection; pancreas allograft; particle; prevent; protein degradation; proteostasis; rational design; response; rituximab; side effect; single-cell RNA sequencing; small molecule; success; targeted treatment; therapeutic target; tool; transcriptomics

PROJECT SUMMARY: Our overarching programmatic goal is to implement rationally-designed therapeutic strategies to improve transplantation rates and reduce allograft loss by reducing or eliminating HLA antibodies (Abs). Patients who have developed HLA Abs as a result of a previous transplant, pregnancy, or blood transfusion may undergo excessive waiting times prior to transplantation because a compatible donor cannot be found. Recent FDA Workshops have been conducted because traditional antihumoral therapies with intravenous immunoglobulin (IVIg) and plasmapheresis do not provide acceptable outcomes for rejection treatment or removal of HLA Abs prior to transplantation (termed desensitization). Our prior studies indicate that bone marrow plasma cells (BMPCs) are the major source of HLA Ab production. BMPCs remain inadequately characterized and exhibit substantial resistance to current therapies in vivo. Several desensitization strategies, e.g., plasmapheresis, steroids, IVIg and rituximab to deplete CD20+ B cells, have been investigated to eradicate Ab-producing PCs, remove preformed anti-HLA Abs or reverse acute Ab-mediated rejection (AMR) with limited success. A plausible explanation for the shortcomings of these strategies is that they do not deplete BMPCs. To achieve our mission and overcome these obstacles, we propose PC targeted therapies as a therapeutic approach that can be enhanced by conducting appropriate basic science studies with subsequent evaluation in preclinical models. In this Project, we will focus on proteasome inhibitor (PI) therapy as the cornerstone of current PC targeted therapies. In Aim 1, we will employ active site-specific inhibitors and covalent probes to quantitate structural and functional adaptations in proteasomes from the BMPCs of HLA-sensitized patients after in vivo PI therapy. Our prior studies indicate that immunoproteasomes, a highly specialized proteasomal variant, are upregulated in BMPCs resistant to standard PI therapy. Importantly, immunoproteasome-specific inhibitors target BMPCs that are resistant to standard PI therapy. In Aim 2, we will employ a novel ex vivo platform that extends the longevity of patient BMPCs through co-culture with BM stromal cell media. We will integrate the results to reveal rationally- designed drug cocktails that synergistically target drug-resistant BMPCs from human patients and that lack broad off-target toxicities. In Aim 3, we will we will deploy cutting-edge genomic and bioinformatic tools to better define the cellular and genetic heterogeneity of BMPCs. Single cell (sc)RNA-Seq transcriptomic profiling will be performed to identify key genomic variations and signatures that define BMPC heterogeneity, reveal rare cell populations and determine the effect of HLA-sensitization and PI therapy on BMPC subsets. Taken together, the results will create tools that can be generalized to improve antihumoral therapies for kidney, heart and liver transplant. Our highly organized, integrated and diverse team works in concert to bring science and medicine together, resulting in basic and clinically-relevant knowledge that can be translated directly to conquer disease.

项目概要： 我们的总体目标是实施合理设计的治疗策​​略以改善 通过减少或消除 HLA 抗体 (Ab)，提高移植率并减少同种异体移植物损失。患者谁 由于先前的移植、怀孕或输血可能会产生 HLA 抗体 由于无法找到相容的供体，移植前的等待时间过长。最近的FDA 举办了研讨会，因为传统的静脉注射免疫球蛋白抗体液疗法 (IVIg) 和血浆置换术不能为排斥反应治疗或 HLA 抗体去除提供可接受的结果 移植前（称为脱敏）。我们之前的研究表明骨髓浆细胞 (BMPC) 是 HLA 抗体生产的主要来源。 BMPC 的表征和表现仍然不充分 对体内现有疗法有显着的耐药性。几种脱敏策略，例如血浆置换、 类固醇、IVIg 和利妥昔单抗可消耗 CD20+ B 细胞，已被研究用于根除产生抗体的 PC， 去除预先形成的抗 HLA 抗体或逆转急性抗体介导的排斥反应 (AMR)，但效果有限。一个看似合理的 对这些策略缺点的解释是它们不会耗尽 BMPC。实现我们的使命 并克服这些障碍，我们提出 PC 靶向治疗作为一种治疗方法，可以 通过进行适当的基础科学研究以及随后的临床前模型评估来增强。在 在这个项目中，我们将重点关注蛋白酶体抑制剂（PI）治疗，作为当前 PC 靶向治疗的基石 疗法。在目标 1 中，我们将采用活性位点特异性抑制剂和共价探针来定量结构和 HLA 致敏患者体内 PI 治疗后 BMPC 中蛋白酶体的功能适应。我们的 先前的研究表明，免疫蛋白酶体（一种高度专业化的蛋白酶体变体）在 BMPC 对标准 PI 治疗有抵抗力。重要的是，免疫蛋白酶体特异性抑制剂针对 BMPC 对标准 PI 治疗有抵抗力。在目标 2 中，我们将采用一种新颖的离体平台来延长寿命 通过与 BM 基质细胞培养基共培养来培养患者 BMPC。我们将整合结果，理性地揭示—— 设计的药物鸡尾酒可以协同靶向人类患者的耐药 BMPC，但缺乏广泛的 脱靶毒性。在目标 3 中，我们将部署尖端的基因组和生物信息学工具来更好地定义 BMPC 的细胞和遗传异质性。单细胞 (sc)RNA-Seq 转录组分析将 用于识别定义 BMPC 异质性的关键基因组变异和特征，揭示稀有细胞 人群并确定 HLA 致敏和 PI 治疗对 BMPC 亚群的影响。综合起来， 结果将创建可推广的工具，以改善肾脏、心脏和肝脏的抗体液治疗 移植。我们高度组织化、综合性和多元化的团队齐心协力，将科学和医学带入 在一起，产生可以直接转化为征服疾病的基础和临床相关知识。