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.

项目摘要： 我们的总体计划目标是实施合理设计的治疗策​​略以改进 通过减少或消除HLA抗体（ABS）来减少移植率并减少同种异体移植损失。患者 由于先前的移植，怀孕或输血而产生了HLA ABS 由于找不到兼容供体，因此在移植前的等待时间过多。最近的FDA 进行研讨会是因为静脉注射免疫球蛋白的传统抗炎性疗法 （IVIG）和血浆置换不提供可接受的拒绝治疗或去除HLA ABS的结果 移植之前（称为脱敏）。我们先前的研究表明骨髓浆细胞 （BMPC）是HLA AB生产的主要来源。 BMPC的表征不足和展示 对当前体内疗法的实质性抗性。几种脱敏策略，例如静脉复置， 已经研究了类固醇，IVIG和利妥昔单抗耗尽CD20+ B细胞，已研究以消除产生AB的PC， 取出预先形成的抗HLA ABS或反向急性AB介导的排斥反应（AMR），并以有限的成功。合理的 这些策略的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，我们将PC靶向疗法作为一种治疗方法，可以是 通过在临床前模型中进行随后的评估进行适当的基础科学研究来增强。在 这个项目，我们将专注于蛋白酶体抑制剂（PI）疗法作为当前PC的基石 疗法。在AIM 1中，我们将采用主动部位特异性抑制剂和共价探针来量化结构和 体内PI治疗后，来自HLA敏化患者的BMPC的蛋白酶体的功能适应。我们的 先前的研究表明，一种高度专业的蛋白酶体变异的免疫蛋白酶体在 BMPC对标准PI治疗有抗性。重要的是，免疫蛋白酶体特异性抑制剂靶向BMPC 对标准PI疗法有抵抗力。在AIM 2中，我们将采用一个延长寿命的新颖的离体平台 通过与BM基质细胞培养基共培养的患者BMPC的患者。我们将整合结果以揭示合理的 设计的药物鸡尾酒可以协同靶向人类患者的耐药性BMPC，并且缺乏广泛 脱靶毒性。在AIM 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转录组分析将是 识别定义BMPC异质性的关键基因组变异和特征的执行，揭示了罕见的细胞 种群并确定HLA敏化和PI治疗对BMPC子集的影响。总的来说， 结果将创建可以推广的工具，以改善肾脏，心脏和肝脏的抗炎性疗法 移植。我们高度有条理，融合和多样化的团队合作，带来科学和医学 一起，可以将基本和临床上的知识直接转化为征服疾病。