Cell specific delivery of novel therapies to enhance glomerular regeneration and repair

细胞特异性递送新疗法以增强肾小球再生和修复

• 批准号: 10247521
• 负责人: Stuart James Shankland
• 金额: $ 81.62万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10247521
• 关键词: Affect; Age; Animal Model; Cells; Cicatrix; Clinical; Clinical Data; Clinical Trials; Deterioration; Disease; Epithelial Cells; Family; Focal Segmental Glomerulosclerosis; Genes; Goals; Human; Innovative Therapy; Kidney; Kidney Diseases; Knowledge; Lead; Libraries; Methods; Molecular; Morphology; Mus; Natural regeneration; Organoids; Parietal; Patients; Peptides; Physiological; Process; Recombinants; Renal function; Renal glomerular disease; Reporter; Research; Research Personnel; Safety; Technology; Testing; Therapeutic; Tissues; Transplantation; Urine; Work; blood filter; cell injury; cell type; design; efficacy evaluation; epithelial stem cell; experimental study; glomerular function; human data; in vivo; in vivo regeneration; injured; innovation; kidney biopsy; kidney cell; migration; nanobodies; novel; novel therapeutics; organoid transplantation; podocyte; preclinical development; prevent; progenitor; regeneration potential; repaired; self-renewal; small molecule; stem cells; stemness; targeted delivery; targeted treatment; therapeutic candidate; therapeutic development; tool

PROJECT SUMMARY/ABSTRACT The goal of this project is to change the treatment paradigm for proteinuric glomerular diseases by combining therapeutics development with cell-specific delivery to enhance podocyte repair and regeneration in vivo. Podocytes, highly specialized terminally differentiated epithelial cells, are injured in the majority of glomerular diseases. As podocytes cannot self-renew, podocyte loss leads to glomerular scarring. A subpopulation of parietal epithelial cells (PECs) can serve as podocyte stem cells (`PEC progenitors'), but their regenerative potential is insufficient to overcome disease-associated glomerular damage. Enhancing productive repair of podocytes thus requires a dual synchronized approach: (i) replacing lost podocytes to increase their number, and (ii) limiting/reversing damage to the remaining podocytes. However, major knowledge gaps prevent us from achieving these goals; these include our limited knowledge on the molecular factors stimulating PEC self-renewal and podocyte regeneration/repair, as well as options methods for delivering these factors to specific kidney cell types in vivo. Our team of four expert investigators will wield complementary tools to close these knowledge gaps and produce innovative therapies. Dr. Wessely will apply Design of Experiment (DoE) approaches to identify novel combinations of molecules that increase PEC progenitors and reduce podocyte loss; Dr. Roberts will conjugate these therapeutics to VHHs (nanobodies) for delivery to PEC progenitors and podocytes; Dr. Freedman will generate gene-edited human kidney organoids to validate effects of VHHs compared to clinical data from patients; Dr. Shankland will use lineage tracing animal models of podocyte depletion and human organoids transplanted into mouse kidneys for in vivo safety and efficacy analysis. This pipeline will ultimately test the hypothesis that targeted delivery of PEC- and podocyte-specific therapeutic cargos can enhance podocyte repair and regeneration in vivo, and restore glomerular function to below the clinical disease threshold. The work will be accomplished through two Specific Aims, each with unique Milestones. The first Aim is to increase glomerular regeneration in vivo by cell targeted delivery of novel combinations of peptides and small molecules to augment podocyte progenitors of parietal epithelial cell origin. The second Aim is to increase productive repair of damaged podocytes by cell-type specific delivery of newly identified therapies. For both aims, we will employ the above pipeline to discover candidate therapeutics by DoE and cross-referenced with glomerular disease signatures from human patients. These will be combined with cell type-specific VHHs from high diversity recombinant VHH libraries to selectively deliver them to human PECs (Aim 1), or podocytes (Aim 2). Enhanced regeneration in vivo will be demonstrated in animal models of FSGS and transplanted human organoids. This process will establish a new paradigm for the treatment of kidney disease, and produce lead therapeutic candidates for further pre-clinical development and ultimately human clinical trials.

项目摘要/摘要 该项目的目的是通过合并来改变蛋白尿肾小球疾病的治疗范例 具有特异性递送的治疗性开发，以增强体内的足细胞修复和再生。 足细胞，高度专业的终末分化上皮细胞，在大多数中受伤 肾小球疾病。由于足细胞不能自我更新，因此足细胞损失会导致肾小球疤痕。一个 顶叶上皮细胞（PEC）的亚群可以用作足细胞干细胞（“ PEC祖细胞”），但它们的 再生潜力不足以克服与疾病相关的肾小球损伤。增强 因此 增加其数量，以及（ii）限制/逆转对剩余足细胞的损害。但是，主要 知识差距阻止我们实现这些目标；这些包括我们对分子的有限知识 刺激PEC自我更新和足细胞再生/维修的因素，以及选择方法 将这些因素输送到体内特定的肾细胞类型中。 我们由四名专家调查人员组成的团队将掌握互补的工具来缩小这些知识差距和 产生创新的疗法。 Wessely博士将采用实验设计（DOE）方法来识别新颖 增加PEC祖细胞并减少足细胞损失的分子组合；罗伯茨博士将连接 这些对VHHS（纳米生物体）的治疗剂，用于递送到PEC祖细胞和足细胞；弗里德曼博士 与来自临床数据相比 患者; Shankland博士将使用Podocyte Deptetion和人类器官的谱系追踪模型 移植到小鼠肾脏中，以进行体内安全性和有效性分析。该管道最终将测试 靶向赋予PEC和足细胞特异性治疗性cargos的假设可以增强Podocyte 体内修复和再生，并将肾小球功能恢复到临床疾病阈值以下。 这项工作将通过两个特定的目标来完成，每个目标都具有独特的里程碑。第一个目的是 通过细胞靶向肽和小型组合的细胞靶向递送，增加体内肾小球再生 分子增强顶皮细胞起源的足细胞祖细胞。第二个目标是增加 通过细胞类型的新鉴定疗法的特异性递送对受损的足细胞的生产性修复。两者 目的，我们将采用上述管道来发现DOE的候选治疗剂，并交叉参考 来自人类患者的肾小球疾病特征。这些将与来自细胞类型的特异性VHH结合 高度多样性重组VHH库有选择地将其传递给人PEC（AIM 1）或足细胞（AIM 2）。在FSG的动物模型中将证明增强体内的再生并移植人 器官。该过程将建立一个新的范式来治疗肾脏疾病，并产生铅 用于进一步临床前发育和最终人类临床试验的治疗候选者。