Application of Progenitor Niche Signals to Ex Vivo Nephrogenesis

祖细胞生态位信号在离体肾发生中的应用

• 批准号: 10260117
• 负责人: Thomas Joseph Carroll
• 金额: $ 53.03万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2022-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10260117
• 关键词: Address; American; Anatomy; Animals; Asses; Biological Assay; Biological Markers; Blood Vessels; Cells; Chronic Kidney Failure; Clinical; Communities; Complex; Dialysis patients; Dialysis procedure; Embryo; End stage renal failure; Engineering; Engraftment; Epithelial; Epithelium; Generations; Goals; Human; Image; Implant; In Vitro; Individual; Injury; Injury to Kidney; Kidney; Kidney Diseases; Kidney Transplantation; Life; Maintenance; Mechanics; Modification; Mus; Nephrons; Organ; Organogenesis; Organoids; Pathway interactions; Patients; Pattern; Pharmacology; Physiological; Protocols documentation; Regenerative response; Renal Replacement Therapy; Renal Tissue; Renal function; Research; Resources; Role; Signal Pathway; Signal Transduction; Site; Source; Structure; Survival Rate; Techniques; Technology; Testing; Time; Tissue Grafts; Tissues; Transplantation; Tubular formation; Urinary system; Vascular Graft; cell type; improved; in vivo; intravital microscopy; nephrogenesis; novel; novel therapeutics; prevent; progenitor; technology development; tool; urinary; Address; American; Anatomy; Animals; Asses; Biological Assay; Biological Markers; Blood Vessels; Cells; Chronic Kidney Failure; Clinical; Communities; Complex; Dialysis patients; Dialysis procedure; Embryo; End stage renal failure; Engineering; Engraftment; Epithelial; Epithelium; Generations; Goals; Human; Image; Implant; In Vitro; Individual; Injury; Injury to Kidney; Kidney; Kidney Diseases; Kidney Transplantation; Life; Maintenance; Mechanics; Modification; Mus; Nephrons; Organ; Organogenesis; Organoids; Pathway interactions; Patients; Pattern; Pharmacology; Physiological; Protocols documentation; Regenerative response; Renal Replacement Therapy; Renal Tissue; Renal function; Research; Resources; Role; Signal Pathway; Signal Transduction; Site; Source; Structure; Survival Rate; Techniques; Technology; Testing; Time; Tissue Grafts; Tissues; Transplantation; Tubular formation; Urinary system; Vascular Graft; cell type; improved; in vivo; intravital microscopy; nephrogenesis; novel; novel therapeutics; prevent; progenitor; technology development; tool; urinary

Summary Approximately 750,000 Americans have end stage renal disease, in which kidney function is insufficient to sustain life. Organ function can be supplemented by dialysis in these individuals, however the 10 year survival rate for individuals on dialysis is just over 10%. Survival rates are much better for patients receiving a kidney transplant, but organ supply does not match demand. Ex vivo organogenesis has the potential to provide functional tissue for renal replacement therapy. Furthermore, defining signals that functionally direct nephrogenesis may identify pathways that can be manipulated to augment the regenerative response of the injured kidney in vivo. Several groups, including our own, have established techniques that allow us to generate cellularly complex kidney organoids derived from human and mouse induced pluripotent cells. These tissues seem an ideal source for generating renal replacement tissue. Theoretically, one would take patient-derived renal organoids and transplant them onto a diseased kidney, where they would integrate with the host urinary system and improve renal function. Although several groups have attempted to perform these types of transplantations, there is no evidence to date that they functionally integrate with the host kidney. In our preliminary studies, we have identified three key obstacles that must be overcome in order to generate ex vivo renal organoids that integrate with the host. First, organoid structure is relatively disorganized, which is in contrast to the precise arrangement of cell types along the cortical-medullary axis of healthy, native kidneys. Second, with current strategies, organoid- derived tubules do not connect with host-derived tubules and the organoid-derived tubules involute over time. Third, we lack robust functional assays to identify experimental modifications that improve organoid function. Each of these barriers must be eliminated to generate functional organoids that can be clinically beneficial to patients. We hypothesize that the best approach to achieve integrated organoid tissue is to selectively generate cell types that match the anatomic site of engraftment. Specifically, we will identify conditions that will allow us to generate proximal nephrons, including glomeruli and proximal tubules with their associated interstitium and vasculature, (herein referred to as cortical organoids) for the purposes of engraftment. To this end, our strategy for ex vivo nephron generation is unique in its emphasis on promoting the anatomically “correct” epithelia and its microenvironment for the site of engraftment. Concurrent to this, we will identify factors and techniques that promote tubule-tubule fusion. Thus, once we have generated cortical organoids, we will utilize this technology to stimulate the tubules of the graft to anastomose with the tubules of the host. Finally, we will use live imaging and well-defined functional assays as a readout of tubular function to continually optimize our strategy. The long-term goal of this proposal is to engineer organized, ex vivo renal tissue that can be induced to form functional nephrons in animal hosts through novel grafting strategies.

概括 大约有75万美国人患有末期肾脏疾病，其中肾脏 功能不足以维持生命。器官功能可以通过透析补充 但是，透析个体的10年生存率刚刚超过10％。 对于接受肾脏移植的患者而言，生存率要好得多，但是器官供应确实 不符合需求。离体器官发生有可能提供功能组织 肾脏替代疗法。此外，定义在功能上引导肾病的信号 可以确定可以操纵的途径，以增强 体内受伤的肾脏。包括我们自己在内的几个小组已经建立了这些技术 允许我们产生源自人和小鼠的细胞复杂的肾脏器官 诱导多能细胞。这些时候似乎是产生肾脏更换的理想来源 组织。理论上，一个人会将患者衍生的肾脏器官移植到一个 患病的肾脏，他们将与宿主尿液系统整合并改善肾脏 功能。尽管几个小组试图执行这些类型的移植，但 迄今为止，尚无证据表明它们在功能上与宿主肾脏合成。在我们的 初步研究，我们已经确定了必须克服的三个关键障碍，以便 生成与宿主集成的离体肾脏器官。首先，器官结构是 相关的混乱，这与沿着细胞类型的精确排列形成鲜明对比 健康的本地肾脏皮质轴轴。第二，有了当前的策略，Organoid- 派生的管不与宿主衍生的管子和类器官衍生的管子连接 随着时间的流逝。第三，我们缺乏可靠的功能测定方法来识别实验修改 改善了器官功能。必须消除这些障碍以产生功能 可能对患者有益的类器官。 我们假设实现整合器官组织的最佳方法是 有选择地生成与植入解剖部位相匹配的细胞类型。具体来说，我们会的 确定将使我们能够产生近端肾词的条件，包括glomerulluli和 近端管及其相关的间质和脉管系统（此处称为皮质 类型）出于植入的目的。为此，我们的离体内肾经马曲的策略 一代是独一无二的，重点是促进解剖上的“正确”上皮及其 植入地点的微环境。同时，我们将确定因素和 促进管管融合的技术。那一旦我们产生了皮质器官， 我们将利用这项技术刺激移植管的管与管子的吻合 主持人。最后，我们将使用实时成像和定义明确的功能测定作为读数 管状功能不断优化我们的策略。该提议的长期目标是 有组织的工程师，可以诱导形成功能性肾脏的实体肾组织 在动物宿主中通过新颖的嫁接策略。