Engineering induction and assembly of human kidney tissue

人体肾脏组织的工程诱导与组装

• 批准号: 10419434
• 负责人: Alex Hughes
• 金额: $ 46万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10419434
• 关键词: Adult; Area; Autologous; Biocompatible Materials; Biological; Cell Lineage; Cells; Chronic Kidney Failure; Coculture Techniques; Complex; Data; Development; Dialysis procedure; Drainage procedure; Drug Targeting; Duct (organ) structure; Economic Burden; End stage renal failure; Engineering; Epithelial; Event; Fetal Kidney; Filtration; Future; Goals; Human; Immunocompromised Host; Invaded; Kidney; Kidney Diseases; Knowledge; Literature; Location; Lung; Mammary gland; Mechanics; Mesenchymal; Mesenchyme; Morphogenesis; Mus; Nephrons; Organ; Organ Transplantation; Organoids; Outcome; Pathology; Patients; Population; Positioning Attribute; Prevalence; Process; Prostate; Public Health; Quality of life; Regenerative Medicine; Renal Tissue; Research; Site; Solid; Surface; System; Technology; Tissue Model; Tissues; To specify; Transplantation; United States National Institutes of Health; Urine; WNT Signaling Pathway; Work; base; biophysical properties; body system; cell assembly; cell community; disability; fetal; human pluripotent stem cell; human tissue; innovation; model building; nephrogenesis; novel strategies; optogenetics; programs; regenerative tissue; screening; spatiotemporal; stem cells; urinary

PROJECT SUMMARY The goal of this proposal is to study and control nephron induction and assembly towards the formation of replacement renal tissue. Kidney organoids re-create an astonishing cellular diversity comparable to the early fetal kidney. However, structural connectivity of urine-producing nephrons and their drainage network formed by ureteric epithelium (UE) is required to avoid rapid pathology, yet has not been achieved. Accordingly, there is an urgent need to achieve connectivity between nephrons and ureteric epithelium before kidney organoids can achieve their potential in regenerative medicine. Our long-term goal is to construct ‘higher-order’ synthetic kidney tissues using human autologous stem cell lineages and assembly technologies that mimic the outcomes of morphogenesis. Our overall objectives at this stage are firstly to gain spatial control over nephron formation by determining how the mechanical microenvironment contributes to their induction sites and maturation. Its second objective is to direct nephron fusion with UE at many spatial sites through a controlled invasive process. Achieving these objectives will mark a transformative advance towards creating replacement kidney tissue. Our central hypothesis is that mechanical compaction of mesenchymal cells during kidney morphogenesis permits nephron induction, and subsequently that tight spatiotemporal control over WNT signaling events is necessary for their efficient fusion with UE. We plan to achieve the objectives through two specific aims. Firstly, we will determine the mechanical basis of nephrogenesis and use it to specify nephron positions. We will study mechanical compaction of the nephrogenic mesenchyme, assess biophysical properties of early nephron cells, and optimize nephrogenesis at specific locations using micropatterning technology. Secondly, we will program WNT-induced fusion of nephrons with ureteric epithelium. We will optimize fusion in nephron-ureteric epithelial co-cultures using optogenetic control over WNT signaling, and then trigger nephron assembly with UE spheroids after transferring them from micropatterned surfaces. The proposed research is innovative because we create fundamental knowledge while creating tissues that are biomaterial- free, human-derived (compatible with patient-derived autologous cell strategies), and therefore open to future development for transplantation. The proposed research is significant because higher-order assembly of human kidney tissue will create a step-change in renal replacement technology beyond dialysis, transplant, and “abiotic” filtration. We expect these efforts to have significant positive impact in the areas of fundamental biological discovery, drug target screening, and regenerative medicine.

项目摘要 该提案的目的是研究和控制肾单位诱导和组装朝向地层 替代肾组织。肾脏器官重新创建了可与 早期胎儿肾脏。但是，产生尿液的肾单位的结构连通性及其排水网络 需要由输尿管上皮（UE）形成以避免快速病理，但尚未实现。根据 在肾脏之前，迫切需要在肾脏和输尿管上皮之间达到连通性 类器官可以在再生医学中发挥潜力。我们的长期目标是构建“高阶” 使用人类自体干细胞谱系和组装技术的合成肾脏组织 形态发生的结果。我们在此阶段的总体目标首先是为了获得对肾统的空间控制 通过确定机械微环境如何促进其感应位点的形成和 成熟。它的第二个目标是通过受控 侵入性过程。实现这些目标将标志着创建更换的变革性进步 肾脏组织。我们的中心假设是肾脏期间间充质细胞的机械压实 形态发生允许肾单位诱导，随后对Wnt的紧密时空控制 信号事件对于它们与UE的有效融合是必需的。我们计划通过两个 具体目标。首先，我们将确定肾病的机械基础，并使用它来指定 肾单位位置。我们将研究肾原性间充质的机械压实，评估生物物理 早期肾单位细胞的特性，并使用微图像在特定位置优化肾脏形成 技术。其次，我们将对Wnt诱导的肾单位融合与输尿管上皮进行编程。我们将 使用对Wnt信号的光遗传学控制，优化肾单位 - 排骨上皮共培养，然后 从微图表表面转移后，用UE球体触发肾单位组件。提议 研究具有创新性，因为我们在创建生物材料的时机的同时创建基本知识 免费，人类衍生（与患者衍生的自体细胞策略兼容），因此与未来开放 移植的发展。拟议的研究很重要，因为人类的高阶组装 肾脏组织将在透析，移植和“非生物”之外创建肾脏替代技术的逐步变化 过滤。我们预计这些努力将对基本生物学领域产生重大积极影响 发现，药物目标筛查和再生医学。