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: 10598587
• 负责人: Alex Hughes
• 金额: $ 44.39万
• 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/10598587
• 关键词: Adult; Area; Autologous; Biocompatible Materials; Biological; Cell Lineage; Cells; Chronic Kidney Failure; Coculture Techniques; Complex; Data; Development; Dialysis procedure; Disease; Donor person; Drainage procedure; Drug Targeting; Duct (organ) structure; Economic Burden; End stage renal failure; Engineering; Epithelium; Event; Fetal Kidney; Filtration; Future; Goals; Human; Immunocompromised Host; Invaded; Kidney; Kidney Diseases; Knowledge; Literature; Location; Lung; Mammary gland; Mechanics; Mesenchymal; Mesenchyme; Morbidity - disease rate; Morphogenesis; Mus; Nephrons; Organ; Organ Transplantation; Organoids; Outcome; Pathology; Patients; Pattern; Population; Positioning Attribute; Prevalence; Process; Prostate; Public Health; Quality of life; Regenerative Medicine; Renal Tissue; Research; Site; Solid; Specific qualifier value; Structure; Surface; System; Technology; Tissue Model; Tissues; Transplantation; United States National Institutes of Health; Urine; WNT Signaling Pathway; Work; biophysical properties; bioprinting; body system; cell assembly; cell community; disability; 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.

项目概要 该提案的目标是研究和控制肾单位的诱导和组装以形成 肾脏替代组织重建了令人惊讶的细胞多样性，可与肾脏替代组织相媲美。 然而，早期胎儿肾脏的产尿肾单位及其排水网络的结构连通性。 需要由输尿管上皮（UE）形成以避免快速病理学，但尚未实现。 迫切需要在肾脏之前实现肾单位和输尿管上皮之间的连接 类器官可以在再生医学中发挥其潜力，我们的长期目标是构建“更高阶”。 使用人类自体干细胞谱系和模仿的组装技术合成肾组织 我们现阶段的总体目标首先是获得对肾单位的空间控制。 通过确定机械微环境如何影响其诱导位点和 其第二个目标是通过受控的方式在许多空间位置引导肾单位与 UE 融合。 实现这些目标将标志着创造替代品的变革性进展。 我们的中心假设是肾脏过程中间充质细胞的机械压缩。 形态发生允许肾单位诱导，以及随后对 WNT 的严格时空控制 信令事件对于它们与UE的有效融合是必要的，我们计划通过两个来实现目标。 首先，我们将确定肾发生的力学基础并用它来指定。 我们将研究肾源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，并使用微图案优化特定位置的肾发生 其次，我们将编程WNT诱导的肾单位与输尿管上皮的融合。 使用对 WNT 信号的光遗传学控制优化肾单位-输尿管上皮共培养物中的融合，然后 从微图案表面转移后触发肾单位与 UE 球体的组装。 研究具有创新性，因为我们在创造生物材料组织的同时创造了基础知识—— 免费、源自人类（与源自患者的自体细胞策略兼容），因此面向未来 所提出的研究具有重要意义，因为人类的高阶组装。 肾组织将为肾脏替代技术带来超越透析、移植和“非生物”的重大变革 我们预计这些努力将在基础生物领域产生重大积极影响。 发现、药物靶点筛选和再生医学。