Engineering induction and assembly of human kidney tissue

项目总结 这项建议的目标是研究和控制肾单位的诱导和组装形成 替换的肾组织。肾脏有机化合物重新创造了惊人的细胞多样性，可与 早期胎儿肾脏。然而，产尿肾单位及其引流网络的结构连通性 形成输尿管上皮(UE)是避免快速病理所需，至今尚未实现。因此， 在肾脏之前，迫切需要实现肾和输尿管上皮之间的连通 有机化合物可以发挥其在再生医学中的潜力。我们的长期目标是建设更高层次的 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。我们在这个阶段的总体目标是首先获得对星云的空间控制 通过确定机械微环境如何对它们的诱导部位和 成熟。它的第二个目标是在许多空间位置通过受控的UE引导肾单位与UE融合 侵入性过程。实现这些目标将标志着创造替代者的变革性进展 肾组织。我们的中心假设是肾脏过程中间充质细胞的机械致密 形态发生允许诱导肾单位，并随后对WNT进行严格的时空控制 信令事件对于它们与UE的有效融合是必要的。我们计划通过两个途径实现这些目标 明确的目标。首先，我们将确定肾脏发生的力学基础，并用它来明确 肾单位位置。我们将研究肾源性间充质的机械压实，评估生物物理学 早期肾单位细胞的特性，并利用微图案化优化特定位置的肾脏发生 技术其次，我们将编程WNT诱导的肾单位与输尿管上皮的融合。我们会 通过光遗传控制WNT信号，优化肾-输尿管上皮共培养中的融合，然后 从微图案化表面转移后，触发带有UE球体的肾单位组件。建议数 研究是创新的，因为我们在创造生物材料组织的同时创造基础知识-- 免费的，人类来源的(与患者来源的自体细胞策略兼容)，因此对未来开放 为移植而开发。这项提出的研究具有重要意义，因为人类的高级组装 肾组织将创造肾脏替代技术的阶段性变化，超越透析、移植和非生物 过滤。我们预计这些努力将在基础生物学领域产生重大积极影响。 发现、药物靶点筛选和再生医学。

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.