Engineering scalable collecting duct networks for functional kidney tissue

项目摘要/摘要 该提案概述了Kayla Wolf博士要完成的综合研究和职业发展计划 在赞助商詹妮弗·刘易斯(ScD)和共同赞助商丽莎·萨特林(Lisa Satlin，MD)的指导下进行博士后培训。这个 拟议项目的总体目标是开发一个功能性收集风管网络，该网络源于 人类多能干细胞，并与单一引流出口相互连接。这个网络可以作为一个 用于假设检验、疾病建模和药物筛选的模型系统，或者可以直接与 富含肾单位的有机体有助于3D肾脏组织的滤液排出。认识到细胞外基质 细胞外基质(ECM)在发育中起着至关重要的作用，本提案既探讨了ECM在集合管中的作用 分化和利用生物材料作为指导组织形态和功能的强大工具。发展中的 收集风道系统将通过三个目标的完成来完成。首先，输尿管芽细胞(UB， 收集导管前体)将得到验证，并且 将测试该方案是否适用于UB表型的可扩展培养方法。第二，政府部门的作用 细胞外基质(ECM)组成对UB形态和功能分化的影响将被测定。 最后，在分支UB网络中构建引流口对转运蛋白表达的影响 和功能将被确定。成功完成这项建议将阐明ECM在驾驶中的作用 收集管道开发，推进生物制造方法，生产急需的工程 用于生成功能性肾组织的解决方案。 Wolf博士(PI)在她的研究生研究期间得到了NIH F31奖学金(F31 CA228317-01)的支持， 她在生物材料、生物工程和机械生物学方面建立了专业知识。在NIH F32的支持下，PI 在哈佛大学接受了广泛的生物制造和肾脏生理学培训，培养了一批优秀的- 成熟的、全球领先的生物医学研究环境。职业发展计划旨在 为PI提供必要的知识和技能，使其成为一名成功的独立学者 研究员。

PROJECT SUMMARY / ABSTRACT The proposal outlines an integrated research and career development plan for Kayla Wolf, PhD, to complete postdoctoral training under the mentorship of sponsor Jennifer Lewis, ScD, and co-sponsor Lisa Satlin, MD. The overarching goal of the proposed project to develop a functional collecting duct network that is derived from human pluripotent stem cells and interconnected with a single drainage outlet. This network could serve as a model system for hypothesis testing, disease modeling, and drug screening or could interconnect directly with nephron-rich organoids to facilitate filtrate drainage in 3D kidney tissues. Recognizing that the extracellular matrix (ECM) plays a crucial role in development, this proposal with both investigate the role of ECM in collecting duct differentiation and leverage biomaterials as a powerful tool for guiding tissue form and function. Development of the collecting duct system will be accomplished by the completion of three aims. First, ureteric bud cells (UB, collecting duct precursors) generated from emerging differentiation protocols will be validated, and the effects of adapting the protocol to scalable culture methods on UB phenotype will be tested. Second, the role of extracellular matrix (ECM) composition on UB morphological and functional differentiation will be determined. Finally, the effects of fabricating a drainage outlet in a branching UB network on transport protein expression and function will be determined. Successful completion of this proposal will elucidate the role of ECM in driving collecting duct development, advance biofabrication methodology, and produce critically needed engineering solutions for generating functional kidney tissue. Dr. Wolf (PI) was supported by an NIH F31 fellowship (F31 CA228317-01) during her graduate research, where she established expertise in biomaterials, bioengineering, and mechanobiology. Under NIH F32 support, the PI receive extensive training in biofabrication and renal physiology at Harvard University, which cultivates a well- established, globally-leading biomedical research environment. The career development plan is designed to equip the PI with the necessary knowledge and skills for a successful career as an independent academic researcher.