CAREER: Synthetic ‘remote control’ of kidney tissue formation towards large-scale models of congenital disease

这个学院早期职业发展(CAREAGE)项目的目标是在模拟人体自然发育的环境中创造肾脏组织。肾脏是出生缺陷的高发区，这些缺陷会影响滤过单位的大小、数量和排列，这些单位被称为肾单位，以及在肾脏内将尿液输送到膀胱的小管。这个项目寻求确定指导肾小管形成的工程规则，然后模拟肾单位与小管网络的融合。总之，这些活动将在培养皿中创造更多真实的肾脏组织，可用于从根本上研究肾脏出生缺陷的来源，并为因此类缺陷而引发的儿童和成人疾病提供新的干预策略。两个综合教育项目旨在让新一代年轻科学家具备为新兴学科“发展工程”做出贡献所需的量化技能。第一个项目是为本科生创造暑期研究体验，通过几个合作伙伴关系，将招聘努力集中在代表性不足的少数族裔、女性和第一代学生，以消除所有人使用和生成技术的障碍，从而扩大参与范围。第二个项目是通过一个名为Quanti.us的大众科学平台创建免费和公开可用的课程/讲座计划模块。这些模块将服务于高中和本科科学教育工作者，旨在激发学生对尖端科学问题的兴趣，同时为他们提供定量解决问题的方法。研究人员的长期研究目标是开发基于组织形态发生控制的先进生物制造策略，期望努力创造出足够复杂的人体组织，作为药物开发的模型或直接用于再生医学。为了实现这一目标，这个职业项目专注于在发育中的肾脏的合成组织模型中模拟肾单位的分化和与输尿管树的融合，这将通过生成大小和连通性可控的人肾小管网络，然后在其中的多个空间位置引导肾单位的形成，以更好地捕捉厘米级的肾脏结构来实现。该项目解决了对体外肾脏发育模型的需求，该模型在长度尺度上具有足够的组织，以适当研究常见的肾脏发育疾病，如CAKUT(肾脏和尿路先天性异常)，其特点是肾脏功能所需的上皮网络组织不完整。研究旨在验证这样一种假设，即在发育过程中，肾上皮芽向周围细胞外基质(ECM)“间充质”分支的机械微环境影响肾小管的伸长和几何间距。该研究计划有两个具体目标：(1)利用组织工程模拟间充质和现有的有限元模型，确定组织微环境对上皮小管引导的力学贡献；(2)通过空间引导分化，利用细胞模式、生化和光遗传学方法，同时指导动态支架内小管网络中许多特定位置的早期肾单位系的分化，将肾单位整合到动态组织支架中。超越单个肾单位或肾小管水平模拟人类肾脏发育的能力最终将为研究和开发先天性肾脏疾病的治疗方法创造机会。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The goal of this Faculty Early Career Development (CAREER) project is to create kidney tissues in an environment mimicking that of natural development in the body. The kidney is a hotspot for birth defects that affect the size, number, and arrangement of filtration units, called nephrons, and the tubules that carry urine within the kidney and out to the bladder. This project seeks determine the engineering rules that guide tubules as they form in the kidney and then to mimic nephron fusion with the tubule network. Together, these activities will create more "true-to-life" kidney tissues in a dish that can be used to study the origin of kidney birth defects at a fundamental level and inform new intervention strategies for childhood and adult diseases that originate from such defects. Two integrated educational projects are designed to equip a new generation of young scientists with the quantitative skills needed to contribute to the emerging discipline of “developmental engineering." The first project is creating a summer research experience for undergraduate students in which participation will be broadened by focusing recruitment efforts on under-represented minority, women, and first-generation students through several partnerships based on the investigator’s commitment to removing barriers to the use and generation of technology by all people. The second project is creating free and publicly available lesson/lecture plan modules through a crowd science platform called quanti.us. These modules will serve high school and undergraduate sciences educators and aim to stoke student interest in cutting-edge scientific problems while equipping them with quantitative problem-solving approaches.The investigator’s long-term research goal is to develop advanced biomanufacturing strategies based on the control of tissue morphogenesis with expectations that efforts will lead to the creation of human tissues with sufficient complexity to be models for drug development or directly in regenerative medicine. Toward this goal, this CAREER project is focused on mimicking nephron differentiation and fusion with the ureteric tree in synthetic tissue models of the developing kidney, which will be accomplished by generating human kidney tubule networks with controlled size and connectivity and then directing nephron formation at many spatial sites within them to better capture kidney structure at cm-scale. The project addresses the need for models of kidney development outside of the body with sufficient organization across length-scales to properly study common development diseases of the kidney such as CAKUT (congenital anomalies of the kidney and urinary tract), which is marked by incomplete organization of epithelial networks necessary for kidney function. Studies are designed to test the hypothesis that the mechanical microenvironment of kidney epithelial bud branching into the surrounding extracellular matrix (ECM) "mesenchyme" during development sculpts tubule elongation and geometric spacing. The research plan is organized under two specific aims: (1) Determine the mechanical contribution of the tissue micro-environment to epithelial tubule guidance using tissue-engineered mimics of the mesenchyme and an existing FEM model and (2) Engineer nephron integration into dynamic tissue scaffolds through spatially guided differentiation, leveraging cell patterning, biochemical, and optogenetic approaches to direct the differentiation of early nephron lineages at many specific sites in tubule networks within dynamic scaffolds at once. The ability to mimic human kidney development beyond the level of a single nephron or tubule will finally create opportunities to study and develop therapies for congenital kidney diseases.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.