Investigating mechanical regulation of nephrogenesis using viscoelastic biomaterials and kidney organoids

项目总结 慢性肾脏疾病(CKD)在美国影响着大约15%的成年人，与不可逆转的肾功能丧失有关 肾单位，构成肾脏的基本功能单位。目前还没有治愈CKD的方法和治疗方法 如肾移植、透析等具有较高的发病率和死亡率。制定战略，以 修复或更换肾脏将通过提供一种替代方案来解决这一重大的公共卫生问题 为患者的治疗和肾脏发育及疾病的新模式进行药物筛选。 细胞外基质的力学性质，如硬度和粘弹性，调节着 体内驱动肾发生的细胞行为，包括增殖、分化和迁移。然而， 虽然推动肾脏形成的分子介质已经被广泛研究，但基质的作用 肾脏形成的机制尚不清楚。除了阐明生物力学在肾脏中的作用之外 发展，了解机械微环境在肾脏形成中的功能作用将有助于 为在体外复制肾脏发生的工程策略提供信息。这项提议的目标是整合 3D粘弹性海藻酸盐水凝胶和肾脏器官类物质测试假说 微环境调控肾的发生。第一个目标是确定基质刚度和 粘弹性在人多能干细胞向多能肾祖细胞分化中的作用 肾脏器官中肾单位随后的细胞组织。第二个目标是调查如何 水凝胶构型影响肾脏类器官的形态和成熟。这些目标将是 通过整合生物工程、生物材料、发育生物学、计算模型和 机械表征技术。 这个项目的完成将加深我们对机械微环境在生物力学中的作用的理解 形成了肾单位，并将填补我们对 体内肾脏发育与干细胞分化。这项工作还将阐明设计原则 设计新的生物材料，支持培养中的肾脏发生和体内肾单位的再生。 培训将在哈佛大学的Mooney实验室与Mahadean实验室合作进行，地址为 哈佛大学和布里格姆妇女医院的邦文垂实验室。该培训计划将加强 申请人在生物材料设计、数量生物学和肾脏器官培养方面的技能，并提供广泛的 对肾脏发育和疾病的了解。

PROJECT SUMMARY Chronic kidney disease (CKD) affects ~15% of adults in the US and is associated with the irreversible loss of nephrons, which form the basic functional unit of the kidney. There is currently no cure for CKD, and treatments such as kidney transplantation and dialysis have a high morbidity and mortality. Developing strategies for repairing or replacing nephrons will address this significant public health problem by providing an alternative treatment for patients and a new model of kidney development and disease for drug screening. Mechanical properties of the extracellular matrix, such as stiffness and viscoelasticity, regulate key aspects of cell behavior that drive nephrogenesis in vivo, including proliferation, differentiation, and migration. However, while the molecular mediators that drive nephrogenesis have been studied extensively, the role of matrix mechanics in nephrogenesis remains unclear. Beyond elucidating the role of biomechanics in kidney development, understanding the functional role of the mechanical microenvironment in nephrogenesis will help to inform engineering strategies to reproduce nephrogenesis in vitro. The goal of this proposal is to integrate 3D viscoelastic alginate hydrogels and kidney organoids to test the hypothesis that the mechanical microenvironment regulates nephrogenesis. The first aim is to determine the role of matrix stiffness and viscoelasticity in the differentiation of human pluripotent stem cells into multipotent nephron progenitor cells and the subsequent cellular organization of nephrons in kidney organoids. The second aim is to investigate how hydrogel architecture affects the morphology and maturation of kidney organoids. These aims will be accomplished by integrating bioengineering, biomaterials, developmental biology, computational modeling, and mechanical characterization techniques. Completion of this project will deepen our understanding of the role of the mechanical microenvironment in the formation of nephrons and will fill a substantial knowledge gap regarding our fundamental understanding of kidney development and stem cell differentiation in vivo. This work will also illuminate design principles for engineering new biomaterials that support nephrogenesis in culture and the regeneration of nephrons in vivo. The training will take place in the Mooney Lab at Harvard University in collaboration with the Mahadevan Lab at Harvard University and the Bonventre Lab at Brigham and Women's Hospital. The training plan will enhance the applicant’s skills in biomaterials design, quantitative biology, and kidney organoid culture and provide a broad understanding of kidney development and disease.