Investigating mechanical regulation of nephrogenesis using viscoelastic biomaterials and kidney organoids

使用粘弹性生物材料和肾类器官研究肾发生的机械调节

• 批准号: 10705067
• 负责人: Bryan Nerger
• 金额: $ 6万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10705067
• 关键词: 3-Dimensional; Address; Adult; Affect; Alginates; Architecture; Biocompatible Materials; Biology; Biomechanics; Biomedical Engineering; Cell Proliferation; Cells; Chronic Kidney Failure; Collaborations; Complex; Computer Analysis; Computer Models; Cues; Dependence; Developmental Biology; Dialysis procedure; Disease Progression; Drug Screening; Elasticity; Embryo; Engineering; Extracellular Matrix; Gene Expression Profile; Goals; Growth; Hospitals; Human; Hydrogels; In Vitro; Kidney; Kidney Diseases; Kidney Transplantation; Knowledge; Mechanics; Mediator; Mesenchymal; Mesenchyme; Modeling; Molecular; Morbidity - disease rate; Morphology; Mus; Natural regeneration; Nephrons; Organoids; Patients; Proliferating; Public Health; Regulation; Renal dialysis; Research; Research Personnel; Role; Structure; Techniques; Testing; Time; Tissues; Training; Universities; Viscosity; Woman; Work; alternative treatment; cell behavior; design; experimental analysis; human pluripotent stem cell; in vivo; insight; interstitial; mechanical behavior; mechanical properties; mechanical signal; migration; mortality; nephrogenesis; nephron progenitor; repair strategy; repaired; response; skills; stem cell differentiation; stem cells; three dimensional cell culture; viscoelasticity

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.

项目摘要 慢性肾脏疾病（CKD）影响美国约15%的成年人，并与不可逆的肾脏损害有关。 肾单位，其形成肾脏的基本功能单位。目前还没有治愈CKD的方法， 如肾移植和透析，发病率和死亡率都很高。拟订战略以 修复或更换肾单位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， 为患者提供治疗，并为药物筛选提供肾脏发育和疾病的新模型。 细胞外基质的机械性质，如刚度和粘弹性，调节细胞外基质的关键方面。 在体内驱动肾发生的细胞行为，包括增殖、分化和迁移。但是，在这方面， 虽然驱动肾发生的分子介质已被广泛研究，但基质的作用 肾发生的机制尚不清楚。除了阐明生物力学在肾脏中的作用外， 了解机械微环境在肾发生中的功能作用将有助于 为体外再生肾的工程策略提供信息。该提案的目标是整合 3D粘弹性藻酸盐水凝胶和肾脏类器官，以测试机械 微环境调节肾发生。第一个目的是确定矩阵刚度的作用， 粘弹性在人多能干细胞分化成多能肾单位祖细胞中的作用， 肾单位在肾类器官中的后续细胞组织。第二个目的是研究如何 水凝胶结构影响肾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，计算建模， 机械表征技术。 该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。 肾单位的形成，并将填补我们对肾单位的基本理解方面的巨大知识空白， 肾脏发育和干细胞分化。这项工作还将阐明设计原则， 设计新的生物材料，支持培养中的肾发生和体内肾单位的再生。 培训将在哈佛大学的穆尼实验室进行， 哈佛大学和布莱根妇女医院的邦文特尔实验室。培训计划将加强 申请人在生物材料设计，定量生物学和肾类器官培养方面的技能，并提供广泛的 了解肾脏发育和疾病。