Rebuilding the glomerular filtration barrier by regenerating adult podocytes

通过再生成年足细胞重建肾小球滤过屏障

• 批准号: 9564892
• 负责人: Stuart James Shankland
• 金额: $ 42.92万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-23 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9564892
• 关键词: 3-Dimensional; Adult; Aging; Albumins; Applications Grants; Biological; Biological Assay; Biological Models; Biological Process; Biomedical Engineering; Blood Proteins; Bone Marrow; Bowman' s space; Candidate Disease Gene; Cell physiology; Cells; Chemotaxis; Clinical; Color; Critical Pathways; Cultured Cells; Development; Endothelial Cells; Epithelial; Epithelial Cells; Exposure to; Extracellular Matrix; Extracellular Matrix Proteins; Genetic; Goals; Grant; Growth Factor; Health; Human; In Vitro; Individual; Injury; Kidney; Knowledge; Label; Measures; Mesenchymal; Mesenchymal Stem Cells; Mus; Natural regeneration; Parietal; Pericytes; Permeability; Phase; Phenotype; Physiology; Production; Proliferating; Renin; Reporter; Reporting; Research; Role; Seminal; Source; Stem cells; System; Testing; Therapeutic; Time; Urine; Validation; Work; cell type; clinically significant; design; experience; genetic manipulation; glomerular basement membrane; glomerular filtration; glomerular function; in vitro Model; in vivo; induced pluripotent stem cell; industry partner; interest; interstitial; kidney cell; mesangial cell; microphysiology system; migration; mouse model; novel; overexpression; podocyte; progenitor; progenitor system; public health relevance; regenerative; repaired; response; self-renewal; stem; stemness; systems research

 DESCRIPTION (provided by applicant): Adult podocytes are terminally differentiated glomerular epithelial cells without regeneration capacity by self-proliferation. Thus, in states of podocyte depletion, rebuilding the kidney at the level of the glomerular filtration barrier is dependent on podocyte regeneration from stem/progenitor cells. Cells of renin lineage (CoRL) and glomerular parietal epithelial cells (PECs) are two recently identified resident progenitors fo adult podocytes in states of podocyte depletion. In this grant proposal we will fully characterize and evaluate their role and function as adult podocyte progenitors, using our bioengineered 3D flow directed microphysiological system (3D MPS) of the glomerular filtration barrier, initially constituting murine podocytes, matrix and endothelial cells. Specific aim (SA)#1 will characterize and evaluate CoRL and PEC progenitor differentiation and biological functions. SA#2 will optimize reprogramming and plasticity of CoRL and PEC progenitors towards a podocyte fate. SA#3 will broaden and validate the scope of podocyte regeneration by additional stem/progenitor cell types such as induced pluripotent stem cells and mesenchymal stem cells. In addition to the availability of cultured CoRL and PECs progenitors, expected deliverables upon completion of the UH2 includes a unique 3D MPS designed to characterize and evaluate CoRL and PEC progenitor viability, differentiation, phenotype, migration and biological function, using several functional assays. Available progenitor cells for study will initially be of mouse origin, taking advantage of their permanent labeling, as they are derived from cell specific reporter mice. The stemness of other candidate podocyte and/or endothelial cell progenitors can also be tested in this 3D MPS by the consortium. Major deliverables for the UH3 are CoRL and PECs that have been reprogrammed to an enhanced podocyte fate through genetic manipulation, changes to the microenvironment, and exposure to specific therapeutics and growth factors. The 3D MPS will be humanized by replacing mouse cells with human cells, and validation of these in vitro studies will be conducted in vivo in cell specific reporter mice where two cell types of interest are simultaneously labeled. Taken together, this grant will provide novel opportunities for the consortia and others to rebuild the kidney at the level of podocytes by resident and non-resident stem/progenitor cells.

 描述（申请人提供）：成体足细胞是终末分化的肾小球上皮细胞，不具有自我增殖的再生能力。因此，在以下状态 足细胞耗竭，在肾小球滤过屏障水平重建肾脏依赖于干/祖细胞的足细胞再生。肾素谱系细胞 (CoRL) 和肾小球壁上皮细胞 (PEC) 是最近鉴定的处于足细胞耗竭状态的成体足细胞的两种常驻祖细胞。在这项拨款提案中，我们将使用我们的生物工程 3D 流动导向微生理系统 (3D MPS) 肾小球滤过屏障（最初由小鼠足细胞、基质和内皮细胞组成）来充分表征和评估它们作为成年足细胞祖细胞的作用和功能。具体目标 (SA)#1 将表征和评估 CoRL 和 PEC 祖细胞分化和生物学功能。 SA#2 将优化 CoRL 和 PEC 祖细胞的重编程和可塑性，以实现足细胞的命运。 SA#3 将通过其他干细胞/祖细胞类型（例如诱导多能干细胞和间充质干细胞）扩大和验证足细胞再生的范围。除了培养的 CoRL 和 PEC 祖细胞的可用性之外，UH2 完成后的预期成果还包括独特的 3D MPS，旨在使用多种功能测定来表征和评估 CoRL 和 PEC 祖细胞的活力、分化、表型、迁移和生物功能。用于研究的可用祖细胞最初将来自小鼠，利用其永久标记，因为它们源自细胞特异性报告小鼠。该联盟还可以在此 3D MPS 中测试其他候选足细胞和/或内皮细胞祖细胞的干性。 UH3 的主要成果是 CoRL 和 PEC，它们已被重新编程，通过基因操作、改变微环境以及接触特定疗法和生长因子来增强足细胞的命运。 3D MPS 将通过用人类细胞替换小鼠细胞来实现人性化，这些体外研究的验证将在细胞特异性报告小鼠体内进行，其中 同时标记两种感兴趣的细胞类型。总而言之，这笔赠款将为该联盟和其他机构提供新的机会，通过以下方式在足细胞水平上重建肾脏： 驻留和非驻留干/祖细胞。