Identifying pathways required for integration of kidney organoid and host epithelia

确定肾类器官和宿主上皮细胞整合所需的途径

基本信息

批准号：
10428383
负责人：
IAIN A. DRUMMOND
金额：
$ 62.78万
依托单位：
MOUNT DESERT ISLAND BIOLOGICAL LAB
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10428383
关键词：
3-Dimensional Adult Basement membrane Bioinformatics Biological Assay Blood Cause of Death Cell Aggregation Cell Culture System Cell Culture Techniques Cell fusion Cells Coupled Data Development Dialysis procedure Distal End stage renal failure Engineering Engraftment Environment Epithelial Epithelial Cells Event Excision Extracellular Matrix Fibroblast Growth Factor Filtration Fishes Genetic Goals Human Immune Implant In Situ Hybridization In Vitro Injury Invaded Investigation Kidney Kidney Diseases Kidney Failure Kidney Transplantation Legal patent Ligands Liquid substance Mammalian Cell Matrix Metalloproteinases Mediating Methodology Modeling Molecular Mus Mutation Natural regeneration Nephrons Organoids Output Pathway interactions Patients Pattern Phenotype Play Plumbing Pluripotent Stem Cells Process Renal function Renal tubule structure Reporting Role Series Signal Pathway Signal Transduction Site Structure System TIMP1 gene Technology Testing Tissue Inhibitor of Metalloproteinases Tissues Urine WNT Signaling Pathway WNT9A gene Work Zebrafish base experimental study genetic analysis glucose uptake in vivo in vivo engraftment insight mutant nephrogenesis nephron progenitor polarized cell prevent programs receptor recruit regenerative therapy renal damage repaired scale up specific biomarkers stem cells three dimensional cell culture tool transcriptomics urinary

项目摘要

Project Summary Kidney disease is the 9th leading cause of death in the U.S. Because few therapies exist to prevent or slow progression, over 700,000 patients have End Stage Renal Disease. These patients are treated with dialysis or renal transplant, the latter resulting in markedly superior survival. However, kidney donors are limited and there is an important unmet need for strategies that enhance renal repair or generate new nephrons for renal replacement. Pluripotent stem cell derived organoids display key features of differentiated kidney tubules and glomerular structures in vitro, and we have shown that they generate patterned nephrons in vivo displaying kidney functions such as filtration and glucose uptake by the proximal tubule. To develop this technology for renal replacement, stem cell derived tubules must be connected to host tubules for urinary output. Our recent work in the zebrafish demonstrated that FGF signaling acts as a chemotactic signal to recruit and polarize cells at sites of new nephron formation and canonical Wnt signaling is required for invasive cell rearrangement to connect tubule lumens. Additional signaling pathways including non-canonical wnt signaling are also likely to play a role in tubule interconnection. To fully explore the requirements for tubule interconnection we have established a synergistic, three-part discovery platform comprising 1) genetic analysis of in vivo new nephron addition in the regenerating zebrafish adult kidney, 2) in vitro 3D cell culture analysis of mammalian epithelial fusion, and 3) in vivo stem cell-derived kidney organoid engraftment to a host mouse collecting system. We will combine these approaches to analyze multiple steps of the tubule fusion process involving 1) recruitment of nephron progenitor cells to target epithelia, 2) removal of intervening ECM/basement membranes, 3) patterned collective cell invasion of target epithelia, and 4) establishment of a continuous patent new lumen to convey the nephron filtrate. These studies will provide important new insights about an essential but understudied cellular mechanism that will be required for in vivo engraftment of new kidney tissue-based renal regeneration therapies.

项目摘要肾脏疾病是美国第九大死亡原因，因为很少有预防或缓慢的疗法进展，超过700,000名患者患有末期肾脏疾病。这些患者接受透析或肾移植，后者导致显着上升的生存。但是，肾脏捐赠者有限，那里是对增强肾脏修复或生成肾脏新肾脏的策略的重要需求替代品。多能干细胞衍生的类器官显示分化肾小管的关键特征和体外肾小球结构，我们已经证明它们在显示体内产生图案化的肾单位肾功能（例如过滤和葡萄糖吸收）近端小管。开发这项技术肾脏更换，干细胞衍生的小管必须连接到宿主小管以进行尿输出。我们最近斑马鱼中的作品表明，FGF信号传导充当趋化信号，可募集和极化细胞在新的Nephron形成和规范的Wnt信号的位置，侵入性细胞重新排列需要连接小管流明。包括非规范Wnt信号在内的其他信号通路也可能在小管互连中发挥作用。为了充分探索小管互连的要求建立了一个协同的三部分发现平台，其中包括1）体内新肾单位的遗传分析在再生斑马鱼成年肾脏中的添加，2）哺乳动物上皮的体外3D细胞培养分析融合和3）体内干细胞衍生的肾脏器官植入到宿主小鼠收集系统中。我们将结合这些方法以分析小管融合过程的多个步骤，涉及1）肾祖细胞靶向上皮，2）去除中间的ECM/地下膜，3）集体细胞入侵目标上皮，4）建立连续的专利新管道，以传达肾脏滤液。这些研究将提供有关基本但经过研究的细胞的重要新见解新的基于肾脏组织的肾脏再生所需的机制疗法。