Investigating organ-specific patterning of the vasculature during human development

研究人类发育过程中脉管系统的器官特异性模式

• 批准号: 10058764
• 负责人: Emily Marie Holloway
• 金额: $ 3.73万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10058764
• 关键词: Address; Adult; Animal Model; Bioinformatics; Biological Assay; Biological Models; Blood Vessels; Candidate Disease Gene; Cell Communication; Cell Line; Cell model; Cell physiology; Cell surface; Coculture Techniques; Data; Development; Diffusion; Embryonic and Fetal Development; Endothelial Cells; Engraftment; Epithelial; Epithelium; Evaluation; Fetal Development; Fetal Kidney; Fetal Lung; Flow Cytometry; Fluorescent in Situ Hybridization; Gene Cluster; Gene Expression; Generations; Genes; Goals; Human; Human Development; In Vitro; Individual; Intestines; Kidney; Knowledge; Liver; Lung; Manuals; Mesenchyme; Mesoderm; Methods; Modeling; Mus; Nutrient; Organ; Organ Specificity; Oxygen; PECAM1 gene; Pattern; Perfusion; Population; Proteins; Regenerative Medicine; Role; Stains; Supporting Cell; System; Testing; Tissue Engineering; Tissues; Umbilical vein; Validation; Vascularization; Work; base; body system; design; endothelial stem cell; epithelial stem cell; experimental study; fetal; gene function; genetic signature; human fetal cells; human fetus tissue; human pluripotent stem cell; human tissue; improved; in vivo; insight; intestinal epithelium; mRNA Expression; mature animal; novel; organ growth; progenitor; protein expression; receptor; single-cell RNA sequencing; stem cells; three dimensional cell culture; transcription factor; transcriptome sequencing

Project Abstract Vasculature is an efficient delivery system for oxygen and nutrients to organs and is essential for organ function. Therefore, vascularization is a critical consideration for regenerative medicine and tissue engineering, since oxygen and nutrient diffusion significantly limits the size of tissues grown in vitro, and perfusion is critical for in vivo engraftment and survival of engineered tissues. Many approaches have been used to generate engineered tissue with a vascular network with or without vascular flow; however, one of the major caveats to these advances is that they often use endothelial cell (EC) lines such as human umbilical vein endothelial cells (HUVECs). While the use of cell lines is ideal for proof-of-concept experiments, there is a significant body of evidence, primarily coming from animal models, demonstrating that ECs have organ-specific gene expression and function. Given that ECs within individual organ systems create organ-specific microenvironments critical for function, organ- specific ECs will be imperative to achieve long-term organ-level function for tissue engineering and regenerative medicine approaches. However, critical gaps in our knowledge exists with respect to human ECs; it is unknown if tissue-specific EC gene expression and function is present during development of human organ systems, and it is further unknown how such expression patterns are established. To begin to address these unknowns I isolated ECs and non-ECs from human fetal kidney, lung and intestine, performed bulk RNA sequencing, and used gene clustering approaches to identify organ-specific EC-enriched genes. My preliminary data has identified over 100 organ-specific EC-enriched genes for each organ analyzed. This is the first evidence to suggest organ-specific EC gene expression is established during embryonic/fetal development. My proposal is designed to: 1) Validate novel organ-enriched EC genes during human development and test organ-specific EC function using in vitro co-culture assays 2) leverage these insights to induce organ-specific patterning of human pluripotent stem cell (hPSC)-derived ECs.

项目摘要 血管系统是向器官输送氧气和营养物质的有效系统，对器官功能至关重要。 因此，血管化是再生医学和组织工程的关键考虑因素，因为 氧气和营养物质的扩散明显限制了体外生长的组织的大小，而灌注对于体内生长至关重要。 体内移植和工程化组织的存活。许多方法已经被用于产生工程化的 具有血管网络的组织，有或没有血管流动;然而，这些进展的主要警告之一 他们经常使用内皮细胞（EC）系，如人脐静脉内皮细胞（HUVEC）。而 细胞系的使用是概念验证实验的理想选择，有大量证据表明， 来自动物模型，证明EC具有器官特异性基因表达和功能。给定 单个器官系统内的EC创造了对功能至关重要的器官特异性微环境， 特定的EC对于实现组织工程和再生的长期器官水平功能是必不可少的， 医学接近。然而，我们对人类内皮细胞的认识存在着重大差距， 如果组织特异性EC基因表达和功能存在于人类器官系统的发育过程中， 还不知道这些表达模式是如何建立的。为了开始解决这些未知数， 从人胎儿肾、肺和肠分离EC和非EC，进行批量RNA测序，和 使用基因聚类方法来识别器官特异性EC富集基因。我的初步数据显示 鉴定了超过100个器官特异性EC富集基因的每个器官分析。这是第一个证据 提示器官特异性EC基因表达在胚胎/胎儿发育期间建立。我的建议是 设计用于：1）在人类发育过程中发现新的器官富集EC基因，并测试器官特异性EC 2）利用这些见解来诱导人类的器官特异性模式化 多能干细胞（hPSC）衍生的EC。