Integration of systems and synthetic biology to advance development of human tissues ex vivo

系统与合成生物学的整合促进人体组织离体发育

基本信息

批准号：
10245099
负责人：
Mo Reza Ebrahimkhani
金额：
$ 41.68万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-30 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10245099
关键词：
Address Adult Advanced Development Animal Experiments Animals Biliary Biology Biomedical Engineering Biotechnology Cells Clustered Regularly Interspaced Short Palindromic Repeats Computational Biology Computer Analysis Data Dependence Development Disease Endothelium Engineered Gene Engineering Epigenetic Process Event FGF2 gene Face Fetal Liver GATA6 transcription factor Gene Activation Genetic Genetic Engineering Genetic Transcription Germ Layers Goals Human Human Development Human Engineering In Vitro Liver Methods Mus Names Organ Organizational Culture Organogenesis Organoids Patients Phenotype Population Regulator Genes Source Stromal Cells Synthetic Genes System Systems Biology Systems Integration Technology Therapeutic Tissue Engineering Tissues Training Transcriptional Activation Transforming Growth Factor beta Vascular Endothelial Cell analog base cell fate specification cell type computational platform drug development fetal genetic signature genome-wide human adult stem cell human disease human model human stem cells human tissue improved in vivo induced pluripotent stem cell insight inter-individual variation intercellular communication liver injury mouse model novel overexpression precision medicine programs self organization spatiotemporal stem cell derived tissues stem cells synthetic biology tool transcription factor transcriptional reprogramming

项目摘要

The rapidly evolving field of stem cell bioengineering and organoids technology face key challenges: a) stem cell derived tissues are stalled developmentally and show fetal-stage phenotypes b) they often lack key subsets of vascular endothelial and stromal cells derived from different germ layers c) There is a lack of toolset to quantitatively assess cell or tissue identity and d) to guide morphogenetic events towards their native adult phenotypes. In this proposal, we will address these issues by undertaking an integrative synthetic biology and systems biology approach. We recently generated novel human fetal liver organoids ex vivo using human iPSCs. Our approach entails genetic engineering of human iPSCs via overexpression of GATA6 transcription factor in pluripotent media with bFGF and TGF-β. Through this strategy, we showed development of human endodermal and mesodermal, intercellular communications, co-differentiation and self-organization of cultures into a multi- cell type fetal liver organoid. We will employ this tissue as a unique testbed to develop and address biotechnology challenges for in vitro maturation, assessment and engineering of organoids. Through aim 1 of this proposal we will develop and validate a set of genetic toolset to drive cell-fate reprogramming of the multicellular tissue. Through aim 2, we will establish a computational platform to quantitatively assess liver organoids and to identify transcriptional regulators of stage specific development IMPACT: We tackle improvement and assessment of human organoids ex vivo, two key challenges in stem cell bioengineering. The successful completion of this study will result in a hypothesis-driven framework for rational engineering and advancement of stem cell-derived tissues ex vivo. Our study will also generate synthetic liver tissues with close proximity to adult human liver. It reduces dependence on animal experiments and increases access to refined human tissues.

干细胞生物工程和器官技术的快速发展的领域面临关键挑战：a）干细胞衍生的组织被停滞不前并显示出胎儿阶段表型b）它们通常缺乏关键子集源自不同细菌层的血管内皮细胞和基质细胞c）缺乏工具集定量评估细胞或组织身份和d）指导形态发生事件对其本地成年表型。在此提案中，我们将通过进行综合合成生物学和系统生物学方法。我们最近使用人IPSC生成了新型的人类胎儿肝癌。我们的方法需要通过GATA6转录因子过表达人IPSC的基因工程具有BFGF和TGF-β的多能培养基。通过这种策略，我们展示了人类内皮层的发展中胚层，细胞间交流，培养物共同分化和自组织细胞型胎儿肝癌。我们将使用该组织作为独特的测试台来开发和解决生物技术体外成熟，器官评估和工程的挑战。通过本提案的目标1我们将开发和验证一组遗传工具集，以驱动多细胞组织的细胞命运重编程。通过AIM 2，我们将建立一个计算平台，以定量评估肝脏器官并确定阶段特定开发影响的转录调节器：我们应对改进和评估人体器官离体，干细胞生物工程的两个主要挑战。成功完成研究将导致一个假设驱动的框架，用于理性工程和干细胞衍生的发展组织离体。我们的研究还将产生与成年人肝密切接近的合成肝组织。它减少对动物实验的依赖，并增加获得精制人体组织的访问。