Programmed Differentiation Circuits for Organoids using Meso-Microfluidics

使用介观微流体对类器官进行编程分化电路

基本信息

批准号：
9896824
负责人：
RON WEISS
金额：
$ 60.13万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-17 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9896824
关键词：
3-Dimensional Achievement Address Adult Affect Albumins Area Automation Automobile Driving Biological Assay Cell Communication Cell Culture Techniques Cell physiology Cells Chemicals Development Elements Engineering Exhibits Generations Genes Genetic Genetic Drift Genetic Engineering Goals Growth Growth Factor Hepatic Hepatocyte Human Lead Liver Manuals Methods MicroRNAs Microfluidics Microscope Monitor Nature Nuclear Receptors Organ Organoids Output Pancreas Patients Physiology Play Population Process Production Property Proteins Regenerative Medicine Role System Testing Time Tissues Transplantation Vascularization Work activating transcription factor body system cancer therapy cell type combinatorial cost design drug development drug testing gene therapy human pluripotent stem cell human tissue humanized mouse improved in vivo induced pluripotent stem cell insight instrumentation liver development miRNA expression profiling novel optogenetics pancreas development personalized medicine programs sensor spatiotemporal stem cell differentiation synthetic biology three dimensional structure transcription factor

项目摘要

We propose a new platform that leverages synthetic biology genetic circuits and micro/mesofluidic instrumentation to rapidly advance the field of organoids. Specifically, we will genetically engineer self- organizing tissues from human pluripotent stem cells into co-developed liver and pancreas organoids possessing vascularization and other mature properties, such as adult level albumin production. The application of synthetic biology to organoid development (programmable organoids) provides an exciting new opportunity for engineering and testing of organoids encoding live cell sensors of cell state and for embedding circuits that express cell-type and cell-state specific transcription factors. We will engineer novel microfluidic and mesofluidic platforms to enable low cost and high throughput development and testing of programmable organoids. Our hypothesis is that co-development of hiPSC-derived liver and pancreas provides cell-cell interactions that contribute to vascularization and other important elements in organoid development that will lead to mature organ formation. To test our hypothesis, we will genetically encode live-cell sensors to monitor liver organoid develop, co-develop liver and pancreas organoids, and create genetic circuits that lead to mature organoid formation. We will use synthetic classifier genetic circuits that evaluate changes in cell state in real time, and generate relevant protein outputs for driving differentiation in a cell-type specific manner. This these circuits will enable autonomous generation of new and improved versions of organoids, including mature liver organoids and co-developed liver/pancreas organoids. Determining the precise spatiotemporal nature of cell state transitions and the relevant transcription factors to drive differentiation is not only essential for creating new and effective organoid developmental programs, but will also provide important scientific insights to understanding the fine aspects of differentiation and co-development. Successful achievement of our aims will have a broad impact in the areas of gene therapy, drug testing, and personalized medicine. For example, the ability to co-develop matched organoid systems will enable patient-specific drug development (e.g. for cancer therapy) that is more accurate than expensive and controversial alternatives, such as the use of humanized mice. This work will also support the long-term goal of producing mature organoids and organ systems suitable for transplantation.

我们提出了一个新平台，该平台利用合成生物学遗传回路和微/中流体的平台仪器以快速前进器官的领域。具体来说，我们将基因工程师自我自我从人多能干细胞中组织组织成共同发育的肝脏和胰腺器官具有血管形成和其他成熟特性，例如成人水平的白蛋白产生。这合成生物学在器官开发中的应用（可编程器官）提供了令人兴奋的新的编码细胞态的活细胞传感器和嵌入的类器官的工程和测试的机会表达细胞类型和细胞态特定转录因子的电路。我们将设计新颖的微流体和中流体平台，以实现低成本和高吞吐量的开发和可编程的测试器官。我们的假设是hipsc衍生的肝脏和胰腺的共同开发提供细胞细胞有助于有助于血管形成和器官发育中其他重要因素的相互作用导致成熟器官形成。为了检验我们的假设，我们将基因编码活细胞传感器来监视肝脏器官发展，共发育肝脏和胰腺器官，并创建导致的遗传回路成熟的器官形成。我们将使用合成分类器遗传回路来评估细胞状态的变化实时，并生成相关的蛋白质输出，以特定于细胞类型的方式驱动分化。这这些电路将能够自动生成新的和改进的器官版本，包括成熟肝脏类器官和联合发育的肝脏/胰腺器官。确定的精确时空性质细胞状态过渡和驱动分化的相关转录因子不仅对创建新的有效的器官发展计划，但也将提供重要的科学见解理解分化和共同开发的精细方面。成功实现我们的目标将对基因疗法，药物测试和个性化医学领域产生广泛的影响。例如，共同开发的匹配器官系统的能力将使患者特定的药物开发（例如癌症治疗）比昂贵且有争议的替代品更准确，例如使用人源化的小鼠。这项工作还将支持生产成熟的器官和器官的长期目标适合移植的系统。