Synthetic gene sensors and effectors to redirect organoid development

合成基因传感器和效应器可重定向类器官的发育

基本信息

批准号：
10155771
负责人：
Calin Belta
金额：
$ 68.26万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-01 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10155771
关键词：
3-Dimensional Blood Vessels Cadherins Cell Communication Cell Differentiation process Cell physiology Cells Classification Communication Complex Computational Technique Computer Models Computer Simulation Development Developmental Process Drug Screening Endoderm Engineering Feedback Gene Expression Generations Genes Genetic Genetic Transcription Growth Factor Guide RNA Hematopoiesis Human Image Individual Investigation Lead Liver Logic Machine Learning Mesoderm Methods Modeling Monitor Organoids Outcome Output Pattern Phenotype Plant Roots Play Population Probability Process Property Proteins Protocols documentation Psychological reinforcement Regenerative Medicine Regulator Genes Reporter Reporting Reproducibility Role Specific qualifier value Stromal Cells Structure Switch Genes Synthetic Genes System Techniques Therapeutic Time Tissue Engineering Tissues Training Tube Visual base cell type computing resources control theory design engineering design extracellular induced pluripotent stem cell information processing innovation insight mathematical model operation personalized medicine precision drugs programs self assembly self organization sensor single-cell RNA sequencing stem cell population synthetic biology three dimensional structure tool

项目摘要

Project Summary Human induced pluripotent stem cell (hiPSC)-derived organoids hold great promise for tissue engineering and personalized drug screening, but obtaining the desired multicellular organization and function from these systems is usually performed in an ad hoc fashion without forward design specification. Recently, we reported successful liver bud formation containing stromal cells, vascular tube-like structures and hematopoiesis-like processes by synthetically inducing diversity in GATA6 expression from a single hiPSC population. This accomplishment suggests that expanding circuit logic operations to artificially control differentiation drivers at particular bifurcations in lineage specification could profoundly impact the complexity and functionality of organoids. In this project, we bring together mathematical modeling, machine learning, optimization, and innovative synthetic biology techniques to elucidate and design fundamental decision and communication rules for guiding cells into complex, heterogeneous tissues. Our overarching hypothesis is that appropriate timing and predictable stochastic control of the expression of intracellular and extracellular factors is critical for redirecting lineage choices in order to elicit desired multicellular organization from a population of differentiating cells. We will develop synthetic tools for sensing differentiation stages of iPSC-derived organoids and construct and characterize a stochastic commitment switch in an inducible reporter system. These tools will be integrated in synthetic gene circuits for engineering emergent multicellular organization through stochastic temporal control of developmental factors. The modular commitment switches developed in this project will be capable of exploring how the degree of subpopulation biasing of cell fate decisions and level of cell fate synchronization at bipotent differentiation stages impacts self-assembly and emergent multicellular organization of an organoid. Our aims - executed through a closed loop of computational and experimental investigations - will shed insight on how generalizable methods of controlled manipulation can elicit desired organoid-level emergent properties.

项目摘要人类诱导的多能干细胞（HIPSC）衍生的类器官对组织工程和个性化的药物筛查，但从中获得所需的多细胞组织和功能系统通常以临时方式执行，而无需前瞻性设计规范。最近，我们报道成功的肝芽形成，含有基质细胞，血管管状结构和造血样通过合成诱导单个HIPSC种群的GATA6表达的多样性来进行的过程。这成就表明，扩大电路逻辑操作以人为地控制差异化驱动程序谱系规范中的特定分叉可能会深刻影响器官。在这个项目中，我们将数学建模，机器学习，优化和创新的合成生物学技术阐明和设计基本决策和交流规则将细胞引导成复杂的异质组织。我们的总体假设是适当的时机和细胞内和细胞外因子表达的可预测随机控制对于重定向至关重要谱系选择是为了从分化细胞的人群中引起所需的多细胞组织。我们将开发合成工具，以感知IPSC衍生的类器官和构造的分化阶段表征可诱导记者系统中的随机承诺开关。这些工具将集成通过随机时间控制，用于工程紧急多细胞组织的合成基因电路发展因素。该项目中开发的模块化承诺开关将能够探索细胞命运决策的亚群偏置程度和细胞命运同步的水平两性分化阶段会影响器官的自组装和新兴的多细胞组织。我们的目标 - 通过计算和实验调查的封闭循环执行 - 将洞察力关于可控操作的可推广方法如何引起所需的类器官级的新兴特性。