Synthetic gene sensors and effectors to redirect organoid development

项目摘要 人诱导多能干细胞(HiPSC)衍生的有机类化合物在组织工程和 个性化药物筛选，但从这些组织和功能获得所需的多细胞组织和功能 系统通常是以特别的方式执行的，没有前向设计规范。最近，我们报道了 含有基质细胞、血管管样结构和造血样结构的成功肝芽形成 通过合成诱导单个HiPSC群体中GATA6表达的多样性来进行处理。这 成果表明，扩展电路逻辑操作以人为控制差异化驱动程序 谱系规范中的特定分支可能会深刻影响 有机化合物。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，用于阐明和设计基本的决策和沟通规则 引导细胞进入复杂的、不同种类的组织。我们的首要假设是，适当的时机和 可预测的随机控制细胞内和细胞外因子的表达是重定向的关键 谱系选择，以便从分化细胞群体中诱导出所需的多细胞组织。我们 将开发用于感知IPSC衍生有机化合物分化阶段的合成工具，并构建和 描述了感应报告器系统中的随机承诺开关。这些工具将集成在 基于随机时间控制的工程多细胞组织合成基因电路 发育因素的影响。在本项目中开发的模块化承诺交换机将能够 探讨细胞命运决定的亚群偏向程度和细胞命运同步化水平如何 二能分化阶段影响有机体的自组装和新出现的多细胞组织。 我们的目标--通过计算和实验研究的闭合循环来执行--将带来洞察力 关于可推广的受控操纵方法如何引出所需的有机体级别的紧急特性。

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.