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

• 批准号: 10571876
• 负责人: Calin Belta
• 金额: $ 64.63万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 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/10571876
• 关键词: 3-Dimensional; Acceleration; Blood Vessels; Cadherins; Calibration; 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; Liver; Logic; Machine Learning; Mesoderm; Methods; Modeling; Monitor; Organoids; Outcome; Output; Pattern; Phenotype; Play; Population; Probability; Process; Property; Proteins; Protocols documentation; Psychological reinforcement; Regenerative Medicine; Reporter; Reporting; Reproducibility; Role; Specific qualifier value; Stromal Cells; Structure; Switch Genes; Synthetic Genes; System; Techniques; Therapeutic; Tissue Engineering; Tissues; Training; Tube; Visual; cell type; computing resources; control theory; design; engineering design; extracellular; gene regulatory network; 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

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 人诱导多能干细胞（hiPSC）衍生的类器官为组织工程和生物医学提供了巨大的希望。 个性化的药物筛选，但获得所需的多细胞组织和功能， 系统通常在没有正向设计规范的情况下以特定方式执行。最近，我们报道了 成功的肝芽形成，包含基质细胞、血管管状结构和造血样细胞。 通过合成诱导来自单个hiPSC群体的GATA 6表达的多样性来进行。这 这一成就表明，扩展电路逻辑操作以人为地控制微分驱动器， 血统规范中的特定分叉可能会深刻影响 类器官在这个项目中，我们将数学建模、机器学习、优化和 创新的合成生物学技术，以阐明和设计基本的决策和通信规则 引导细胞进入复杂的异质组织。我们的首要假设是，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 谱系选择，以便从分化细胞群体中引出所需的多细胞组织。我们 将开发用于感测iPSC衍生的类器官的分化阶段的合成工具，并构建和 表征诱导型报告系统中的随机承诺开关。这些工具将被整合到 通过随机时间控制工程涌现多细胞组织的合成基因电路 的发展因素。在这个项目中开发的模块化承诺交换机将能够 探索细胞命运决定的亚群偏向程度和细胞命运同步化水平， 双能分化阶段影响类器官的自组装和涌现的多细胞组织。 我们的目标-通过计算和实验研究的闭环执行-将揭示洞察力 关于控制操作的可推广方法如何引发所需的类器官水平的涌现特性。