Elucidating ECM Signaling in Cardiac Organoids with Machine Learning and Single-cell Multiomics

项目摘要 细胞外基质(ECM)是体内含量最丰富的生物材料。在心脏发育过程中，ECM 在心脏的形状和图案的形成中起着关键作用，如通过 与分化细胞的复杂相互作用。尽管ECM-细胞相互作用中的问题可能会导致心脏 疾病，由特定的ECM组分激活的信号通路仍然知之甚少。我们最近 成功开发出人诱导多能干细胞来源的心脏器官(IPSC-Cos)，可以 概括地说，心脏发生。在这份多PI R01提案中，我们的团队将进一步阐明ECM的机制- 影响心脏分化和形态发生的细胞相互作用。我们将应用机器学习和 新的IPSC双报告系(Tbx5-Clover2-NKX2.5-TagRFP)阐明细胞组成对TagRFP的影响 IPSC-Cos的形态发生(目标1)。之后，我们将筛选36种不同的ECM组合 这可以可靠地诱导IPSC-CO的形成，使用8个额外的IPSC品系进行验证(目标2)。通过严格地 利用弹性性能测量和单细胞多组学分析优化的ECM制备的IPSC-CoS， 我们将阐明与ECM信号转导和机械转导相关的生物和物理效应 单元格分辨率(目标3)。总而言之，了解ECM-细胞相互作用的确切作用和机制 可能有助于寻找新的生物材料或治疗心脏病的方式。

Project Summary Extracellular matrix (ECM) is the most abundant biomaterial in the body. During cardiac development, the ECM plays critical roles in the formation of shapes and patterns of the heart such as chambers and trabeculae through elaborate interactions with differentiating cells. Although problems in ECM-cell interactions can lead to heart diseases, signaling pathways activated by the specific ECM components are still poorly understood. We recently succeeded in developing human induced pluripotent stem cell-derived cardiac organoids (iPSC-COs) that can recapitulate cardiogenesis. In this multi-PI R01 proposal, our team will further elucidate the mechanisms of ECM- cell interactions that influence cardiac differentiation and morphogenesis. We will apply machine learning and novel iPSC double reporter lines (Tbx5-Clover2-Nkx2.5-TagRFP) to elucidate the effect of cell composition on morphogenesis of iPSC-COs (Aim 1). Afterwards, we will screen 36 different combinations of ECM compositions that can reliably induce iPSC-CO formation using 8 additional iPSC lines for validation (Aim 2). By rigorously analyzing iPSC-COs made with optimized ECM using elastic property measurement and single-cell multiomics, we will elucidate the biological and physical effects associated with ECM signaling and mechanotransduction at single-cell resolution (Aim 3). In summary, understanding the exact role and mechanism of ECM-cell interactions may contribute to finding new biomaterials or therapeutic modalities for treatment of heart diseases.