Molecular profiling of global tissue dynamics at sub cellular resolution

结构和功能之间的关系是理解有多少生物和 从小分子化学物质到大体解剖学，化学过程的长度跨度很大。穿过 新技术的爆炸性发展，包括单细胞RNA测序(ScRNAseq)和多路组织 成像(MTI)，组织现在可以以令人难以置信的分子和细胞细节可视化。然而，如此富有的 组织结构图谱通常是来自fiX样本的静态快照，并且缺乏重要信息 关于组织的实际功能--细胞、flUID和生物分子如何动态地相互作用来管理 多细胞行为。我们的项目旨在通过构建一个集成的计算系统来克服这个限制 以及用于将组织内的功能动力学定量地链接到高分辨率的实验平台 其分子和细胞组成的空间图谱。因此，该项目的目标是生产分子 组织动力学作为一种将结构与功能联系起来的通用方法--ProProfiLing(MOTID) 多细胞社区，旨在适用于不同的模型、组织类型和动态 读数。在这个项目中，我们的目标是MOTID能够同时监测动态 组织区域内相当一部分细胞的形态和迁移，结合fluid. 特定分子从微血管循环通过间质的相运动。高度 由互补基因组的交互统计挖掘指导的同一组织的多路成像 数据，将揭示与观察到的动态相对应的免疫学上的defiNed细胞类型身份 行为。因此，MOTID将提供一个功能图谱，将细胞和fl的uid动力学与分子联系起来 细胞状态的标记。作为验证技术的原则应用的证据，我们将 检测小鼠心脏不停跳缺血/再灌流损伤模型的动态行为，以及 恶性黑色素瘤的基因工程模型。为了完成MOTID的成功开发， 这个项目建立在我们团队在活体显微镜方面的专业知识和广泛的初步数据基础上， 活组织内单细胞动力学的分割、高度多路数据的解释和构建 用于系统级分析的综合实验/计算平台。

The relationship between structure and function is central to understanding how many biological and chemical processes operate, across length scales from small molecule chemicals to gross anatomy. Through an explosion in new technologies, including single-cell RNA sequencing (scRNAseq) and multiplexed tissue imaging (MTI), tissues can now be visualized with incredible molecular and cellular detail. However, such rich atlases of tissue structure are typically static snapshots from a fixed sample, and lack important information about how the tissue actually functions — how cells, fluids, and biomolecules dynamically interact to govern multicellular behaviors. Our project aims to overcome this limitation by building an integrated computational and experimental platform for quantitatively linking functional dynamics within tissue to a high-resolution spatial map of its molecular and cellular composition. As a result, the project aims to produce Molecular profiling of Tissue Dynamics (MOTID) as a generalizable method that links structure with function in multicellular communities, designed to be applicable across diverse models, tissue-types, and dynamic readouts. In this project, we aim for MOTID to be capable of simultaneously monitoring the dynamic morphology and migration of a substantial fraction all cells within a tissue region, combined with the fluid- phase movement of particular molecules moving from microvascular circulation through interstitium. Highly multiplexed imaging of the same tissue, guided by interactive statistical mining of complementary genomic data, will reveal immunologically-defined cell-type identities that correspond to the observed dynamic behavior. Thus, MOTID will provide a functional atlas that correlates cellular and fluid dynamics with molecular markers of cell state. As proof of principle applications upon which to validate the technology, we will examine dynamic behaviors in a mouse model of ischemia/reperfusion injury in the beating heart, and a genetically engineered model of malignant melanoma. To accomplish the successful development of MOTID, this project builds upon our team's expertise and extensive preliminary data in intravital microscopy, segmentation of single-cell dynamics within live tissues, interpretation of highly multiplexed data, and building integrated experimental/computational platforms for systems-level analysis.