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 从小分子化学品到大体解剖学的长度尺度上，化学过程都在进行。通过 新技术的爆炸式发展，包括单细胞RNA测序（scRNAseq）和多重组织 随着MTI成像技术的发展，组织现在可以以令人难以置信的分子和细胞细节进行可视化。然而，如此富有 组织结构图谱通常是固定样本的静态快照，缺乏重要信息 关于组织实际上是如何运作的--细胞、体液和生物分子如何动态地相互作用， 多细胞行为我们的项目旨在通过建立一个集成的计算模型来克服这一限制。 和实验平台，用于定量地将组织内的功能动力学与高分辨率 其分子和细胞组成空间图。因此，该项目旨在生产分子 组织动力学（MOTID）作为一种将结构与功能联系起来的可推广方法， 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 组织区域内的大部分所有细胞的形态和迁移，与细胞液结合， 从微血管循环中通过微循环的特定分子的相位运动。高度 同一组织的多重成像，由互补基因组的交互式统计挖掘引导 数据，将揭示免疫学定义的细胞类型的身份，对应于观察到的动态 行为因此，MOTID将提供一个将细胞和体液动力学与分子生物学相关联的功能图谱。 细胞状态的标志物。作为验证该技术的原理应用的证明，我们将 在跳动的心脏中检查缺血/再灌注损伤小鼠模型的动力学行为， 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的专业知识和大量的活体显微镜初步数据的基础上， 活组织内单细胞动力学的分割，高度多路复用数据的解释， 用于系统级分析的集成实验/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.