Molecular profiling of global tissue dynamics at sub cellular resolution

亚细胞分辨率下整体组织动力学的分子分析

• 批准号: 10706567
• 负责人: Miles A Miller
• 金额: $ 33.29万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10706567
• 关键词: 3-Dimensional; Address; Algorithms; Anatomy; Atlases; Behavior; Binding; Biological; Blood Vessels; Cells; Cellular Morphology; Cellular Structures; Characteristics; Chemicals; Circulation; Complex; Computer Analysis; Computing Methodologies; Confocal Microscopy; Data; Development; Disease; Disease Progression; Excision; Explosion; Face; Genetic Engineering; Goals; Health; Heart; Human; Image; Imaging technology; Immune; Immunologics; In Vitro; Infiltration; Ischemia; Knowledge; Length; Leukocytes; Link; Liquid substance; Macrophage; Maps; Measures; Methods; Microscopy; Mining; Modeling; Molecular; Molecular Profiling; Monitor; Morphology; Movement; Myeloid Cells; Myocardial Infarction; Neoplasm Metastasis; Nucleic Acids; Operative Surgical Procedures; Pericytes; Pharmaceutical Preparations; Pharmacodynamics; Phase; Physiological; Population; Process; Proteins; Proteomics; Reperfusion Injury; Resolution; Sampling; Stains; Structure; System; Techniques; Technology; Tissue atlas; Tissue imaging; Tissues; Translating; Vascular Endothelial Growth Factors; Vascular Permeabilities; Vesicle; Visualization; biological systems; cancer cell; cell behavior; cell community; cell motility; cell type; cellular imaging; computational pipelines; computational platform; design; genomic data; improved; in vivo; in vivo imaging; intravital microscopy; mast cell; melanoma; migration; molecular marker; molecular subtypes; mouse model; multiplexed imaging; neutrophil; new technology; novel strategies; response; sample fixation; single-cell RNA sequencing; small molecule; technology validation; tool; tumor

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.

结构和功能之间的关系对于理解有多少生物和功能至关重要。 化学过程的运作范围涵盖从小分子化学品到大体解剖学。 新技术的爆炸式增长，包括单细胞 RNA 测序 (scRNAseq) 和多重组织 成像（MTI），现在可以以令人难以置信的分子和细胞细节来可视化组织。 组织结构图谱通常是固定样本的静态快照，缺乏重要信息 关于组织的实际功能——细胞、液体和生物分子如何动态相互作用以进行控制 我们的项目旨在通过构建集成的计算来克服这一限制。 以及将组织内的功能动态定量关联到高分辨率的实验平台 因此，该项目旨在制作分子和细胞组成的空间图。 组织动力学 (MOTID) 的分析作为一种将结构与功能联系起来的通用方法 多细胞群落，旨在适用于不同的模型、组织类型和动态 在这个项目中，我们的目标是让 MOTID 能够同时监测动态。 组织区域内大部分细胞的形态和迁移，与流体相结合 特定分子从微血管循环穿过间质的相运动。 在互补基因组的交互式统计挖掘的指导下，对同一组织进行多重成像 数据，将揭示与观察到的动态相对应的免疫学定义的细胞类型身份 因此，MOTID 将提供将细胞和流体动力学与分子相关联的功能图谱。 作为验证该技术的原理应用的证明，我们将 检查跳动心脏缺血/再灌注损伤小鼠模型的动态行为，并 恶性黑色素瘤基因工程模型的成功开发， 该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的基础上， 活组织内单细胞动力学的分割、高度多重数据的解释以及构建 用于系统级分析的集成实验/计算平台。