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了解多少生物学和 化学过程在从小分子化学物质到总体解剖结构的长度范围内运作。通过 新技术的爆炸，包括单细胞RNA测序（SCRNASEQ）和多重组织 成像（MTI），现在可以用令人难以置信的分子和细胞细节来可视化组织。但是，如此富有 组织结构的图谱通常是固定样本的静态快照，并且缺乏重要信息 关于组织的实际功能 - 细胞，流体和生物分子如何动态相互作用以控制 多细胞行为。我们的项目旨在通过构建集成的计算来克服这一限制 和实验平台，用于定量将组织中的功能动力学连接到高分辨率 其分子和细胞组成的空间图。结果，该项目旨在产生分子 组织动力学（MOTID）作为一种可推广的方法，将结构与功能联系起来 多细胞社区，旨在适用于潜水员模型，组织类型和动态 读数。在这个项目中，我们的目标是使Motid能够简单地监视动态 在组织区域内所有细胞的大量分数的形态和迁移，并结合流体 特定分子从微血管循环到间质的相位运动。高度 同一组织的多路形成像，以完整基因组的互动统计挖掘为指导 数据将揭示与观察到的动态相对应的免疫学细胞类型身份 行为。那就是Motid将提供一个功能性地图集，将细胞和流体动力学与分子相关 细胞状态的标记。作为验证技术的原则申请的证明，我们将 在跳动心脏中缺血/再灌注损伤的小鼠模型中检查动态行为， 恶性黑色素瘤的基因工程模型。为了完成Motid的成功发展， 该项目基于我们团队的专业知识和插入显微镜的广泛初步数据， 活组织中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。