Spatial genetics investigation of multinucleated cells

项目概要/摘要 器官系统由大量的细胞亚群组成， 特定组织内的细胞与其功能和体内平衡控制密切相关。 同样，在单细胞的空间内，生物化学环境也是异质的 分子间的动态相互作用决定了细胞的适应性、行为和命运。 我们对细胞相互作用和生物分子相互作用的理解， 被革命性的单细胞基因组学和单分子成像技术所改变。 然而，这些技术的开发和应用一直是集中在 关于单核细胞的背景。隐藏在盲点中的是多核细胞类型， 包括肌纤维，心肌细胞，合体滋养层，某些癌细胞，所有这些， 出差错时会引起毁灭性的人类疾病。多核细胞的合胞体性质， 具有多倍性和巨大的胞质体积， 关于细胞-细胞相互作用和生物分子分布的空间组织。 基本的但在很大程度上未知的问题包括：1）组织的异质性如何 合胞体细胞周围的微环境2)共用细胞体的细胞核 协调基因表达以响应外部刺激和细胞间通讯？第三章 什么是控制mRNAs在细胞中转运和定位的机制？ 多核细胞？我们的研究计划利用了特殊细胞类型的独特功能 作为理解各种发展系统的机械基础的一种手段。这 建议利用肌纤维作为合适的模型来研究上述问题， 一种空间上确定的方式。首先，我们将开发和部署新的方法来探测 多种类型的合胞体组织的空间转录组学。第二，我们将制定 反向工程策略组装人肌肉单合胞体基因 跟踪性，然后将它们移植到活体动物中，用于细胞间通讯的体内研究。 和合胞体内mRNA分布。第三，深入开展基因功能研究， 机制研究，以揭示细胞间通讯和细胞内 mRNA运输。总的来说，这项研究计划依赖于我们在基因组学，细胞， 这样我们就可以创造一个创新和发现的良性循环 在MIRA奖的过程中。我们预计，知识和技术将受益于 更大的生物界，包括遗传学，细胞生物学家和发育生物学家。

Project Summary/Abstract Organ systems are composed of a wealth of cellular subpopulations whose spatial organization within a given tissue are deeply intertwined with their functions and homeostasis controls. Similarly, within the space of a single-cell, the biochemical environment is also heterogeneous where the dynamic interactions of molecules determine the fitness, behavior and fate of the cell. Our understanding of the cell interactions and the biomolecule interactions has recently been transformed by the revolutionary single-cell genomics and single-molecule imaging technologies. However, the development and application of these technologies have been exclusively centered on the context of mononuclear cellularity. Hiding in the blind spot are multinucleated cell-types, including myofibers, cardiomyocytes, syncytiotrophoblasts, certain cancer cells, all of which cause devasting human disease when go awry. The syncytial nature of the multinucleated cells, which possess the polyploidy and often the vast cytosolic volumes, raises fascinating questions with respect to the spatial organizations of the cell-cell interactions and biomolecule distributions. The fundamental yet largely unknown questions include: 1) How heterogeneous is the tissue microenvironment that surrounds the syncytial cells? 2) Do nuclei from the shared cell body coordinate gene expression in response to the external stimuli and cell-cell communications? 3) What is the mechanism that governs the transports and localizations of mRNAs in the multinucleated cells? Our research program exploits the unique features of specialized cell-types as a means to understand mechanistic underpinnings of various developmental systems. This proposal leverages myofiber as the uniquely-suited model to investigate the above questions in a spatially defined manner. First, we will develop and deploy new methodologies to probe the spatial transcriptomics for the multiple types of syncytial tissues. Second, we will devise the reverse engineering strategy to assemble the single-syncytium of human muscle with genetic trackabilities and later graft them to live animals for the in vivo study of the cell-cell communication and intra-syncytium mRNA distributions. Third, we will conduct the in-depth gene function and mechanism studies to unveil new paradigms of intercellular communication and the intracellular mRNA trafficking. Broadly, this research program relies on our diverse expertise in genomics, cell engineering and computation such that we can create a virtuous cycle of innovation and discovery over the course of the MIRA award. We anticipate that the knowledge and techniques will benefit the greater biological community, including genetics, cell biologists and developmental biologists.