SUBSTRATE RIGIDITY AND GENE EXPRESSION: Role of Nuclear Tension

基质刚性和基因表达：核张力的作用

• 批准号: 9238291
• 负责人: Tanmay P. Lele
• 金额: $ 46.49万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9238291
• 关键词: 3-Dimensional; Address; Adhesions; Area; Biocompatible Materials; Biological Assay; Biology; Biomechanics; Biomedical Engineering; Cell Culture Techniques; Cell Nucleus; Cell physiology; Cells; Cellular biology; Characteristics; Chemicals; Chromatin Structure; Collaborations; Complex; Cues; Cytoskeleton; Development; Electron Microscope; Engineering; Fibroblasts; Florida; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genome; Goals; Hydrogels; Intermediate Filaments; Laboratories; Lamin Type B; Massachusetts; Mechanics; Mediating; Messenger RNA; MicroRNAs; Microtubules; Modeling; Molecular; Molecular Biology; Motor; Myosin ATPase; Nuclear; Nuclear Export; Nuclear Structure; Optics; Performance; Property; Proteins; Regulation; Research; Research Personnel; Resources; Role; Schools; Solid; Techniques; Technology; Testing; Time; Tissue Engineering; Tissues; Transcript; Universities; Work; base; biomaterial development; genome-wide; histone modification; improved; interest; medical schools; novel; professor; research study; scaffold; tissue repair; tissue support frame

PROJECT SUMMARY/ABSTRACT The use of solid scaffolds that provide cells with mechanical and chemical cues is a promising approach for guiding tissue repair and promoting tissue-scaffold integration. It is becoming increasingly clear that tuning scaffold rigidity is a powerful way to control cell function, but how scaffold rigidity regulates gene expression is not well-understood. Previously, we tested the hypothesis that substrate rigidity controls gene expression by tuning nuclear tension. We took advantage of the fact that the LINC (linker of nucleoskeleton-to-cytoskeleton) complex is a known molecular linker of the nucleus to the cytoskeleton, and asked how it regulates the sensitivity of genome-wide transcription to substrate rigidity. Our results were the first to show that the LINC complex facilitates mechano-regulation of expression across the genome. Combined with myosin inhibition studies, we were able to identify genes that depend on nuclear tension for their mechanosensitivity. In this continuation, we propose to identify molecular mechanisms for these highly novel findings. Two specific aims are proposed: Aim 1: To identify the nuclear molecular linkers necessary for rigidity-mediated control of gene expression. Aim 2: To determine the mechanisms by which nuclear-cytoskeletal linkage regulates gene mechanosensitivity. Scientifically, this work addresses important and longstanding questions about the mechanisms by which the cell microenvironment controls gene expression. Clinically, the long-term impact of this work is to promote the rational development of new biomaterials with mechanical properties tuned for tissue engineering and repair. An additional benefit is the development of an integrated approach using both technologies from engineering and techniques from molecular cell biology for addressing a fundamental question in rigidity sensing. This work is of fundamental interest to diverse fields including cell-biomaterial interactions, nuclear and cell mechanics and molecular and cell biology of gene regulation. The project builds collaboration between the groups of Lele (University of Florida), Nickerson (University of Massachusetts Medical School) and Roux (Sanford Research). Each laboratory brings unique resources to this research including access and expertise in using sophisticated optical and electron microscopes, strong expertise with wet-bench cell and molecular biology experiments specifically related to the nucleus, and expertise in cell and nuclear mechanosensing and biomaterial development and characterization. The team will also benefit from the support of well-known molecular and cell biologists and bioengineers who have pioneered experimental techniques in a broad number of areas in LINC complex biology, nuclear/chromatin structure and function and tissue biomechanics.

项目摘要/摘要 使用固体支架为细胞提供机械和化学线索是一种很有前途的方法 引导组织修复，促进组织-支架一体化。越来越清楚的是，调整 支架刚性是控制细胞功能的一种有效方法，但支架刚性如何调控基因表达 不是很好理解。此前，我们测试了底物刚性通过以下方式控制基因表达的假设 调谐核张力。我们利用了LINC(核骨架到细胞骨架的连接物)这一事实。 复合体是已知的细胞核与细胞骨架的分子连接物，并被问及它如何调节敏感性 从全基因组转录到底物刚性。我们的结果首次表明LINC复合体 促进整个基因组表达的机械调节。结合肌球蛋白抑制研究，我们 能够识别依赖于核张力的机械敏感性基因。在这个续篇中，我们 建议确定这些高度新颖的发现的分子机制。提出了两个具体目标：目标 1：鉴定刚性介导的基因表达调控所必需的核分子连接物。目标2：实现 确定核-细胞骨架连锁调节基因机械敏感性的机制。 从科学上讲，这项工作解决了重要的和长期存在的问题，即 细胞微环境控制基因表达。在临床上，这项工作的长期影响是促进 合理开发用于组织工程和修复的机械性能可调的新型生物材料。 另一个好处是开发了一种使用工程中的两种技术的集成方法 以及来自分子细胞生物学的技术，用于解决刚性传感中的一个基本问题。这部作品 对包括细胞-生物材料相互作用、核和细胞力学在内的不同领域具有根本意义 基因调控的分子和细胞生物学。该项目建立了乐乐集团之间的合作 (佛罗里达大学)、尼克森(马萨诸塞大学医学院)和Roux(桑福德研究)。 每个实验室都为这项研究带来了独特的资源，包括使用尖端技术的途径和专业知识 光学和电子显微镜，对湿法细胞和分子生物学实验有很强的专业知识 特别与核有关，以及细胞和核机械传感和生物材料方面的专业知识 开发和表征。该团队还将受益于知名分子和细胞的支持 在LINC的许多领域开创了实验技术的生物学家和生物工程师 复杂的生物学、核/染色质结构和功能以及组织生物力学。