Substrate Rigidity and Long Non-Coding RNA Expression: Role of TRPV4 Ion Channel

细胞生长的有机材料（基质）的硬度改变了许多细胞功能，包括迁移，增殖和分化。使用为细胞提供正确的机械提示的基底是用于增强组织修复/再生和促进组织-生物材料整合的有前途的方法。长链非编码核糖核酸（LncRNA）是一种信使分子，是细胞分化、发育的有力调节因子，在癌症的发展过程中起着重要作用。 由于基质硬度改变细胞功能，该项目将测试硬度是否调节LncRNA的产生。 我们认为与LncRNA产生相关的一个重要细胞变化是上皮-间充质转化（EMT）。 这是发育、组织修复、瘢痕形成和癌症发展中必不可少的细胞过程，改变了细胞是静止还是迁移。该项目将识别在导致EMT的条件下以基质刚度依赖性方式打开或关闭的LncRNA。该研究计划的总体目标是为目前对基质刚度信号在LncRNA表达和细胞分化中的作用的理解做出变革性贡献。通过增强我们对基质性质如何调节细胞中基因表达的理解，这项研究的结果将在细胞-生物材料相互作用、细胞机械转导以及基因调控的分子和细胞生物学等领域产生广泛的影响。 PI将为本科生和研究生提供从事独立项目的机会，并将与马里兰州大学合作，为来自代表性不足群体的高中生开设“细胞生物力学”暑期课程。该项目的目标是使用全局表达分析来识别和表征基质刚度敏感的LncRNA，并确定TRPV 4在基质硬度诱导的LncRNA表达和EMT中的作用。基于初步数据，中心假设是基质硬度以依赖于TRPV 4的方式控制LncRNA表达，从而控制EMT。Two Specific Aims将通过利用涉及人和小鼠正常原代细胞、TRPV 4缺失和同源野生型小鼠、LncRNA阵列以及EMT的体外和体内模型的分子策略来检验这一假设。具体目标1将使用体外和体内模型系统鉴定和表征基质硬度敏感性LncRNA。具体目标2将描绘TRPV 4、基质硬度敏感性LncRNA和EMT调节之间的关联。当完成时，我们期望本研究的结果将提供关于介导LncRNA表达和EMT的分子机制的新信息和见解。

The stiffness of the organic material (the matrix) that cells grow on changes numerous cellular functions including migration, proliferation, and differentiation. Using a substrate that provides the correct mechanical cues to cells is a promising approach for enhancing tissue repair/regeneration and facilitating tissue-biomaterials integration. Long non-coding ribonucleic acids (LncRNAs) are messenger molecules that are powerful regulators of cell differentiation, development, and important during the progress of cancers. Since matrix stiffness changes cell functions, the project will test whether stiffness regulates LncRNA production. One important cell change that we think is associated with LncRNA production is the epithelial-mesenchymal-transition (EMT). This is an essential cellular process in development, tissue repair, scar formation, and cancer development that changes whether the cells are quiescent or migratory. This project will identify LncRNAs that are turned on or off in a matrix stiffness-dependent manner under conditions that cause EMT. The overall goal of this research program is to produce transformative contributions to the current understanding of the role of matrix stiffness signals in LncRNA expression and cell differentiation. By enhancing our understanding of how matrix properties regulate gene expression in cells, results of this research will have broad-ranging impacts in fields as diverse as cell-biomaterial interactions, cell mechanotransduction, and molecular and cell biology of gene regulation. The PI will will provide opportunities for undergraduate and graduate students to work on independent projects, and will create a summer course on 'Cell Biomechanics' in collaboration with the University of Maryland's Upward Bound Math-Science program for high school students from underrepresented groupsThe objective of this project is to identify and characterize matrix stiffness-sensitive LncRNAs using global expression analysis, and to determine the role of TRPV4 in matrix stiffness-induced LncRNA expression and EMT. Based on the preliminary data, the central hypothesis is that matrix stiffness controls LncRNA expression, and thereby EMT in a manner dependent on TRPV4. Two Specific Aims will test this hypothesis by utilizing molecular strategies involving human and mouse normal primary cells, TRPV4 null and congenic wild-type mice, LncRNA arrays, and in vitro and in vivo models of the EMT. Specific Aim 1 will identify and characterize matrix stiffness-sensitive LncRNAs using in vitro and in vivo model systems. Specific Aim 2 will delineate the association between TRPV4, matrix stiffness-sensitive LncRNAs, and modulation of EMT. When completed, we expect that the results of this study will provide novel information and insight regarding the molecular mechanisms mediating LncRNA expression and EMT.