Cell junction and nuclear forces as mediators of epithelial cell homeostasis

细胞连接和核力作为上皮细胞稳态的介质

• 批准号: 10206611
• 负责人: Daniel E Conway
• 金额: $ 41.93万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10206611
• 关键词: 3-Dimensional; Actomyosin; Adherens Junction; Basic Science; Biosensor; Cell Adhesion; Cell Cycle; Cell Nucleus; Cell Proliferation; Cell-Cell Adhesion; Cell-Matrix Junction; Cells; Chromatin; Chronic; Complex; Cytoskeleton; Desmosomes; Development; Disease; Duct (organ) structure; Epithelial; Epithelial Cells; Epithelial Physiology; Fibrosis; Fluorescence Resonance Energy Transfer; Functional disorder; Goals; Homeostasis; Inflammation; Inflammatory; Intercellular Junctions; Malignant Neoplasms; Measures; Mechanical Stress; Mediator of activation protein; Mesenchymal Stem Cells; Morphogenesis; Nuclear; Nuclear Lamin; Nuclear Lamina; Nuclear Pore; Nuclear Pore Complex; Organ; Permeability; Process; Proteins; Research; Research Proposals; Role; Sepsis; Structure; Tight Junctions; Tissues; Tube; Work; base; cell dimension; cell type; cohesion; epithelial to mesenchymal transition; experience; experimental study; in vitro Model; in vivo; insight; mechanical force; migration; monolayer; mouse model; new therapeutic target; stem cell differentiation; tumor progression; wound healing

Epithelial cells, which line both the inside cavities and outside of the body, exist in tissues as monolayers, multilayers of cells, and three dimensional tube/duct structures. Proper formation and homeostasis of the epithelium is critical for tissue and organ function; dysregulation of the epithelium is associated with epithelial barrier loss (including sepsis), defective wound healing, and development and progression of cancer. Strong cell-cell junctions are critical to the integrity of the epithelium, including cell cohesion, barrier function, and ability to resist mechanical stress. Loss of junctions is associated with epithelial dysfunction including inflammatory-induced increases in permeability and epithelial to mesenchymal transition (EMT). Although formation cell-cell adhesions have been shown to be critical regulators of cell proliferation, migration, and tissue organization, very little is known how cell-cell junction forces contribute to these processes. In addition, the nucleus, which is physically connected to the cytoskeleton by the LINC (Linker of Nucleoskeleton and Cytoskeleton) complex also experiences mechanical force resulting from both actomyosin contractility and externally applied forces across cell-cell contacts and cell-matrix adhesions. Nuclear forces have been shown to regulate the cell cycle, nuclear pore complex, and chromatin. Recent work by my group has also shown that the LINC complex is a critical structure for epithelial homeostasis. This proposal examines the role of force across proteins in both cell-cell junctions and the nucleus as mediators of epithelial homeostasis. The major research goals of this R35 MIRA renewal are to 1) examine how mechanical forces regulate epithelial homeostasis and morphogenesis, 2) identify the role of the LINC complex in epithelial homeostasis and mesenchymal stem cell differentiation, and 3) investigate how forces across nuclear lamins and nuclear pore complexes regulate epithelial physiology. Proposed experiments include in vitro models of ductal/glandular epithelium using FRET-based tension biosensors to directly measure forces across tight junctions, adherens junctions, and desmosomes, as well as the LINC complex, nuclear pores, and the nuclear lamina. New and existing technical approaches will be used to modulate these structures, with the objective of identifying both the upstream regulators of force and the downstream processes regulated by force. Additionally, in vivo mouse models will be used to assess the role of the LINC complex and desmosomes in 3D epithelial tissue homeostasis. This comprehensive study of cell adhesion and nuclear forces will greatly advance the understanding of how epithelial homeostasis is regulated, which is relevant to the processes of wound repair, inflammation, and epithelial tissue development and organization, as well as epithelial diseases, including cancer, fibrosis, and chronic inflammation. Furthermore, results from this study concerning the role of forces on the nuclear lamina and nuclear pore complexes will be relevant to nearly all cell types and tissues.

排列在体内腔和体外的上皮细胞以单层的形式存在于组织中， 多层单元格和三维管道/管道结构。正确的形成和动态平衡 上皮细胞对组织和器官的功能至关重要；上皮细胞的失调与上皮细胞 屏障丧失(包括败血症)，伤口愈合缺陷，以及癌症的发展和进展。强壮 细胞-细胞连接对上皮的完整性至关重要，包括细胞凝聚力、屏障功能和 抵抗机械应力的能力。连接缺失与上皮功能障碍有关，包括 炎症引起的通透性增加和上皮向间充质转化(EMT)。虽然 细胞间黏附的形成已被证明是细胞增殖、迁移和 在组织结构方面，人们对细胞-细胞连接力如何对这些过程做出贡献知之甚少。此外, 细胞核，它通过LINC(核骨架连接物)与细胞骨架物理连接 细胞骨架)复合体也经历由肌动蛋白收缩和 细胞-细胞接触和细胞-基质粘连的外加力。核力量已经被展示出来 调节细胞周期、核孔复合体和染色质。我的团队最近的研究也表明， LINC复合体是上皮细胞动态平衡的关键结构。这项提议考察了武力的作用。 跨越细胞间连接和细胞核中的蛋白质，作为上皮细胞动态平衡的中介。少校 R35 Mira更新的研究目标是1)检查机械力是如何调节上皮细胞的 动态平衡和形态发生，2)确定LINC复合体在上皮动态平衡和形态发生中的作用 间充质干细胞分化，以及3)研究核板和核孔之间的作用力 复合体调节上皮生理学。建议的实验包括导管/腺体的体外模型。 使用基于FRET的张力生物传感器直接测量跨越紧密连接的力的上皮细胞，黏附 连接、桥粒以及LINC复合体、核孔和核层。新的和 将使用现有的技术方法来调整这些结构，目的是确定两者 FORCE的上游调节器和FORCE调节的下游过程。此外，在体内 将使用小鼠模型来评估LINC复合体和桥粒在3D上皮组织中的作用 动态平衡。这种对细胞黏附和核力的综合研究将极大地推动 了解与伤口修复过程相关的上皮动态平衡是如何调节的， 炎症、上皮组织的发育和组织，以及上皮性疾病，包括 癌症、纤维化和慢性炎症。此外，这项关于部队作用的研究结果 在核层和核孔复合体上，几乎所有的细胞类型和组织都是相关的。