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: 9142466
• 负责人: Daniel E Conway
• 金额: $ 36.71万
• 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 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9142466
• 关键词: Adherens Junction; Affect; Basic Science; Biosensor; Cell Nucleus; Cell Proliferation; Cell-Cell Adhesion; Cell-Matrix Junction; Cells; Chronic; Complex; Cytoskeleton; Desmosomes; Development; Disease; Duct (organ) structure; Epithelial; Epithelial Cells; Epithelium; Fibrosis; Fluorescence Resonance Energy Transfer; Focal Adhesions; Functional disorder; Geometry; Goals; Homeostasis; Inflammation; Inflammatory; Intercellular Junctions; Malignant Neoplasms; Measures; Mechanical Stress; Mechanics; Mediator of activation protein; Nature; Nuclear; Nuclear Matrix; Nuclear Proteins; Organ; Organelles; Permeability; Positioning Attribute; Process; Protein Isoforms; Proteins; Regulation; Research; Research Proposals; Role; Sepsis; Structure; Tight Junctions; Tissues; Tube; Wound Healing; base; cohesion; epithelial to mesenchymal transition; insight; migration; monolayer; mutant; new therapeutic target; novel; tumor progression

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. Although mechanical forces on epithelial cells have been shown to influence cell organization, proliferation, and migration, it is not known the mechanisms by which cells respond to force. This proposal examines the role of force across proteins in both cell-cell junctions and the nuclear linker of nucleoskeleton to cytoskeleton (LINC) complex as mediators of epithelial homeostasis. 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 to altering junction forces, externally applied forces are likely transmitted inside the cell, across the cytoskeleton, and onto organelles. The nucleus, which is physically connected to the cytoskeleton by the LINC complex, is likely affected by external forces. Nuclear forces have been suggested to regulate nuclear geometry and nuclear positioning, both of which are altered in a variety of diseases, including cancer. The major research goals of this MIRA proposal are to examine how forces across cell-cell junction proteins and the nuclear LINC complex regulate epithelial proliferation, migration, junction stability, and 3D organization. Novel FRET-based tension biosensors will be used to directly measure forces across tight junctions, adherens junctions, and desmosomes at cell-cell junctions and specific isoforms of nesprin at the LINC complex. Treatments or mutants for which junction or nuclear force is perturbed will be used to assess the causal nature of force in regulation of the epithelium. An additional goal of this proposal is to identify the relationship between forces across cell-cell junctions, cell-matrix adhesions, and the nuclear LINC complex, identifying how forces are transmitted from one region of the cell to another. This comprehensive study of cell-cell, cell-matrix, and nuclear forces will greatly advance the understanding of epithelial homeostasis, which includes the processes of wound repair, inflammation, and epithelial tissue development and organization, as well as epithelial diseases, including cancer, fibrosis, and chronic inflammation. Junction and nuclear forces may represent a universal mechanism to regulate proliferation, migration, and organization; therefore, results from this study may also be relevant to a large number of non- epithelial cells and tissues.

上皮细胞排列在体内腔和体外，以单层形式存在于组织中， 多层细胞和三维管/导管结构。的正确形成和稳态 上皮对于组织和器官功能至关重要；上皮细胞的失调与上皮细胞的 屏障丧失（包括败血症）、伤口愈合缺陷以及癌症的发生和进展。虽然 上皮细胞上的机械力已被证明会影响细胞组织、增殖和 迁移时，尚不清楚细胞对力做出反应的机制。该提案审查了 细胞与细胞连接处以及核骨架与细胞骨架的核连接物 (LINC) 中蛋白质的作用力 复合物作为上皮稳态的介质。强大的细胞与细胞连接对于细胞的完整性至关重要 上皮，包括细胞凝聚力、屏障功能和抵抗机械应力的能力。结点损失为 与上皮功能障碍相关，包括炎症引起的通透性和上皮细胞的增加 间质转化（EMT）。尽管细胞间粘附的形成已被证明是至关重要的 细胞增殖、迁移和组织组织的调节剂，但人们对细胞与细胞连接的机制知之甚少 力量有助于这些过程。除了改变连接力之外，外部施加的力也可能 在细胞内传递，穿过细胞骨架，到达细胞器上。原子核，物理上的 通过 LINC 复合物与细胞骨架相连，很可能受到外力的影响。核力量有 有人建议调节核几何形状和核定位，这两者都在各种情况下发生改变 疾病，包括癌症。 MIRA 提案的主要研究目标是研究力量如何跨越 细胞-细胞连接蛋白和核 LINC 复合物调节上皮增殖、迁移、连接 稳定性和 3D 组织。基于 FRET 的新型张力生物传感器将用于直接测量力 穿过细胞与细胞连接处的紧密连接、粘附连接和桥粒以及特定的异构体 LINC 综合体的 Nesprin。连接或核力受到干扰的治疗或突变体将是 用于评估上皮调节力的因果性质。该提案的另一个目标是 确定跨细胞-细胞连接、细胞-基质粘附和核的力之间的关系 LINC 复合体，识别力如何从细胞的一个区域传递到另一区域。这 对细胞-细胞、细胞-基质和核力的全面研究将极大地促进对细胞-细胞、细胞-基质和核力的理解 上皮稳态，包括伤口修复、炎症和上皮组织的过程 发育和组织，以及上皮疾病，包括癌症、纤维化和慢性病 炎。联合力量和核力量可能代表了调节扩散的通用机制， 迁移和组织；因此，这项研究的结果也可能与大量非 上皮细胞和组织。