Mitosis in Confining Microenvironments

限制性微环境中的有丝分裂

• 批准号: 10719384
• 负责人: Taeyoon Kim
• 金额: $ 26.21万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-25 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10719384
• 关键词: 3-Dimensional; Adult; Alginates; Automobile Driving; Behavior; Biocompatible Materials; Biological Process; CRISPR/Cas technology; Cell Culture Techniques; Cell Cycle; Cell Line; Cell Mobility; Cell Separation; Cell division; Cell membrane; Cell-Matrix Junction; Cells; Characteristics; Collagen; Complex; Computer Models; Cultured Cells; Development; Disease; Elasticity; Embryo; Engineering; Experimental Models; Extracellular Matrix; Feedback; G1 Phase; Generations; Goals; Growth; Homeostasis; Human; Ion Channel; Knock-out; Knowledge; Location; Malignant Neoplasms; Mammalian Cell; Measurement; Mechanics; Mediating; Mission; Mitosis; Mitotic; Modeling; Morphology; Movement; Myosin ATPase; Natural regeneration; Osmotic Pressure; Physical Examination; Physiological; Process; Property; Public Health; Relaxation; Research; Research Project Grants; Role; Stress; Testing; United States National Institutes of Health; cell growth; cell osmotic pressure; cell type; defined contribution; disability; experimental study; extracellular; force feedback; innovation; insight; novel; novel therapeutics; regenerative; three dimensional cell culture; tumor; viscoelasticity; wound healing

PROJECT SUMMARY Cell division underlies the development of humans from embryos to full-grown adults, regenerative processes such as wound healing, and diseases such as cancer. While much is known about the intracellular aspects of mammalian cell division, less is known about the extracellular aspects of cell division. In many physiological contexts, cells divide in mechanically confining microenvironments, including dense extracellular matrices (ECMs) and growing tumors. Cell division requires extensive morphological changes, including significant growth during the G1 phase of the cell cycle and elongation along the mitotic axis during mitosis, or mitotic elongation. Both growth and mitotic elongation are strictly required for successful cell division. A mechanically confining microenvironment provides a physical barrier to both cell growth and mitotic elongation, and cells must overcome this confinement for successful cell division. Our recent studies have shown that single dividing cells in three- dimensional (3D) matrices generate protrusive forces along the mitotic axis to drive mitotic elongation via a combination of interpolar spindle elongation and cytokinetic ring contraction. We have also found that cell growth during the G1 phase is mediated by outward force generation. However, it remains unclear how these forces and their underlying mechanisms adapt to confining microenvironments with a wide range of stiffness and viscoelasticity. In this project, we will determine how cells tune extracellular forces to sustain cell division in highly confining microenvironments, using a powerful combination of rigorous agent-based modeling and experiments with engineered biomaterials for 3D cell culture. We hypothesize that in microenvironments with increased confinement, i) protrusive activity increases to make space and activate mechanosensitive channels for driving G1 phase cell growth via increased osmotic pressure, and ii) enhanced cytokinetic ring contraction drives mitotic elongation. The main hypothesis will be tested by pursuing the following three aims: (1) Determine how mitotic elongation of isolated cells within highly confining microenvironments is accomplished via a novel force feedback mechanism; (2) Define how isolated cells achieve G1 phase cell growth in highly confining microenvironments; and (3) Establish how growth and mitotic elongation of cells in growing spheroids induce overall expansion of spheroids in highly confining microenvironments. The proposed research project is significant because it will reveal how cells modulate their force generation, to drive cell growth and mitotic elongation for cell division in physiologically relevant microenvironments, and also elucidate the role of matrix remodeling and multicellular cooperation in cell division. The approach is innovative because of i) the development and use of agent-based models that can rigorously capture the most important aspects of cell growth, mitotic elongation, and confining microenvironments with complex rheological properties, ii) the focus on extracellular aspects of cell division, iii) the role of matrix viscoelasticity in cell division, and iv) the examination of the physical basis for spheroid growth.

项目概要 细胞分裂是人类从胚胎到成熟成人的发育、再生过程的基础 例如伤口愈合和癌症等疾病。虽然人们对细胞内方面了解很多 哺乳动物细胞分裂，但人们对细胞分裂的细胞外方面知之甚少。在许多生理 在这种情况下，细胞在机械限制的微环境中分裂，包括致密的细胞外基质 （ECM）和生长中的肿瘤。细胞分裂需要广泛的形态变化，包括显着的生长 细胞周期的G1期和有丝分裂期间沿有丝分裂轴的伸长，或有丝分裂伸长。 生长和有丝分裂伸长都是细胞成功分裂所必需的。机械限制 微环境为细胞生长和有丝分裂伸长提供了物理障碍，细胞必须克服 这种限制是细胞成功分裂的前提。我们最近的研究表明，三个细胞中的单个分裂细胞 三维（3D）矩阵沿着有丝分裂轴产生突出力，通过a驱动有丝分裂伸长 极间纺锤体伸长和细胞因子环收缩的组合。我们还发现细胞生长 G1 阶段是由外向力的产生介导的。然而，目前尚不清楚这些力量如何 及其潜在机制适应具有广泛刚度和范围的限制微环境 粘弹性。在这个项目中，我们将确定细胞如何调节细胞外力以高度维持细胞分裂。 限制微环境，使用严格的基于代理的建模和实验的强大组合 使用用于 3D 细胞培养的工程生物材料。我们假设在微环境中 限制，i）增加突出活动以腾出空间并激活机械敏感通道以进行驾驶 G1 期细胞通过增加渗透压生长，ii) 增强的细胞因子环收缩驱动有丝分裂 伸长。主要假设将通过追求以下三个目标来检验：（1）确定有丝分裂如何进行 通过一种新颖的力反馈实现高度限制的微环境中分离细胞的伸长 机制; (2) 定义分离的细胞如何在高度限制的微环境中实现G1期细胞生长； (3) 确定生长球体中细胞的生长和有丝分裂伸长如何诱导细胞的整体扩张 高度限制的微环境中的球体。拟议的研究项目意义重大，因为它将 揭示细胞如何调节其力的产生，以驱动细胞生长和有丝分裂伸长以进行细胞分裂 生理相关的微环境，并阐明基质重塑和多细胞的作用 细胞分裂中的合作。该方法具有创新性，因为 i) 基于代理的开发和使用 可以严格捕获细胞生长、有丝分裂伸长和限制的最重要方面的模型 具有复杂流变特性的微环境，ii) 关注细胞分裂的细胞外方面，iii) 基质粘弹性在细胞分裂中的作用，以及 iv) 检查球体生长的物理基础。