Investigating the mechanobiology of ER stress in the context of cell proliferation

研究细胞增殖背景下内质网应激的力学生物学

• 批准号: 10246471
• 负责人: Vladislav Belyy
• 金额: $ 10万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10246471
• 关键词: 3-Dimensional; Adhesions; Apoptotic; Autoimmune Diseases; Award; Binding Proteins; Cardiovascular system; Cell Death; Cell Line; Cell Proliferation; Cells; Cellular Mechanotransduction; Cellular biology; Cessation of life; Chemicals; Communication; Complement; Coupled; Coupling; Cues; Defect; Dependence; Development; Diabetes Mellitus; Disease; Embryo; Endoplasmic Reticulum; Engineering; Environment; Enzymes; Exhibits; Extracellular Matrix; Fibroblasts; Foundations; Future; Genes; Growth; Health; Human; Impairment; Individual; Inositol; Investigation; Joints; Laboratories; Lead; Learning; Life; Link; Location; Malignant Neoplasms; Mammalian Cell; Maps; Measures; Mechanics; Mediating; Membrane; Mentors; Molecular; Monitor; Morphogenesis; Multiple Myeloma; Mus; Nature; Nerve Degeneration; Neurons; Outcome; Output; Pathway interactions; Patients; Pattern; Phase; Phosphotransferases; Play; Porosity; Postdoctoral Fellow; Process; Property; Proteins; Publishing; Quantitative Microscopy; Regulation; Research; Resolution; Resources; Ribonucleases; Role; Scientist; Signal Pathway; Signal Transduction; Stress; Therapeutic; Tissues; Vascularization; Vertebrates; Virus Diseases; Work; Zebrafish; biological adaptation to stress; cell growth; cell motility; cell type; collaborative environment; endoplasmic reticulum stress; experimental study; extracellular; human disease; in vivo; inhibitor/antagonist; mechanotransduction; migration; optogenetics; pleiotropism; programs; protein folding; rapid technique; response; scaffold; sensor; skills; spatiotemporal; three dimensional cell culture; tool; transmission process

Project summary The unfolded protein response (UPR) is a critically important signaling network that is responsible for maintaining the health of the endoplasmic reticulum (ER). While the typical outcome of UPR activation is cytoprotective, prolonged or excessive UPR activity can drive cells towards apoptotic death. The UPR serves as a potent controller of cell fate, and its dysregulation is known to be implicated in a broad range of human diseases such as diabetes, neurodegeneration, autoimmune disorders, and cancer. The UPR comprises three interconnected branches that exhibit spatiotemporally distinct patterns of activation both in normal development and in disease. A lack of understanding of how the UPR is differentially regulated in different cell types and tissues has so far precluded it from being successfully targeted in human patients. The best-studied branch of the UPR is mediated by the ER membrane-resident bifunctional kinase-RNase IRE1 (inositol-requiring enzyme 1). Recent work demonstrated that IRE1 signaling is closely tied to adhesion, cell migration, and cells’ ability to receive and respond to external cues. Cell signaling is often coupled to mechanical and chemical changes in the local microenvironment, but the involvement of IRE1 and the UPR in this coupling is only beginning to be revealed. Since UPR components are ubiquitously expressed in nearly all cell types, context-dependent regulation offers an attractive potential explanation for the observed large variances in UPR signaling across tissues. However, it remains to be determined what properties of the local environment cause cells to rely on UPR signaling and how this information is communicated. I propose to build on the tools I developed as a postdoc and on the unique combined resources of my two co-mentors to answer these challenging and exciting questions. I will engineer precisely defined growth substrates of varying chemical composition, porosity, stiffness, and 3-dimensional organization. I will then use chemical inhibitors and optogenetic activators of IRE1 to identify which substrate properties render cells reliant on IRE1 signaling and which properties render IRE1 dispensable. Substrate dependence of IRE1 signaling will be functionally separated from general UPR activation and mapped to specific nodes within the UPR. Finally, a targeted approach will identify the specific molecular players responsible for the information flow between ER stress sensors and the extracellular environment. Throughout the mentored phase of the award, I will continue to hone my skills and qualifications as an independent scientist. Working closely with the lab of Dr. Valerie Weaver will provide me with the expertise in cellular mechanobiology and substrate engineering, complementing the deep background of my primary mentor, Dr. Peter Walter, in UPR signaling. Learning how information flows between the ER and the extracellular matrix will reveal exciting new cell biology, result in possible therapeutic applications, and serve as a strong foundation for an independent research program in my future laboratory.

项目摘要 展开的蛋白质反应（UPR）是一个至关重要的信号网络，负责 保持内质网（ER）的健康状况。虽然UPR激活的典型结果是 细胞保护性，延长或过量的UPR活性可以将细胞驱动到凋亡死亡。 UPR用作 细胞命运的有效控制器及其功能失调是隐含在广泛的人类疾病中的 例如糖尿病，神经变性，自身免疫性疾病和癌症。 UPR包括三个 在正常发展中，在空间上存在激活模式的互连分支 和疾病。缺乏对不同细胞类型和 到目前为止，组织无法成功地针对人类患者。 UPR的最佳研究分支是由ER膜居民双功能激酶-RNase介导的 IRE1（肌醇征用酶1）。最近的工作表明，IRE1信号传导与粘合剂紧密相关， 细胞迁移以及细胞接收和响应外部线索的能力。细胞信号通常耦合到 局部微环境的机械和化学变化，但IRE1和UPR的参与 这种耦合才刚刚开始揭示。由于UPR组件几乎无处不在 细胞类型，与上下文相关的调节为观察到的大型说明提供了有吸引力的潜在解释 UPR信号跨组织的方差。但是，尚待确定本地的特性 环境会导致单元格依靠UPR信号传导以及如何传达此信息。 我建议以我作为博士后开发的工具以及我两者的独特资源建立基础 联合官员回答这些挑战和令人兴奋的问题。我将精确地设计增长 不同化学成分，孔隙率，刚度和三维组织的底物。然后我会使用 IRE1的化学抑制剂和光遗传激活剂，以识别哪些底物特性使细胞可靠 在IRE1信号传导上以及哪些属性使IRE1可置。 IRE1信号的底物依赖性将 在功能上与一般UPR激活分离，并映射到UPR内的特定节点。最后，一个 有针对性的方法将确定负责ER之间信息流的特定分子参与者 应力传感器和细胞外环境。 在整个奖项的整个阶段，我将继续磨练自己的技能和资格 独立科学家。与Valerie Weaver博士的实验室紧密合作将为我提供专业知识 细胞机理生物学和底物工程，完成了我主要心理的深层背景 彼得·沃尔特（Peter Walter）博士，在UPR信号传导中。了解信息如何在ER和细胞外基质之间流动 将揭示令人兴奋的新细胞生物学，从而导致可能的治疗应用，并成为强大的基础 对于我未来的实验室中的独立研究计划。