Probing ER stress signaling with orthogonal control of receptor oligomerization

通过受体寡聚化的正交控制探索内质网应激信号传导

• 批准号: 10665915
• 负责人: Vladislav Belyy
• 金额: $ 24.9万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10665915
• 关键词: 3-Dimensional; Adhesions; Apoptotic; Autoimmune Diseases; Award; Binding Proteins; Cardiovascular system; Cell Death; Cell Line; Cells; 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; mechanotransduction; migration; optogenetics; pleiotropism; programs; protein folding; rapid technique; receptor; 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 未折叠蛋白反应（UPR）是一个非常重要的信号网络，负责 维持内质网（ER）的健康。虽然UPR激活的典型结果是 细胞保护性的、延长的或过度的UPR活性可驱使细胞走向凋亡性死亡。普遍定期审议作为 细胞命运的有力控制者，已知其失调与多种人类疾病有关 如糖尿病、神经变性、自身免疫性疾病和癌症。普遍定期审议由三个部分组成 相互连接的分支，在正常发育中表现出时空上不同的激活模式， 和疾病。缺乏对UPR如何在不同细胞类型中差异调节的理解， 组织迄今为止阻止了它在人类患者中的成功靶向。 研究最多的UPR分支是由ER膜驻留双功能激酶-RNase介导的 IRE 1（肌醇需要酶1）。最近的研究表明，IRE 1信号传导与粘附密切相关， 细胞迁移以及细胞接收和响应外部信号的能力。细胞信号通常与 机械和化学变化的地方微环境，但参与IRE 1和普遍定期审议， 这种联系才刚刚开始显露出来。由于UPR组分在几乎所有的细胞中都普遍表达， 细胞类型，上下文相关的调节提供了一个有吸引力的潜在解释所观察到的大 不同组织中UPR信号的差异。然而，仍有待确定的是， 环境导致细胞依赖于UPR信号以及如何传递此信息。 我建议建立在我作为博士后开发的工具和我的两个独特的综合资源上。 共同导师来回答这些具有挑战性和令人兴奋的问题。我将设计精确定义的增长 不同化学成分、孔隙率、硬度和三维组织的基底。然后我将使用 IRE 1的化学抑制剂和光遗传学激活剂，以确定哪些底物特性使细胞依赖于 IRE 1信号和哪些属性使IRE 1无效。IRE 1信号传导的底物依赖性将 在功能上与一般的UPR激活分开，并映射到UPR内的特定节点。最后 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， 压力传感器和细胞外环境。 在整个指导阶段的奖励，我将继续磨练我的技能和资格， 独立科学家与瓦莱丽·韦弗博士的实验室密切合作将为我提供以下方面的专业知识 细胞机械生物学和基质工程，补充了我的主要导师的深厚背景， 博士彼得·沃尔特，在UPR信号。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，导致可能的治疗应用，并作为一个强大的基础， 在我未来的实验室里进行独立的研究项目