Mechanisms restraining the accumulation of antibody secreting cells

抑制抗体分泌细胞积累的机制

• 批准号: BB/W015242/1
• 负责人: Martin Turner
• 金额: $ 95.83万
• 依托单位: Babraham Institute
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FW015242%2F1
• 关键词: Mechanisms; restraining; accumulation; antibody; secreting

The B lymphocyte is the only immune cell type capable of development into the antibody secreting cell (ASC) a cell type commonly referred to as the plasma cell. Defining the genes and cellular processes that regulate ASC development, lifespan and what regulates the amount of antibody secreted by an ASC are fundamental questions -with answers that are likely to be relevant to many other dynamic cell systems in the body. The practical application of this knowledge is of direct relevance to the development of vaccines, interventions for autoimmune/allergic conditions, the impact on healthy ageing of abnormally increased numbers of ASC and the production of synthetic biological therapeutics by mammalian cells. B lymphocyte activation and the pathway of ASC development is underpinned by dynamic changes in gene transcription coupled to post-transcriptional regulation of mRNA localisation, translation and stability. These act together to determine, where, when and in what amounts proteins are produced. The mechanisms of translational control of ASC function and survival is an area ripe for further discoveries in the emerging and important areas of frontier bioscience: these include linking the molecular biology of stress responses to cell differentiation; the avoidance of proteotoxicity; and the determinants of cellular longevity. Knowledge of these may have relevance to almost all biological systems and practical application in biotechnology or medical contexts.This project builds on a new genetic screen for RNA binding proteins (RBP) that regulate the development and survival of ASC. The screen identified genes that were required for the accumulation of ASC and also genes which, when deleted, led to increased ASC formation and survival. The findings of this screen have been published on a preprint server, and this grant proposal is seeking to understand the in vivo immunology, the cell biology and molecular mechanism of modules within two multiprotein complexes [CCR4NOT and EIF3] known to be fundamental for transcription, RNA decay and translation.Do these modules limit ASC development or survival, or do they affect both in vivo? Are the CCR4NOT and EIF3 modules coupled physically and functionally to each other in ASC to form a continuum that connects transcription mRNA stability and translation? Are they regulating homeostatic stress responses such as ER and oxidative stress that is fundamental for the secretory output of ASC? Do they mediate a mechanism to interlink proteostasis with nutrient sensing and oxidative sensing pathways? The module components are encoded by genes only found in higher eukaryotes and thus likely to be important for the unique biology of multicellularity.We propose to study these genes using methods that evaluate quantitatively ASC formation and function in vivo as this is the powerful physiological system of choice. We propose to combine this with in vitro studies of cell biology informed by integrative data-driven transcriptomics methods that take into account the abundance and turnover of mRNA and its translation efficiency. Furthermore, we will determine the composition and location of the multiprotein complexes in which the protein encoded by the genes that limit ASC function reside. This combination of approaches can provide mechanistic insight into how the different processes which control gene expression are coupled to regulate cell function and homeostasis.

B淋巴细胞是唯一能够发育成抗体分泌细胞（ASC）的免疫细胞类型，该细胞类型通常被称为浆细胞。定义调控ASC发育、寿命的基因和细胞过程以及调控ASC分泌的抗体量的因素是基本问题-答案可能与体内许多其他动态细胞系统相关。这些知识的实际应用与疫苗的开发、自身免疫/过敏性疾病的干预、异常增加的ASC数量对健康衰老的影响以及哺乳动物细胞合成生物治疗剂的生产直接相关。B淋巴细胞活化和ASC发展的途径是由基因转录的动态变化以及mRNA定位、翻译和稳定性的转录后调节所支持的。它们共同作用决定蛋白质在何处、何时以及以何种数量产生。ASC功能和存活的翻译控制机制是一个成熟的领域，在新兴的和重要的前沿生物科学领域的进一步发现：这些包括连接应激反应的分子生物学细胞分化;避免蛋白毒性;和细胞寿命的决定因素。这些知识可能与几乎所有的生物系统和生物技术或医学context.This项目的实际应用相关的RNA结合蛋白（RBP），调节ASC的发展和生存的一个新的遗传筛选。筛选鉴定了ASC积累所需的基因，以及当缺失时导致ASC形成和存活增加的基因。这项筛选的结果已经发表在预印本服务器上，这项拨款申请旨在了解体内免疫学，细胞生物学和两种多蛋白复合物[CCR 4 NOT和EIF 3]中模块的分子机制，已知这两种多蛋白复合物是转录，RNA降解和翻译的基础。这些模块是否限制ASC的发育或存活，或者它们是否影响体内两者？CCR 4 NOT和EIF 3模块是否在ASC中物理和功能上相互耦合，形成连接转录mRNA稳定性和翻译的连续体？它们是否调节稳态应激反应，如ER和氧化应激，这是ASC分泌输出的基础？它们是否介导了一种机制，将蛋白质稳态与营养感测和氧化感测途径联系起来？该模块组件是由基因编码的，只发现在高等真核生物，因此可能是重要的独特的生物学multicellularity.We建议研究这些基因的方法，定量评估ASC的形成和功能在体内，因为这是强大的生理系统的选择。我们建议联合收割机将其与细胞生物学的体外研究结合起来，这些研究由综合数据驱动的转录组学方法提供信息，该方法考虑了mRNA的丰度和周转率及其翻译效率。此外，我们将确定多蛋白复合物的组成和位置，其中限制ASC功能的基因编码的蛋白质驻留。这种方法的组合可以提供对控制基因表达的不同过程如何耦合以调节细胞功能和稳态的机制见解。