Decoding and functional characterization of the tonic B cell Antigen receptor signaling Network in human B cells

人 B 细胞中补性 B 细胞抗原受体信号网络的解码和功能表征

• 批准号: 406068353
• 负责人: Dr. Michael Engelke
• 金额: --
• 依托单位: Institut für Zelluläre und Molekulare Immunologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/406068353?language=en
• 关键词: Decoding; functional; characterization; tonic; cell

Beyond the development, activation, and differentiation of B cells the B cell antigen receptor (BCR) controls their survival. The BCR-mediated survival signals are referred to as tonic BCR signals because they occur in the absence of antigen binding. Despite substantial evidence for an importance of the phosphatidyl-3-kinase signaling pathway the molecular details of the tonic BCR signal transduction remain poorly understood. A functional characterization of the underlying signaling processes is, however, important to improve our understanding of survival signals in B cells and their dysregulation in B cell neoplasia. Hence, the overall aim of this project is the elucidation of tonic BCR signal transduction in primary human B cells to identify and functionally characterize key processes with a particular focus on the functional interplay of cytoskeleton-related processes and BCR signaling events that we will study in a comparative manner between normal human B cells and lymphoma cells. We will address this aim with three subprojects that will be carried out in close collaboration between our laboratories at the Universities of Göttingen and Frankfurt.Based on our previous studies of tonic BCR signaling in BL cells, our first subproject will aim at investigating tonic BCR signaling in normal human B cells by phosphoproteomics. By using specific small molecule inhibitors, we will address the role of the BCR-proximal effectors as well as the cytoskeleton in the context of tonic BCR signaling. We will systematically compare these results with the phosphoproteome data that we have obtained for tonic BCR signaling in lymphoma cells.In the second subproject, we will, based on our preliminary data, mechanistically characterize tonic BCR signaling processes on a molecular level. We will generate B cell lines that are deficient for expression of certain signal effectors. Because of the pivotal role of the actin cytoskeleton in controlling BCR signals, we will mainly focus on candidate proteins that are putative regulators of the cytoskeleton. The cell lines will allow us to analyze the role of certain cytoskeletal regulators in tonic BCR signal transduction and will be used to perform comparative analyses in primary human B cells.In the third subproject we will study spacial aspects of tonic BCR signaling. Here, we will focus on the processes that stabilize preformed signaling complexes to control tonic BCR signaling with a particular focus on the role of the actin-based cytoskeleton.

除了B细胞的发育、活化和分化之外，B细胞抗原受体（BCR）控制它们的存活。BCR介导的存活信号被称为紧张性BCR信号，因为它们在不存在抗原结合的情况下发生。尽管有大量证据表明磷脂酰-3-激酶信号通路的重要性，但对强直性BCR信号转导的分子细节仍知之甚少。然而，潜在的信号传导过程的功能表征对于提高我们对B细胞中的存活信号及其在B细胞瘤形成中的失调的理解是重要的。因此，本项目的总体目标是阐明原代人B细胞中的强直BCR信号转导，以识别和功能表征关键过程，特别关注细胞因子相关过程和BCR信号事件的功能相互作用，我们将在正常人B细胞和淋巴瘤细胞之间进行比较研究。我们将通过哥廷根大学和法兰克福大学的实验室密切合作开展三个子项目来实现这一目标。基于我们先前对BL细胞中的强直性BCR信号转导的研究，我们的第一个子项目将旨在通过磷酸化蛋白质组学研究正常人B细胞中的强直性BCR信号转导。通过使用特定的小分子抑制剂，我们将解决BCR近端效应器以及细胞骨架在强直BCR信号传导背景下的作用。我们将系统地比较这些结果与磷酸化蛋白质组的数据，我们已经获得了紧张BCR信号在淋巴瘤cells.在第二个子项目，我们将根据我们的初步数据，在分子水平上的紧张BCR信号过程的机械特征。我们将产生某些信号效应子表达缺陷的B细胞系。由于肌动蛋白细胞骨架在控制BCR信号中的关键作用，我们将主要关注被认为是细胞骨架调节因子的候选蛋白。这些细胞系将使我们能够分析某些细胞骨架调节因子在强直性BCR信号转导中的作用，并将用于在原代人类B细胞中进行比较分析。在这里，我们将集中在稳定预形成的信号复合物，以控制紧张BCR信号的过程中，特别侧重于肌动蛋白为基础的细胞骨架的作用。