Large-scale functional interrogation of evolutionary forces in pre-germinal center B cell lymphoma development

对生发前中心 B 细胞淋巴瘤发展中进化力量的大规模功能询问

• 批准号: 468687596
• 负责人: Professor Dr. Roland Rad
• 金额: --
• 依托单位: Institut für Molekulare Onkologie und Funktionelle Genomik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/468687596?language=en
• 关键词: Large; scale; functional; interrogation; evolutionary

Mantle cell lymphoma (MCL) is an aggressive malignancy, characterized by Cyclin D1 overexpression through a chromosomal translocation acquired during early B cell development. Genome sequencing has given important insights into the mutational landscapes of MCL, but also revealed many layers of unexplored complexity, which is far from being understood. Gene dysregulation by non-genetic mechanisms, including epigenetic and transcriptional changes, is widespread, and discriminating the relevant drivers is challenging. Identifying the key regulatory nodes, positioning them in pathways, networks and processes, understanding genetic interactions and pathway rewiring, dissecting the extensive context-dependency of signaling outputs and understanding cell/cell communication in the tumor microenvironment are only some of the major unresolved challenges. Building on mouse models, genetic tools, methodologies and data resources developed by us, we propose to address these challenges at different levels. In AIM-1 and 2 we will perform genome wide screens and comprehensive large-scale surveys to characterize cell-intrinsic and extrinsic processes underlying lymphoma evolution. By combining in vivo functional screening with high-resolution chromatin profiling and super-enhancer mapping, we aim to discover core cell-intrinsic regulatory circuitries driving different stages of tumor evolution in mice (AIM-1). We will also characterize the composition and architecture of the lymphoma microenvironment and monitor its changes longitudinally during lymphoma initiation and progression (AIM-2). Single cell sequencing and multispectral imaging will aid systematic surveys for cellular and molecular players involved in cell/cell communication within the MCL niche. In AIM-3 we will functionally characterize key discoveries emanating from aim-1/2. One focus is triggered by our recent work on CD40 activation, a hallmark event in MCL, which discovered an unexpected role of IL9/IL9R mediated cell/cell communication in lymphoma pathogenesis. Our data suggest consecutive feed-forward loops involving multiple molecular and cellular constituents. We will functionally interrogate this model in mice by characterizing the involved cell types, genes, signaling pathways, as well as organismal phenotypes related to their perturbation.The systematic discovery and mechanistic characterization of key regulatory nodes, processes and dependencies in MCL evolution is promising to inform precision oncology efforts in MCL. Moreover, the rich data resource created here will drive hypothesis generation and downstream functional studies far beyond the framework of this proposal.

套细胞淋巴瘤 (MCL) 是一种侵袭性恶性肿瘤，其特征是通过早期 B 细胞发育过程中获得的染色体易位导致 Cyclin D1 过度表达。基因组测序为 MCL 的突变景观提供了重要的见解，但也揭示了许多尚未探索的复杂性，而这些复杂性还远未被理解。非遗传机制（包括表观遗传和转录变化）引起的基因失调非常普遍，区分相关驱动因素具有挑战性。识别关键的调控节点，将它们定位在通路、网络和过程中，了解遗传相互作用和通路重新布线，剖析信号输出的广泛上下文依赖性以及了解肿瘤微环境中的细胞/细胞通信只是一些尚未解决的主要挑战。基于我们开发的小鼠模型、遗传工具、方法和数据资源，我们建议在不同层面应对这些挑战。在 AIM-1 和 2 中，我们将进行全基因组筛选和全面的大规模调查，以表征淋巴瘤进化背后的细胞内在和外在过程。通过将体内功能筛选与高分辨率染色质分析和超级增强子图谱相结合，我们的目标是发现驱动小鼠肿瘤进化不同阶段的核心细胞内在调节电路（AIM-1）。我们还将描述淋巴瘤微环境的组成和结构，并在淋巴瘤发生和进展过程中纵向监测其变化（AIM-2）。单细胞测序和多光谱成像将有助于对 MCL 生态位内参与细胞/细胞通讯的细胞和分子参与者进行系统调查。在AIM-3中，我们将从功能上描述aim-1/2的关键发现。我们最近对 CD40 激活的研究引发了一个焦点，CD40 激活是 MCL 的一个标志性事件，它发现了 IL9/IL9R 介导的细胞/细胞通讯在淋巴瘤发病机制中的意外作用。我们的数据表明连续的前馈循环涉及多个分子和细胞成分。我们将通过表征所涉及的细胞类型、基因、信号通路以及与其扰动相关的有机体表型，对小鼠模型进行功能性研究。MCL 进化中关键调控节点、过程和依赖性的系统发现和机制表征有望为 MCL 的精准肿瘤学工作提供信息。此外，这里创建的丰富数据资源将推动假设生成和下游功能研究远远超出本提案的框架。