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，其特征是在早期B细胞发育过程中通过染色体易位获得的Cyclin D1过表达。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，这还远远没有被理解。非遗传机制（包括表观遗传和转录变化）引起的基因失调很普遍，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，理解遗传相互作用和通路重新布线，剖析信号输出的广泛背景依赖性，以及理解肿瘤微环境中的细胞/细胞通信，这些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，以表征淋巴瘤演变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定位相结合，我们的目标是发现驱动小鼠（AIM-1）肿瘤演变不同阶段的核心细胞内在调控回路。我们还将描述淋巴瘤微环境的组成和结构，并监测其在淋巴瘤发生和进展（AIM-2）过程中的纵向变化。单细胞测序和多光谱成像将有助于系统调查的细胞和分子的球员参与细胞/细胞内的MCL利基通信。在AIM-3中，我们将从功能上描述来自AIM-1/2的关键发现。一个焦点是由我们最近关于CD 40活化的工作触发的，CD 40活化是MCL中的标志性事件，其发现了IL 9/IL 9 R介导的细胞/细胞通讯在淋巴瘤发病机制中的意想不到的作用。我们的数据表明，连续前馈循环涉及多个分子和细胞成分。我们将在功能上询问这个模型在小鼠中，通过表征所涉及的细胞类型，基因，信号通路，以及与它们的扰动相关的有机体表型，系统地发现和机制表征MCL进化中的关键调控节点，过程和依赖关系，有望为MCL的精确肿瘤学研究提供信息。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。