Using mouse models to identify actionable molecular liabilities in Cluster 5 DLBCL

使用小鼠模型识别 Cluster 5 DLBCL 中可采取行动的分子责任

• 批准号: 442814095
• 负责人: Professor Dr. Christian Reinhardt
• 金额: --
• 依托单位: Klinik für Hämatologie und Stammzelltransplantation
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2020
• 资助国家: 德国
• 起止时间: 2019-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/442814095?language=en
• 关键词: Using; mouse; models; identify; actionable

DLBCL remains a clinical challenge, as relapsed and refractory disease is very difficult to treat. However, recent genomics efforts have revealed the landscape of recurrent aberrations in DLBCL. While this area of investigation is still in flux, a rather clear picture of ABC-DLBCL, and particularly C5 DLBCL lymphomagenesis, is emerging: A hallmark feature of C5 DLBCL is inappropriate NFB activation, which is achieved through recurrent mutations within the BCR (CD79B) and/or TLR (MYD88) pathways. A further hallmark of C5 DLBCL are BCL2 copy number gains. Lastly, C5 DLBCL is characterized by recurrent inactivating aberrations in PRDM1, which lead to a block in plasma cell differentiation. These recurrent mutations may offer opportunities for targeted therapeutic interventions, which may allow chemotherapy-free treatment approaches for this disease. However, there is currently a lack of suitable in vivo experimental platforms that faithfully mimic the genomic landscape of human C5 DLBCL. Here, we set out to expand on our existing efforts on C5 DLBCL modeling to develop a comprehensive and versatile in vivo preclinical platform that employs a combination of large-scale in vitro vulnerability screening, and an in vivo validation tool box to develop innovative, chemotherapy-free and genomics-guided therapeutic approaches for the treatment of C5 DLBCL patients. Our proposal builds on substantial preliminary data and we have successfully established a whole range of innovative technologies to fully capture the biology of C5 DLBCL, including longitudinal in vivo imaging, high throughput immuno-histochemistry, CRISPR/Cas9 drop out screening, CyTOF mass cytometry, 3'-RNA sequencing, whole exome and whole genome sequencing, single cell RNA sequencing and cytokine profiling. Our systems can further be exploited to accelerate the search for resistance-mediating genes and pathways, once viable targeted treatment approaches have been identified. Based on these considerations, we have formulated three specific aims:Aim 1: Generation and characterization of an inducible autochthonous mouse model harboring the dominant aberrations defining C5 DLBCLAim 2: Systematic CRISPR/Cas9 loss of function screening to identify genotype-specific vulnerabilities in C5 DLBCLAim 3: Assess the efficacy of combined BCL2-, BTK-, CD20- and PD-1/PD-L1 targeting in autochthonous C5 DLBCL models in vivoThese aims test the hypotheses that 1) oncogenic Myd88 mutations cooperate with Bcl2, Prdm1 and Cd79b aberrations to promote C5 DLBCL, 2) autochthonous mouse models of C5 DLBCL and cell line models derived thereof may serve as experimental platforms to develop and refine novel chemotherapy-free treatment algorithms of C5 DLBCL, and 3) MYD88-driven C5 DLBCL may display an PD-L1 surface expression. The proposed experiments will pave the way for the clinical development of genomics-guided precision medicine approaches to C5 DLBCL.

由于复发性和难治性疾病的治疗非常困难，DLBCL仍然是一个临床挑战。然而，最近的基因组学研究揭示了DLBCL中复发性畸变的情况。虽然这一研究领域仍在不断变化，但ABC-DLBCL，特别是C5 DLBCL淋巴瘤发生的相当清晰的图像正在出现：C5 DLBCL的一个标志性特征是不适当的NF B激活，这是通过BCR（CD 79 B）内的复发突变实现的和/或TLR（MYD 88）途径。C5 DLBCL的另一个标志是BCL 2拷贝数增加。最后，C5 DLBCL的特征在于PRDM 1中的复发性失活畸变，其导致浆细胞分化阻滞。这些复发性突变可能为靶向治疗干预提供机会，这可能使这种疾病的无化疗治疗方法成为可能。然而，目前缺乏忠实地模拟人C5 DLBCL的基因组景观的合适的体内实验平台。在这里，我们着手扩大我们在C5 DLBCL建模方面的现有努力，以开发一个全面和通用的体内临床前平台，该平台采用大规模体外脆弱性筛选和体内验证工具箱的组合，以开发创新的，无化疗和基因组学指导的治疗方法，用于治疗C5 DLBCL患者。我们的提案建立在大量的初步数据基础上，我们已经成功建立了一整套创新技术，以充分捕获C5 DLBCL的生物学，包括纵向体内成像，高通量免疫组织化学，CRISPR/Cas9脱落筛选，CyTOF质谱仪，3 '-RNA测序，全外显子组和全基因组测序，单细胞RNA测序和细胞因子分析。一旦确定了可行的靶向治疗方法，我们的系统可以进一步用于加速寻找耐药介导基因和途径。基于这些考虑，我们制定了三个具体目标：目标1： 产生和表征具有定义C5 DLBCLAim 2的显性畸变的可诱导的本地小鼠模型： 系统性CRISPR/Cas9功能丧失筛查以识别C5 DLBCLAim 3中的基因型特异性漏洞：评估BCL 2-、BTK-、CD 20-和PD-1/PD-L1组合靶向在体内自体C5 DLBCL模型中的功效。这些目的测试以下假设：1）致癌Myd 88突变与Bcl 2、Prdm 1和Cd 79 b畸变协同促进C5 DLBCL，2）C5 DLBCL的自体小鼠模型及其衍生的细胞系模型可以用作开发和改进C5 DLBCL的新的无化疗治疗算法的实验平台，和3）MYD 88驱动的C5 DLBCL可以显示PD-L1表面表达。拟议的实验将为基因组学指导的C5 DLBCL精准医学方法的临床开发铺平道路。