Cooperative role of TET2 and IDH2 mutations in angioimmunoblastic T-cell lymphomagenesis

TET2 和 IDH2 突变在血管免疫母细胞 T 细胞淋巴瘤发生中的协同作用

• 批准号: 10299140
• 负责人: Wing C. Chan
• 金额: $ 50.08万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10299140
• 关键词: Affect; Aggressive Clinical Course; Arginine; B-Lymphocytes; CD28 gene; CD4 Positive T Lymphocytes; CRISPR/Cas technology; Cell Differentiation process; Cell Line; Cell model; Cells; Characteristics; Clinical; Clonal Expansion; DNA; DNA Methylation; Development; Diagnosis; Disease; Enzymes; Epigenetic Process; Event; Family member; Gene Expression; Gene Expression Profiling; Gene Mutation; Genes; Genetic; Genomics; Glutamine; Goals; Helper-Inducer T-Lymphocyte; Hematologic Neoplasms; Hematopoiesis; Hematopoietic stem cells; Human; Immunoblastic Lymphadenopathy; Immunophenotyping; In Vitro; Induced Mutation; Knockout Mice; Lead; Lesion; Lymphoma; Lymphomagenesis; Malignant - descriptor; Malignant lymphoid neoplasm; Mediating; Metabolic; Modification; Molecular; Mus; Mutation; Myeloid Cells; Myeloproliferative disease; Oncogenic; Outcome; Pathogenesis; Pathogenicity; Pathologic; Pathway interactions; Patients; Peripheral; Phenotype; Play; Pre-Clinical Model; Preclinical Testing; Production; Prognosis; RHOA gene; Research; Role; Sampling; Specimen; Stromal Cells; System; T cell differentiation; T-Cell Activation; T-Cell Development; T-Cell Lymphoma; T-Cell Transformation; T-Lymphocyte; Testing; Therapeutic Intervention; Transgenic Organisms; Validation; accurate diagnosis; cell transformation; drug efficacy; effective therapy; efficacy study; efficacy testing; epigenome; epigenomics; genome-wide; histone demethylase; human model; inhibitor/antagonist; insight; loss of function mutation; mouse model; mutant; mutant mouse model; neoplastic; neoplastic cell; novel; novel therapeutic intervention; novel therapeutics; patient derived xenograft model; polarized cell; pre-clinical; programs; targeted treatment; therapeutic evaluation; treatment response; tumor; tumor microenvironment

Abstract Angioimmunoblastic T-cell lymphoma (AITL) is the most common subtype of peripheral T-cell lymphoma (PTCL) with distinct pathological and clinical characteristics. It is a difficult-to-diagnose and lacks effective therapies due to poor understanding of the molecular pathogenesis. Using genome-wide gene expression profiling (GEP), we have defined a robust molecular classifier for accurate diagnosis, identified oncogenic pathways, discovered its cell-of-origin, and deciphered the role of the tumor milieu in disease prognosis. Using high-throughput genomic analysis, we also identified frequent mutations in epigenetic regulators (TET2, IDH2 and DNMT3A) and regulators of T-cell activation (RHOA and CD28) in AITL. Co-occurrence of TET2 mutations with IDH2 mutation (hotspot: arginine-172; IDH2R172) is unique in AITL compared to other hematological malignancies where they are mutually exclusive, suggesting their cooperative role in T-cell lymphomagenesis. We have generated unique murine models with conditional loss of Tet2 (Tet2-/-) and double mutant (Tet2-/- /IDH2R172K) in CD4+ T-cells, and murine lymphomas with follicular helper T-cell (TFH) phenotype were observed. In addition, we generated several patient-derived xenografts (PDX) of AITL with IDH2R172 and/or TET2 mutations. We hypothesize that TET2 deficiency in T cells alters the DNA methylation profile leading to an altered genetic program favoring TFH cell differentiation and clonal expansion, facilitating transformation with subsequent hits, including IDH2R172 mutation, which will further alter the epigenome and oncogenic pathways in TFH -cell transformation. We will define the precise role of these genetic lesions in T-cell differentiation and in AITL pathogenesis. We also edited normal human CD4+ T-cells using CRISPR/CAS9 to generate TET2-/- T-cells for functional analysis and cross-validation of murine models. SinceTET2 mutations are in hematopoietic stem/progenitor cells (HSPC) in some AITLs, interactions between TET2 deficient T- and stromal cells during lymphomagenesis will be explored. We will test the efficacy of therapeutic interventions, including demethylating agents for TET2 deficiency and IDH2 inhibitors and/or glutamine depletion for TET2/IDH2 double mutant tumors in murine models and PDX of AITL. To accomplish the objective, three aims are planned: Specific Aim 1: To determine the mechanisms by which Tet2 deficiency mediates T-cell transformation and the role of Tet2 mutation in stromal cells in lymphomagenesis. Specific Aim 2: To delineate the mechanisms of combined Tet2 and IDH2R172K mutations in AITL pathogenesis. Specific Aim 3: To evaluate therapies targeting oncogenic mechanisms mediated by Tet2 and IDH2R172 mutations. Our long-term research goal is to define the pathobiology of AITL, through integrated functional epigenomic approaches using in vitro modified human T-cells, patient samples and relevant murine models, thus identifying promising novel targets for treatment that may be tested pre-clinically in PDX models.

抽象的 血管免疫母细胞性 T 细胞淋巴瘤 (AITL) 是外周 T 细胞淋巴瘤最常见的亚型 （PTCL）具有独特的病理和临床特征。该病诊断困难，缺乏有效的治疗方法 由于对分子发病机制了解不足而无法进行治疗。使用全基因组基因表达 分析（GEP），我们定义了一个强大的分子分类器，用于准确诊断，识别致癌基因 途径，发现其细胞起源，并破译肿瘤环境在疾病预后中的作用。使用 通过高通量基因组分析，我们还发现了表观遗传调节因子（TET2、IDH2）的频繁突变 和 DNMT3A）以及 AITL 中 T 细胞激活的调节因子（RHOA 和 CD28）。 TET2 突变同时发生 与其他血液学相比，具有 IDH2 突变（热点：精氨酸-172；IDH2R172）的 AITL 是独一无二的 它们相互排斥的恶性肿瘤，表明它们在 T 细胞淋巴瘤发生中的合作作用。 我们已经生成了独特的小鼠模型，具有条件性丢失 Tet2 (Tet2-/-) 和双突变体 (Tet2-/-) 观察到 CD4+ T 细胞中的 /IDH2R172K）以及具有滤泡辅助 T 细胞（TFH）表型的小鼠淋巴瘤。 此外，我们还生成了几个具有 IDH2R172 和/或 TET2 突变的 AITL 患者来源的异种移植物 (PDX)。 我们假设 T 细胞中 TET2 缺陷会改变 DNA 甲基化谱，从而导致 有利于 TFH 细胞分化和克隆扩增的遗传程序，促进转化 随后的打击，包括 IDH2R172 突变，这将进一步改变表观基因组和致癌性 TFH-细胞转化的途径。我们将定义这些基因损伤在 T 细胞中的精确作用 分化和 AITL 发病机制。我们还使用 CRISPR/CAS9 编辑了正常人类 CD4+ T 细胞 生成 TET2-/- T 细胞，用于小鼠模型的功能分析和交叉验证。由于TET2突变位于 一些 AITL 中的造血干/祖细胞 (HSPC)，TET2 缺陷的 T 细胞和基质细胞之间的相互作用 将探索淋巴瘤发生过程中的细胞。我们将测试治疗干预措施的功效，包括 用于 TET2 缺陷的去甲基化剂和用于 TET2/IDH2 双的 IDH2 抑制剂和/或谷氨酰胺消耗 小鼠模型中的突变肿瘤和 AITL 的 PDX。为了实现这一目标，计划实现三个目标： 具体目标 1：确定 Tet2 缺陷介导 T 细胞转化的机制以及 基质细胞中 Tet2 突变在淋巴瘤发生中的作用。 具体目标 2：阐明 Tet2 和 IDH2R172K 联合突变在 AITL 发病机制中的机制。 具体目标 3：评估针对 Tet2 和 IDH2R172 介导的致癌机制的疗法 突变。 我们的长期研究目标是通过综合功能表观基因组学来定义 AITL 的病理学 使用体外修饰的人类 T 细胞、患者样本和相关小鼠模型的方法，从而确定 有前途的新治疗靶标可在 PDX 模型中进行临床前测试。