Non-coding RNAs at the interface of aberrant NF-kB signals and lymphomagenesis

异常 NF-kB 信号与淋巴瘤发生界面的非编码 RNA

• 批准号: 8974297
• 负责人: Ricardo C Aguiar
• 金额: --
• 依托单位: SOUTH TEXAS VETERANS HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8974297
• 关键词: 3' Untranslated Regions; Address; Adult; B lymphoid malignancy; B-Cell Development; B-Lymphocytes; Balkans; Binding Sites; Biological Models; Biology; Blood Cells; Cell Line; Chemical Exposure; Clinical; Code; Collection; Data; Development; Diagnosis; Disease; Early Diagnosis; Ectopic Expression; Epigenetic Process; Epithelial; Family; Feedback; Functional disorder; Gene Expression; Gene Targeting; Genes; Genetic; Genome; Goals; Health; Heterogeneity; Human; In Vitro; Infectious Agent; Knowledge; Lymphocyte; Lymphocyte Biology; Lymphoma; Lymphomagenesis; MAP3K7 gene; MS4A1 gene; Malignant Neoplasms; Malignant lymphoid neoplasm; Maps; Mature B-Lymphocyte; Measurement; Mediating; MicroRNAs; Military Personnel; Modeling; Molecular; Monoclonal Antibody Therapy; NF-kappa B; Oncogenic; Pathogenesis; Pathway interactions; Patients; Play; Population; Porifera; Primary Neoplasm; RNA; RNA-Binding Proteins; Research; Research Proposals; Retroviridae; Risk; Role; Sampling; Signal Transduction; Somatic Mutation; Study models; System; TNF receptor-associated factor 5; TRAF2 gene; Testing; Therapeutic; Translational Repression; Tumor Suppressor Genes; Untranslated RNA; Vietnam; Western World; Xenograft Model; base; biological systems; cancer type; gain of function; genome-wide; improved; in vivo; knock-down; large cell Diffuse non-Hodgkin' s lymphoma; loss of function; malignant lymphocyte; member; non-invasive imaging; novel; novel marker; novel therapeutic intervention; patient population; prediction algorithm; screening; small hairpin RNA; transcriptome; tumor

DESCRIPTION (provided by applicant): Diffuse Large B-cell lymphoma (DLBCL) is the most common lymphoid malignancy in adults. Despite recent progress, a large fraction of these patients still die of their disease. The hallmar of the most aggressive subtype of DLBCL, Activated B-cell (ABC)-like DLBCL, is constitutive activation of the NF-¿B pathway. In approximately half of these cases, aberrant NF-¿B activation results from somatic mutations in negative and positive regulators of this pathway. However, in a large proportion of NF-¿B-addicted DLBCLs the molecular basis for these abnormalities is unknown. MicroRNAs (miRNAs) are non-protein coding genes that regulate the human transcriptome by pairing to the 3'UTR of target genes, inducing RNA cleavage and/or translational inhibition. MiRNAs are frequently disrupted in DLBCL, suggesting that they may play a role in constitutively activating NF-kB signals in these tumors. Concordantly, evidence from other biological systems indicates that there is an extensive interplay between miRNAs and the NF-¿B pathway. The overarching objective of this proposal is to systematically test the novel hypothesis that genetic and epigenetic disruption of the interactions between miRNAs and members of the NF-¿B pathway contributes to the constitutive activation of these signals in the fatal subtypes of DLBCL. More specifically, we will: 1. Characterize and functionally validate the direct interaction between the DLBCL-relevant miR-125a/b and miR-18a, miR-19a/b with the tumor suppressor gene, and negative NF-¿B regulator, TNFAIP3; 2. Identify and characterize the miRNAs that directly target the positive NF-¿B regulators CARD11, MYD88, CD79B, TRAF2, TRAF5 and TAK1; 3. Define the role of the Lin28b and the let-7 miRNA family in an epigenetically-driven self-sustained loop that could constitutively activate NF-¿B in DLBCL. To achieve the goals of this research proposal, we will make extensive use of genetically modified DLBCL cell lines, in vitro and in vivo, and primary lymphoma samples. In brief, miRNA expression and function will be modulated with stable expression of their precursor sequence via a retrovirus system (gain-of-function), or with "miRNA sponges", constructs that sequester the miRNAs away from endogenous binding sites resulting in a significant functional knockdown (loss-of-function). Furthermore, stable ectopic expression and shRNA-mediated knockdown of LIN28B will be used in complementary fashion to define the presence of an epigenetically-driven positive feedback loop that maintains NF-¿B constitutive active in DLBCL. Throughout this study, the models that yield relevant result in vitro will be validated in a xenograft model o human lymphoma with non-invasive imaging capability. Finally, the relevance of these results will be confirmed in the most critical setting, primary tumors, via measurement of miRNAs, target genes expression and characterization of NF-¿B activity. The identification of miRNAs that when disrupted activate NF-¿B signals will clarify the function of multiple oncogenic and tumor suppressive miRNAs and advance our understanding of normal and malignant lymphocyte biology. These findings will elucidate the molecular basis for a substantial fraction of DLBCLs that are "addicted" to NF-kB signals, an important knowledge gap to be addressed as it relates to the fatal subsets of DLBCLs. Further, since high NF-kB activity is not exclusive of ABC-DLBCL, but contributes to the pathogenesis of multiple mature B-cell malignancies as well as epithelial cancer, our data is likely to have broad relevance and eventually benefit a large patient population. Finally, as the potential of inhibiting and restoring miRNA levels with therapeutic intent is becoming a reality, our studies may provide a blueprint for testing this concept in cancer.

描述（由申请人提供）： 弥漫性大 B 细胞淋巴瘤 (DLBCL) 是成人中最常见的淋巴恶性肿瘤，尽管最近取得了进展，但其中很大一部分患者仍然死于该病，这是 DLBCL 最具侵袭性的亚型——激活的 B 细胞 (ABC) 的标志。 ) 类似 DLBCL，是 NF-¿ 的组成型激活在大约一半的病例中，NF-¿ B 激活是由该途径的负调节因子和正调节因子的体细胞突变引起的，但是，在很大一部分 NF-¿ B 依赖性 DLBCL 的这些异常的分子基础尚不清楚，MicroRNA (miRNA) 是通过与靶基因 3'UTR 配对、诱导 RNA 裂解和/或翻译抑制来调节人类转录组的基因。在 DLBCL 中经常被破坏，这表明它们可能在这些肿瘤中持续激活 NF-kB 信号中发挥作用，来自其他生物系统的证据表明， miRNA 和 NF-¿ 之间存在广泛的相互作用该提案的首要目标是系统地测试新的假设，即 miRNA 与 NF-¿ 成员之间相互作用的遗传和表观遗传破坏。 B 途径有助于 DLBCL 致命亚型中这些信号的组成型激活。更具体地说，我们将： 1. 表征并功能验证 DLBCL 相关 miR-125a/b 和 miR-18a、miR-19a 之间的直接相互作用。 /b 带有抑癌基因，且 NF-¿ 阴性B 调节因子，TNFAIP3；2. 识别并表征直接靶向阳性 NF-¿ B 调节因子 CARD11、MYD88、CD79B、TRAF2、TRAF5 和 TAK1；3. 定义 Lin28b 和 let-7 miRNA 家族在表观遗传驱动的自我维持环中的作用，该环可以组成性激活 NF-¿ B 在 DLBCL 中的应用 为了实现本研究计划的目标，我们将在体外和体内广泛使用转基因 DLBCL 细胞系以及原发性淋巴瘤样本。简而言之，将通过稳定表达来调节 miRNA 的表达和功能。它们的前体序列通过逆转录病毒系统（功能获得）或“miRNA海绵”构建，将miRNA与内源性结合位点隔离开，从而导致显着的功能敲低（功能丧失）。 LIN28B的稳定异位表达和shRNA介导的敲低将以互补的方式用于定义维持NF-¿的表观遗传驱动的正反馈环的存在在整个研究中，在体外产生相关结果的模型将在具有非侵入性成像能力的人类淋巴瘤异种移植模型中得到验证。 、原发性肿瘤，通过测量 miRNA、靶基因表达和 NF-¿ 的表征B 活性的鉴定，当被破坏时，会激活 NF-¿ B 信号将阐明多种致癌和肿瘤抑制 miRNA 的功能，并增进我们对正常和恶性淋巴细胞生物学的理解。这些发现将阐明大部分的分子基础。 DLBCL 对 NF-kB 信号“上瘾”，这是一个需要解决的重要知识空白，因为它与 DLBCL 的致命亚型有关。此外，因为高 NF-kB 活性并不排除 ABC-DLBCL，而是有助于发病机制。多种成熟 B 细胞恶性肿瘤以及上皮癌的研究，我们的数据可能具有广泛的相关性，并最终使大量患者受益，因为具有抑制和恢复 miRNA 水平的潜力。治疗意图正在成为现实，我们的研究可能为在癌症中测试这一概念提供蓝图。