Investigating the role of epitranscriptomic A-to-I RNA editing in T-cell acute lymphoblastic leukemia

研究表观转录组 A-to-I RNA 编辑在 T 细胞急性淋巴细胞白血病中的作用

• 批准号: 10220900
• 负责人: Qingfei Jiang
• 金额: $ 19.12万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10220900
• 关键词: 14 year old; Ablation; Acute Lymphocytic Leukemia; Acute T Cell Leukemia; Address; Adenosine; Adult; Adult Precursor T Lymphoblastic Leukemia; Age; Apoptosis; B-Cell Development; Basic Science; Biological Assay; Biology; Bone Marrow; CD34 gene; Cancer Patient; Cell Maintenance; Cell Survival; Cell physiology; Cells; Characteristics; Childhood; Clinical; Data; Deaminase; Detection; Development; Development Plans; Diagnostic; Disease; Disease Progression; Drug resistance; Engraftment; Event; Future; Gene Expression; Gene Expression Profile; Generations; Genes; Goals; Grant; Hematologic Neoplasms; Human; IL7 gene; Immunocompromised Host; Impairment; Inflammation; Inflammatory; Inosine; Investigation; JAK2 gene; Janus kinase; Maintenance; Malignant - descriptor; Malignant Childhood Neoplasm; Malignant Neoplasms; Malignant lymphoid neoplasm; Mediating; Medical; Messenger RNA; MicroRNAs; Modeling; Mus; Mutation; NOTCH1 gene; Notch Signaling Pathway; Oncogenic; Pathway interactions; Patients; Peripheral; Pharmacology; Phenotype; Population; RNA; RNA Editing; RNA I; Regulatory Pathway; Relapse; Research; Research Ethics; Resistance; Risk; Role; Sampling; Signal Pathway; Signal Transduction; Specimen; System; T-Lymphocyte; Testing; Therapeutic; Training; Transcript; Translational Research; Umbilical Cord Blood; Validation; Xenograft Model; Xenograft procedure; adenosine deaminase; base; biobank; cancer stem cell; cancer type; career development; cell transformation; chemotherapy; cytokine; data management; epitranscriptomics; gamma secretase; human model; in vivo; inhibitor/antagonist; knock-down; leadership development; leukemia initiating cell; mouse model; mutant; new therapeutic target; notch protein; novel; novel therapeutics; overexpression; premalignant; prevent; progenitor; self-renewal; small hairpin RNA; stem cell genes; stem cells; therapeutic target; therapy development; therapy resistant; tool; transcriptome; transcriptome sequencing

Project Summary The ultimate goal of this proposal is to address a compelling unmet medical need to identify novel epitranscriptomic mechanisms of oncogenic transformation that will guide development of diagnostic and therapeutic strategies capable of predicting and preventing progression of T-cell acute lymphoblastic leukemia (T-ALL). Acute lymphoblastic leukemia (ALL) is the most prevalent hematological cancer in children younger than 14 years of age. Despite progress in intensive chemotherapy, 20-25% of pediatric and over 50% of adult patients show resistance to therapy and relapse. Widespread aberrant epitranscriptomic ADAR1-mediated adenosine-to-inosine (A-to-I) RNA editing has been associated with clinical characteristics of several cancer types and generation of leukemia initiating cells (LICs) with enhanced pro-survival and self-renewal capacity. Interestingly, activation of janus kinase (JAK)/STAT signaling by interleukin-7 (IL-7) drives expression of key stem cell regulatory pathway ADAR1-LIN28 and pro-survival factors in hematological malignancies including T-ALL. Our central hypothesis is that mutational pro-inflammatory signals, such as NOTCH and JAK/STAT signaling pathway, induce aberrant RNA editing driven by ADAR1 activation in T-ALL-initiating cells that accentuated by enhanced survival and self-renewal capacity. The three specific aims will be (1) examine if the ADAR1-mediated RNA editing promotes oncogenic transformation of normal T cell progenitor to T-ALL LICs by enhancing T-ALL LIC survival and self-renewal pathways; (2); examine whether ADAR1 activity is enhanced in T-ALL LIC cells due to NOTCH or JAK/STAT signaling pathway activation, and lastly (3) determine whether direct inhibition of ADAR1 activity by shRNA strategy impairs the survival and self-renewal impairs T-ALL LIC maintenance. This proposal will utilize established biorepository of primary T-ALL patient specimen, along with age-matched healthy control samples. In addition, lentiviral-based tools and established robust in vivo human xenograft T-ALL mouse models will be used for selective ablation of ADAR1 transcripts to thoroughly investigate the relationship between ADAR1 expression and generation of drug-resistant T- ALL-initiating LIC cells. Detailed functional and mechanistic lentiviral-directed transcript overexpression and knockdown studies will validate sensitive whole transcriptome RNA-Sequencing that correlate ADAR1 expression with LIC survival and self-renewal signaling pathway and stem cell gene expression signatures. This research strategy is uniquely integrated into my career development plan, including additional trainings in data management, basic and translational research, research ethics, and professional and leadership development. By providing a more mechanistic understanding of the role of ADAR1 in cancer, this grant will inform future RNA editase detection and inhibition strategies that may help to obviate cancer resistance and relapse.

项目摘要 该提案的最终目标是解决一个迫切的未满足的医疗需求，以确定新的 致癌转化的表观转录组学机制，将指导诊断和 能够预测和预防T细胞急性淋巴细胞性白血病进展的治疗策略 白血病（T-ALL）。急性淋巴细胞白血病（ALL）是世界上最常见的血液系统癌症。 14岁以下的儿童。尽管强化化疗取得了进展，但20-25%的儿童 超过50%的成年患者表现出对治疗的抵抗和复发。广泛性畸变 ADAR 1介导的腺苷-肌苷（A-to-I）RNA编辑与 几种癌症类型的临床特征和白血病起始细胞（LIC）的产生， 增强有利于生存和自我更新的能力。有趣的是，Janus激酶（JAK）/STAT的激活 通过白细胞介素-7（IL-7）的信号传导驱动关键干细胞调节途径ADAR 1-LIN 28的表达， 包括T-ALL在内的血液恶性肿瘤中的促生存因子。我们的核心假设是， 突变的促炎信号，如NOTCH和JAK/STAT信号通路，诱导异常的炎症反应。 在T-ALL起始细胞中由ADAR 1激活驱动的RNA编辑，通过增强存活率而加剧 自我更新的能力。本研究的三个具体目标是：（1）检测ADAR 1介导的RNA编辑是否 通过增强T-ALL LIC促进正常T细胞祖细胞向T-ALL LIC的致癌转化 生存和自我更新途径;（2）;检查T-ALL LIC细胞中ADAR 1活性是否增强 由于NOTCH或JAK/STAT信号通路激活，最后（3）确定是否直接 通过shRNA策略抑制ADAR 1活性损害存活和自我更新损害T-ALL LIC 上维护本提案将利用已建立的原代T-ALL患者标本生物储存库， 沿着与年龄匹配的健康对照样品。此外，基于慢病毒的工具和已建立的稳健的 体内人异种移植T-ALL小鼠模型将用于选择性消除ADAR 1转录物， 深入研究ADAR 1表达与耐药T细胞产生的关系， ALL启动LIC细胞。详细的功能性和机制性慢病毒定向转录物过表达 敲除研究将验证敏感的全转录组RNA测序， ADAR 1表达与LIC生存和自我更新信号通路及干细胞基因表达 签名.这一研究策略被独特地融入了我的职业发展计划，包括 在数据管理、基础和转化研究、研究伦理方面的额外培训，以及 专业和领导力发展。通过提供一个更机械的理解的作用， ADAR 1在癌症中的作用，这项资助将为未来的RNA编辑酶检测和抑制策略提供信息， 有助于抑制癌症抵抗和复发。