LIN28 and RNA Uridylation in Breast Cancer: Mouse Models and Molecular Analyses

乳腺癌中的 LIN28 和 RNA 尿苷化：小鼠模型和分子分析

• 批准号: 9398268
• 负责人: John Patrick Hagan
• 金额: $ 36.07万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9398268
• 关键词: Address; Adult; Adverse effects; Affect; Animals; Antineoplastic Agents; Biogenesis; Breast; Breast Cancer Model; Breast Cancer therapy; Cancer Etiology; Cancer Patient; Cancer Prognosis; Carcinoma; Cells; Cellular biology; Colon Adenocarcinoma; Data; Disease; Disease Progression; Drug Design; E2F1 gene; ERBB2 gene; Family; Fertility; Foundations; Gene Expression Regulation; Generations; Genes; Genetic Transcription; Genetically Engineered Mouse; Goals; Growth; Human; Immunocompetent; Investigation; Ionizing radiation; Knock-out; Knockout Mice; Malignant Neoplasms; Mammals; Mammary Neoplasms; Mammary gland; Mediating; Messenger RNA; Metabolism; MicroRNAs; Modeling; Molecular Analysis; Molecular Target; Morbidity - disease rate; Multiple Myeloma; Mus; Non-Insulin-Dependent Diabetes Mellitus; Oncogenes; Oncogenic; Organism; Pathology; Pathway interactions; Patients; Perlman syndrome; Phenocopy; Phenotype; Publishing; RNA; RNA-Binding Proteins; Recurrence; Regulator Genes; Reporting; Research; Resistance; Resources; Role; Signal Transduction; Tamoxifen; Testing; Therapeutic; Transcript; Transferase; Treatment Efficacy; Tumor Subtype; Tumor Suppressor Proteins; Validation; Warburg Effect; Work; cancer cell; cancer therapy; cancer type; chemotherapy; embryonic stem cell; field study; gene discovery; genome-wide analysis; glucose metabolism; human disease; improved; induced pluripotent stem cell; inhibitor/antagonist; innovation; malignant breast neoplasm; mortality; mouse model; new therapeutic target; novel; novel drug class; outcome forecast; overexpression; polyadenylated messenger RNA; pre-clinical; prevent; restoration; stem cell biology; tissue repair; triple-negative invasive breast carcinoma; tumor; tumor growth; tumor progression; tumor xenograft; tumorigenesis; uridylate

PROJECT SUMMARY / ABSTRACT A major limitation in finding more effective cancer therapies is the identification and exploitation of novel targets for rational drug design. Recent and provocative evidence implicates a novel form of post-transcriptional gene regulation involving 3’ RNA uridylation mediated by Terminal Uridylyl Transferases (TUTases) as a critical gene regulator and driver of tumorigenesis. These data indicate that dysregulated TUTase activity alone and in concert with the onco-fetal LIN28/let-7 pathway are hallmarks of poor prognosis in multiple cancer types. Approximately 15% of all cancers reactivate oncogenic LIN28 expression (either LIN28A or LIN28B) and these cancers are typically characterized by poor prognosis. Functionally, LIN28 directly blocks the biogenesis of the tumor suppressor let-7 microRNA family. Recently, we discovered the mechanism behind this blockade. Notably, the TUTase ZCCHC11 through an association with LIN28A inactivates let-7 by preventing the conversion of precursor let-7 into its mature, tumor suppressor form. In addition, overexpression of the TUTase ZCCHC11 is implicated in poor prognosis breast cancer independently of the LIN28/let-7 pathway. Strikingly, a recent report has shown that many polyadenylated mRNAs are frequently uridylated at their 3’ ends in a gene-specific manner and targeted for degradation; however, this observation’s relevance to cancer is currently unknown. Therefore, our research has three major objectives. The first goal is to develop novel preclinical mouse models for LIN28-positive breast cancers. These mouse models will elucidate how LIN28 modifies disease progression and will be an invaluable resource to evaluate new drugs targeted to this cancer type. The second goal is to determine whether organism-wide loss of Zcchc11 in adult mice has phenotypic consequences using a conditional mouse knockout. Our published work on conditional inactivation of either Lin28A or Lin28B in adult mice demonstrate that both genes are dispensable in adults, suggesting that inhibiting either gene is unlikely to cause deleterious side effects in patients. The last goal is to determine if loss of the TUTase Zcchc11 inhibits Lin28A+/Her2+ and Lin28A-/Her2+ mouse mammary tumors. This research will be the first proof-of-principle that TUTase loss affects tumors in an immunocompetent mammal and addresses possible side-effects in cancer patients treated with specific inhibitors of the TUTase ZCCHC11. This work lays the foundation for new fields of cancer investigations by dramatically shifting the current paradigm to include TUTases as cancer gene regulators and explores the feasibility of a novel drug class targeted at inhibition of LIN28 or TUTases. In addition to cancer, our investigation of regulators of the LIN28/let-7 pathway holds significance for stem cell biology, tissue repair, fertility, growth, glucose metabolism, Type 2 diabetes, and the generation of induced pluripotent stem cells. Moreover, our genome-wide analysis of 3’ RNA uridylation has implications for other human diseases where dysfunctional RNA turnover contributes to pathology such as multiple myeloma and Perlman syndrome.

项目摘要/摘要 寻找更有效的癌症治疗方法的一个主要限制是识别和开发新的靶点 合理的药物设计。最新的和具有挑衅性的证据暗示了一种新的转录后基因调控形式 参与末端尿酰转移酶(TUTase)介导的3‘RNA尿苷基化作为关键基因调节因子 肿瘤发生的驱动力。这些数据表明，异常的TUTase活性单独和与肿瘤胎儿 LIN28/let-7通路是多种肿瘤预后不良的标志。在所有癌症中约有15% 重新激活致癌的LIN28表达(Lin28a或Lin28b)，这些癌症的典型特征是 预后。在功能上，LIN28直接阻断肿瘤抑制基因let-7microRNA家族的生物发生。最近， 我们发现了这种封锁背后的机制。值得注意的是，通过与Lin28a的关联，TUTase ZCCHC11 通过阻止前体let-7转化为其成熟的肿瘤抑制形式来使let-7失活。此外, TUTase ZCCHC11的过度表达独立于LIN28/let-7与乳腺癌预后不良有关 路径。令人惊讶的是，最近的一份报告显示，许多多腺化的mRNAs的3‘端经常是尿苷化的 以基因特有的方式并以降解为目标；然而，这一观察结果与癌症的相关性目前 未知。因此，我们的研究有三大目标。第一个目标是开发新的临床前小鼠模型 用于LIN28阳性乳腺癌。这些小鼠模型将阐明LIN28如何改变疾病的进展和将 是评估针对这种癌症类型的新药的宝贵资源。第二个目标是确定是否 在成年小鼠中，Zcchc11在整个生物体中的丢失在有条件的小鼠基因敲除中具有表型后果。我们的 在成年小鼠中发表的关于Lin28a或Lin28b条件失活的研究表明，这两个基因都是 对成年人来说是可有可无的，这表明抑制这两种基因中的任何一种不太可能对患者造成有害的副作用。这个 最后一个目标是确定TUTase Zcchc11的缺失是否抑制了Lin28a+/Her2+和Lin28a-/Her2+小鼠的乳房 肿瘤。这项研究将是TUTase丢失影响免疫活性肿瘤的第一个原理证明 并解决了使用TUTase ZCCHC11的特定抑制剂治疗癌症患者可能出现的副作用。 这项工作为癌症研究的新领域奠定了基础，将当前的范式戏剧性地转变为 包括TUTase作为癌症基因调节因子，并探索一种新的药物类别的可行性 LIN28或TUTASES。除了癌症，我们对LIN28/let-7通路调节因子的研究对 干细胞生物学，组织修复，生育，生长，葡萄糖代谢，2型糖尿病，以及诱导的 多能干细胞。此外，我们对3‘RNA尿酸化的全基因组分析对其他人类也有意义 如多发性骨髓瘤和帕尔曼综合征等导致RNA转换功能障碍的疾病。