Regulation of the LIN28/let-7 pathway in cancer

癌症中 LIN28/let-7 通路的调节

• 批准号: 9215116
• 负责人: Richard I. Gregory
• 金额: $ 40.49万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-12-19 至 2021-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9215116
• 关键词: Address; Affinity; Apoptosis; Beckwith-Wiedemann Syndrome; Binding; Binding Sites; Biochemical; Biogenesis; Biological Assay; Bladder; Breast; CRISPR/Cas technology; Cancer cell line; Cell Line; Cell Proliferation; Cell physiology; Cloning; Complementary DNA; Complex; Data; Development; Disease; Embryo; Enzymes; Exonuclease; Gene Expression; Gene Targeting; Genetic; Germ-Line Mutation; H19 RNA; H19 gene; Hepatoblastoma; Human; Lead; Link; Lung; Malignant Childhood Neoplasm; Malignant Neoplasms; Maps; Measures; Mediating; Messenger RNA; Methods; MicroRNAs; Microprocessor; Molecular; Monitor; Mus; Mutation; Nephroblastoma; Neuroblastoma; Oncogenes; Oncogenic; Pathway interactions; Patients; Pediatric Neoplasm; Perlman syndrome; Porifera; Precursor RNA; Predisposition; Proteins; RNA; RNA Decay; RNA Degradation; RNA Interference; RNA Precursors; RNA-Binding Proteins; Recruitment Activity; Regulation; Risk; Role; Sampling; Somatic Mutation; TYRP1 gene; Testing; Transferase; Tumor Suppressor Genes; Tumor Suppressor Proteins; Tumorigenicity; Untranslated RNA; Xenograft procedure; cancer cell; cell growth; embryonic stem cell; experimental study; fetal; gain of function; in vitro Assay; insight; knock-down; loss of function; metaplastic cell transformation; miRNA expression profiling; migration; mouse model; novel; novel strategies; novel therapeutics; overexpression; protein B; protein expression; stoichiometry; transcriptome; transcriptome sequencing; tumor; tumorigenesis

Regulation of the LIN28/let-7 pathway in cancer The RNA-binding proteins LIN28A/B control the biogenesis of let-7 tumor suppressor miRNAs. Reactivation of this embryonic pathway is sufficient to promote cellular transformation and drive tumorigenesis in mouse models. Notably, LIN28B functions as an oncogene in neuroblastoma, Wilms tumors (nephroblastoma), hepatoblastoma, and others. At the molecular level, LIN28A recruits the Terminal Uridyl Transferases (TUTases), ZCCHC6/11, to let-7 precursor RNAs targeting them for destruction by the DIS3L2 exonuclease. Germline DIS3L2 mutations cause Perlman syndrome - a rare developmental and cancer predisposition disorder with ~6,000-fold elevated risk of Wilms tumorigenesis. Somatic mutations in DIS3L2 have also been found in ~30% sporadic Wilms tumors. Despite this genetic evidence that DIS3L2 functions as a tumor suppressor its molecular and cellular function remains unaddressed. Similarly, the Beckwith-Wiedemann syndrome (BWS) of fetal overgrowth is associated with ~600-fold elevated risk of developing Wilms tumors, yet the molecular and cellular function of the H19 long-non-coding RNA, a likely tumor suppressor gene in BWS, remains unknown. Considering the strong links between miRNA, LIN28, DIS3L2, and Wilms as well as preliminary data linking H19 to the LIN28/let-7 pathway, leads us to hypothesize that dysregulation of let-7 biogenesis is a unifying mechanism in Wilms tumorigenesis. Loss-of-function studies will be performed to address the possible redundant role of ZCCHC6/11 in LIN28B regulation of let-7, and biochemical approaches will be employed to elucidate the mechanism by which LIN28B suppresses let-7 in human cancer cells. Relative levels of let-7, H19, LIN28A/B, and DIS3L2 will be measured in a panel of cell lines and primary human Wilms tumors and RNA sequencing analysis will be performed on a subset. H19- and DIS3L2 loss- and gain-of-function experiments will be performed in human cancer cells and the effects on gene expression, cell growth, and tumorigenicity will be measured. Novel approaches will be applied to identify new DIS3L2 target RNAs. This will provide important mechanistic insight into the LIN28/let-7 pathway in human cancer cells, uncover the tumor suppressive role of H19 in pediatric cancer, and highlight the relevance of RNA decay mechanisms in cancer by elucidating DIS3L2 tumor suppressive function.

癌症中LIN 28/let-7通路的调节 RNA结合蛋白LIN 28 A/B控制let-7肿瘤抑制miRNA的生物发生。重新启动 这种胚胎途径足以促进小鼠的细胞转化并驱动肿瘤发生 模型值得注意的是，LIN 28 B在神经母细胞瘤、肾母细胞瘤（肾母细胞瘤）、 肝母细胞瘤等。在分子水平上，LIN 28 A募集末端尿苷基转移酶 （TUTases），ZCCHC 6/11，以使靶向它们的let-7前体RNA被DIS 3L 2核酸外切酶破坏。 生殖系DIS 3L 2突变导致Perlman综合征-一种罕见的发育和癌症易感性 肾母细胞瘤（Wilms tumor genesis）风险增加约6,000倍。DIS 3L 2中的体细胞突变也已被发现。 在约30%的散发性肾母细胞瘤中发现。尽管有遗传证据表明DIS 3L 2作为肿瘤发挥作用， 其分子和细胞功能仍然没有得到解决。类似地，Beckwith-Wiedemann 胎儿过度生长综合征（BWS）与发生肾母细胞瘤的风险增加约600倍有关， H19长链非编码RNA的分子和细胞功能，可能是BWS中的肿瘤抑制基因， 仍然未知。考虑到miRNA、LIN 28、DIS 3L 2和Wilms之间的紧密联系， H19与LIN 28/let-7通路相关的初步数据，使我们假设let-7的失调 生物发生是肾母细胞瘤发生的统一机制。将进行功能丧失研究， 阐明ZCCHC 6/11在let-7的LIN 28 B调节中可能的冗余作用，以及生物化学方法 将用于阐明LIN 28 B抑制人癌细胞中let-7的机制。 将在一组细胞系和原代细胞系中测量let-7、H19、LIN 28 A/B和DIS 3L 2的相对水平。 人肾母细胞瘤和RNA测序分析将在一个子集上进行。H19-和DIS 3L 2损失-和 将在人癌细胞中进行功能获得实验，并研究对基因表达、细胞增殖和细胞凋亡的影响。 生长和致瘤性。新的方法将被应用于识别新的DIS 3L 2靶标 RNA。这将为人类癌细胞中的LIN 28/let-7通路提供重要的机制见解， 揭示H19在儿科癌症中的肿瘤抑制作用，并强调RNA衰变的相关性 通过阐明DIS 3L 2肿瘤抑制功能来阐明癌症中的机制。