Project 3: lncRNA SNHG1 and ATG7 in Basal-subtype Muscle-invasive Bladder Tumorigenesis

项目3：lncRNA SNHG1和ATG7在基底亚型肌侵袭性膀胱肿瘤发生中的作用

• 批准号: 10229414
• 负责人: DIANE M SIMEONE
• 金额: $ 33.52万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-12 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10229414
• 关键词: Ablation; Autophagocytosis; BIRC4 gene; Biological; Biological Markers; Biological Process; Bladder; CD44 gene; CRISPR/Cas technology; Carcinogens; Catabolism; Cell model; Cells; Characteristics; Chemicals; Data; Ectopic Expression; Exposure to; Funding; Genes; Goals; Human; In Vitro; Invaded; Knock-in Mouse; Knock-out; Knockout Mice; MMP2 gene; MMP9 gene; Malignant Neoplasms; Malignant neoplasm of urinary bladder; Mediating; Mediator of activation protein; Molecular; Mus; Muscle; Neoplasm Metastasis; Nitrosamines; Pathway interactions; Peroxisome Proliferator-Activated Receptors; Phenotype; Play; Prognostic Marker; Resistance; Resources; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Small Nucleolar RNA; Stratification; System; Testing; Transgenic Mice; Transgenic Organisms; Untranslated RNA; Up-Regulation; Urothelial Cell; Variant; Xenograft Model; base; cancer biomarkers; cancer cell; cancer subtypes; carcinogenesis; effective therapy; in vivo; inhibitor/antagonist; knock-down; molecular subtypes; mortality; muscle invasive bladder cancer; new therapeutic target; non-muscle invasive bladder cancer; novel; overexpression; promoter; small hairpin RNA; tumor progression; tumorigenesis

PROJECT 3: SUMMARY The major goal of this project is to study the molecular mechanisms that underlie the critical steps of bladder cancer (BC) progression: invasion and metastasis. Specifically, we will focus on how autophagy-related gene 7 (ATG7)-mediated autophagy signaling drives BC cell invasion in vitro and metastasis in vivo. During the last funding period, we found that basal-subtype muscle-invasive bladder cancer (MIBC) in mice induced by bladder carcinogen N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) markedly overexpress ATG7 and long non-coding RNA (lncRNA) small nucleolar RNA host gene 1 (SNHG1), and have significantly upregulated autophagy. In stark contrast, knockin mice lacking the RING domain of XIAP, which are completely resistant to BBN-induced basal MIBCs, have markedly reduced autophagy. We also found that, during BBN-mediated bladder tumorigenesis, the RING domain of XIAP is essential for SNHG1 overexpression, and that ectopic expression of SNHG1 in vitro induces autophagy and promotes BC cell invasion accompanied by upregulated ATG7, MMP2 and MMP9. Furthermore, we showed that knockdown of ATG7 strongly inhibits autophagy, abolishes BC cell invasion and reduces the expression of basal MIBC marker KRT14. These data reveal a heretofore unknown role of autophagy in basal MIBC formation. Based on these data, we hypothesize that the upregulation of SNHG1 and ATG7 by the RING domain of XIAP, and the autophagic signaling that these molecules trigger play critical roles in the genesis and progression of basal MIBC. We will test this hypothesis in three Specific Aims. Aim 1 will define the regulatory circuitry in the SNHG1/ATG7/autophagy signaling axis that is operative in basal MIBC in vitro. Aim 2 will determine the biological effects of the SNHG1/ATG7/autophagy signaling on BC cell invasion in vitro and tumorigenesis and metastasis in vivo. Aim 3 will test the hypotheses that overexpression of SNHG1 in basal urothelial cells of transgenic mice promotes basal MIBC formation, and that ablation of ATG7 in these cells of knockout mice renders mice resistant to basal MIBC formation and progression. These complementary approaches will provide definitive evidence regarding the in vivo roles of SNHG1 and ATG7 in the formation and progression of basal MIBC. While invasion and metastasis are the main reasons of the high mortality caused by MIBC, very little is known about the principal molecules or pathways that drive these crucially important biological processes. Our proposed studies that are highly focused on an important, but poorly understood signaling pathway comprising SNHG1/ATG7/autophagy should yield critical information on not only the underlying mechanisms, but also novel prognostic biomarkers to differentiate MIBC subtypes and new druggable targets to treat this aggressive form of BC.

项目3：摘要 该项目的主要目标是研究膀胱关键步骤的分子机制 癌症（BC）进展：入侵和转移。具体而言，我们将重点介绍自噬相关基因7 （ATG7）介导的自噬信号传导在体外驱动BC细胞侵袭，并在体内转移。最后 资助期，我们发现膀胱诱导的小鼠中的基底肌肉肌肉侵入性膀胱癌（MIBC） 致癌物N-丁基-N-（4-羟基丁基）亚硝胺（BBN）明显过表达ATG7和长的非编码 RNA（lncRNA）小核仁RNA宿主基因1（SNHG1），并具有显着上调的自噬。在 鲜明的对比，缺少XIAP的环域的敲击蛋白小鼠完全抵抗BBN诱导的 基底MIBC显着降低了自噬。我们还发现，在BBN介导的膀胱期间 肿瘤发生，XIAP的环域对于SNHG1的过表达至关重要，并且异位表达 SNHG1体外诱导自噬并促进BC细胞侵袭，并伴有上调ATG7，MMP2 和mmp9。此外，我们表明ATG7的敲低强烈抑制自噬，废除了BC细胞 入侵并降低基础MIBC标记KRT14的表达。这些数据揭示了迄今未知 自噬在基础MIBC形成中的作用。基于这些数据，我们假设SNHG1的上调 和XIAP环域的ATG7，以及这些分子触发的自噬信号发出关键 基础MIBC的起源和进展中的作用。我们将以三个具体目标来检验这一假设。目标1 将定义在基底MIBC中可操作的SNHG1/ATG7/自噬信号轴中的调节电路 体外。 AIM 2将确定SNHG1/ATG7/自噬信号对BC细胞入侵的生物学作用 体内体外和肿瘤发生和转移。 AIM 3将检验SNHG1过表达的假设 在转基因小鼠的基底尿路上皮细胞中，促进了基底MIBC的形成，并且在这些中烧蚀了ATG7 基因敲除小鼠的细胞使小鼠具有抗基础MIBC形成和进展的小鼠。这些补充 方法将提供有关SNHG1和ATG7在形成中的体内作用的明确证据 基础MIBC的进展。而入侵和转移是由 MIBC，关于驱动这些至关重要的生物学的主分子或途径知之甚少 过程。我们提出的研究高度集中于重要但知之甚少的信号传导 包括SNHG1/ATG7/自噬的途径不仅应提供有关基础的关键信息 机制，以及将MIBC亚型和新靶标区分开的新型预后生物标志物 治疗这种侵略性的卑诗省。