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

基本信息

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
项目类别：
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

项目摘要

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：总结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：侵袭和转移。具体来说，我们将重点关注自噬相关基因7如何 (ATG7) 介导的自噬信号驱动 BC 细胞体外侵袭和体内转移。最后期间资助期间，我们发现膀胱诱发小鼠基底亚型肌层浸润性膀胱癌（MIBC）致癌物 N-丁基-N-(4-羟丁基)亚硝胺 (BBN) 显着过度表达 ATG7 和长非编码 RNA (lncRNA) 小核仁 RNA 宿主基因 1 (SNHG1)，并且具有显着上调的自噬。在形成鲜明对比的是，缺乏 XIAP RING 结构域的敲入小鼠，它们对 BBN 诱导的完全抵抗基础 MIBC 的自噬显着减少。我们还发现，在 BBN 介导的膀胱在肿瘤发生过程中，XIAP 的 RING 结构域对于 SNHG1 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，同时上调 ATG7、MMP2 和MMP9。此外，我们发现 ATG7 的敲低会强烈抑制自噬，消除 BC 细胞侵袭并降低基础 MIBC 标记物 KRT14 的表达。这些数据揭示了一个迄今为止未知的自噬在基础 MIBC 形成中的作用。基于这些数据，我们假设 SNHG1 的上调和 ATG7 由 XIAP 的 RING 结构域组成，这些分子触发的自噬信号发挥着至关重要的作用基础 MIBC 发生和进展中的作用。我们将在三个具体目标中检验这一假设。目标1 将定义在基底 MIBC 中起作用的 SNHG1/ATG7/自噬信号轴中的调节电路体外。目标 2 将确定 SNHG1/ATG7/自噬信号传导对 BC 细胞侵袭的生物学效应体外和体内肿瘤发生和转移。目标 3 将检验 SNHG1 过度表达的假设转基因小鼠基底尿路上皮细胞中的 ATG7 促进基底 MIBC 形成，并且这些细胞中 ATG7 的消除基因敲除小鼠的细胞使小鼠能够抵抗基础 MIBC 的形成和进展。这些互补的方法将提供关于 SNHG1 和 ATG7 在形成和形成中的体内作用的明确证据。基础 MIBC 的进展。而侵袭和转移是造成高死亡率的主要原因 MIBC，对于驱动这些极其重要的生物的主要分子或途径知之甚少。流程。我们提出的研究高度关注重要但知之甚少的信号传导包含 SNHG1/ATG7/自噬的通路不仅应该产生关于潜在的关键信息机制，以及区分 MIBC 亚型的新预后生物标志物和新的药物靶标治疗这种侵袭性的 BC。