Coordinated Actions of SNAI1 and AR in Prostate Cancer Dissemination and Metastatic Outgrowth

SNAI1 和 AR 在前列腺癌播散和转移生长中的协调作用

• 批准号: 10387135
• 负责人: Jack Tran
• 金额: $ 4.12万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10387135
• 关键词: Address; Androgen Antagonists; Androgen Receptor; Binding; Biological Assay; Breast Cancer Model; CCND1 gene; Cancer Patient; Cell Count; Cell Cycle; Cell Proliferation; Cells; Cessation of life; Clinical; Cyclin D1; Data; Development; Distant; Down-Regulation; Doxycycline; Elements; Epithelial; Epithelial Cells; Event; Frequencies; Future; Genetic Transcription; Goals; Growth; Human; In Vitro; Injections; Lead; Malignant neoplasm of prostate; Measures; Mediating; Mesenchymal; Messenger RNA; Metastatic Prostate Cancer; Modeling; Monitor; Morphology; Mus; NPR2 gene; Nature; Neoplasm Circulating Cells; Neoplasm Metastasis; Organ; Patients; Physiological; Process; Property; Prostate; Proteins; RNA Interference; Receptor Signaling; Regulation; Reporting; Repression; Research; Resistance development; Risk; Role; Sampling; Site; Stains; System; Testing; Therapeutic; Tissue Microarray; United States; Vimentin; anticancer research; cancer cell; cancer seeding; cancer type; cell growth; chromatin immunoprecipitation; deep learning algorithm; design; in vivo; innovation; insight; knock-down; migration; neoplastic cell; overexpression; prevent; programs; promoter; prostate cancer cell; prostate cancer metastasis; prostate cancer progression; receptor binding; tumor; virtual; wound healing

SUMMARY Late stages of cancer metastasis involves the seeding of cancer cells at distant organs and the lethal outgrowth of seeded cells. In 2019, virtually all 31,620 prostate cancer (PCa) deaths in the United States were attributed to metastasis. Epithelial Mesenchymal Transition (EMT) is important for physiological development; however, EMT promotes migration, invasion, and seeding in multiple cancer types. In contrast, EMT inhibits proliferation and tumor outgrowth. SNAI1 and Twist1 are major EMT drivers that promote cancer cell seeding and inhibit cell proliferation and tumor outgrowth, in non-PCa. SNAI1 expression is detected in invasive primary and metastatic PCa. In contrast, the androgen receptor (AR) is a major driver of PCa cell proliferation and tumor outgrowth. Of note, AR activates, but SNAI1 represses, transcription of Cyclin D1 (a cell cycle driver). Both bind to an overlapping region of the ccnd1 (Cyclin D1) proximal promoter, suggesting direct competitive regulation of Cyclin D1 transcription. Given that outgrowth occurs at the end of the metastasis cascade, EMT is likely to occur transiently, early in PCa patients. In fact, vimentin (a marker of EMT) is highly expressed in PCa circulating tumor cells; however, vimentin is lowly expressed once seeded PCa cells grow into detectable metastases. While EMT and AR signaling have been independently investigated in PCa, how EMT and AR coordinate PCa metastasis remains unclear. Adhering to the transient nature of EMT in clinical PCa, a TetON-SNAI1 expression system was created, whereby transient expression of SNAI1 can be turned ON/OFF with doxycycline. Preliminary data shows that SNAI1 induces a mesenchymal morphology in PCa cells, suppresses cell growth, and upregulates mRNA of EMT markers such as vimentin and NPR2. However, AR signaling suppresses SNAI1’s ability to upregulate vimentin and NRP2 and downregulates Twist1 mRNA, independent of SNAI1 expression. Altogether, these data suggest that AR and SNAI1 have opposing roles in regulating EMT and proliferation; and are consistent with the late sequence of events that occur during PCa metastasis: i.e., seeding followed by outgrowth. Therefore, I hypothesize that SNAI1 mediates an anti-proliferative EMT-like invasion program to promote PCa seeding; subsequently, AR antagonizes SNAI1-mediated cyclin D1 repression to restore proliferation and promote metastatic outgrowth. The following aims are designed to address the central hypothesis. Aim1: Determine how AR impacts SNAI1’s ability to promote PCa invasion and metastatic seeding. Aim2: Determine how SNAI1 and AR regulate PCa proliferation. Aim3: Determine extent of AR and SNAI1 co- expression and activity in clinical PCa samples. Patients with aggressive PCa are treated with standard anti AR- signaling therapies; but will develop resistance within 2 years. Given the limited treatment options available for this group of patients, further study of SNAI1 and AR will help develop research models that are more clinically accurate and provide deeper insights for future therapeutic strategies.

总结 癌症转移的晚期阶段涉及癌细胞在远处器官的种植和致命的转移。 种子细胞的生长。2019年，美国几乎所有31，620例前列腺癌（PCa）死亡病例均为 归因于转移。上皮间充质转化（EMT）是重要的生理发育; 然而，EMT促进多种癌症类型的迁移、侵袭和播种。相反，EMT抑制 增殖和肿瘤生长。SNAI 1和Twist 1是促进癌细胞接种的主要EMT驱动因素 并抑制非PCa细胞增殖和肿瘤生长。SNAI 1表达在侵袭性原发性 和转移性前列腺癌相比之下，雄激素受体（AR）是PCa细胞增殖和肿瘤发生的主要驱动因素。 结果值得注意的是，AR激活，但SNAI 1抑制，细胞周期蛋白D1（细胞周期驱动程序）的转录。均结合 ccnd 1（细胞周期蛋白D1）近端启动子的重叠区域，表明直接竞争性调节 细胞周期蛋白D1转录。考虑到肿瘤生长发生在转移级联反应的末端， 在PCa患者中为一过性早期。事实上，波形蛋白（EMT的标志物）在PCa循环肿瘤中高度表达。 然而，一旦接种的PCa细胞生长成可检测的转移灶，波形蛋白就低表达。当EMT 和AR信号已经在PCa中独立研究，EMT和AR如何协调PCa转移 仍不清楚坚持临床PCa中EMT的短暂性，TetON-SNAI 1表达系统 由此可以用多西环素打开/关闭SNAI 1的瞬时表达。初步数据 显示SNAI 1诱导PCa细胞中的间充质形态，抑制细胞生长，并上调 EMT标志物如波形蛋白和NPR 2的mRNA。然而，AR信号抑制SNAI 1的能力， 上调波形蛋白和NRP 2，下调Twist 1 mRNA，与SNAI 1表达无关。总的来说， 这些数据表明AR和SNAI 1在调节EMT和增殖中具有相反的作用; 与PCa转移期间发生的晚期事件序列一致：即，播种后， 结果因此，我假设SNAI 1介导了一个抗增殖的EMT样侵袭程序， 促进PCa播种;随后，AR拮抗SNAI 1介导的细胞周期蛋白D1抑制，以恢复 增殖并促进转移性生长。以下目标旨在解决 假说.目的1：确定AR如何影响SNAI 1促进PCa侵袭和转移播种的能力。 目的2：确定SNAI 1和AR如何调节PCa增殖。目的3：确定AR和SNAI 1共同作用的程度。 表达和活性。患有侵袭性PCa的患者用标准抗AR治疗， 信号疗法;但将在2年内产生耐药性。鉴于治疗选择有限， 对于这组患者，SNAI 1和AR的进一步研究将有助于开发更临床化的研究模型。 准确，并为未来的治疗策略提供更深入的见解。