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: 10622314
• 负责人: Jack Tran
• 金额: $ 4.28万
• 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/10622314
• 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 Cells; Epithelium; Event; Frequencies; Future; Genetic Transcription; Goals; Growth; Human; In Vitro; Inhibition of Cell Proliferation; Injections; Invaded; Malignant neoplasm of prostate; Measures; Mediating; Mesenchymal; Messenger RNA; Metastatic Prostate Cancer; Micrometastasis; Modeling; Monitor; Morphology; Mus; NPR2 gene; Nature; Neoplasm Circulating Cells; Neoplasm Metastasis; Organ; Patients; Physiological; Process; Proliferating; Property; Prostate; Proteins; RNA Interference; Receptor Signaling; Regulation; Reporting; Repression; Research; Resistance development; Risk; Role; Sampling; Site; Stains; System; Testing; Therapeutic; Tissue Microarray; Tumor Promotion; 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.

摘要 晚期癌症转移包括癌细胞在远处器官的种植和致命的 种子细胞的生长。2019年，美国几乎所有31,620例前列腺癌(PCA)死亡病例都是 归因于转移。上皮间质转化(EMT)对生理发育具有重要意义； 然而，EMT促进多种癌症类型的迁移、侵袭和播种。相反，EMT抑制了 增殖和肿瘤生长。Snai1和Twist1是促进癌细胞种植的主要EMT驱动因素 并在非前列腺癌中抑制细胞增殖和肿瘤生长。Snai1在侵袭性原发肿瘤中的表达 和转移性前列腺癌。相反，雄激素受体(AR)是前列腺癌细胞增殖和肿瘤的主要驱动力 外延生长。值得注意的是，AR激活了Cyclin D1(细胞周期驱动因子)的转录，但SNAI1抑制了转录。两个都绑定 CCND1(Cyclin D1)近端启动子的重叠区域，提示直接竞争调控 细胞周期蛋白D1转录。鉴于外延生长发生在转移级联的末端，很可能会发生EMT。 一过性的，在PCa患者的早期。事实上，波形蛋白(EMT的标志物)在PCa循环肿瘤中高表达 然而，一旦播种的PCa细胞成长为可检测到的转移瘤，波形蛋白的表达就会降低。而EMT 以及AR信号在前列腺癌中的独立研究，EMT和AR如何协调前列腺癌的转移 目前仍不清楚。Teton-SNAI1表达系统在临床前列腺癌中的应用 从而可以用多西环素开启/关闭SNAI1的瞬时表达。初步数据 显示SNAI1诱导PCa细胞间充质形态，抑制细胞生长，并上调 EMT标志物如Vimentin和NPR2的mRNA。然而，AR信号抑制了SNAI1的S能力 上调Vimentin和NRP2，下调Twist1mRNA，不依赖于SNAI1的表达。总而言之， 这些数据表明，AR和SNAI1在调节EMT和增殖方面具有相反的作用；并且 与在PCa转移过程中发生的晚期事件序列一致：即，播种之后是 外延生长。因此，我假设SNAI1介导了一个抗增殖的EMT样侵袭程序，以 促进PCA播种；随后，AR拮抗SNAI1介导的细胞周期蛋白D1抑制恢复 增殖并促进转移性生长。以下目标旨在解决中央 假设。目的：研究AR如何影响SNAI1促进前列腺癌侵袭和转移的S能力。 目的：研究SNAI1和AR对PCa增殖的调控作用。AIM3：确定AR和SNAI1协同作用的程度 PCa临床标本的表达和活性。侵袭性前列腺癌患者接受标准的抗AR- 信号疗法；但将在2年内产生抗药性。考虑到可供选择的治疗方法有限 这组患者，对SNAI1和AR的进一步研究将有助于开发更具临床意义的研究模型 准确，并为未来的治疗策略提供更深入的见解。