Neuroendocrine differentiation post anti-androgenic therapy: Role of Tribbles 2

抗雄激素治疗后的神经内分泌分化：Tribbles 2 的作用

• 批准号: 10659525
• 负责人: JAGADANANDA GHOSH
• 金额: $ 34.77万
• 依托单位: HENRY FORD HEALTH SYSTEM
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-20 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10659525
• 关键词: Address; Adenocarcinoma; Age; Androgen Antagonists; Androgen Receptor; Apoptosis; Binding; Bypass; Cancer Cell Growth; Castrate sensitive prostate cancer; Castration; Cell Culture Techniques; Cell Line; Cells; Cessation of life; Characteristics; Chromogranin A; Clinic; Clinical; Development; Distant Metastasis; Dose; EZH2 gene; Environment; Epithelium; Exons; FDA approved; Family; Gene Chips; Gene Expression; Gene Expression Profiling; Gene Targeting; Generations; Goals; Health; Image; Immunoprecipitation; In Vitro; Injections; Kidney; Knock-out; Knockout Mice; Knowledge; LNCaP; Link; Liver; Luciferases; Lung; Malignant neoplasm of prostate; Mass Spectrum Analysis; Measures; Medical; Metastatic to; Modeling; Molecular; Molecular Target; Monitor; Movement; Mus; Neoplasm Metastasis; Neuroendocrine Therapy; Neuron-Specific Enolase; Neurons; Neurosecretory Systems; Pathologic; Patients; Phenotype; Play; Population; Prevention; Prostate; Prostate Neuroendocrine Neoplasm; Prostatic Neoplasms; Proteins; Public Health; Regimen; Regulation; Research; Resistance; Role; SCID Mice; Sampling; Seminal; Synaptophysin; TP53 gene; Testing; Therapeutic; Transgenic Mice; Transgenic Organisms; Up-Regulation; Work; Xenograft procedure; abiraterone; androgenic; bone; cell killing; cell type; clinically relevant; design; effective therapy; enzalutamide; in vivo; in vivo Model; inhibitor; knock-down; knockout gene; male; member; molecular marker; molecular targeted therapies; mouse model; multicatalytic endopeptidase complex; neuroendocrine differentiation; neuroendocrine phenotype; new therapeutic target; novel; overexpression; pre-clinical; profiles in patients; prostate cancer cell; prostate cancer cell line; prostate cancer model; prostate cancer progression; receptor function; therapy development; therapy resistant; transcription factor; transdifferentiation; tumor; tumor growth; tumor progression

Anti-androgenic therapy is the mainstay for both primary and disseminated forms of prostate cancer. FDA- approved enzalutamide (Xtandi) is at the forefront of anti-androgens with superior patient profile and is the one most prescribed. However, enzalutamide resistant prostate cancer (ERPC) invariably develops, which is incurable and responsible for most of the prostate cancer-related deaths, underscoring that management of ERPC is an unmet and urgent medical need. Development of an effective therapy against ERPC is suffering from lack of proper understanding about critical molecular targets to effectively kill ERPC cells. To identify potential new targets, we developed an ERPC model which mimics the clinical conditions in patients undergoing standard enzalutamide therapy. We treated androgen-sensitive prostate cancer cells with enzalutamide in long- term culture to generate cell lines (LNCaP-ENR, PCa-2B-ENR) which are no longer sensitive to clinically relevant doses of enzalutamide. Comprehensive gene expression analysis revealed that the ERPC cells overexpress Tribbles 2 (Trib2), a member of the tribbles pseudokinase family. Overexpression of Trib2 was also found in PDX and patient prostate tumors after enzalutamide treatment. Forced overexpression Trib2 results in enhanced prostate cancer cell growth and resistance to enzalutamide, apalutamide, darolutamide and abiraterone. Inhibition of Trib2 re-sensitizes resistant cells to enzalutamide and decreases their viability, indicating a possible direct link between Trib2 and development of enzalutamide resistance. Interestingly, Trib2 downregulates Rb1 and p53, while induces the neuronal transcription factors (N-Myc, BRN2) and the neuroendocrine (NE) markers (Chromogranin A, Neuron-specific enolase and Synaptophysin). Inhibition of N-Myc or BRN2 re-sensitizes resistant cells to enzalutamide. These findings suggest that Trib2 drives cellular trans-differentiation from luminal to NE phenotype to pose resistance to anti-androgens. Thus, Trib2 emerges as a novel, promising molecular target for therapy of ERPC-NE. However, the mechanism and role of Trib2 in NE differentiation needs to be determined using appropriate in vitro and in vivo models. Thus, this proposal has been designed which will characterize how Trib2 induces NE differentiation (Aim 1), determine the impact of Trib2 inhibition in enhanced ERPC tumor growth and distant metastasis (Aim 2), and test the impact of Trib2 gene-targeting on prostate tumor progression and NE differentiation using transgenic Trib2 knockout mouse models (Aim 3). Accomplishing these goals will establish Trib2 as a molecular driver for treatment-induced NE differentiation and will help develop a new targeted therapeutic strategy for enzalutamide resistant, NE type, lethal prostate cancer.

抗雄激素治疗是前列腺癌原发和播散性治疗的主要手段。FDA- 批准的苯扎鲁胺(Xtandi)是抗雄激素药物的最前沿，具有更好的患者资料，是 大多数是处方药。然而，对苯扎鲁胺耐药的前列腺癌(ERPC)总是发生的，这是 无法治愈，并对大多数前列腺癌相关死亡负有责任，强调了 ERPC是一种未得到满足的紧急医疗需求。针对ERPC的有效治疗方法的开发正在受挫 从缺乏对关键分子靶点的正确理解到有效地杀死ERPC细胞。要确定 潜在的新靶点，我们开发了一种ERPC模型，它模拟了患者在接受 标准的苯扎鲁胺疗法。我们用苯扎鲁胺治疗雄激素敏感的前列腺癌细胞。 长期培养以产生对临床不再敏感的细胞系(LNCaP-ENR，PCA-2B-ENR) 剂量的苯扎鲁胺。综合基因表达分析显示，ERPC细胞过度表达 TRIBBLES 2(Trib2)，属于TRIBBLES假激酶家族的成员。在PDX中也发现了Trib2的过表达 以及接受苯扎鲁胺治疗后的前列腺癌患者。强制过度表达Trib2导致增强 前列腺癌细胞生长及对苯扎鲁胺、阿帕鲁胺、达鲁他胺和阿比特龙的耐药性。 抑制Trib2使耐药细胞对苯扎鲁胺重新敏感，并降低其存活率，表明可能存在 Trib2与苯扎鲁胺耐药性的发生有直接联系。有趣的是，Trib2下调了Rb1的表达 和P53，同时诱导神经转录因子(N-Myc，Brn2)和神经内分泌(NE)标志物 (嗜铬粒素A、神经元特异性烯醇化酶和突触素)。N-Myc或Brn2复敏剂的抑制作用 对苯扎鲁胺耐药的细胞。这些发现表明，Trib2促进了细胞从腔到腔的转分化 转化为NE表型，对抗雄激素产生抵抗。因此，trib2作为一种新颖的、有前途的分子应运而生。 ERPC-NE的治疗靶点。然而，Trib2在NE分化中的机制和作用尚需进一步研究。 采用适当的体外和体内模型测定。因此，这项建议的设计将 鉴定Trib2如何诱导NE分化(目标1)，确定抑制Trib2对增强的 ERPC肿瘤生长和远处转移(AIM 2)，并检测Trib2基因靶向对前列腺癌的影响 利用转基因Trib2基因敲除小鼠模型研究肿瘤进展和NE分化(目标3)。完成 这些目标将使Trib2成为治疗诱导NE分化的分子驱动因素，并将有助于 开发对苯扎鲁胺耐药的NE型致死性前列腺癌的新的靶向治疗策略。