权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

The role for alcohol-induced Golgi disorganization in the progression of prostate cancer

酒精引起的高尔基体紊乱在前列腺癌进展中的作用

基本信息

批准号：
10223172
负责人：
Armen Petrosyan
金额：
$ 34.31万
依托单位：
UNIVERSITY OF NEBRASKA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-02 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10223172
关键词：
ATF6 gene Ablation Address Alcohol abuse Alcohol consumption Alcoholism Alcohols Androgen Receptor Animal Model Attenuated Biology Cancer Etiology Cancer Patient Carcinogenesis Mechanism Case-Control Studies Cell Survival Cell surface Cells Cessation of life Chronic Cleaved cell Clinical Coat Protein Complex I Consensus Cytoplasm Data Development Diagnosis Dimerization Docking Endoplasmic Reticulum Enzymes Ethanol Event Glycogen Synthase Kinases Goals Golgi Apparatus Growth HDAC6 gene Heat-Shock Proteins 90 Heavy Drinking Hepatocyte Impairment In Vitro Incidence Integrins LNCaP Lead Light Link Literature Malignant Neoplasms Malignant neoplasm of prostate Mediating Molecular Motor Mus Neoplasm Metastasis Nonmuscle Myosin Type IIA Oncogenic Organ Outcome Study Pathway interactions Peptide Hydrolases Phenotype Phosphorylation Phosphotransferases Physiological Polysaccharides Population Study Prostate Prostatic Neoplasms Proteins Reporting Research Risk Factors Role Sampling Signal Pathway Stress Testing Therapeutic Therapeutic Intervention Transactivation Tumor Promotion United States Vesicle Xenograft procedure advanced prostate cancer alcohol abstinence alcohol abuse therapy alcohol effect alcohol prevention androgen sensitive base biological adaptation to stress carcinogenesis castration resistant prostate cancer chronic alcohol ingestion endoplasmic reticulum stress epidemiology study experimental study glycosylation glycosyltransferase in vivo innovation insight knock-down macrogolgin matriptase men migration misfolded protein mortality non-muscle myosin novel overexpression prevent prostate cancer cell prostate cancer cell line prostate cancer progression protein transport response sensor tumor tumor initiation tumor progression

项目摘要

ABSTRACT Chronic alcohol abuse and alcoholism are considered risk factors for prostate cancer (PCa) progression, but the mechanism is unknown. The current project will address an important question raised by clinicians: whether alcohol abstinence is an important therapeutic intervention in PCa. Previously, we found that: (1) fragmentation of the Golgi complex correlates with the progression of PCa; (2) ethanol (EtOH) induces Golgi disorganization that is caused by the impaired dimerization of the largest Golgi matrix protein giantin, which, in turn, alters intra-Golgi localization of some Golgi proteins. Indeed, we recently observed that in androgen-responsive PCa cells, EtOH- induced Golgi fragmentation leads to translocation of glycogen synthase kinase β (GSK3β) from the Golgi to the cytoplasm, followed by the activation of HDAC6-HSP90-AR pathway. Additionally, we reported that non-muscle myosin IIA (NMIIA) motor protein forces EtOH-induced Golgi disruption. Also, alcohol induces endoplasmic reticulum (ER) stress and unfolded protein response (UPR), which are the known drivers of PCa advancement; however, the mechanism of EtOH-induced UPR in cancer remains largely uncovered. Here, we found that in low passage LNCaP cells, one of the UPR sensor, ATF6, is cleaved in Golgi by S1P and S2P proteases; however, EtOH treatment alters the Golgi localization of S1P and S2P, trapping them in the ER and facilitating ATF6 transactivation. Further, EtOH induces translocation of the glycosyltransferase MGAT3 from the Golgi to the cytoplasm followed by its degradation. However, MGAT5, the enzyme that competes with MGAT3 for N-glycan branching, is still retained in the Golgi. This results in MGAT5-mediated glycosylation of pro-metastatic proteins (including matriptase and integrins) and their overexpression at the cell surface. Finally, we detected that loss of NMIIA function prevents alcohol-induced Golgi fragmentation in PCa cells. This, in turn, recovers MGAT3’s intra- Golgi localization and reduces the expression of integrins. In light of these facts, we propose to test the hypothesis that alcohol accelerates PCa progression through Golgi fragmentation, which: (a) enhances ER stress response via induction of ATF6-mediated UPR, and (b) stimulates expression of pro-metastatic proteins over their abnormal MGAT5-mediated glycosylation. Hence, targeting alcohol-induced Golgi fragmentation is therapeutically important. The three specific aims of the proposed study are to: 1) elucidate the mechanism of alcohol-induced activation of ER stress by determining how alcohol metabolites induce translocation of Golgi localized S1P and S2P proteases to the ER; 2) examine the role of EtOH-induced MGAT5-mediated glycosylation in the progression of PCa; and 3) examine whether inhibition or knockdown of NMIIA prevents EtOH-induced Golgi fragmentation and attenuates the aggressive phenotype of PCa cells. The approach will include a variety of in vitro and in vivo experiments using EtOH-treated PCa cell lines, animal models and clinical samples from alcohol consuming PCa patients. The accomplishment of the goal of the proposed study would expand our understanding of the basic biology of PCa carcinogenesis, and elucidate the mechanisms of alcohol's tumor promotion action.

抽象的长期酗酒和酗酒被认为是前列腺癌 (PCa) 进展的危险因素，但是其机制尚不清楚。当前的项目将解决临床医生提出的一个重要问题：是否戒酒是前列腺癌的重要治疗干预措施。之前我们发现：（1）碎片化高尔基复合体与 PCa 的进展相关； (2) 乙醇 (EtOH) 引起高尔基体解构，即由最大的高尔基体基质蛋白巨蛋白的二聚化受损引起，这反过来又改变了高尔基体内部一些高尔基体蛋白的定位。事实上，我们最近观察到，在雄激素反应性 PCa 细胞中，EtOH- 诱导的高尔基体断裂导致糖原合成酶激酶 β (GSK3β) 从高尔基体易位到细胞质，随后激活 HDAC6-HSP90-AR 通路。此外，我们报告称，非肌肉肌球蛋白 IIA (NMIIA) 运动蛋白迫使乙醇诱导的高尔基体破坏。此外，酒精还会诱发内质网状 (ER) 应激和未折叠蛋白反应 (UPR)，这是 PCa 进展的已知驱动因素；然而，EtOH 诱导的 UPR 在癌症中的作用机制在很大程度上仍不清楚。在这里，我们发现在低 LNCaP细胞传代，UPR传感器之一ATF6在高尔基体中被S1P和S2P蛋白酶切割；然而， EtOH 处理改变了 S1P 和 S2P 的高尔基体定位，将它们困在 ER 中并促进 ATF6 反式激活。此外，EtOH 诱导糖基转移酶 MGAT3 从高尔基体易位至细胞质随后降解。然而，MGAT5（与 MGAT3 竞争 N-聚糖的酶）分支，仍保留在高尔基体中。这导致 MGAT5 介导的促转移蛋白糖基化（包括基质酶和整合素）及其在细胞表面的过度表达。最后，我们检测到丢失 NMIIA 功能可防止 PCa 细胞中酒精诱导的高尔基体断裂。这反过来又恢复了 MGAT3 的内部高尔基体定位并减少整合素的表达。鉴于这些事实，我们建议检验假设酒精通过高尔基体断裂加速 PCa 进展，这：(a) 通过以下方式增强 ER 应激反应 ATF6 介导的 UPR 的诱导，以及 (b) 刺激促转移蛋白的异常表达 MGAT5 介导的糖基化。因此，针对酒精诱导的高尔基体断裂在治疗上很重要。该研究的三个具体目标是：1）阐明酒精诱导的激活机制通过确定酒精代谢物如何诱导高尔基体局部 S1P 和 S2P 蛋白酶的易位来确定 ER 应激去急诊室； 2) 检查EtOH诱导的MGAT5介导的糖基化在PCa进展中的作用；和 3) 检查 NMIIA 的抑制或敲低是否可以防止 EtOH 诱导的高尔基体断裂并减弱 PCa 细胞的侵袭性表型。该方法将包括使用各种体外和体内实验乙醇处理的 PCa 细胞系、动物模型和来自饮酒 PCa 患者的临床样本。这拟议研究目标的实现将扩大我们对 PCa 基础生物学的理解致癌作用，并阐明酒精促癌作用的机制。