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.

抽象的慢性酒精滥用和酒精中毒被认为是前列腺癌（PCA）进展的危险因素，但该机制未知。当前的项目将解决临床医生提出的一个重要问题：是否是戒酒是PCA中重要的治疗干预措施。以前，我们发现：（1）高尔基复合物与PCA的进展相关。（2）乙醇（ETOH）引起高尔基的混乱由最大的高尔基体基质蛋白蛋白的二聚化受损而导致，从而改变高尔基体内某些高尔基蛋白的定位。确实，我们最近观察到，在雄激素响应的PCA细胞中，EtOH- 诱导的高尔基碎裂导致糖原合酶激酶β（GSK3β）从高尔基体转移到细胞质，然后激活HDAC6-HSP90-AR途径。此外，我们报告了非肌肉肌球蛋白IIA（NMIIA）运动蛋白力EtOH诱导的高尔基损伤。此外，酒精会诱导内质网状（ER）应激和展开的蛋白质反应（UPR），这是PCA进步的已知驱动因素；然而，癌症中EtOH诱导的UPR的机制仍然在很大程度上发现。在这里，我们发现很低通过S1P和S2P蛋白酶在Golgi中裂解通道LNCAP细胞，其中一种UPR传感器ATF6。然而， ETOH处理改变了S1P和S2P的高尔基体定位，将它们捕获到ER中并支持ATF6 反式激活。此外，ETOH诱导糖基转移酶MGAT3从高尔基体转运细胞质随后降解。但是，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介导的前转移蛋白的糖基化（包括基质酶和整联蛋白）及其在细胞表面的过表达。最后，我们检测到了 NMIIA功能可防止酒精诱导的PCA细胞中的高尔基体碎片化。反过来，这恢复了MGAT3的内部 - 高尔基体定位并降低整联蛋白的表达。鉴于这些事实，我们建议检验假设酒精通过高尔基分裂加速了PCA的进展，这是：（a）通过 ATF6介导的UPR的诱导，（b）刺激促蛋白异常的表达 MGAT5介导的糖基化。因此，靶向酒精诱导的高尔基体破裂在治疗上很重要。拟议研究的三个特定目的是：1）阐明酒精诱导的激活机理通过确定酒精代谢产物如何诱导高尔基体局部S1P和S2P蛋白酶的易位，可以通过确定质地抗应力。到急诊室； 2）检查ETOH诱导的MGAT5介导的糖基化在PCA进展中的作用； 3）检查NMIIA的抑制或敲低是否可以防止EtOH诱导的高尔基体破裂并减弱 PCA细胞的侵略性表型。该方法将包括多种使用的体外和体内实验来自ETOH处理的PCA细胞系，动物模型和饮酒饮酒患者的临床样品。这实现拟议研究的目标将扩大我们对PCA基本生物学的理解致癌，并阐明了酒精肿瘤促进作用的机制。