The GABA-B receptor is a novel drug target for pancreatic cancer

GABA-B受体是胰腺癌的新型药物靶点

• 批准号: 8252196
• 负责人: Hildegard M. Schuller
• 金额: $ 26.45万
• 依托单位: UNIVERSITY OF TENNESSEE KNOXVILLE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8252196
• 关键词: Adenocarcinoma; Adenocarcinoma Cell; Adenylate Cyclase; Adrenergic Agents; Adrenergic beta-Antagonists; Agonist; Aminobutyric Acids; Angiogenic Factor; Apoptotic; Area; Biological; Breast; Butanones; CREB1 gene; Cancer Etiology; Cancer Prognosis; Cell Line; Cell Proliferation; Cells; Cessation of life; Cleaved cell; Clinical Trials; Colon; Complex; Country; Coupled; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Data; Development; Diabetes Mellitus; Diagnosis; Disease; Drug Delivery Systems; EGF gene; Effectiveness; Enzymes; Epidermal Growth Factor Receptor; Epinephrine; Epithelial Cells; G-Protein-Coupled Receptors; Gefitinib; Gene Silencing; Genes; Glutamate Decarboxylase; Goals; Growth; Growth Factor; Guide prevention; Hamsters; Human; Human Cell Line; Hypermethylation; Immunohistochemistry; In Vitro; Laboratories; Lead; MAPK3 gene; Malignant Neoplasms; Malignant neoplasm of pancreas; Mus; National Cancer Institute; Neoplasm Metastasis; Neurotransmitters; Nicotine; Nicotinic Receptors; Nitrosamines; Norepinephrine; Nude Mice; Ovary; PTGS2 gene; Pancreas; Pancreatic Ductal Adenocarcinoma; Pancreatic duct; Pancreatitis; Pathway interactions; Prevention; Prevention therapy; Production; Propranolol; Prostate; Publishing; Receptor Activation; Regulation; Research; Resistance; Risk Factors; Role; Signal Transduction; Smoking; Staging; Starvation; Stomach; Stress; Testing; Time; Tissues; Transactivation; Translating; Woman; Xenograft procedure; adrenergic; base; celecoxib; cell motility; combat; effective therapy; gamma-Aminobutyric Acid; gemcitabine; improved; in vivo; inhibitor/antagonist; men; migration; mortality; novel; overexpression; pre-clinical; promoter; receptor; response; treatment strategy; tumorigenic; Adenocarcinoma; Adenocarcinoma Cell; Adenylate Cyclase; Adrenergic Agents; Adrenergic beta-Antagonists; Agonist; Aminobutyric Acids; Angiogenic Factor; Apoptotic; Area; Biological; Breast; Butanones; CREB1 gene; Cancer Etiology; Cancer Prognosis; Cell Line; Cell Proliferation; Cells; Cessation of life; Cleaved cell; Clinical Trials; Colon; Complex; Country; Coupled; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Data; Development; Diabetes Mellitus; Diagnosis; Disease; Drug Delivery Systems; EGF gene; Effectiveness; Enzymes; Epidermal Growth Factor Receptor; Epinephrine; Epithelial Cells; G-Protein-Coupled Receptors; Gefitinib; Gene Silencing; Genes; Glutamate Decarboxylase; Goals; Growth; Growth Factor; Guide prevention; Hamsters; Human; Human Cell Line; Hypermethylation; Immunohistochemistry; In Vitro; Laboratories; Lead; MAPK3 gene; Malignant Neoplasms; Malignant neoplasm of pancreas; Mus; National Cancer Institute; Neoplasm Metastasis; Neurotransmitters; Nicotine; Nicotinic Receptors; Nitrosamines; Norepinephrine; Nude Mice; Ovary; PTGS2 gene; Pancreas; Pancreatic Ductal Adenocarcinoma; Pancreatic duct; Pancreatitis; Pathway interactions; Prevention; Prevention therapy; Production; Propranolol; Prostate; Publishing; Receptor Activation; Regulation; Research; Resistance; Risk Factors; Role; Signal Transduction; Smoking; Staging; Starvation; Stomach; Stress; Testing; Time; Tissues; Transactivation; Translating; Woman; Xenograft procedure; adrenergic; base; celecoxib; cell motility; combat; effective therapy; gamma-Aminobutyric Acid; gemcitabine; improved; in vivo; inhibitor/antagonist; men; migration; mortality; novel; overexpression; pre-clinical; promoter; receptor; response; treatment strategy; tumorigenic

Project Summary. Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of cancer mortality in Western countries and smoking, diabetes, and pancreatitis are risk factors. The prognosis of this cancer is extremely poor due to its resistance to available therapies and extensive metastasis. New strategies to combat this deadly disease are thus urgently needed and are among priority areas of research identified by the National Cancer Institute. The long-term goal of this project is to develop novel and effective strategies for the treatment and prevention of PDAC. The current project takes advantage of our discovery that beta-adrenoreceptors regulate the growth of human PDAC cell lines and their cells of origin, pancreatic duct epithelial cells. Stimulation of these G- protein coupled receptors by agonists induced signaling via adenylyl cyclase=>cAMP=>PKA=>CREB and transactivated the EGF pathway in a PKA-dependent manner. Isoroterenol additionally stimulated cell migration and had strong anti-apoptotic effects as evidenced by suppression of starvation-induced cleaved casapse 3. GABA and baclophen had strong inhibiting effects on all these responses and additionally reduced proliferation, migration of untreated cells. In support of these in vitro findings, nicotine-induced increase in systemic stress neurotransmitters adrenaline and noradrenaline strongly stimulated the growth of PDAC xenografts, induced p-CREB and p-ERK1/2 in xenograft cells while suppressing the GABA synthesizing enzyme GAD65 and GABA. Treatment of the mice with GABA completely blocked xenograft growth while returning levels of p-CREB, p-ERK1/2, GAD and GABA to normal levels. These data suggest the GABAB receptor as a novel drug target for the treatment and prevention of PDAC. To test this hypothesis we propose four specific aims: Specific aim 1: To evaluate the anti-tumorigenic effects of GABA and the GABAB receptor agonist baclophen on early and advanced stages of PDAC xenograft development in nude mice in the presence and absence of stress neurotransmitter stimulation in response to nicotine and NNK. Specific aim 2: To test the hypothesis that the observed suppression of GAD65 and GABA in nicotine and NNK-treated PDAC xenografts is caused by gene promoter hypermethylation of GAD65, that GABA reverses these effects and to verify these mechanism of gene silencing and reversal in vitro in pancreatic duct epithelial cells. Specific aim 3: To test the hypothesis that the antitumorigenic effects of gefitinib and gemcitabine on PDAC cells are reduced in the presence of stress neurotransmitter stimulation in vitro and in PDAC xenografts and that combination treatments with either agent plus GABA or baclophen improves their effectiveness. Specific aim 4: To assess the effects of the beta-blocker propranolol or the COX-2 inhibitor celecoxib in vitro and in PDAC xenografts with and without stimulation by stress neurotransmitters, and to compare their effectiveness to that of GABA and baclophen. Data to be generated by this project may lead to the successful prevention and treatment of smoking- associated PDAC in a marker-guided fashion with GABA-ergic agents and generate a better understanding of the complex mechanisms of action of stimulating and inhibiting neurotransmitters in the regulation of PDAC. Since GABA and baclophen are already approved for the treatment of non-cancerous conditions in humans, results of this research can be rapidly translated into clinical trials.

项目摘要。 胰腺导管腺癌（PDAC）是西方国家癌症死亡率的第四个主要原因 吸烟，糖尿病和胰腺炎是危险因素。由于 它抵抗可用的疗法和广泛的转移。打击这种致命疾病的新策略 因此，迫切需要，并且是国家癌症研究所确定的研究领域。 该项目的长期目标是制定新颖有效的治疗和预防策略 PDAC。当前的项目利用了我们发现β-肾上腺肾上腺素受体调节增长的发现 人类PDAC细胞系及其起源细胞的胰管上皮细胞。刺激这些g- 激动剂通过腺苷酸环酶诱导的信号传导=> camp => camp => pka => creb和creb，并且 以PKA依赖性方式对EGF途径进行反式激活。异戊烯醇另外刺激细胞 迁移并具有强烈​​的抗凋亡作用，抑制饥饿诱导的裂解证明了 casapse3。GABA和BACLOPHEN对所有这些反应具有强大的抑制作用，并降低了 增殖，未处理细胞的迁移。为了支持这些体外发现，尼古丁诱导的增加 系统性应激神经递质肾上腺素和去甲肾上腺素强烈刺激PDAC的生长 异种移植物，异种移植细胞中的P-CREB和P-ERK1/2，同时抑制GABA合成 酶GAD65和GABA。用GABA治疗小鼠完全阻断了异种移植的生长 P-CREB，P-ERK1/2，GAD和GABA的返回水平为正常水平。这些数据表明GABAB 受体是治疗和预防PDAC的新型药物靶标。为了检验这一假设，我们提出了 四个具体目标： 特定目的1：评估GABA和GABAB受体激动剂Baclophen的抗肿瘤效应 在存在和不存在的情况下 响应尼古丁和NNK的应力神经递质刺激。 特定目的2：检验以下假设：尼古丁中观察到的GAD65和GABA的抑制 NNK处理的PDAC异种移植是由GAD65的基因启动子高甲基化引起的，即GABA逆转 这些影响并验证这些基因沉默和体外逆转胰管逆转的机制 细胞。 特定目的3：测试吉非替尼和吉西他滨对PDAC的抗肿瘤作用的假设 在体外和PDAC异种移植物中存在应力神经递质刺激的情况下，细胞会降低。 与代理，GABA或BACLOPHEN的组合处理可提高其有效性。 特定目的4：评估β受体阻滞剂普萘洛尔或COX-2抑制剂Celecoxib在体外的影响 在PDAC异种移植物中，有压力神经递质刺激和没有刺激，并比较其 GABA和BACLOPHEN的有效性。 该项目要生成的数据可能会导致成功预防和治疗吸烟 - 与GABA-ARGIC代理商以标记引导方式相关的PDAC，并更好地理解 PDAC调节中刺激和抑制神经递质的动作的复杂机制。 由于GABA和BACLOPHEN已经被批准用于治疗人类的非癌状况，因此 这项研究的结果可以迅速转化为临床试验。