Targeting GCNT3 for Pancreatic Cancer

靶向 GCNT3 治疗胰腺癌

• 批准号: 10260098
• 负责人: Chinthalapally V. Rao
• 金额: --
• 依托单位: OKLAHOMA CITY VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-10-01 至 2025-09-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10260098
• 关键词: Address; Adenocarcinoma; Anabolism; Binding; Carcinoma; Cell physiology; Chemoresistance; Clinical; Clinical Research; Clustered Regularly Interspaced Short Palindromic Repeats; Combined Modality Therapy; Development; Disease; Docking; Dose; Drug Delivery Systems; Duct (organ) structure; Enzymes; Family; Female; Future; Genes; Genetic; Genetic Engineering; Growth; Health; Human; Impairment; In Vitro; Incidence; Investigation; KPC model; Knock-out; Lead; Lesion; Light; MUC5AC gene; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Methods; Molecular; Mucin 1 protein; Mucin-2 Staining Method; Mucins; Mus; N-Acetylglucosaminyltransferases; Nature; Neoplasm Metastasis; Neoplasms; Oncogenic; Outcome; Paclitaxel; Pancreas; Pancreatic Intraepithelial Neoplasia; Pathogenesis; Patients; Pharmaceutical Preparations; Pharmacology; Pharmacotherapy; Positron-Emission Tomography; Production; Property; Research; Resistance; Role; Signal Pathway; Signal Transduction; Site; Slice; Small Interfering RNA; Source; Survival Analysis; Survival Rate; Testing; Therapeutic; Therapeutic Effect; Tissues; Treatment Efficacy; Tumor Tissue; Tumor Weights; Up-Regulation; Veterans; Xenograft procedure; cancer therapy; chemotherapy; clinically relevant; effective therapy; efficacy evaluation; gemcitabine; gene synthesis; human data; improved; in silico; in vivo; inhibitor; knock-down; male; mortality; neoplastic; neoplastic cell; next generation sequencing; novel; novel strategies; novel therapeutic intervention; novel therapeutics; overexpression; pancreatic cancer cells; pancreatic cancer patients; pancreatic neoplasm; patient derived xenograft model; preclinical study; response; simulation; targeted treatment; therapeutic target; transcriptome sequencing; translational approach; treatment response; treatment strategy; tumor; tumor growth; tumor progression; tumor xenograft; tumorigenesis; uptake; whole genome

Summary Pancreatic cancer (PC) has the highest mortality of any cancer, with a 5-year survival rate of less than 10%. Despite progress in the development of targeted therapies, the survival rate is still unacceptably low. It is, therefore, important to identify novel therapeutic approaches that could lead to a more effective treatment for this malignant disease. This proposal explores the novel concept that a core 2 mucin synthesis gene, GCNT3, regulates PC growth and survival, ascribing a new and critical role to GCNT3. We have shown that GCNT3 is overexpressed in PC patients and contributes to PC pathogenesis, indicating that GCNT3 is a promising therapeutic target. However, how GCNT3 regulates PC is not clear. Using in silico small molecular docking simulation approaches, we discovered that talniflumate is a novel inhibitor that selectively binds to GCNT3. Examining the molecular mechanisms of GCNT3-mediated PC growth, we found that siRNA-mediated knockdown of GCNT3 or treatment with talniflumate, inhibited xenograft tumors. Talniflumate also reduced GCNT3 expression, leading to reduced production of mucins in vivo and in vitro, and improved cellular gemcitabine uptake. Taken together, our findings strongly suggest that targeting mucin biosynthesis through GCNT3 may improve responsiveness to drug treatment. We, therefore, hypothesize that aberrant expression of GCNT3 will lead to excessive mucin synthesis during PC development and cause chemoresistance. Hence, the inhibition of aberrant mucin synthesis is sufficient to disrupt the barrier properties of the mucin mesh to efficiently increase the access of chemotherapeutics to target sites. Three independent, yet interrelated, specific aims are proposed to address this hypothesis. Aim 1: Generate GCNT3-deficient KPC mice and determine whether targeting GCNT3 ablates the mucin barrier and inhibits PC progression/metastasis by increasing gemcitabine and nab-paclitaxel (NPT) efficacy. Aim 2: Determine whether combinational therapy of talniflumate plus NPT are effective in treating PC by a) evaluating the efficacy and dose-response effects of the combination administered in vivo in a clinically relevant KPC model of spontaneous PC and b) determining mucin disruption and gemcitabine or NPT uptake in combination-treated tumors. c) We will establish the GCNT3 and NPT levels in tumor tissue and their correlation with mucin subtypes (Mucs) and PC treatment efficacy via whole genome transcriptome sequencing and PET imaging. Aim 3: We will investigate the role of GCNT3 in gemcitabine resistance, mucin overexpression and PC growth. a) For clinical relevance, we will assess the inhibition of human patient- derived xenografts (PDX) and PC slice cultures by combinational therapy. b) Evaluate the efficacy of combination against GCNT3 overexpressing human PC PDX tumors in vivo. Veterans Health Relevance: Pancreatic cancer major health problem and many veterans and their families suffered from this neoplastic disease. The results obtained from this study will have high impact, since they will shed light on future development of novel therapeutic strategies for PC. Thus, this proposal is highly translational in nature

摘要 胰腺癌(PC)是所有癌症中死亡率最高的，5年生存率不到10%。 尽管靶向治疗的发展取得了进展，但存活率仍然低得令人无法接受。它是, 因此，重要的是确定新的治疗方法，可以导致更有效的治疗 这种恶性疾病。这一建议探索了一个新的概念，即核心2粘蛋白合成基因GCNT3， 调节PC的生长和生存，将一个新的关键角色归因于GCNT3。我们已经证明了GCNT3 在PC患者中过表达，并在PC的发病机制中起作用，表明GCNT3是一种有前途的 治疗靶点。然而，GCNT3如何调控PC尚不清楚。在电子小分子对接中的应用 模拟方法，我们发现他尼氟酸酯是一种新型的选择性结合GCNT3的抑制剂。 研究GCNT3介导的PC生长的分子机制，我们发现siRNA介导的 阻断GCNT3或用他尼氟酸盐治疗，均可抑制异种移植瘤。他尼氟米特也降低了 GCNT3的表达，导致体内和体外粘蛋白的产生减少，并改善细胞 吉西他滨摄取。综上所述，我们的发现强烈表明，通过 GCNT3可提高对药物治疗的反应性。 因此，我们假设GCNT3的异常表达将导致粘蛋白合成过多 在PC开发过程中，并导致化疗耐药性。因此，对异常粘蛋白合成的抑制是 足以破坏粘蛋白网状物的阻隔性能，从而有效地增加 化疗药物进入靶点。提出了三个独立但又相互关联的具体目标来解决 这个假说。目的1：建立GCNT3缺陷的KPC小鼠，并确定靶向GCNT3是否能消除 粘蛋白屏障通过增加吉西他滨和nab-紫杉醇(NPT)抑制PC进展/转移 功效。目的2：确定他尼氟酸钠联合NPT治疗前列腺癌是否有效 通过在临床上评估体内给药的疗效和剂量-反应效应 自发性PC的相关KPC模型和b)确定粘蛋白破坏和吉西他滨或NPT摄取 联合治疗的肿瘤。C)我们将建立肿瘤组织中GCNT3和NPT的水平以及它们的 全基因组转录组分析粘蛋白亚型与PC疗效的相关性 测序和正电子发射计算机断层扫描。目的3：我们将研究GCNT3在吉西他滨耐药、粘蛋白中的作用 过度表达与PC生长。A)对于临床意义，我们将评估人类患者的抑制- 衍生异种移植物(PDX)和PC切片联合培养。B)评价以下项目的成效 联合治疗体内高表达人PC PDX肿瘤的GCNT3。 退伍军人健康相关性：胰腺癌是许多退伍军人和他们的主要健康问题 许多家庭都患有这种肿瘤疾病。这项研究的结果将产生很大的影响，因为 它们将为PC的新治疗策略的未来发展提供线索。因此，这项建议具有很高的 翻译性