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

概括 胰腺癌（PC）是所有癌症中死亡率最高的，5年生存率不到10%。 尽管靶向治疗的开发取得了进展，但生存率仍然低得令人无法接受。这是， 因此，重要的是要找到新的治疗方法，以提供更有效的治疗方法 这种恶性疾病。该提案探讨了核心 2 粘蛋白合成基因 GCNT3 的新概念， 调节 PC 的生长和存活，赋予 GCNT3 一个新的关键作用。我们已经证明 GCNT3 在 PC 患者中过度表达并有助于 PC 发病机制，表明 GCNT3 是一种有前途的 治疗目标。然而，GCNT3如何调控PC尚不清楚。使用计算机模拟小分子对接 通过模拟方法，我们发现他尼氟酯是一种选择性结合 GCNT3 的新型抑制剂。 检查 GCNT3 介导的 PC 生长的分子机制，我们发现 siRNA 介导的 敲低 GCNT3 或用他尼氟酯治疗可抑制异种移植肿瘤。他尼氟酯也减少了 GCNT3 表达，导致体内和体外粘蛋白的产生减少，并改善细胞 吉西他滨的摄取。综上所述，我们的研究结果强烈表明，通过靶向粘蛋白生物合成 GCNT3 可以改善对药物治疗的反应。 因此，我们假设 GCNT3 的异常表达将导致粘蛋白合成过多 在 PC 发育过程中并导致化学耐药性。因此，抑制异常粘蛋白合成是 足以破坏粘蛋白网的屏障特性，从而有效地增加 针对靶点进行化疗。提出了三个独立但相互关联的具体目标来解决 这个假设。目标 1：生成 GCNT3 缺陷的 KPC 小鼠并确定靶向 GCNT3 是否会消融 通过增加吉西他滨和白蛋白结合型紫杉醇 (NPT) 的浓度来破坏粘蛋白屏障并抑制 PC 进展/转移 功效。目标 2：确定他尼氟酯联合 NPT 联合治疗是否能有效治疗 PC [0102] a)评估在临床试验中体内施用的组合的功效和剂量反应效应 自发 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 在联合治疗的肿瘤中。 c) 我们将建立肿瘤组织中的GCNT3和NPT水平及其 通过全基因组转录组研究与粘蛋白亚型 (Mucs) 和 PC 治疗效果的相关性 测序和 PET 成像。目标 3：我们将研究 GCNT3 在吉西他滨耐药性、粘蛋白中的作用 过度表达和 PC 生长。 a) 对于临床相关性，我们将评估人类患者的抑制作用- 通过联合治疗衍生异种移植物 (PDX) 和 PC 切片培养物。 b) 评估效果 体内针对 GCNT3 过度表达的人 PC PDX 肿瘤的组合。 退伍军人健康相关性：胰腺癌是主要的健康问题，许多退伍军人及其家属 家庭患有这种肿瘤疾病。这项研究获得的结果将产生重大影响，因为 他们将为 PC 新型治疗策略的未来发展提供线索。因此，这个提议非常具有针对性 本质上是翻译性的