Discovery of Novel Inhibitors Targeting trans-Golgi Network Acidification in Pancreatic Cancer

发现针对胰腺癌跨高尔基体网络酸化的新型抑制剂

基本信息

批准号：
10563637
负责人：
Cosimo Commisso
金额：
$ 64.74万
依托单位：
SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-22 至 2027-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10563637
关键词：
Address Area Biological Assay Biological Models Biosensor Cancer Etiology Cell Death Cell membrane Cells Cessation of life Chemicals Clinical Clinical Trials Complement Cytosol Data Analyses Death Rate Development Disease Diversity Library Ensure Future Glycolysis Golgi Apparatus Grant Ion Transport Ions Libraries Malignant Neoplasms Malignant neoplasm of pancreas Maps Metabolic Metabolism Modality Normal Cell Oncogenic Organelles Outcome Pancreatic Ductal Adenocarcinoma Pathway interactions Patients Pharmaceutical Preparations Play Process Production Protons Research Role Stress Fibers Structure-Activity Relationship Survival Rate Testing Therapeutic Therapeutic Agents United States Validation Warburg Effect Work analog antiporter cancer cell cancer therapy cancer type cell transformation cytotoxicity design extracellular fitness high throughput screening improved in vivo inhibitor innovation kinase inhibitor lead optimization miniaturize mouse model mutant neoplastic cell novel novel therapeutic intervention novel therapeutics overexpression pH Homeostasis pancreatic ductal adenocarcinoma cell pancreatic ductal adenocarcinoma model prevent scaffold screening small molecule small molecule inhibitor therapeutically effective tool trans-Golgi Network tumor metabolism

项目摘要

PROJECT SUMMARY Pancreatic ductal adenocarcinoma (PDAC) is projected to be the second leading cause of cancer-related deaths in the United States by 2030. With a 5-year relative survival rate of just 10%, PDAC has the highest death rate among the most common cancers, underscoring the urgent need for new therapeutic strategies that improve clinical outcomes. Recent years have witnessed a growing appreciation of the role that metabolic reprogramming plays in conferring survival advantages to PDAC cells, and the targeting of metabolic processes is considered a promising area for the development of novel therapies. In contrast to normal cells, cancer cells favor glycolysis for energy production, a phenomenon called the Warburg effect. As a glycolytic byproduct, transformed cells produce an excessive amount of H+ ions. This cytosolic accumulation of H+ would be detrimental to cell fitness; therefore, tumor cells have evolved mechanisms to achieve pH homeostasis. Using PDAC as a model system, we discovered that cytosolic pH in cancer cells is regulated by organellar sequestration of H+ ions via compartmental ion transport and that the trans-Golgi network (TGN) can act as a “sink” for cytosolic H+. Importantly, normal cells do not employ this homeostatic mechanism. Using the NHE7 Na+/H+ antiporter, which is primarily localized to the TGN, we have shown that targeting NHE7 causes acidification of the cytosol and subsequent cell death in PDAC cells but not normal cells. We have further validated these findings in mouse models of PDAC. Patient-centric data analyses show that TGN localized transporters are frequently overexpressed in PDAC and their elevated expression is correlated with worse overall survival. To robustly quantify pH in the lumen of the TGN, we have developed an innovative TGN-targeted pH biosensor assay, which has been optimized for high-throughput (HT) screening of small molecule compound libraries to identify novel TGN-selective acidification inhibitors. We have validated our TGN pH biosensor HT assay in a set of clinical trial and approved drugs, as well as kinase inhibitors. To complement our HT screen, we have designed a cutting- edge secondary and tertiary assay pipeline to further validate TGN-selective compounds. We hypothesize that novel Golgi–specific small molecule inhibitors of organelle acidification will selectively prevent cancer cells from achieving proper pH homeostasis, resulting in acidic cytosolic pH and cell death. In this grant we will: 1) perform a large-scale HT screen to identify inhibitors of TGN acidification and confirm the hits, 2) validate the confirmed hits using secondary assays and prioritize hit scaffolds, and 3) map compound effects to TGN components and select and characterize final probe(s). Successful completion of this work will provide novel small molecule chemical probes validated to interfere with TGN acidification and cause cancer-selective cell death, serving as starting points to be further developed into safe and effective therapeutic agents for PDAC.

项目概要胰腺导管腺癌 (PDAC) 预计将成为癌症相关的第二大原因到 2030 年，美国的死亡人数。PDAC 的 5 年相对生存率仅为 10%，死亡率最高最常见癌症中的发病率，强调迫切需要新的治疗策略来改善近年来，人们越来越认识到代谢重编程的作用。赋予 PDAC 细胞生存优势，代谢过程的靶向被认为是开发新疗法的前景广阔的领域与正常细胞相比，癌细胞有利于糖酵解。对于能量生产，这种现象称为瓦伯格效应作为糖酵解副产物，转化细胞。产生过量的 H+ 离子，这种 H+ 的胞质积累对细胞健康有害；因此，肿瘤细胞已经进化出以 PDAC 作为模型系统实现 pH 稳态的机制。我们发现癌细胞中的细胞质 pH 值是通过细胞器对 H+ 离子的螯合来调节的区室离子运输以及跨高尔基体网络 (TGN) 可以充当胞质 H+ 的“接收器”。重要的是，正常细胞不采用 NHE7 Na+/H+ 逆向转运蛋白的这种稳态机制。主要定位于 TGN，我们已经证明靶向 NHE7 会导致胞质溶胶酸化，随后，PDAC 细胞发生细胞死亡，而正常细胞则不然。我们在小鼠中进一步验证了这些发现。以患者为中心的数据分析模型表明，TGN 局部转运蛋白经常出现。 PDAC 中过度表达，其表达升高与较差的总体生存率密切相关。为了量化 TGN 管腔中的 pH 值，我们开发了一种创新的 TGN 靶向 pH 生物传感器测定法，已针对小分子化合物库的高通量 (HT) 筛选进行了优化，以识别新的我们在一系列临床试验中验证了 TGN pH 生物传感器 HT 测定。和批准的药物，以及激酶抑制剂，为了补充我们的 HT 筛选，我们设计了一种切割- 边缘二级和三级检测管道，以进一步验证 TGN 选择性化合物。新型高尔基体特异性细胞器酸化小分子抑制剂将选择性地阻止癌细胞实现适当的 pH 稳态，导致细胞质 pH 呈酸性和细胞死亡。在这笔资助中，我们将：1) 执行。大规模 HT 筛选，以识别 TGN 酸化抑制剂并确认命中，2) 验证已确认的结果使用二次分析和优先命中支架进行命中，以及 3) 将化合物效应映射到 TGN 成分和选择并表征最终探针。这项工作的成功完成将提供新颖的小分子。经验证可干扰 TGN 酸化并导致癌症选择性细胞死亡的化学探针进一步开发为安全有效的 PDAC 治疗剂。