Targeting Cancer through Suppressing Stress Induction of GRP78/BiP

通过抑制 GRP78/BiP 的应激诱导来靶向癌症

基本信息

批准号：
10310435
负责人：
AMY S LEE
金额：
$ 36.99万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
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：
2019-12-01 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10310435
关键词：
3-Dimensional Acute Affect Affinity Antineoplastic Agents Apoptosis Biological Markers Biological Models Breast Breast Cancer cell line Cancer Model Cardiac Glycosides Cardiac development Cell Line Cell Proliferation Cell Survival Cell model Cell surface Cells Chromatin Clinical Clinical Trials Colon Colon Carcinoma Colorectal Cancer Combined Modality Therapy DNA Damage Digoxin Drug resistance Endoplasmic Reticulum Endothelial Cells Equilibrium Family Future GRP78 gene Growth Heart failure Hematopoietic Neoplasms Homeostasis Hormonal Human Hypoxia In Vitro Malignant Neoplasms Mediating Messenger RNA Molecular Chaperones Monitor Na(+)-K(+)-Exchanging ATPase Neoplasm Circulating Cells Neoplasm Metastasis Oncogenic Organoids Oxygen PI3K/AKT Pathway interactions Patient Selection Patients Pharmaceutical Preparations Phenotype Plasma Pre-Clinical Model Proliferating Property Protein Isoforms Proteins Radiation therapy Resistance Signal Pathway Signal Transduction Small Interfering RNA Solid Stress Testing Time Translating Translations Treatment Protocols Tumor Tissue Up-Regulation anti-cancer antiangiogenesis therapy cancer cell cancer survival clinically relevant colon cancer cell line colon cancer patients combat design dosage drug-sensitive efficacy evaluation efficacy testing endoplasmic reticulum stress glucose-regulated proteins high-throughput drug screening imaging platform in vivo in vivo Model inhibitor knock-down malignant breast neoplasm member neoplastic cell novel novel therapeutic intervention overexpression patient biomarkers patient derived xenograft model patient population patient stratification polysome profiling pre-clinical preclinical evaluation protein folding refractory cancer response stemness targeted treatment therapy resistant three-dimensional modeling transcriptome sequencing treatment response tumor tumor growth tumor hypoxia tumor microenvironment

项目摘要

Aggressive cancer cells often upregulate GRP78, a major endoplasmic reticulum (ER) chaperone and key regulator of the unfolded protein response (UPR) to augment protein folding capacity and maintain ER homeostasis, thereby promoting survival and acquiring therapeutic resistance. A high-throughput drug screen revealed that a class of compounds referred to as cardiac glycosides (CGs) such as digoxin (DIG), in current use to treat heart failure, as novel inhibitors of GRP78 stress induction in a wide range of human cancer. The discovery that CGs can suppress stress induction of GRP78, which is pivotal for cancer survival, invasion and oncogenic signaling, opens up a new mechanism for the antineoplastic action of the CGs. Among the CGs, oleandrin (OLN) with 100-fold higher affinity for the α3 versus the α1 subunit of Na-K ATPase and high tolerability, is most potent compared to DIG and other CGs. Here we propose to test the efficacy of OLN to suppress GRP78 and cell viability, taking advantage of unique sets of preclinical patient-derived breast and colon cancer models that closely recapitulate the in vivo milieu developed by our collaborative team. We hypothesize that OLN, acting through the α3 subunit of the Na-K ATPase, acutely suppresses stress induction of both intracellular and the cell surface form of GRP78 through blocking its acute translation following ER stress. OLN may also preferentially suppress translation of other proteins critical for survival under stress. As a consequence, ER homeostasis is perturbed and ER-stress mediated apoptosis is triggered. These contribute majorly to the antineoplastic mechanism of OLN. In this proposal, we will: 1) establish the efficacy of OLN and for comparison, DIG, in suppressing GRP78 expression, cell viability, stemness and invasion in patient-derived breast circulating tumor cells both in vitro and in vivo; 2) establish the efficacy of OLN and DIG in lowering GRP78 expression and viability of patient-derived colorectal cancer cells and enhancing standard therapy in organoids. Promising treatment schedules will be validated in PDX models in vivo. The GRP78 expression levels, the status of the α3 subunit of Na-K ATPase and the OLN/DIG concentration in the plasma and tumor in the model systems will be monitored; and 3) examine the mechanisms whereby OLN and DIG suppress stress induction of GRP78 and its impact on the UPR and apoptosis as well as the importance of GRP78 in the anti-cancer effect of OLN/CG. In summary, our studies will provide critical preclinical evaluation of the efficacy of OLN/DIG in cancer leveraging novel patient-derived breast and colon cancer model systems, and a high content imaging platform that mimics the hypoxic tumor microenvironment for monitoring the response. Our studies will also explore the utility of GRP78 and α3 subunit of Na-K ATPase as biomarkers for stratifying patients for OLN/DIG therapy and drug response, and provide proof-of-concept for further development of CG agents in combination or targeted therapy settings in suppressing GRP78, impacting the UPR and apoptosis. Our results will have far-reaching impact as they are also applicable to other solid and blood cancers dependent on GRP78 for growth, invasion and resistance.

侵袭性癌细胞通常上调 GRP78，这是一种主要的内质网 (ER) 伴侣和关键未折叠蛋白反应 (UPR) 的调节因子，以增强蛋白折叠能力并维持 ER 体内平衡，从而促进生存并获得治疗耐药性。揭示了一类被称为强心苷（CG）的化合物，例如地高辛（DIG），目前在用于治疗心力衰竭，作为多种人类癌症中 GRP78 应激诱导的新型抑制剂。发现 CG 可以抑制 GRP78 的应激诱导，而 GRP78 对于癌症的生存、侵袭和扩散至关重要致癌信号，开辟了 CG 抗肿瘤作用的新机制。夹竹桃苷 (OLN) 对 Na-K ATP 酶的 α3 亚基的亲和力比 α1 亚基高 100 倍，并且具有高耐受性，与 DIG 和其他 CG 相比最有效，在此我们建议测试 OLN 抑制 GRP78 的功效。和细胞活力，利用独特的临床前患者衍生的乳腺癌和结肠癌模型紧密地概括了我们的合作团队开发的体内环境，我们概括了 OLN 的作用。通过 Na-K ATP 酶的 α3 亚基，急性抑制细胞内和细胞内的应激诱导 GRP78 的表面形式也可能通过在内质网应激后阻断其急性翻译而优先发生。抑制对应激下生存至关重要的其他蛋白质的翻译，因此，内质网稳态受到影响。扰动和内质网应激介导的细胞凋亡被触发，这些主要有助于抗塑性作用。 OLN 的机制在本提案中，我们将： 1）建立 OLN 的功效并进行比较，DIG，在抑制患者来源的乳腺循环肿瘤中的 GRP78 表达、细胞活力、干性和侵袭体外和体内细胞；2) 确定 OLN 和 DIG 在降低 GRP78 表达和活力方面的功效患者来源的结直肠癌细胞和增强类器官的标准治疗有希望的治疗。方案将在体内 PDX 模型中验证 GRP78 的表达水平、α3 亚基的状态。将监测模型系统中血浆和肿瘤中的Na-K ATP酶和OLN/DIG浓度； 3) 检查 OLN 和 DIG 抑制 GRP78 应激诱导的机制及其对 UPR 和细胞凋亡以及 GRP78 在 OLN/CG 抗癌作用中的重要性综上所述，我们的研究将为 OLN/DIG 在癌症中的功效提供关键的临床前评估，利用新的源自患者的乳腺癌和结肠癌模型系统，以及模仿癌症的高内容成像平台我们的研究还将探索 GRP78 的效用。 Na-K ATPase 的 α3 亚基作为生物标志物，用于对 OLN/DIG 治疗和药物反应患者进行分层，并为联合或靶向治疗环境中 CG 药物的进一步开发提供概念验证抑制 GRP78、影响 UPR 和细胞凋亡，我们的结果将产生深远的影响。也适用于依赖 GRP78 生长、侵袭和抵抗的其他实体癌和血液癌。