Therapeutic targets and novel anticancer agents for endocrine cancers

内分泌癌的治疗靶点和新型抗癌药物

• 批准号: 10702517
• 负责人: Naris Nilubol
• 金额: $ 136.02万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10702517
• 关键词: Adjuvant; Adrenocortical carcinoma; Antineoplastic Agents; Apoptosis; Auranofin; BRAF gene; Biological Assay; Blood Vessels; Cancer Patient; Cancer cell line; Caspase; Cell Cycle Arrest; Cell Proliferation; Cells; Client; Clinical; Clinical Trials; Combination Drug Therapy; Control Groups; Cyclin-Dependent Kinases; Cytoplasm; Cytotoxic agent; Data; Data Analyses; Distant Metastasis; Dose; Drug Combinations; Drug Controls; Drug Delivery Systems; Enzymes; Exhibits; External Beam Radiation Therapy; Extracellular Matrix; Extracellular Matrix Proteins; Extravasation; FDA approved; FOXM1 gene; Family; Gene Expression Profiling; Goals; Gold; Heat-Shock Proteins 90; Human; ITGB3 gene; In Vitro; Iodine; LOX gene; Laboratories; Libraries; Malignant Neoplasms; Malignant neoplasm of adrenal cortex; Malignant neoplasm of thyroid; Matrix Metalloproteinases; Microtubule Depolymerization; Mitochondria; Molecular; Morbidity - disease rate; Mus; Mutate; Mutation; Neoadjuvant Therapy; Operative Surgical Procedures; Oral; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Pre-Clinical Model; Process; Protein Isoforms; Proteins; Quality of life; Radiation therapy; Radiation-Sensitizing Agents; Radiosensitization; Recurrence; Refractory; Reporting; Resistance; Rheumatoid Arthritis; Role; Selenium; Strategic Planning; Survivors; Systemic Therapy; Systemic disease; TGFB1 gene; TIMP1 gene; TNF gene; Testing; Toxic effect; Translating; Treatment Protocols; Treatment outcome; Vascular Endothelial Cell; Vascular Endothelial Growth Factors; Vascular Endothelium; Vascular Permeabilities; Xenograft procedure; anaplastic thyroid cancer; base; beta catenin; cancer care; cancer cell; cancer initiation; cancer recurrence; cell motility; chemotherapy; clinically relevant; computerized; cyclin A2; cytokine; differential expression; drug repurposing; drug use screening; effective therapy; experimental study; free radical oxygen; high-throughput drug screening; improved; in vivo; inhibitor; insight; interstitial; malignant endocrine gland neoplasm; molecular targeted therapies; nanomedicine; novel; novel anticancer drug; overexpression; particle; pressure; prognostic significance; public database; screening; synergism; targeted treatment; therapeutic target; treatment group; tumor; tumor microenvironment; tumor progression

There are three main projects: I. Novel synergistic drug combination therapy for adrenocortical cancer (ACC) 1.1 Synergy of MELK and CDK inhibitors targeting multiple clinically-relevant molecules in ACC 1.2 Novel synergistic combination of PI3K and HSP90 inhibitors in ACC 1.3 Drug repurposing study identified auranofin as a novel radiosensitizer in ACC II. Targeted therapy focusing on the effects of TNF-a on tumor microenvironment to enhance drug delivery III. Quantitative high-throughput drug screening in BRAF V600E WT anaplastic thyroid cancer (ATC) 1.1 Synergy of MELK and CDK inhibitors targeting multiple clinically-relevant molecules in ACC Aim: To identify the effective novel synergistic drug combinations against ACC of OTS167 (MELK inhibitor) and RGB286638 (multi-CDK inhibitor) and to study the molecular mechanisms of synergy. Summary: The in vitro synergistic efficacy of OTS-167 and RGB286638 was confirmed by our laboratory. The combination effectively caused G2M cell cycle arrest, induced caspase-dependent apoptosis, inhibited b-catenin via FOXM1, and pSTMN1, resulting in increased depolymerized microtubule and decreased cell invasion. The additional data from the last report showed that the combination treatment downregulated cyclins A2, B1, and B2, which are the partnering proteins of CDKs 1 and 2 with similar prognostic significance in patients with ACC to MELK and CDKs 1 and 2. In vivo studies showed the anti-tumor efficacy of the combination treatment is higher than that of single-drug and control groups. The analyses of xenografts from mice in the combination treatment group confirmed the effects seen in vitro. 1.2 The novel synergistic combination of PI3K and HSP90 inhibitors in ACC Aim: To identify and validate the effective novel synergistic drug combinations using PI3K inhibitor and Heat-shock protein 90 inhibitors and to study the molecular mechanisms of synergy Summary: The most effective synergistic combinations identified by the computerized drug combination matrix screening in ACC cells are one of the three HSP90 inhibitors and PIK-75, a selective p110a inhibitor of PI3K. We analyzed publicly available databases and found that ACC overexpressed several isoforms of HSP90 and HSP-90 clients involving in ACC initiation and progression. We validated the synergy in two ACC cell lines in a cell proliferation assay. The ACC tumor spheroid assays showed increased efficacy in the combination treatment group as compared to the single-drug and control groups. Because PIK-75 is not yet available for a clinical trial, we studied clinically available GDC-0077 which selectively targets p110a isoform of PI3K in combination with STA9090 (HSP90 inhibitor). We did not find synergistic activity at clinically achievable doses. Next, we are testing GDC-0032 which is a new clinically available selective p110a isoform inhibitor of PI3K with a much higher potency than GDC-0077. We found that the combination treatments (colony formation, cell migration) and reduced PI3K pathway. 1.3 Drug repurposing study identified auranofin as a novel radiosensitizer in ACC Aim: To evaluate the effects and mechanism of action of auranofin in combination with radiotherapy (RT) in ACC preclinical models Summary: Despite a complete surgery, 50-80% of patients with ACC develop locoregional recurrence with or without distant metastasis. Mitotane is still the only FDA-approved systemic therapy for ACC, and it imparts no survival benefit but has significant toxicities. Thus, the role of adjuvant or neoadjuvant treatment using in mitotane, chemotherapy, and/or RT in ACC is not known. Because an effective treatment regimen is urgently needed to reduce ACC recurrence and the associated morbidity, we analyzed the data on a quantitative high-throughput drug screening in two ACC cell lines and identified auranofin as a novel radiosensitizer that also exhibited a potent antiproliferative effect in ACC. In addition to locoregional control when used with RT, we anticipate that auranofin can control the systemic disease at a clinically achievable dose. Auranofin was selected because of its high anti-neoplastic efficacy, compared to the current cytotoxic agents used in ACC (Figure 7), and radiosensitization effect by inhibiting TXNRD. Auranofin is an FDA-approved, orally-administered, gold particle-based drug with low toxicity for rheumatoid arthritis. The analyses of the independent, publicly available databases showed several enzymes in the TXNRD pathway, targetable by auranofin, are differentially expressed in ACC, suggesting that the combination of auranofin and RT can be effective. TXNRD is the pro-survival reducing enzyme of TXN, which cleans up ROS and oxygen free radicals created in abundance in both the cytoplasm and mitochondria during external beam radiation therapy. Although the radiosensitizing effects were promising, we found that the doses of auranofin and RT were higher than expected. We discovered that selenium supplement in the culture medium was likely the culprit that partially reversed auranofin-induced TXNRD inhibition. Thus, the current experiments are performed without selenium supplements. II. Targeted therapy focusing on the effects of TNF-a on tumor microenvironment to enhance drug delivery Aim: to elucidate the mechanism of action of TNF-a in reducing tumor interstitial pressure leading to improved drug delivery efficiency. I. Summary: Because the effects of TNF-a on expression and functions of several intratumoral extracellular matrix proteins and cytokines and the roles of these proteins and cytokines on intratumoral vascular permeability are not fully understood. The insight of this process can be used to improve drug delivery in TNF-a insensitive tumors by targeting the downstream targets of TNF-a. To understand the molecular mechanism of TNF-a-induced vascular leakage which results in decreased tumor IFP and increased intratumoral drug delivery, we performed ECM, human matrix metalloproteinase (MMP) arrays, and gene expression profiling of iodine-resistant thyroid cancer cells treated with TNF-a, we identified and validated that TNF-a down-regulated TGFB and LOX, and upregulated MMPs, TIMP1, and ITGB3 which have roles in vascular permeability in TME. siLOX treated iodine-refractory thyroid cancer xenograft recapitulated the intratumoral vascular leakage seen in xenografts treated with nanomedicine carrying TNF-a. Next, we validated the effects of TNF-a on vascular endothelium using vascular endothelial cells and found that TNF-a treatment downregulated TGFB1, Lox, and VEGF-R (a key regulator of vascular permeability). III. Quantitative high-throughput drug screening in BRAF V600E WT anaplastic thyroid cancer (ATC) Aim: To identify novel synergistic combinations of drugs in BRAF V600E WT ATC and to study the mechanism of synergy. Summary: ATC is a rare and one of the most aggressive malignancy. The median survival of patients with ATC is 6 months prior to the discovery of the combination of trametinib and dabrafenib in BRAF V600E mutated ATC that effectively improves treatment outcomes. Unfortunately, patients with BRAF V600E WT ATC remain without effective treatment options. Thus, it is critically important to identify the effective drug combinations that can be readily translated to a clinical trial. We performed a quantitative high-throughput drug screening using a pharmaceutical library of over 5,000 drugs on three ATC cell lines with RAS, BRAF V600E, and non-RAS, non-BRAF mutations. Ninety drugs were effective in RAS a *TRUNCATED*

主要有三个项目：1.新型协同药物联合治疗肾上腺皮质癌（ACC） 1.1针对ACC多种临床相关分子的MELK和CDK抑制剂的协同作用1.2 ACC中PI3K和HSP90抑制剂的新型协同作用1.3药物重新利用研究确定了金糠蛋白作为ACC II的新型放射增敏剂。靶向治疗关注TNF-a对肿瘤微环境的影响以增强药物递送1.1针对ACC多临床相关分子的MELK和CDK抑制剂协同作用目的：寻找针对ACC的有效新型协同药物组合OTS167 （MELK抑制剂）和RGB286638（多重CDK抑制剂），并研究协同作用的分子机制。摘要：本实验室证实OTS-167与RGB286638具有体外协同作用。联合有效阻滞G2M细胞周期，诱导caspase依赖性细胞凋亡，通过FOXM1和pSTMN1抑制b-catenin，导致解聚微管增加，降低细胞侵袭。上一篇报道的额外数据显示，联合治疗下调了细胞周期蛋白A2， B1和B2，它们是cdk1和2的伴侣蛋白，在ACC患者中与MELK和cdk1和2具有相似的预后意义。体内研究表明，联合治疗的抗肿瘤效果高于单药组和对照组。对联合治疗组小鼠异种移植物的分析证实了在体外观察到的效果。1.2 ACC中PI3K与HSP90抑制剂的新型协同组合目的：鉴定并验证PI3K抑制剂与热休克蛋白90抑制剂有效的新型协同药物组合，并研究协同作用的分子机制计算机化药物联合基质筛选在ACC细胞中发现的最有效的协同组合是三种HSP90抑制剂之一和PIK-75 （PI3K的选择性p110a抑制剂）。我们分析了公开可用的数据库，发现ACC过度表达了几种HSP90和HSP-90客户端亚型，这些亚型与ACC的发生和发展有关。我们在细胞增殖试验中验证了两种ACC细胞系的协同作用。与单药组和对照组相比，ACC肿瘤球体检测显示联合治疗组的疗效增加。由于PIK-75尚未用于临床试验，我们研究了临床可用的GDC-0077，它选择性地靶向PI3K的p110a亚型，与STA9090 （HSP90抑制剂）联合使用。在临床可达到的剂量下，我们没有发现协同作用。下一步，我们正在测试GDC-0032，这是一种新的临床可用的选择性p110a亚型PI3K抑制剂，其效力比GDC-0077高得多。我们发现联合处理（菌落形成、细胞迁移）和减少PI3K通路。1.3药物再利用研究发现金糠蛋白是一种新型的ACC放射增敏剂目的：评价金糠蛋白联合放疗（RT）在ACC临床前模型中的作用及机制摘要：尽管进行了完全手术，50-80%的ACC患者仍发生局部复发并伴有或不伴有远处转移。米托坦仍然是fda批准的唯一一种用于ACC的全身疗法，它没有带来生存益处，但具有显著的毒性。因此，辅助或新辅助治疗使用米托坦、化疗和/或RT在ACC中的作用尚不清楚。由于迫切需要一种有效的治疗方案来减少ACC复发和相关的发病率，我们分析了两种ACC细胞系的定量高通量药物筛选数据，并确定了金蜡蛋白是一种新型的放射增敏剂，在ACC中也表现出有效的抗增殖作用。除了与RT联合使用时的局部控制外，我们预计金糠蛋白可以在临床可达到的剂量下控制全体性疾病。与目前用于ACC的细胞毒性药物相比，选择Auranofin是因为其具有较高的抗肿瘤功效（图7），并且通过抑制TXNRD具有放射增敏作用。Auranofin是一种fda批准的口服金颗粒药物，具有低毒性，用于治疗类风湿性关节炎。对独立的、公开的数据库的分析显示，在ACC中，一些TXNRD通路中被金糠蛋白靶向的酶是差异表达的，这表明金糠蛋白和RT联合使用是有效的。TXNRD是TXN的促生存还原酶，可清除体外放射治疗期间细胞质和线粒体中大量产生的ROS和氧自由基。虽然放射线致敏效果很有希望，但我们发现金磺芬和RT的剂量比预期的要高。我们发现，在培养基中添加硒可能是部分逆转auranofin诱导的TXNRD抑制的罪魁祸首。因此，目前的实验是在不添加硒的情况下进行的。二世。靶向治疗关注TNF-a对肿瘤微环境的影响，增强给药目的：阐明TNF-a降低肿瘤间质压力，提高给药效率的作用机制。1 .摘要：由于TNF-a对肿瘤内几种细胞外基质蛋白和细胞因子的表达和功能的影响以及这些蛋白和细胞因子对肿瘤内血管通透性的作用尚不完全清楚。这一过程的见解可用于通过靶向TNF-a的下游靶点来改善TNF-a不敏感肿瘤的药物递送。为了了解TNF-a诱导的血管渗漏导致肿瘤IFP降低和肿瘤内药物传递增加的分子机制，我们对TNF-a治疗的碘耐药甲状腺癌细胞进行了ECM、人基质金属蛋白酶（MMP）阵列和基因表达谱分析，我们发现并验证了TNF-a下调TGFB和LOX，上调MMPs、TIMP1和ITGB3，这些基因在TME中与血管通透性有关。siLOX治疗的碘难治性甲状腺癌异种移植物再现了用携带TNF-a的纳米药物治疗的异种移植物所见的瘤内血管渗漏。接下来，我们利用血管内皮细胞验证了TNF-a对血管内皮的影响，发现TNF-a治疗下调了TGFB1、Lox和VEGF-R（血管通透性的关键调节因子）。三世。BRAF V600E WT间变性甲状腺癌（ATC）的定量高通量药物筛选目的：在BRAF V600E WT间变性甲状腺癌（ATC）中寻找新的协同药物组合，并研究协同作用机制。摘要：ATC是一种罕见且最具侵袭性的恶性肿瘤之一。在发现曲美替尼和达非尼联合治疗BRAF V600E突变的ATC有效改善治疗结果之前，ATC患者的中位生存期为6个月。不幸的是，BRAF V600E WT ATC患者仍然没有有效的治疗选择。因此，确定可以轻易转化为临床试验的有效药物组合是至关重要的。我们使用超过5000种药物的药物文库对三种具有RAS、BRAF V600E和非RAS、非BRAF突变的ATC细胞系进行了定量高通量药物筛选。90种药物在RAS中有效*TRUNCATED*