Notch-1 and IGF-1 crosstalk: new therapeutic strategies for NSCLC

Notch-1 和 IGF-1 串扰：NSCLC 的新治疗策略

• 批准号: 8256620
• 负责人: MAURIZIO BOCCHETTA
• 金额: $ 30.09万
• 依托单位: LOYOLA UNIVERSITY CHICAGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-11 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8256620
• 关键词: 1-Phosphatidylinositol 3-Kinase; Acute T Cell Leukemia; Affect; Alzheimer' s Disease; Amyloid; Antibodies; Antineoplastic Agents; Apoptosis; Area; Biological; Biological Preservation; Breast Cancer Cell; Cancer Biology; Cancer Etiology; Cell Death; Cell Line; Cell Nucleus; Cells; Cessation of life; Chemicals; Cisplatin; Clinic; Clinical; Clinical Trials; Data; Development; Diagnosis; Disease; Doctor of Philosophy; Early Diagnosis; Family; Feedback; Genetic Transcription; Health; Human; Hypoxia; In Vitro; Incidence; Insulin-Like Growth Factor I; Life; Life Expectancy; Link; Lung; Lung Adenocarcinoma; Maintenance; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Malignant neoplasm of lung; Mediating; Methods; Molecular; Mus; Non-Small-Cell Lung Carcinoma; Outcomes Research; PTEN gene; Pathway interactions; Patients; Pharmaceutical Preparations; Phosphoric Monoester Hydrolases; Positive Reinforcements; Pre-Clinical Model; Process; Proteins; Refractory; Regulation; Repression; Role; Sampling; Screening procedure; Second Messenger Systems; Signal Pathway; Signal Transduction; Smoke; Specific qualifier value; Staging; Stem cells; Testing; Therapeutic; Treatment Efficacy; Tumorigenicity; cancer cell; cancer stem cell; chemotherapy; cigarette smoking; genetic manipulation; human FRAP1 protein; improved; in vivo; inhibitor/antagonist; killings; malignant phenotype; mouse model; notch protein; novel therapeutics; presenilin; receptor; research study; second messenger; secretase; small molecule; stem; therapeutic target; treatment strategy; tumor

DESCRIPTION (provided by applicant): Lung cancer is the leading cause of cancer related deaths in the USA. The most frequent form of lung cancer is non small cell lung cancer (NSCLC). Among the three major histological types of NSCLC, adenocarcinoma of the lung (ACL) is the most prevalent, and its incidence is steadily raising. ACL is relatively common (from 15 to 20% of cases) in people who have never smoked, therefore ACL will remain a major health problem even after cigarette smoking eradication. ACL is normally diagnosed at advanced stages because early detection methods remain problematic. Median survival after advanced ACL diagnosis is only 5 months, and chemotherapy only slightly improves it. It is therefore imperative to find novel therapeutic strategies to treat this deadly disease. Our preliminary data indicate that the evolutionarily conserved Notch-1 receptor provides critical survival signals to ACL cells in hypoxia, a condition that best recapitulates ACL microenvironment. Hypoxic conditions are those that interfere with standard chemotherapy and favor the preservation of niches for cancer stem cells. Therefore, targeting a signaling pathway that favor survival of cancer cells in hypoxia represents a particularly promising therapeutic strategy for the treatment of ACL. To be activated, Notch-1 requires a number of proteolitic cleavages, including the participation of a ?-secretase. Since this protein also participates in the accumulation of ¿-amyloid in Alzheimer's Disease, a great effort has been spent to synthesize specific small molecules that inhibit ?-secretase (?-secretase inhibitors or GSI). Therefore, GSI are indirect inhibitors of Notch signaling because, in the absence of the activating proteolitic cleavage catalyzed by ?-secretase, Notch-1 (as well as other Notch receptors) cannot translocate in the nucleus and affect transcription. We used as GSI a compound developed by Merck (MRK-003). This compound proved very efficient in killing ACL cells specifically under hypoxia. Our hypothesis is that Notch-1 provides survival signals to ACL cells mediating an intricate network of signaling pathways that includes PTEN, STAT, IGF-1, IGF-1R, Akt and its downstream targets. We also hypothesize that targeting Notch signaling in vivo is a valid therapeutic strategy for the treatment of ACL. We want to verify our hypotheses according to the following Specific Aims: (1) Verify whether there is interplay and a positive feedback loop between Notch-1 and the IGF-1 signaling pathway and what are the biological consequences of such interaction in hypoxia. (2) Study the molecular mechanisms through which hypoxia strengthen Notch-1 signaling and how Notch-1 protects ACL cells from apoptosis specifically under hypoxia. We will focus on Notch-1 regulation of PTEN and the PI3 kinase/PDK-1/Akt/mTOR axis. (3) Verify the therapeutic efficacy of MRK-003, alone or in combination with cisplatin, or with an inhibitory antibody for the IGF-1R (MK-0646), or with an Akt specific inhibitor (MK-2206) in a mouse preclinical model of advanced ACL. We will use cisplatin in combination with MRK-003 to simultaneously target non-hypoxic and hypoxic tumor areas. MK-0646 and MK-2206 will be also used in separate experiments because these compounds may have synergistic effects with MRK-003 because of our preliminary results sustaining Aims 1 and 2. We will dissect the contribution of Notch inhibition mechanistically using ACL cell lines in which we can artificially downregulate Notch-1. The results obtained in Aim 1 and 2 will be verified in vivo in the mouse model and in human clinical samples. The outcome of the research proposed here will provide a better understanding of the molecular processes that contribute to the maintenance of the malignant phenotype in ACL. Furthermore, the study will test the clinical usefulness of targeting Notch signaling to treat NSCLC using GSI. Finally, these experiments will elucidate the role of Notch-1 in lung tumorigenicity in vitro and in vivo. PUBLIC HEALTH RELEVANCE: Lung cancer is the most frequent cause of cancer related death in the U.S.A. The studies proposed here will allow a better understanding of lung cancer biology and will test novel therapeutic strategies that could be introduced in the clinic rapidly.

描述（由申请人提供）：肺癌是美国癌症相关死亡的主要原因。最常见的肺癌是非小细胞肺癌（NSCLC）。在NSCLC的三种主要组织学类型中，肺腺癌（ACL）是最常见的，并且其发病率正在稳步上升。 ACL 在从未吸烟的人群中相对常见（占 15% 至 20% 的病例），因此即使在根除吸烟后，ACL 仍将是一个主要的健康问题。 ACL 通常在晚期才被诊断出来，因为早期检测方法仍然存在问题。晚期 ACL 诊断后的中位生存期仅为 5 个月，化疗只能略微改善这一情况。因此，迫切需要找到新的治疗策略来治疗这种致命的疾病。我们的初步数据表明，进化上保守的 Notch-1 受体在缺氧（最能概括 ACL 微环境的条件）下为 ACL 细胞提供关键的生存信号。缺氧条件会干扰标准化疗并有利于癌症干细胞的生存环境。因此，针对有利于癌细胞在缺氧条件下存活的信号通路代表了治疗 ACL 的一种特别有前景的治疗策略。为了被激活，Notch-1 需要许多蛋白水解，包括 β-分泌酶的参与。由于该蛋白还参与阿尔茨海默氏病中β-淀粉样蛋白的积累，因此人们付出了巨大的努力来合成抑制β-分泌酶的特定小分子（β-分泌酶抑制剂或GSI）。因此，GSI 是 Notch 信号传导的间接抑制剂，因为在缺乏 β-分泌酶催化的激活蛋白水解裂解的情况下，Notch-1（以及其他 Notch 受体）不能在细胞核中易位并影响转录。我们使用 Merck 开发的化合物 (MRK-003) 作为 GSI。事实证明，这种化合物在缺氧条件下能够非常有效地杀死 ACL 细胞。我们的假设是，Notch-1 向 ACL 细胞提供生存信号，介导复杂的信号通路网络，包括 PTEN、STAT、IGF-1、IGF-1R、Akt 及其下游靶标。我们还假设体内靶向 Notch 信号传导是治疗 ACL 的有效治疗策略。我们希望根据以下具体目标来验证我们的假设：（1）验证Notch-1和IGF-1信号通路之间是否存在相互作用和正反馈循环，以及这种相互作用在缺氧条件下的生物学后果是什么。 (2)研究缺氧增强Notch-1信号传导的分子机制以及Notch-1如何在缺氧下特别保护ACL细胞免于凋亡。我们将重点关注 PTEN 和 PI3 激酶/PDK-1/Akt/mTOR 轴的 Notch-1 调节。 (3) 在小鼠晚期 ACL 临床前模型中验证 MRK-003 单独或与顺铂、IGF-1R 抑制性抗体 (MK-0646) 或 Akt 特异性抑制剂 (MK-2206) 联合使用的治疗效果。我们将使用顺铂与MRK-003组合，同时靶向非缺氧和缺氧肿瘤区域。 MK-0646 和 MK-2206 也将用于单独的实验，因为我们的初步结果维持了目标 1 和 2，这些化合物可能与 MRK-003 具有协同作用。我们将使用 ACL 细胞系，在其中人为下调 Notch-1，从机制上剖析 Notch 抑制的贡献。目标 1 和 2 中获得的结果将在小鼠模型和人类临床样本中进行体内验证。这里提出的研究结果将有助于更好地理解有助于维持 ACL 恶性表型的分子过程。此外，该研究将测试使用 GSI 靶向 Notch 信号传导治疗 NSCLC 的临床有效性。最后，这些实验将阐明Notch-1在体外和体内肺致瘤性中的作用。公共卫生相关性：肺癌是美国癌症相关死亡的最常见原因。此处提出的研究将有助于更好地了解肺癌生物学，并将测试可快速引入临床的新治疗策略。