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: 8456065
• 负责人: MAURIZIO BOCCHETTA
• 金额: $ 28.29万
• 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 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8456065
• 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; 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; screening; second messenger; secretase; small molecule; stem; therapeutic target; treatment strategy; tumor; tumor microenvironment

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.

描述（由申请人提供）：肺癌是美国癌症相关死亡的主要原因。非小细胞肺癌（NSCLC）是一种常见的恶性肿瘤。在NSCLC的三种主要组织学类型中，肺腺癌（ACL）是最常见的，并且其发病率稳步上升。ACL在从不吸烟的人群中相对常见（15%至20%的病例），因此即使在戒烟后，ACL仍将是一个主要的健康问题。ACL通常在晚期诊断，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只有5个月，化疗只能略微改善其生存期，因此必须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受体提供了关键的生存信号，ACL细胞在缺氧，最好的概括ACL微环境的条件。低分化条件是那些干扰标准化疗并有利于保留癌症干细胞的小生境的条件。因此，靶向有利于癌细胞在缺氧中存活的信号通路代表了治疗ACL的特别有前途的治疗策略。为了被激活，Notch-1需要大量的蛋白水解裂解，包括？分泌酶由于这种蛋白质也参与阿尔茨海默病中淀粉样蛋白的积累，因此人们花费了很大的努力来合成抑制淀粉样蛋白的特定小分子。分泌酶（？-分泌酶抑制剂或GSI）。因此，GSI是Notch信号传导的间接抑制剂，因为在不存在由？-分泌酶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。该化合物被证明在缺氧条件下特异性地杀死ACL细胞是非常有效的。我们的假设是Notch-1为ACL细胞提供生存信号，介导包括PTEN、STAT、IGF-1、IGF-1 R、Akt及其下游靶点在内的复杂信号通路网络。我们还假设，靶向Notch信号在体内是一种有效的治疗策略，用于治疗ACL。我们希望通过以下具体目的来验证我们的假设：（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回路，以及这种相互作用在缺氧中的生物学后果是什么。(2)研究缺氧增强Notch-1信号通路的分子机制，以及Notch-1在缺氧条件下如何特异性地保护ACL细胞免于凋亡。我们将重点关注Notch-1对PTEN和PI 3激酶/PDK-1/Akt/mTOR轴的调节。(3)在晚期ACL小鼠临床前模型中，验证MRK-003单独给药或与顺铂、IGF-1 R抑制性抗体（MK-0646）或Akt特异性抑制剂（MK-2206）联合给药的疗效。我们将使用顺铂与MRK-003联合，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，因为这些化合物可能与MRK-003具有协同效应，因为我们的初步结果支持目的1和2。我们将使用ACL细胞系从机制上剖析Notch抑制的贡献，在ACL细胞系中，我们可以人工下调Notch-1。目标1和2中获得的结果将在小鼠模型和人体临床样本中进行体内验证。这里提出的研究结果将提供一个更好的理解的分子过程，有助于维持ACL的恶性表型。此外，该研究将测试靶向Notch信号传导以使用GSI治疗NSCLC的临床有用性。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。

项目成果

Notch signaling in lung cancer.

• DOI: 10.1586/era.10.158
• 发表时间: 2011-04
• 期刊: Expert review of anticancer therapy
• 影响因子: 3.3
• 作者: Galluzzo P;Bocchetta M
• 通讯作者: Bocchetta M

Depletion of Amyloid Precursor Protein (APP) causes G0 arrest in non-small cell lung cancer (NSCLC) cells.

• DOI: 10.1002/jcp.24875
• 发表时间: 2015-06
• 期刊: JOURNAL OF CELLULAR PHYSIOLOGY
• 影响因子: 5.6
• 作者: Sobol, Anna;Galluzzo, Paola;Weber, Megan J.;Alani, Sara;Bocchetta, Maurizio
• 通讯作者: Bocchetta, Maurizio

Amyloid precursor protein (APP) affects global protein synthesis in dividing human cells.

• DOI: 10.1002/jcp.24835
• 发表时间: 2015-05
• 期刊: JOURNAL OF CELLULAR PHYSIOLOGY
• 影响因子: 5.6
• 作者: Sobol, Anna;Galluzzo, Paola;Liang, Shuang;Rambo, Brittany;Skucha, Sylvia;Weber, Megan J.;Alani, Sara;Bocchetta, Maurizio
• 通讯作者: Bocchetta, Maurizio

Multimodality Approaches to Treat Hypoxic Non-Small Cell Lung Cancer (NSCLC) Microenvironment.

• DOI: 10.1177/1947601912457025
• 发表时间: 2012-02-01
• 期刊: Genes & cancer
• 作者: Liang, Shuang;Galluzzo, Paola;Bocchetta, Maurizio
• 通讯作者: Bocchetta, Maurizio

Deubiquitinase OTUD6B Isoforms Are Important Regulators of Growth and Proliferation.

• DOI: 10.1158/1541-7786.mcr-16-0281-t
• 发表时间: 2017-02
• 期刊: Molecular cancer research : MCR
• 作者: Sobol A;Askonas C;Alani S;Weber MJ;Ananthanarayanan V;Osipo C;Bocchetta M
• 通讯作者: Bocchetta M