Molecular Intervention in Thoracic Malignancies

胸部恶性肿瘤的分子干预

• 批准号: 6558691
• 负责人: DAVID SCHRUMP
• 金额: --
• 依托单位: DIVISION OF CLINICAL 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/6558691
• 关键词: apoptosis; cell cycle; clinical research; clinical trial phase I; deoxycytidine; drug screening /evaluation; enzyme inhibitors; enzyme therapy; gene expression; gene therapy; human subject; human therapy evaluation; lung neoplasms; mesothelioma; microarray technology; neoplasm /cancer chemotherapy; neoplasm /cancer genetics; neoplasm /cancer immunotherapy; neoplastic cell; nucleic acid amplification techniques; peptides; protein kinase; thorax neoplasm; tissue /cell culture; tumor antigens

Pulmonary, esophageal, and pleural malignancies arise via multistep mechanisms in cancerization fields. Although their etiologies may differ, these malignancies exhibit universal molecular themes including aberrant signal transduction via ras pathways, inactivation of the Rb and p53 tumor suppressor pathways, and expression of telomerase, all of which appear critical for malignant transformation. In addition to disrupting cell cycle regulation, oncogene and tumor suppressor gene mutations influence the metastatic phenotype of thoracic neoplasms by modulating expression of adhesion molecules, matrix metalloproteinases, as well as proangiogenic ligands such as vascular endothelial growth factor. Global derangements in chromatin structure and DNA methylation which silence a variety of tumor suppressor genes facilitate de novo expression of several cancer testis antigens in thoracic malignancies. Mutations involving erbB1/2, cyclin D, Rb, p16, p14/ARF, and p53 are attractive targets for intervention in thoracic malignancies and their precursor lesions; SV40 oncoproteins may be unique targets for molecular intervention in malignant pleural mesotheliomas. In published studies we have demonstrated that abrogation of erbB signal transduction by 17-AAG and related compounds markedly enhances sensitivity to chemotherapeutic agents and inhibits the metastatic phenotype of lung and esophageal cancer cells. Additional laboratory efforts have focused on the analysis of gene induction in thoracic malignancies mediated by DNA demethylating agents and histone deacetylase (HDAC) inhibitors. We have observed that under exposure conditions that facilitate expression of tumor suppressor genes such as p16, 5 aza 2' deoxycytidine (DAC) mediates induction of MAGE-3 and NY-ESO-1 cancer testis antigens. In addition we have demonstrated that DAC-mediated target gene induction can be augmented by the HDAC inhibitor Depsipeptide FR901228 (DP). Furthermore, we have shown that following exposure to DAC, or sequential DAC/DP, cancer cells can be recognized by cytolytic T lymphocytes specific for NY-ESO-1, and that sequential DAC/DP treatment mediates profound proapoptotic effects in cancer cells but not cultured normal human bronchial epithelial cells. These data have provided the preclinical rationale for two active clinical trials examining gene induction mediated by DAC and DP, as well as another protocol evaluating sequential DAC/DP infusion in thoracic oncology patients scheduled to commence in the near future. Having observed that DAC, DP, and sequential DAC/DP mediate growth arrest irrespective of histology or tumor suppressor gene status, we have utilized cDNA microarray techniques to examine the mechanisms by which these agents mediate apoptosis preferentially in cancer cells. Thus far, this analysis has revealed highly complex effects of these agents in cancer cells without obvious patterns of induction relative to genotype. Several novel mechanisms have been identified, and results pertaining to some of these experiments have been submitted for publication. In addition to our in vitro experiments, we have used cDNA microarray techniques to analyze gene expression in tissue biopsy specimens following Decitabine, or Depsipeptide treatment of lung cancer patients. We can reliably amplify mRNA from FNA and bronchoscopic biopsy specimens from patients on these trials. Because the evaluation of gene induction in cancer specimens may be adversely affected by stromal contamination, current efforts are underway to optimize laser capture microdissection and RNA amplification techniques in order to enhance the specificity of genetic analysis.Thus far, evidence of target gene induction has been observed in several patients on these trials. Nearly one-third of all patients dying from extrathoracic malignancies suffer from pulmonary metastases. In preclinical studies, we have evaluated the pharmacokinetics and acute toxicity of paclitaxel administered by retrograde isolated lung perfusion techniques, and have observed significant enhancement of paclitaxel cytotoxicity in cancer cells by moderate hyperthermia. These experiments have provided the preclinical rationale for an ongoing Phase I study. Furthermore, we have extended our efforts pertaining to the regional treatment of pulmonary malignancies by examining the toxicity and potential efficacy of cytotoxic agents administered via novel aerosolization techniques. The following clinical protocols submitted by the Thoracic Oncology Section are open for patient accrual: 1) Phase I Study of Decitabine Mediated Induction of Tumor Antigen and Tumor Suppressor Gene Expression in Patients with Cancers Involving the Lungs, Esophagus, or Pleura. 2) Phase II Study of Gene Induction Mediated by Four-Hour Depsipeptide Infusion in Lung Cancer Patients. 3) Phase I Study of Retrograde Isolated Lung Perfusion with Paclitaxel and Moderate Hyperthermia in Patients with Unresectable Pulmonary Malignancies. 4) Phase I and Clinical Pharmacologic Study of Inhaled Doxorubicin in Adults with Advanced Solid Tumors Affecting the Lungs.

肺，食管和胸膜恶性肿瘤是通过癌化场中的多步骤机制而产生的。尽管它们的病因可能有所不同，但这些恶性肿瘤表现出通用的分子主题，包括通过RAS途径的异常信号转导，RB和p53肿瘤抑制途径的失活以及端粒酶的表达，所有这些都对恶性转化至关重要。除了破坏细胞周期调节外，癌基因和肿瘤抑制基因突变通过调节粘附分子的表达，基质金属蛋白酶以及血管生成的配体（如血管内皮生长因子）来影响胸腔肿瘤的转移表型。染色质结构和DNA甲基化中的全球危险使各种肿瘤抑制基因静音促进了胸腔恶性肿瘤中几种癌症抗原的从头表达。 涉及ERBB1/2，Cyclin D，RB，P16，P14/ARF和P53的突变是干预胸腔恶性肿瘤及其前体病变的有吸引力的靶标。 SV40癌蛋白可能是恶性胸膜间皮瘤分子干预的独特靶标。在已发表的研究中，我们已经证明，17-AAG和相关化合物对ERBB信号转导显着提高了对化学治疗剂的敏感性，并抑制了肺和食管癌细胞的转移表型。 额外的实验室工作重点是分析由DNA脱甲基剂和组蛋白脱乙酰基酶（HDAC）抑制剂介导的胸部恶性肿瘤的基因诱导。我们已经观察到，在暴露条件下，促进肿瘤抑制基因的表达，例如p16、5 AZA 2'DEXYCYTIDINE（DAC）介导了MAGE-3和NY-ESO-ESO-1癌症睾丸抗原的诱导。此外，我们已经证明了DAC介导的靶基因诱导可以通过HDAC抑制剂Depspyphetide FR901228（DP）增强。此外，我们已经表明，在暴露于DAC或顺序DAC/DP之后，癌细胞可以通过针对NY-ESO-1的细胞溶解T淋巴细胞识别，并且顺序DAC/DP治疗介导了癌细胞中的深刻凋亡效应，但未培养的正常人支气管人体支气管上皮细胞。这些数据为两项活跃的临床试验提供了临床前原理，该试验研究了由DAC和DP介导的基因诱导，以及另一种评估胸腔肿瘤学患者的顺序DAC/DP输注的方案，该患者计划在不久的将来开始。 在观察到DAC，DP和顺序DAC/DP介导了生长停滞，而与组织学或肿瘤抑制基因状态无关，我们已经利用cDNA微阵列技术来检查这些试剂在癌细胞中优先介导凋亡的机制。到目前为止，该分析揭示了这些药物在癌细胞中的高度复杂作用，而没有明显的诱导模式相对于基因型。已经确定了几种新型机制，并且与其中一些实验有关的结果已提交出版。 除了我们的体外实验外，我们还使用了cDNA微阵列技术来分析解替滨后组织活检标本中的基因表达，或对肺癌患者的二肽治疗。我们可以可靠地从患者的FNA和支气管镜活检标本中可靠地放大这些试验中的MRNA。由于对癌症标本中基因诱导的评估可能会受到基质污染的不利影响，因此目前正在努力优化激光捕获的显微解剖和RNA扩增技术，以增强遗传分析的特异性，因此，在这些试验中，已经在几个患者中观察到了靶基因诱导的证据。 几乎三分之一因牙外恶性肿瘤而死的患者患有肺转移。在临床前研究中，我们已经评估了通过逆行分离的肺灌注技术给予紫杉醇的药代动力学和急性毒性，并观察到通过中度热心理中癌细胞中紫杉醇细胞毒性的显着增强。这些实验为正在进行的I期研究提供了临床前原理。此外，我们通过检查通过新型的雾化技术施用的细胞毒性剂的毒性和潜在疗效，扩大了与肺部恶性肿瘤区域治疗有关的努力。 胸部肿瘤科提交的以下临床方案开放供患者应计： 1）第一阶段的研究替替滨介导的肿瘤抗原和肿瘤抑制基因表达的诱导患者，涉及肺，食道或胸膜的癌症。 2）在肺癌患者中由四小时二肽输注介导的基因诱导的II期研究。 3）I阶段的研究对无法切除的肺部恶性肿瘤的患者逆行分离的肺灌注和中等温度。 4）在影响肺的晚期实体瘤的成年人中吸入的阿霉素的I期和临床药理学研究。