Structure-Functions Studies of Twist1-induced Radioresistance in Lung Cancer

Twist1 诱导的肺癌放射抗性的结构功能研究

• 批准号: 8658053
• 负责人: Phuoc T. Tran
• 金额: $ 33.47万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8658053
• 关键词: Bioinformatics; Biology; Blood Vessels; Cancer Etiology; Cancer Model; Cell Cycle; Cells; Cessation of life; ChIP-on-chip; Chest; Collaborations; Colon Carcinoma; DNA Repair; Development; Diagnosis; Disease; Embryo; Epithelial Cells; Epithelium; Etiology; Fibroblasts; Future; Gene Expression; Gene Targeting; Helix-Turn-Helix Motifs; Histologic; Human; Hypoxia; In Vitro; Knowledge; Lung; Lung Neoplasms; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of prostate; Medical; Methods; Modeling; Molecular; Mus; Non-Small-Cell Lung Carcinoma; Outcome; Pathway interactions; Patients; Phenotype; Platinum; Primary Neoplasm; Radiation; Radiation Oncologist; Radioresistance; Recurrence; Research; Research Design; Role; Staging; Structure; Structure-Activity Relationship; TWIST1 gene; Tertiary Protein Structure; Testing; Therapeutic; Tissues; Transgenic Organisms; Treatment outcome; Unresectable; Veterinarians; Work; base; cancer care; cancer cell; cancer therapy; cell type; chemotherapy; genome-wide; in vivo; insight; malignant breast neoplasm; mouse model; mutant; neoplastic cell; non-invasive imaging; novel; overexpression; pre-clinical; public health relevance; radiation resistance; response; therapeutic target; transcription factor; tumor

DESCRIPTION (provided by applicant): Unresectable locally advanced non-small cell lung cancer treated with concurrent platinum-based chemotherapy and thoracic radiation results in primary tumor recurrence in >35-80% of cases from radioresistant tumors. Therefore, research aimed at understanding mechanisms of radioresistance and strategies to overcome this problem are paramount to better lung cancer treatment outcomes. We found TWIST1 was commonly overexpressed in human lung cancers and Twist1 could confer radiation resistance to lung epithelial cells, lung cancer cells and autochthonous mouse lung tumors in vivo. Twist1 is a basic helix- loop-helix transcriptional factor. The domains of Twist1 and transcriptional targets of Twist1 that are required for radioresistance are unknown. In order to maximize the therapeutic potential of inhibiting Twist1 for lung cancer, we must precisely define the protein domains of Twist1 and/or the downstream transcriptional targets of Twist1 required for radioresistance. In the present proposal we have three specific aims to test two hypotheses that will provide more mechanistic insight into the structure-function relationships of Twist1 and lung cancer radioresistance. One central hypothesis of this proposal is that specific protein domains of Twist1 are required for radioresistance. We have developed a novel lung epithelium specific Twist1 mouse model and various mutant versions of Twist1 to facilitate these studies. Our second testable hypothesis is there is a defined set of "core" transcriptional targets of Twist1 required for radioresistance. The studies outlined in our proposal will provide valuable knowledge to the pathophysiologic role of Twist1 in lung cancer and Twist1- induced radioresistance. Finally, validating a novel preclinical inducible lung cancer model of radioresistance will create a model to test new treatments for this fatal disease in combination with radiation. Specific Aim #1: Characterize the domains of Twist1 required for radioresistance in vitro. We will use mouse embryo fibroblasts (MEFs) and isogenic human lung cancer cells expressing Twist1 and Twist1 mutants to examine which domains of Twist1 are necessary for a radioresistant phenotype. We will determine the cell autonomous mechanisms of Twist1-induced radioresistance in vitro. Specific Aim #2: Characterize the domains of Twist1 required for radioresistance of KrasG12D-induced autochthonous lung tumors. We will test this hypothesis with serial non-invasive imaging and confirm our findings with standard molecular-histologic methods to follow the response to radiation in our novel Twist1 transgenic lung tumor model. We will also examine the dominant mechanisms for Twist1-dependent radiation response modulation in vivo. Specific Aim #3: Determine Twist1 transcriptional targets correlated with Twist1-induced radioresistance using gene expression and ChIP-chip profiling. We will use integrative genome-wide bioinformatic approaches on MEFs and human lung cancer cells to isolate the "core" gene targets required for Twist1-induced radioresistance.

描述（由申请人提供）：通过基于铂的化学疗法和胸腔放射治疗的不可切除的局部晚期非小细胞肺癌导致原发性肿瘤复发，> 35-80％的辐射疗法肿瘤中的病例中，有35-80％。因此，旨在理解放射性机制和克服该问题的策略机制的研究至关重要。我们发现Twist1通常在人类肺癌中过表达，Twist1可以在体内对肺上皮细胞，肺癌细胞和自围候小鼠肺肿瘤赋予辐射抗性。 Twist1是基本的螺旋环螺旋转录因子。辐射抗性所需的Twist1和转录靶标的域是未知的。为了最大化抑制肺癌扭曲1的治疗潜力，我们必须精确地定义Twist1和/或放射性抗抑郁所需的Twist1的下游转录靶标的蛋白质结构域。在本提案中，我们有三个特定的目的，旨在检验两个假设，这些假设将为Twist1和肺癌放射线的结构功能关系提供更多的机械洞察力。该提议的一个中心假设是散热器需要特定的蛋白质结构域。我们已经开发了一种新型的肺上皮特异性扭曲1小鼠模型和扭曲的各种突变版本，以促进这些研究。我们的第二个可检验的假设是，辐射抗性所需的一组定义的“核心”转录靶标。我们的提案中概述的研究将为Twist1在肺癌和Twist1诱导的放射线抗性中的病理生理作用提供宝贵的知识。最后，验证一种新型的放射线诱导型诱导肺癌模型将创建一个模型，以测试这种致命疾病与辐射结合使用的新治疗方法。特定目标＃1：表征体外放射线所需的扭曲1的域。我们将使用小鼠胚胎成纤维细胞（MEF）和表达Twist1突变体的等源性人肺癌细胞来检查twist1的哪些结构域对于放射性表型是必需的。我们将在体外确定扭曲1诱导的辐射抗性的细胞自主机制。具体目标＃2：表征KRASG12D诱导的自动肺肺肿瘤所需的Twist1域。我们将通过串行非侵入性成像检验该假设，并使用标准分子 - 历史方法确认我们的发现，以遵循我们新型Twist1转基因肺肿瘤模型的辐射反应。我们还将检查体内依赖于扭曲的辐射反应调制的主要机制。特定目标＃3：确定使用基因表达和芯片芯片分析与扭曲1诱导的放射线相关的Twist1转录靶标。我们将在MEF和人类肺癌细胞上使用全基因组生物信息学方法来隔离扭曲1诱导的放射线所需的“核心”基因靶标。