(PQC1) PLK1 And EPHX3 Promotion Of Genomic Instability In Bronchial Dysplasia

(PQC1) PLK1 和 EPHX3 促进支气管发育不良的基因组不稳定性

基本信息

批准号：
8928116
负责人：
Daniel Thomas Merrick
金额：
$ 17.27万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-16 至 2018-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8928116
关键词：
Affect American Apoptosis Apoptotic Area Aromatic Polycyclic Hydrocarbons Biopsy Breast Cancerous Carcinogen exposure Carcinoma Cell Array Analyses Cell Culture Techniques Cell Cycle Arrest Cell Cycle Progression Cell Line Cell physiology Cells Cervical Chemoprevention Chemopreventive Agent Clinical Research Clone Cells Colorectal Coupled DNA Damage DNA Sequence Alteration Development Diagnosis Diagnostic Dysplasia EPHX1 gene Early Diagnosis Early Intervention Enzyme Inhibition Enzyme Inhibitor Drugs Enzyme Inhibitors Enzymes Epigenetic Process Epithelial Epithelial Cells Epithelium Epoxide hydrolase Excision Frequencies Gene Expression Gene Expression Profiling Genes Genetic Genomic Instability Genomics Goals Health In Vitro Incidence Individual Induction of Apoptosis Learning Lesion Lung Malignant - descriptor Malignant Epithelial Cell Malignant Neoplasms Malignant neoplasm of lung Measures Mediating Mediator of activation protein Metabolic Marker Methylation Molecular Abnormality Mutagenesis Mutation Natural History Operative Surgical Procedures Outcome PLK1 gene Pancreas Pathway Analysis Patients Phosphorylation Premalignant Premalignant Cell Preventive Process Property Prostate Proteins Publications Refractory Repetitive Sequence Reporting Resources Risk Role Sampling Site Squamous Cell Lung Carcinoma Squamous cell carcinoma System Testing Therapeutic Intervention Time Tobacco Tobacco smoke airway epithelium body system bronchial epithelium cancer prevention cancer risk chemotherapy cigarette smoking cost differential expression enzyme activity genome sequencing genome-wide high risk human PLK1 protein inhibitor/antagonist interest killings medical complication mortality overexpression prevent prognostic targeted treatment

项目摘要

DESCRIPTION (provided by applicant): Bronchial dysplasia (BD) is a precancerous lesion that can progress to squamous cell carcinoma (SCC). Identification of lesions with high risk for progression is important to help identify cellular mechanisms underlying malignant transformation and to help identify patients that would benefit from preventive measures. We have compared high risk persistent BD to regressive BD by gene expression array analysis and identified more than 300 genes that are differentially expressed in persistent BD. Using a pathway analysis to identify genes with a strong relationship to persistence, we have identified two factors, polo-like kinase (PLK1) and epoxide hydrolase 3 (EPHX3), which demonstrate a synergistic relationship with persistence and suggest a potential interaction in promoting genomic instability in BD. EPHX3 converts cigarette smoke associated pro- carcinogenic polycyclic aromatic hydrocarbons (PAHs) to mutagenic metabolites. PLK1 promotes cell cycle progression through the G2-M checkpoint. This checkpoint normally operates to arrest the cell cycle when genetic damage is present with the subsequent induction of apoptosis in cells with extensive damage. We have hypothesized that the combination of increased mutational capacity coupled with G2-M checkpoint abrogation and subsequent cell cycle progression promotes and establishes genetic damage that mediates progression to invasive lung cancer. Furthermore, specific inhibitors of these factors are available suggesting a potential mechanism for preventing the development of lung SCC. We propose studies to establish the effect of PLK1 and EPHX3 inhibition on progression associated cellular activities and to assess the role of PLK1 and EPHX3 overexpression in promoting genomic instability using bronchial cell cultures established from persistent BD and normal bronchial epithelium. The first aim of the proposal will expand on preliminary studies that indicate PLK1 inhibition with Volasertib (Boehringer-Ingelheim) significantly reduces proliferation and induces apoptotic activity in cultured persisten BD derived epithelial cells without affecting these properties in normal epithelial cultures. Similarly, we have shown that the EPHX3 inhibitor, AUDA, reduces EPHX3 activity in SCC cell lines and will study its effects in BD derived cultures. The second aim will evaluate the role of genomic instability in persistent BD by characterizing the types of alterations and quantifying the amount of damage that are associated with carcinogen exposure. Genetic instability induced by tobacco smoke condensate treatment of persistent BD and normal bronchial cultures will be measured by genome-wide sequencing and methylation array analyses of cell lines cultured in the presence and absence of PLK1 and EPHX inhibitors to test our hypothesis. Long term goals that would be facilitated by the demonstration of PLK1 and EPHX3 dependent genomic instability include the establishment of an important cellular mechanism through which progression associated activities are mediated and the provision of rationale for the development of potentially effective chemopreventive therapy targeting these enzymes.

描述（由申请人提供）：支气管发育不良（BD）是一种癌前病变，可以发展为鳞状细胞癌（SCC）。鉴定进展高风险的病变对于帮助识别恶性转化的细胞机制很重要，并帮助识别将受益于预防措施的患者。我们通过基因表达阵列分析比较了高风险持续的BD与回归BD，并确定了300多个在持续BD中差异表达的基因。使用途径分析来鉴定与持久性有很强关系的基因，我们已经确定了两个因素，分别是类似polo样激酶（PLK1）和环氧水解酶3（EPHX3），它们证明了与持久性的协同关系，并暗示了BD中促进基因组不稳定性的潜在相互作用。 EPHX3将香烟烟与致癌的多环芳烃（PAHS）转化为诱变代谢物。 PLK1通过G2-M检查点促进细胞周期进程。当存在遗传损伤时，该检查点通常会在遗传损伤中造成细胞周期，随后在具有广泛损害的细胞中凋亡的后续诱导。我们假设，增加的突变能力与G2-M检查点废除和随后的细胞周期进展相结合会促进并建立遗传损害，从而介导进展为侵入性肺癌。此外，可以使用这些因素的特定抑制剂，这表明了防止肺SCC发展的潜在机制。我们提出的研究是建立PLK1和EPHX3抑制对相关细胞活性的影响，并评估PLK1和EPHX3过表达在使用持续性BD和正常支气管上皮的支气管细胞培养物中使用支气管细胞培养物来促进基因组不稳定性的作用。该提案的第一个目的将扩大初步研究，该研究表明PLK1抑制volasertib（Boehringer-ingelheim）可显着减少增殖，并诱导培养的持久性BD衍生的上皮细胞中的凋亡活性，而不会影响正常上皮培养物中这些特性。同样，我们已经表明，EPHX3抑制剂AUDA降低了SCC细胞系中的EPHX3活性，并将研究其在BD衍生培养物中的作用。第二个目标将通过表征变化的类型并量化基因组不稳定性在持久性BD中的作用与致癌物暴露有关的损害量。烟草烟雾凝结物对持续性BD和正常支气管培养物的治疗引起的遗传不稳定性将通过在存在和不存在PLK1和EPHX抑制剂的存在下培养的细胞系的细胞系来衡量，以测试我们的假设。 PLK1和EPHX3依赖性基因组不稳定性的演示将促进的长期目标包括建立重要的细胞机制，通过该机制介导了相关活性，并为开发具有潜在有效的化学预防疗法的基本原理提供了靶向这些酶的靶向。