SPRL--A MOLECULAR MARKER FOR BRONCHIAL PRENEOPLASIA

SPRL--支气管癌前病变的分子标记

• 批准号: 2895441
• 负责人: DERICK LAU
• 金额: $ 8.65万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-07-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2895441
• 关键词: DNA footprinting; biomarker; bronchus neoplasm; cell transformation; cellular oncology; expression cloning; gel mobility shift assay; gene expression; genetic regulatory element; human tissue; immunocytochemistry; in situ hybridization; metastasis; molecular cloning; nuclear runoff assay; oncoproteins; preneoplastic state; proline; respiratory epithelium; squamous cell carcinoma; tissue /cell culture; transcription factor

DESCRIPTION (Applicant's Description): This program is designed to establish the applicant as an independent investigator of biomarker research in bronchial carcinogenesis, and in early detection and chemoprevention of cancers of the upper aerodigestive tract. The applicant has previous research experience in molecular and cellular pharmacology and has recently become an established medical oncologist specializing in the treatment of lung cancer. The sponsor is an accomplished pulmonary physiologist and molecular biologist in the field of development and differentiation of bronchial epithelium, and in the area of regulation of biomarkers of the bronchial epithelium in response to environmental toxins such as ozone and tobacco smoke. The plans of the sponsor are to provide supervision to the applicant to acquire expertise in molecular techniques, in the design and execution of experiments to elucidate the mechanisms of regulation of a molecular marker, spr1, associated with bronchial metaplasia, and in the application of this biomarker for early detection of lung cancer. The sponsor has cloned a small proline-rich protein (spr-1) which is overexpressed in tracheobronchial epithelium undergoing metaplastic change from a normal mucociliary phenotype to a squamous appearance. The spr1 expression is down-regulated by vitamin A and up-regulated by tumor promoters. Therefore, it appears that spr1 is a potential early biomarker for preneoplastic transformation of the bronchial epithelium. In their laboratory, there is a series of human bronchial epithelial cells displaying varied degree of spr1 expression and tumorigenicity potential. This proposal is, thus, intended to use this cell-culture model to elucidate the mechanism of regulation of spr1 expression in the multistep carcinogenesis of the bronchial epithelium, and to use archival specimens to map spr1 expression in the field of cancerization of lung tissues from patients with lung cancer. The first aim is to test the hypothesis that there is an increasing expression of spr1 in the field of cancerization extending from a focus of bronchial squamous carcinoma to the surrounding dysplastic, metaplastic and normal bronchial epithelia as determined by immunohistochemical staining and in situ hybridization of archival human squamous lung carcinoma. The second aim is to use a human tracheobronchial cell model to test the hypothesis that the cell type-specific expression of spr1 is regulated at the level of transcription by using a nuclear run-on transcriptional assay and a transfection study with chimeric constructs of the spr1 promoter region attached to a reporter gene. The third aim is to identify the cis-elements and the transcriptional factors involved in the regulation of spr1 gene by employing the techniques of DNA-binding mobility shift gel assay, DNA footprinting and expression cloning. These studies are to set the stage for using spr1 as a biomarker for identifying the early steps of multistep carcinogenesis, for early detection of lung cancer, and for testing the efficacy and mechanisms of chemopreventive agents in reversing malignant transformation of the bronchial epithelium.

描述（申请人的描述）：该计划旨在 使申请人成为生物标志物研究的独立研究者 支气管癌的发生以及早期检测和化学预防 上呼吸消化道癌症。 申请人曾有过 分子和细胞药理学方面的研究经验，最近 成为一名资深的肿瘤内科医师，专门从事以下疾病的治疗 肺癌。 申办者是一位卓有成就的肺生理学家，并且 发育和分化领域的分子生物学家 支气管上皮，以及生物标志物的调节领域 支气管上皮对臭氧等环境毒素的反应 烟草烟雾。 发起人的计划是向 申请人获得分子技术、设计和 进行实验以阐明调节机制 分子标记 spr1 与支气管化生相关，并且在 该生物标志物在肺癌早期检测中的应用。 申办者克隆了一种富含脯氨酸的小蛋白（spr-1），它是 在经历化生改变的气管支气管上皮中过度表达 从正常的粘膜纤毛表型变为鳞状外观。 spr1 表达被维生素 A 下调，并被肿瘤上调 发起人。 因此，spr1 似乎是一个潜在的早期生物标志物 用于支气管上皮的癌前转化。 在他们的 实验室中，有一系列人类支气管上皮细胞显示 spr1表达程度和致瘤性潜力不同。 这 因此，该提案旨在利用这种细胞培养模型来阐明 spr1表达在多步致癌过程中的调控机制 支气管上皮，并使用档案标本来绘制 spr1 患者肺组织癌化领域的表达 肺癌。 第一个目的是检验以下假设： spr1在癌化领域的表达从焦点延伸 支气管鳞癌向周围的不典型增生、化生和 通过免疫组织化学染色确定正常支气管上皮细胞 档案人类鳞状肺癌的原位杂交。 第二个 目的是使用人类气管支气管细胞模型来检验该假设 spr1 的细胞类型特异性表达在以下水平受到调节 通过使用核连续转录测定和 spr1启动子区嵌合构建体的转染研究 附着在报告基因上。 第三个目标是识别顺式元素 以及参与spr1基因调控的转录因子 采用 DNA 结合迁移率位移凝胶测定技术，DNA 足迹和表达克隆。 这些研究旨在为使用 spr1 作为生物标记物奠定基础 确定多步致癌的早期步骤，以便早期发现 肺癌，并测试其功效和机制 化学预防剂逆转肿瘤恶变 支气管上皮。