GRSA Proteins in the Diagnosis of Breast Cancer

GRSA 蛋白在乳腺癌诊断中的应用

• 批准号: 6515212
• 负责人: WILLIAM G NORTH
• 金额: $ 15.9万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6515212
• 关键词: MCF7 cell; athymic mouse; breast neoplasm /cancer diagnosis; breast neoplasms; diagnosis design /evaluation; early diagnosis; glycopeptides; human tissue; immunocytochemistry; molecular cloning; monoclonal antibody; neoplasm /cancer immunodiagnosis; neoplasm /cancer relapse /recurrence; polymerase chain reaction; surface antigens; tumor antigens; vasopressins; western blottings

DESCRIPTION (provided by applicant): The objective of this project is to test the hypothesis that improved diagnostic techniques for breast cancer can be generated with agents recognizing GRSA proteins. Such agents comprise monoclonal antibodies and engineered single-chain variable region (scFv) forms of these antibodies. GRSA (Glycopeptide-Related Cell Surface Antigen) proteins are specific tumor products that result from the expression of the vasopressin (VP) gene. They are produced by seemingly all breast cancer and all ductal carcinoma in Situ (DCIS). Alternatively, VP gene expression and GRSA proteins are absent from normal breast tissue and from all breast fibrocystic conditions including atypical ductal hyperplasia (ADH). A confirmation of this distinction should provide a method for the early detection of initial and recurrent disease. Preliminary studies indicate GRSA proteins could represent a target for immunodiagnosis and for future therapeutic interventions. Therefore a principal focus of research over the funding period is to generate and evaluate ScFv GRSA-directed molecules from available antibodies for these purposes. Specific Aims are directed at: (i) assessing new immunohistochemical and RT-PCR diagnostic procedures for breast cancer and DCIS that feature monoclonal antibodies against GRSA proteins or primers for VPmRNA; (ii) manufacturing single-chain scFv diabodies and triabodies representing our monoclonal antibodies that recognize GRSA proteins on breast cancer; (iii) appraising these scFv molecules as useful clinical agents in vitro based on their ability to provide positive immunostaining of all breast carcinoma in an archival library, and to bind to cancer cells in culture, and; (iv)appraising these scFv molecules as useful clinical agents in vivo based on their ability to selectively target GRSA proteins on mouse tumor xenografts. Studies will employ RT-PCR, ligation, and cloning; ABC immunohistochemistry, Western analysis; flow cytometry; RIA; radiometric analysis of tissues; and gamma-camera scintigraphy. Expected outcomes of this research are the development of simple, sensitive, reliable, and universally available diagnostic methods that distinguish DCIS and breast cancer from atypical hyperplasia in tissue biopsies, and that can detect and localize recurrent disease in patients through radioimmunoimaging. These methods could be applicable to all patients with breast cancer or suspected breast cancer.

描述（由申请人提供）：该项目的目的是测试 改善乳腺癌诊断技术的假设可以是 由识别GRSA蛋白的药物产生。这样的代理人包括 单克隆抗体和工程单链变量区域（SCFV）形式 这些抗体。 GRSA（与糖肽相关的细胞表面抗原）蛋白 是由加压素表达产生的特定肿瘤产物 （VP）基因。它们是由所有乳腺癌和所有导管产生的 原位癌（DCIS）。另外，VP基因表达和GRSA蛋白 正常乳房组织和所有乳腺纤维囊性条件不存在 包括非典型导管增生（ADH）。对这种区别的确认 应该提供一种早期检测初始和经常性的方法 疾病。初步研究表明GRSA蛋白可以代表目标 用于免疫诊断和未来的治疗干预措施。因此 资金期间研究的主要重点是生成和评估 出于这些目的，SCFV GRSA指导的分子。 具体目标针对：（i）评估新的免疫组织化学和RT-PCR 具有单克隆的乳腺癌和DCI的诊断程序 针对GRSA蛋白的抗体或VPMRNA的引物； （ii）制造 代表我们的单克隆的单链SCFV糖尿病生和Triabodies 识别乳腺癌上GRSA蛋白的抗体； （iii）评估 这些SCFV分子是根据其能力作为有用的临床剂 提供档案中所有乳腺癌的阳性免疫染色 图书馆，并与培养中的癌细胞结合； （iv）评估这些SCFV 分子作为体内有用的临床剂，基于它们的能力 选择性地靶向小鼠肿瘤异种移植物上的GRSA蛋白。研究将采用 RT-PCR，连接和克隆； ABC免疫组织化学，西方分析；流动 细胞仪；瑞亚组织的辐射分析；和伽玛相机闪烁显像。 这项研究的预期结果是简单，敏感， 可靠且普遍可用的诊断方法，可区分DCIS 以及组织活检中非典型增生的乳腺癌，这可以 通过放射免疫成像检测并定位患者的复发性疾病。 这些方法可能适用于所有乳腺癌患者或 怀疑的乳腺癌。