Protein Profiling in Nipple Aspirate Fluid

乳头抽吸液中的蛋白质分析

• 批准号: 6948527
• 负责人: EDWARD R SAUTER
• 金额: $ 20.32万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-10 至 2007-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6948527
• 关键词: biomarker; biopsy; body fluids; breast neoplasm /cancer diagnosis; breast neoplasms; clinical research; cytology; diagnosis design /evaluation; early diagnosis; female; fine needle aspiration; gene expression; human subject; liquid chromatography mass spectrometry; noninvasive diagnosis; proteomics; women' s health

DESCRIPTION (provided by applicant) Currently available breast cancer screening tools such as mammography and breast examination miss up to 40% of early breast cancers, are least effective in detecting cancer in young women whose tumors are often more aggressive, and requires that an invasive needle or surgical biopsy be performed when an area of suspicion is identified to confirm the presence of malignancy. Our goal is to develop a noninvasive method to assist in the early detection of breast cancer or precancerous changes. The primary focus of this study is to detect promising markers in nipple aspirate fluid (NAF) by performing a comprehensive analysis of the proteins that are expressed (protenomics). SELDI (Surface Enhanced Laser Desorption/Ionization) mass spectrometry methods will be used for the analyses. We have compelling preliminary data using protenomic analysis of NAF that proteins of a given molecular weight can be identified which are highly expressed in women with breast cancer and are expressed little or not at all in women without disease. These analyses can be performed using one microliter of NAF. We wish to study women scheduled to undergo diagnostic breast biopsy. This population would be well served if we could determine whether someone has breast cancer without submitting them to an invasive procedure. Specifically, in the feasibility aspect of the study (R21 component, Phase I), we will: 1) determine the protein expression pattern in NAF among women with and without breast cancer using SELDI and 2) identify promising candidate markers in NAF that can be used in screening for the presence of breast cancer. We will establish quantifiable milestones which must be attained in order to proceed to further development of a predictive model among women scheduled to undergo breast biopsy. Specifically, in the second stage of the study (R33 component, Phase II), we will use the markers identified in Phase I, along with a number of other clinical measures, to predict the presence of both precancer and invasive breast cancer. In addition, we will accurately quantitate and identify the sequences of the proteins identified by SELDI and the majority of proteins present in NAF using tandem mass spectrometry (MS/MS) methods in combination with the use of isotope-coded affinity tags (ICATs) and microbore liquid chromatography/MS/MS. Our long term goal is to develop and validate the use of markers in NAF to noninvasively evaluate the breast for cancer, which we hope will serve as an adjunct to currently available methods of risk assessment and disease detection.

描述（由申请人提供） 目前可用的乳腺癌筛查工具，例如乳房X线摄影和 乳房检查错过了多达40％的早期乳腺癌，最不可能有效 在发现肿瘤通常更具侵略性的年轻女性的癌症时， 需要在区域时进行侵入性针头或手术活检 识别怀疑以确认存在恶性肿瘤。我们的目标是 开发一种无创方法来协助乳房早期检测 癌症或癌前变化。这项研究的主要重点是检测 乳头抽吸流体（NAF）中有前途的标记 分析表达的蛋白质（蛋白质组学）。 Seldi（表面 将使用增强激光解吸/电离）质谱法 用于分析。我们使用Protenomic拥有引人入胜的初步数据 可以鉴定出给定分子量的蛋白质的NAF分析 在乳腺癌女性中高度表达并表达 在没有疾病的女性中，几乎没有或根本很少。这些分析可以进行 使用NAF的一个微量柱。我们希望研究计划接受的女性 诊断乳房活检。如果我们可以 确定某人是否患有乳腺癌而不将其提交给 侵入性程序。具体而言，在研究的可行性方面（R21 组件，I阶段），我们将：1）确定蛋白质表达模式 使用SELDI和患有乳腺癌和没有乳腺癌的女性中的NAF，2）确定 NAF中有希望的候选标记，可用于筛选 乳腺癌的存在。我们将建立可量化的里程碑 为了进一步发展预测性，必须达到 计划进行乳房活检的女性中的模型。具体来说，在 研究的第二阶段（R33组件，第二阶段），我们将使用标记 在第一阶段确定，以及许多其他临床指标 预测前癌和侵入性乳腺癌的存在。在 此外，我们将准确量化并确定 SELDI鉴定的蛋白质和NAF中存在的大多数蛋白质使用 串联质谱法（MS/MS）方法与使用 同位素编码的亲和力标签（ICAT）和微孔液相色谱/MS/MS。 我们的长期目标是开发和验证NAF中使用标记的使用 非侵入性评估癌症的乳腺癌，我们希望这将成为当前可用的风险评估和疾病方法的辅助 检测。