A biosensor to detect ERE-binding proteins in cancer

描述（由申请人提供）：生物传感器提供了包含分子相互作用信息的快速数据，这些信息不容易使用平衡测定或组织化学方法获得。这项研究将是一系列长期目标中的第一项，即开发应用于乳腺癌活检提取物的快速生物传感器方法，用于（1）预测淋巴结阴性和淋巴结阳性乳腺癌的复发风险，（2）识别肿瘤中先前未识别的ERE结合组分的存在，以及（3）获得导致选择对于给定个体的乳腺癌具有高成功可能性的有效治疗性治疗方式的信息。作为该系列的第一项研究，本研究主要集中在建立新的生物传感器方法，该方法与肿瘤组织活检提取物中代表的癌症的预后信息相关。所提出的方法利用IA，Inc.的生物传感器技术与广泛的肿瘤银行和相关的记录，并在路易斯维尔大学的受体制备和测定资源。具体目标是：（1）使用基于抗体的传感器来评估乳腺癌活检提取物中已建立的肿瘤标志物，（2）评估高风险和低风险淋巴结阴性和淋巴结阳性活检的所选肿瘤提取物与标准化hER制剂竞争结合至携带各种ERE的传感器的能力。将使用已知与肿瘤发生过程相关的基因的启动子区上游发现的ERE序列。包括乳腺癌中发现的蛋白质基因，这些蛋白质与患者对他莫昔芬治疗的反应的预后相关，如组织蛋白酶D和pS2，以及转录因子h-fos和jun的基因。（3）将生物传感器结果与电泳迁移率变化和超迁移分析的数据以及患者特征和临床随访的回顾性分析相关联。(4)比较有和没有4-OH-他莫昔芬的肿瘤提取物与ERE纤维的结合，并将结果与治疗结果数据相关联。这种采用微型生物传感器技术的新方法有望快速评估肿瘤标志物，并发现具有临床意义的新的癌症相关调节蛋白。它可以揭示肿瘤中以前未被识别的ERE结合蛋白，并将导致生物传感器产品，这是一种改进的方法，用于识别最有可能对个体癌症产生积极结果的治疗方式。 II期研究将扩大样本基础，并将包括更广泛的用于肿瘤标志物鉴定的抗体。将包括共激活子和共阻遏物相互作用以及ERE和其他反应元件相互作用。将对先前未识别的ERE结合组分进行表征。如果4-OH-他莫昔芬传感器数据与治疗结果相关性良好，则将对其他治疗药物进行类似评估。

DESCRIPTION (provided by applicant): Biosensors provide rapid data containing information on molecular interactions that are not easily available using equilibrium assays or histochemical methods. This study will be the first of a series with long-term objectives of developing rapid biosensor methods applied to extracts of breast cancer biopsies for (1) predicting risk of recurrence in both node negative and node positive breast cancers, (2) recognizing the presence of previously unidentified ERE-binding components in tumors, and (3) obtaining information leading to selection of effective therapeutic treatment modalities having a high likelihood of success for a given individual's breast cancer. As the first of the series, this study focuses primarily on establishing novel biosensor methods, which correlate with prognostic information for the cancer represented in extracts of tumor tissue biopsies. The proposed methods utilize IA, Inc.'s biosensor technology in conjunction with the extensive tumor bank and associated records, and resources for receptor preparation and assay at the University of Louisville. Specific aims are: (1) Use antibody-based sensors to assess established tumor markers in breast cancer biopsy extracts, (2) Assess capacity of selected tumor extracts of high risk and low risk node negative and node positive biopsies to compete with standardized hER preparation for binding to sensors bearing various EREs. ERE sequences found upstream of the promotor region of genes having known relevance to processes of tumorigenesis will be used. Included will be genes for proteins found in breast cancers, which have been associated with prognosis for patient response to Tamoxifen treatment, such as cathepsin D and pS2, and genes for transcription factors h-fos and jun. (3) Correlate the biosensor results with data from electrophoretic mobility shift and supershift assays and with retrospective analysis of patient characteristics and clinical follow-up. (4) Compare binding of tumor extract to ERE fibers with and without 4-OH-tamoxifen and correlate results with treatment outcome data. This novel approach with miniaturized biosensor techniques holds promise for rapid assessment of tumor markers and discovering new cancer associated regulatory proteins of clinical significance. It could reveal previously unrecognized ERE-binding proteins in tumors, and will lead to a biosensor product, which is an improved method for identifying treatment modalities most likely to produce a positive outcome for an individual cancer. Phase II studies will expand the sample base and will include a broader range of antibodies for tumor marker identifications. Co-activator and co-repressor interactions as well as ERE and other response element interactions will be included. Previously unidentified ERE binding-components will be characterized. If the 4-OH-tamoxifen sensor data correlates well with treatment outcome, additional therapeutic agents will be similarly assessed.