Core D: Molecular Diagnostics

核心 D：分子诊断

• 批准号: 7728770
• 负责人: Dave S B Hoon
• 金额: $ 21.03万
• 依托单位: SAINT JOHN'S CANCER INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7728770
• 关键词: Aliquot; Back; Biological Assay; Biological Markers; Blood; Cryopreservation; DNA; Data; Databases; Diagnosis; Diagnostic; Drops; Freezing; Funding; GPR-9-6 receptor; Guidelines; Hemorrhage; Histopathology; Immunohistochemistry; Individual; International; Label; Lasers; Light; Malignant Neoplasms; Messenger RNA; Metastatic Melanoma; Microdissection; Molecular; Molecular Genetics; Monitor; Neoplasm Metastasis; Nucleic Acids; Operative Surgical Procedures; Paraffin Embedding; Patients; Polymerase Chain Reaction; Primary Neoplasm; Process; Production; Publishing; Qualifying; Quality Control; RNA; RNA analysis; Randomized; Recombinants; Records; Resource Development; Reverse Transcriptase Polymerase Chain Reaction; Running; Sampling; Serum; Shipping; Ships; Site; Specimen; System; Tissues; Tumor Tissue; Validation; Visual; editorial; genetic analysis; improved; melanoma; programs; tumor; virtual

The Core, Aims I/I I, III are involved in the accrual of patients with paraffin-embedded (PE) SLNs from the individual multicenter sites. To date, we have received 1,143 patients who have entered into the trial that were qualified for RT-PCR randomization of which 29 were ineligible. Of these 274 were PCR+ (26%) and 794 were PCR- (74%). Forty patients were ineligible and 21 are still pending results. The PE tissue sections prepared as documented in the SOPs were sent from different sites in the USA or around the world to the core and been processed for PCR analysis. The studies have been running accordingly as planned. All centers are receiving results in a timely manner. Overall, there have been no major problems in the study workflow. Discussions have been carried out with each center to improve processing and shipping of tissue specimens. For centers which have not complied with our guidelines nor have poor quality specimens, were dropped from the PCR randomized study. For Aim IV, we are collecting PE and/or IHC positive SLN blocks from JWCI and from the MLST-I trial centers. The blocks have been sectioned for immunohistochemistry (IHC) and RNA isolation. The sections are being used to assess new melanoma IHC and RT-PCR biomarkers in Project II. For Aim V, we have been isolating and quantifying RNA from SLN, non-SLN and metastatic melanoma tissues. The RNA is isolated and cryopreserved for Project II. Over 200 tissue specimens have been processed. These specimens have served as a valuable resource for development and validation of new mRNA markers. In addition, we have received over 250 primary and metastatic tumor specimens from the Sydney Melanoma Unit for RNA analysis. The blocks have been sent to us, cut into sections, processed and sent back to them. These specimens have been important for studies we do not have particular types of melanomas or metastasis to specific sites. Aim VI, we have continued to improve the SOPs and streamline operations. We have expanded the ItemTracker program for all our specimen banking. Microsoft SQL Server serves as the database management engine, which enables ItemTracker to input and store millions of records. The system's intuitive visually-driven user interface incorporates functionality for barcodes, label making, data filtering, data fields customization, visual representation of storage layouts, and much more. All specimens are barcoded prior to freezing. We have a virtual specimen tracker in each freezer to monitor storage. In Aim VII, we have accrued 121 primary melanomas. Centers have provided tissue blocks whereby the Core cuts the tissues for IHC, and DNA/RNA analysis. The blocks are returned back to the center within a month. Specific subtypes of melanoma are being collected. These specimens are being used in Project II. DMA analysis and IHC have been performed on the samples and have shown good quality results. Studies such as the CCR9 receptor mRNA analysis was performed on these specimens. This was recently published in Clin Cancer Res and received a special editorial review. The primary tumor tissue was also used for assessment of methylated tumor-related DMA markers. The core performs laser microdissection using the Veritas Arcturus system equipped with both UV and laser light. For Aim VII, serum has been collected from the patients entered into the MSLT-II trial. The serum has been processed for molecular/genetic analysis. Bloods are being collected for molecular studies from selected MLST sites. The blood is received and processed under a specific molecular SOP and then cryostored. We have 1,836 bleeds from the multicenter sites. The bloods' serum is processed and will be used for DMA studies in Project II. Aim VIII will not be performed any longer as the need for this is not a major priority any longer due to funds available.

核心、目标 I/I I、III 涉及石蜡包埋 (PE) SLN 患者的累积 单个多中心站点。迄今为止，我们已接收了 1,143 名参加试验的患者，这些患者 符合 RT-PCR 随机化资格，其中 29 名不符合资格。其中 274 例为 PCR+ (26%)，794 例为 PCR+ PCR-（74%）。 40 名患者不符合资格，21 名患者仍在等待结果。 PE 组织切片 按照 SOP 中记录的方式准备的材料从美国或世界各地的不同地点发送到 核心并进行 PCR 分析处理。这些研究已按计划进行。全部 中心正在及时收到结果。总体来说，研究中没有出现大的问题 工作流程。已与每个中心进行讨论，以改善组织的加工和运输 标本。对于不遵守我们的指南或样本质量较差的中心， 从 PCR 随机研究中退出。 对于 Aim IV，我们正在从 JWCI 和 MLST-I 试验中收集 PE 和/或 IHC 阳性 SLN 块 中心。这些块已被切片用于免疫组织化学 (IHC) 和 RNA 分离。各部分 正在用于评估项目 II 中新的黑色素瘤 IHC 和 RT-PCR 生物标志物。 对于 Aim V，我们一直在分离和定量来自 SLN、非 SLN 和转移性黑色素瘤的 RNA 组织。为项目 II 分离并冷冻保存 RNA。已采集200多份组织样本 已处理。这些标本已成为开发和验证新产品的宝贵资源。 mRNA 标记。此外，我们还收到了超过 250 个来自该中心的原发性和转移性肿瘤标本。 悉尼黑色素瘤 RNA 分析中心。这些块已发送给我们，切割成段，加工并 送回给他们。 这些标本对于研究非常重要，我们没有特定类型的黑色素瘤或 转移到特定部位。 目标六，我们持续改进SOP并简化运营。我们已经扩大了 适用于我们所有样本银行的 ItemTracker 程序。 Microsoft SQL Server 作为数据库 管理引擎，使 ItemTracker 能够输入和存储数百万条记录。系统直观 视觉驱动的用户界面整合了条形码、标签制作、数据过滤、数据字段的功能 定制、存储布局的可视化表示等等。所有样本在检测前均已打上条形码 冷冻。我们在每个冰箱中都有一个虚拟样本跟踪器来监控存储情况。 在目标 VII 中，我们积累了 121 个原发性黑色素瘤。各中心提供了组织块，使 Core 切割组织以进行 IHC 和 DNA/RNA 分析。块在一个时间内返回到中心 月。正在收集黑色素瘤的特定亚型。这些标本正用于项目 II。 已对样品进行 DMA 分析和 IHC，并显示出良好的质量结果。研究 例如对这些标本进行CCR9受体mRNA分析。这是最近发布的 在临床癌症研究中，并接受了特别编辑评论。原发肿瘤组织也用于 甲基化肿瘤相关 DMA 标记物的评估。核心使用激光显微切割 Veritas Arcturus 系统配备紫外线和激光。 对于目标 VII，已从参加 MSLT-II 试验的患者中收集了血清。血清有 已进行分子/遗传分析处理。正在从选定的人身上采集血液用于分子研究 MLST 网站。血液在特定的分子 SOP 下被接收和处理，然后冷冻保存。我们 多中心站点有 1,836 次出血。血液的血清经过处理后将用于 DMA 项目II的研究。 目标 VIII 将不再执行，因为这不再是主要优先事项，因为 可用资金。