miRNA Profiling in Fixed Cancer Samples

固定癌症样本中的 miRNA 分析

• 批准号: 7544746
• 负责人: DAVID M BROWN
• 金额: $ 60.76万
• 依托单位: AMBION DIAGNOSTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7544746
• 关键词: Adult; Alpha 1 Casein Kinase 1; Animals; Applications Grants; Archives; Binding; Biological Assay; Biological Models; Bladder; Blinded; Brain; Breast; Burkitt Lymphoma; Cancer Detection; Cancer Diagnostics; Cancer Patient; Cell Culture System; Cell physiology; Cells; Chronic Lymphocytic Leukemia; Clinical; Clinical Trials; Colon; Colon Adenocarcinoma; Cultured Cells; Data; Development; Diagnosis; Diagnostic; Disease; Evaluation; Exhibits; Eye; Formalin; Freezing; Funding; Gene Expression; Genes; Genus Cola; Goals; Grant; Heart; Hospitals; Human; Intervention; Label; Lead; Link; Lung; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of pancreas; Measures; Methods; MicroRNAs; Microarray Analysis; Molecular Profiling; Mus; Normal tissue morphology; Oligonucleotides; Oncogenes; Oncogenic; Organ; Paraffin Embedding; Patients; Performance; Phase; Plant Genome; Procedures; Process; Prognostic Marker; Prostate; Proteins; RNA; Reagent; Regulation; Relative (related person); Research; Research Project Grants; Sampling; Screening for cancer; Series; Site; Skeletal Muscle; Small Business Funding Mechanisms; Small Business Innovation Research Grant; Small Interfering RNA; Source; Staging; Stratification; Structure of parenchyma of lung; System; TP53 gene; Technology; Testing; Therapeutic; Therapeutic Agents; Therapeutic Intervention; Thymus Gland; Tissue Sample; Tissues; Translations; Tumor Tissue; United States; Work; assay development; base; calin; cancer type; cost; human disease; human tissue; inhibitor/antagonist; interest; mammalian genome; method development; outcome forecast; prognostic; programs; tissue fixing; tumor; tumorigenesis

DESCRIPTION (provided by applicant): During Phase I of our proposed research, we will develop and validate procedures for recovering, labeling, and analyzing miRNAs from fixed tissue samples. The procedures will be based on the miRNA microarray and fixed tissue RNA isolation systems that we developed in other SBIR-funded programs. The development of our miRNA isolation and labeling procedures will be accomplished using a model system wherein mouse organs will be split and half is flash-frozen and the other half formalin-fixed using a procedure that is commonly employed in hospitals. The frozen and fixed samples will be processed to recover the miRNAs. The miRNAs from the fixed and frozen tissues will be independently labeled and analyzed using miRNA microarrays. The isolation, labeling, and hybridization procedures will be varied until the fixed samples yield the same miRNA expression profiles as the equivalent frozen samples. The fixed sample procedures will) then be used to analyze formalin fixed, human tissue samples to analyze miRNA profiles from multiple organs. The fixed tissue miRNA profiles will be compared to the profiles generated from frozen samples to verify that the fixed tissue miRNA profiling process can be used for stored, human fixed tissue samples. During Phase II of our research project, we will use the miRNA isolation, labeling and microarray analysis procedures developed during Phase I to analyze archived, fixed human cancer tissues to identify miRNAs with expression profiles that are significantly different from equivalent, normal tissues. The most interesting miRNAs or miRNA signatures might provide opportunities for diagnostic/prognostic assay development or even an intervention point for therapeutic agents.

描述（由申请人提供）： 在我们提出的研究的第一阶段，我们将开发和验证从固定组织样品中恢复，标记和分析miRNA的程序。该程序将基于我们在其他SBIR资助的程序中开发的miRNA微阵列和固定组织RNA隔离系统。将使用模型系统来完成我们的miRNA隔离和标记程序的开发，其中小鼠器官将被拆分，一半是闪烁的，而另一半则使用医院中通常采用的程序进行了福尔马林固定。将处理冷冻和固定样品以恢复miRNA。来自固定组织和冷冻组织的miRNA将使用miRNA微阵列独立标记和分析。隔离，标记和杂交程序将变化，直到固定样品产生与等效冷冻样品相同的miRNA表达曲线为止。然后将使用固定的样品程序来分析福尔马林固定的人体组织样品，以分析来自多个器官的miRNA谱。固定组织miRNA轮廓将与从冷冻样品产生的曲线进行比较，以验证固定组织miRNA分析过程可用于存储的人类固定组织样品。 在我们的研究项目的第二阶段中，我们将使用在I期期间开发的miRNA隔离，标记和微阵列分析程序来分析存档的，固定的人类癌组织，以鉴定具有与等效的正常组织明显不同的表达谱的miRNA。最有趣的miRNA或miRNA签名可能为诊断/预后测定开发甚至治疗剂的干预点提供机会。