EDRN Biomarker Reference Laboratory

EDRN 生物标志物参考实验室

• 批准号: 8565914
• 金额: $ 45.67万
• 依托单位: NATIONAL CANCER INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8565914
• 关键词: Benign; Biological Assay; Biological Markers; Blinded; Cell Count; Cells; Collaborations; DNA Sequence Rearrangement; Detection; Development; ETV1 gene; Early Detection Research Network; Epithelial Cells; Frequencies; Genetic Research; Health Sciences; Human; Individual; Laboratories; Malignant Neoplasms; Malignant neoplasm of prostate; Mass Spectrum Analysis; Microdissection; Molecular Genetics; National Cancer Institute; Normal Cell; Oncogenes; One-Step dentin bonding system; Pacific Northwest; Prostate; Prostatectomy; Prostatic Diseases; Proteins; Proteomics; Research; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Services; Specimen; Tissues; Transcript; Universities; Validation; base; cancer cell; cell stroma; laser capture microdissection; protein expression; research study; tumor

Uniformed Services University of the Health Sciences (USU)-Center for Prostate Disease Research (CPDR) will serve as National Cancer Institute (NCI)-Early Detection Research Network (EDRN)-Biomarker Reference Laboratory (BRL). CPDR will provide support for current NCI-EDRN efforts focused on the development of a multiplex quantitative mass spectrometry-based detection of biomarkers by using normal and cancer cells and adjacent stroma cells enriched by laser capture microdissection (LCM). Our focus on LCM based CaP biomarker discovery and validation in relation to key observations in CaP molecular genetics research. Enrichment of prostate cancer epithelial cells by Laser Capture Microdissection (LCM) for development of a SRM-MS based assay for detection of truncated ERG protein. -The initial detection of the biomarkers will focus on human prostate tissue with the ERG oncogene rearrangements. The CPDR will enrich the fraction of prostate cancer cells by microdissection of cancer epithelial cells and matched benign epithelial cells from 23 prostatectomy specimens. - Detection of aberrant ERG rearrangements and expression in prostatectomy specimens by IHC and RT-PCR assay: Prostatectomy specimens will be pre-screened for ERG protein expression by IHC using ERG-MAb and for ERG fusion transcripts by RT-PCR. - Enrichment of benign and cancer epithelial cells from whole mount prostatectomy sections: For pilot experiments, three prostatectomy specimens will be used for LCM of 100,000 ERG positive cancer cells and matched benign epithelial cells. This number of cells should be sufficient to optimize detection of the truncated ERG by SRM-MS at the Pacific Northwest National Laboratory (PNNL), DOE. As ERG expression is relatively homogenous in individual tumor foci, entire ERG positive focus (based on ERG IHC of consecutive tissue section) can be dissected from FFPE-whole mounted prostate sections. Additional 20 pairs of cancer and benign epithelial cells ((50,000 cells/specimen; 10 ERG positive and 10 ERG negative) will be obtained which will be sent for blinded examination at PNNL. Enrichment of prostate cancer epithelial cells by Laser Capture Microdissection (LCM) for development of a SRM-MS based assay for detection of truncated ETV1 protein. The purpose is identical to the previous one and the steps and task requirements will be identical. This study will be performed in collaboration with PNNL and other proteomic EDRN centers. Since the frequency of ETV1 is much lower (>5%), assistance from other EDRN investigators will also be sought in obtaining such specimens.

统一的卫生科学大学（USU）前列腺疾病研究中心（CPDR）将担任国家癌症研究所（NCI） - 早期检测研究网络（EDRN） - Biomarker参考实验室（BRL）。 CPDR将为当前的NCI-EDRN努力提供支持，这些努力专注于通过使用正常和癌细胞以及通过激光捕获显微解剖（LCM）富含的癌细胞以及相邻的基质细胞来开发基于多重定量质谱的检测。 我们关注基于LCM的CAP生物标志物发现和与CAP分子遗传学研究中的关键观察有关的验证。 通过激光捕获微分解（LCM）富集前列腺癌上皮细胞，以开发基于SRM-MS的测定法，以检测截短的ERG蛋白。 - 对生物标志物的初始检测将集中在ERG癌基因重排的人类前列腺组织上。 CPDR将通过对癌细胞上皮细胞的显微解剖并与来自23个前列腺切除术标本的良性上皮细胞相匹配。 - IHC和RT-PCR分析中的异常ERG重排和表达的异常ERG重排和表达：前列腺切除术标本将被预先筛选，用于使用ERG-MAB和RT-PCR进行ERG-MAB和ERG融合转录的IHC表达ERG蛋白。 - 整个前列腺切除术切片中良性和癌症上皮细胞的富集：对于试点实验，将使用三个前列腺切除术标本，用于100,000个ERG阳性癌细胞的LCM，并匹配良性上皮细胞。这一数量的细胞应足以优化SRM-MS在DOE太平洋国家实验室（PNNL）对截短的ERG的检测。由于ERG表达在单个肿瘤灶中相对均匀，因此可以从FFPE - 整个固定的前列腺段中阐述整个ERG阳性焦点（基于连续组织的ERG IHC）。另外20对癌症和 将获得良性上皮细胞（（50,000个细胞/样品； 10 ERG阳性和10 ERG阴性），将在PNNL上发送盲目检查。 通过激光捕获微分解（LCM）富集前列腺癌上皮细胞，以开发基于SRM-MS的测定法，以检测截短的ETV1蛋白。目的与上一个相同，步骤和任务要求将相同。这项研究将与PNNL和其他蛋白质组学EDRN中心合作进行。由于ETV1的频率要低得多（> 5％），因此也将寻求其他EDRN研究者的援助来获得此类 标本。