Secondary analysis of Human Mammary Tumor Virus (HMTV) in breast cancer.

人类乳腺肿瘤病毒（HMTV）在乳腺癌中的二次分析。

• 批准号: 9113513
• 负责人: Stella Maris Melana
• 金额: $ 8.48万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-20 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9113513
• 关键词: Abdomen; American; B-Lymphocytes; Betaretrovirus; Biological Assay; Breast Cancer Cell; Breast Epithelial Cells; Cell Line; Cell Nucleus; Cells; Characteristics; Chromosomes; Chronic Fatigue Syndrome; Communities; DNA; DNA Sequence; Data; Dendritic Cells; Detection; E-Cadherin Staining Method; Educational process of instructing; Endogenous Retroviruses; Epithelial; Epithelial Cells; Fluorescent in Situ Hybridization; Genes; Geographic Locations; Health; Human; Human Genome; In Vitro; Laboratories; MCF10A cells; Malignant neoplasm of prostate; Mammary Neoplasms; Mesenchymal; Metaphase; Metastatic breast cancer; Mouse Mammary Tumor Virus; Murine leukemia virus; Mus; Participant; Pathogenesis; Patients; Peripheral Blood Mononuclear Cell; Play; Pleural; Population; Process; Publishing; RNA Sequences; Reporting; Retroviridae; Role; Specimen; Structure; System; T-Lymphocyte; Techniques; Vimentin; Viral; Virus; Virus Integration; Woman; Work; deep sequencing; effusion; experience; human disease; integration site; interest; leukemia; malignant breast neoplasm; mammary tumor virus; nano-string; neoplastic; particle; protein expression; research study; virus envelope

 DESCRIPTION (provided by applicant): Since the discovery of Mouse Mammary Tumor Virus (MMTV) as the etiological agent of mammary tumors in mice there have been numerous efforts to determine if a similar agent might be involved in the pathogenesis of human breast cancer. Renewed interest in this issue began in the middle 1990's when we reported that a 660 bp sequence, 9098% homologous to the MMTV envelope (env) gene, with no significant homology to any human gene or human endogenous retrovirus (HERV) nor to any other virus, was detected in 38% (226 of 314) of American women's breast cancers specimens. Similar results have been found across populations from different geographic regions by other laboratories as well as our own. Subsequently a 9.9 kb proviral sequence detected in human breast tumors, amplified, and found to be 95% homologous to MMTV was designated as HMTV. HMTV viral particles with betaretrovirus characteristics have been isolated from primary cultures of metastatic breast cancer cells (MMSM cells) established from pleural and abdominal effusions of patients with metastatic breast cancer. The RNA sequence of these viral particles was 8595% homologous to MMTV. Recently we have shown that HMTV from MSSM cells is able to infect and replicate in primary cultures of human mammary epithelial cells (HMEC), peripheral blood mononuclear cells (PBMC), dendritic cells of healthy donors and the following cell lines: MCF10A, MCF10F, 293T, Ramos (B cells), Jurkat (T cells). In the infected MCF10F cells, expression of proteins characteristic of epithelial cells such as Ecadherin decreased and the expression of proteins characteristic of mesenchymal cells such as Vimentin increased. This phenomenon is typical of the process of Epithelial Mesenchymal Transition observed when healthy epithelial cells become neoplastic. Despite the substantial evidence supporting the presence of HMTV in human breast cancer, controversy continues concerning its relevance and veracity. In addition, the study of murine like viruses in human disease is currently under intense scrutiny and criticism, mainly due to the finding that Xenotropic Murine Leukemia virusrelated virus (XMRV) which was thought to play a role in human prostate cancer and in chronic fatigue syndrome (CFS) now appears to be a mouse contaminant. In this application we propose to demonstrate that HMTV is present in human breast cancer using techniques that can guarantee results free of contamination. If HMTV in breast cancer were the result of murine DNA contamination, it could be determined by this proposed study. Preliminary data using sensitive assays show no detection of murine DNA contamination when HMTV is found. In addition, HMTV has been visualized in the nucleus of breast cancer specimens as well as integrated in chromosomes of MSSM cells. Expansion of these data can provide convincing evidence to the scientific community that HMTV is not a contaminant

 描述（由适用提供）：由于发现小鼠乳腺肿瘤的病因学药物的发现小鼠乳腺肿瘤病毒（MMTV），因此已经做出了许多努力来确定是否可能涉及人类乳腺癌发病机理。对这个问题的重新兴趣始于1990年代中期，当时我们报告说，与MMTV信封（ENK）基因同源的660 bp序列（9098％）在38％的38％（226 of 226 of 314 hommers Cancers of American Womens cancers Cancers Cancers of American anders Cancers of Amery norde cans of nose Gene或Human Gene或Human Gene或Human Gene Gene或Human Gene forovirus（HERV）均无重要同源。其他实验室以及我们自己的不同地理区域的人群中也发现了类似的结果。 随后，在人类乳腺肿瘤中检测到的9.9 Kb病毒序列被放大并发现与MMTV同源95％被指定为HMTV。已从转移性乳腺癌患者的胸膜和腹部狂热者建立的转移性乳腺癌细胞（MMSM细胞）的原发性培养物中分离出具有β病毒特征的HMTV病毒颗粒。这些病毒颗粒的RNA序列与MMTV同源8595％。最近，我们表明，来自MSSM细胞的HMTV能够在人类乳腺上皮细胞（HMEC）（HMEC），周围血液单核细胞（PBMC），健康供体的树突细胞，健康供体细胞和以下细胞系的树突状细胞和以下细胞系：MCF10A，MCF10A，MCF10A，293T，Ramos（293T，Ramos（B Ramos）（b cillkat），Yerkkat（t））中（T）。在感染的MCF10F细胞中，蛋白质特征上皮细胞（如eCadherin）的表达以及在波形蛋白等间充质细胞（如波形蛋白）中的蛋白质表达增加。这种现象是当健康上皮细胞变成肿瘤时观察到的上皮间质转变过程的典型特征。尽管有大量证据支持HMTV在人类乳腺癌中的存在，但关于其相关性和真实性的争议仍在继续。此外，目前对人类疾病中的鼠类病毒之类的研究正受到严格​​的审查和批判主义的审查，这主要是由于发现异形鼠白血病病毒相关病毒（XMRV）被认为在人类前列腺癌中起作用，在人类前列腺癌和慢性疲劳综合征（CFS）中起着作用。在此应用中，我们建议使用可以保证没有污染结果的技术证明HMTV存在于人类乳腺癌中。如果乳腺癌中的HMTV是鼠DNA污染的结果，则可以通过这项拟议的研究确定。使用敏感测定的初步数据显示，当发现HMTV时未发现鼠DNA污染。此外，HMTV已在乳腺癌标本的核中可视化，并整合在MSSM细胞的染色体中。这些数据的扩展可以为科学界提供令人信服的证据，即HMTV不是污染物