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(由申请人提供)：自从发现小鼠乳腺肿瘤病毒(MMTV)是小鼠乳腺肿瘤的病原体以来，人们进行了大量的努力，以确定类似的病原体是否可能与人类乳腺癌的发病机制有关。人们对这一问题的兴趣始于20世纪90年代中期的S，当时我们报道在38%(226/314)的美国女性乳腺癌标本中检测到一个660bp的序列，与MMTV包膜基因(Env)9098%的同源性，而与任何人类基因或人类内源性逆转录病毒(HERV)或任何其他病毒没有显著同源性。其他实验室和我们自己的实验室也在不同地理区域的人群中发现了类似的结果。 随后，在人乳腺肿瘤中检测到的9.9kb的前病毒序列，经扩增，发现与MMTV有95%的同源性，被命名为HMTV。从转移性乳腺癌患者胸腔积液和腹部积液中建立的转移性乳腺癌细胞(MMSM细胞)的原代培养中分离出具有β-病毒特征的HMTV病毒颗粒。这些病毒颗粒的RNA序列与MMTV的同源性为8595%。最近，我们发现MSSM细胞来源的HMTV能够在原代培养的人乳腺上皮细胞(HMEC)、外周血单核细胞(PBMC)、健康供者树突状细胞以及以下细胞系中感染和复制：MCF10A、MCF10F、293T、Ramos(B细胞)、Jurkat(T细胞)。在感染的MCF10F细胞中，Ecadherin等上皮细胞特有的蛋白质表达减少，Vimentin等间充质细胞特有的蛋白质表达增加。这种现象是当健康的上皮细胞癌变时观察到的上皮间质转化的典型过程。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，类鼠病毒在人类疾病中的研究目前正受到严格的审查和批评，主要是因为发现异嗜性小鼠白血病病毒相关病毒(XMRV)被认为在人类前列腺癌和慢性疲劳综合征(CFS)中发挥作用，现在似乎是一种小鼠污染物。在这项申请中，我们建议使用可以保证结果无污染的技术来证明人类乳腺癌中存在HMTV。如果乳腺癌中的HMTV是小鼠DNA污染的结果，这项拟议的研究可能会确定这一点。使用敏感分析的初步数据显示，当发现HMTV时，没有检测到小鼠DNA污染。此外，HMTV已在乳腺癌标本的细胞核中可见，并整合在MSSM细胞的染色体中。这些数据的扩展可以向科学界提供令人信服的证据，证明HMTV不是污染物