Persistent microRNA changes in serum of cancer-free breast cancer patients

无癌乳腺癌患者血清中持续的 microRNA 变化

• 批准号: 8104694
• 负责人: Harikrishna Nakshatri
• 金额: $ 20.1万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-09 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8104694
• 关键词: Accounting; Adipose tissue; Anabolism; Animal Model; Animals; Biological Markers; Blood; Blood Circulation; Breast Cancer Cell; Breast Cancer Treatment; Cancer Control; Cancer Etiology; Cancer Patient; Cancer Survivor; Cell Count; Cell Death; Cells; Chronic; Clinical; Complex; DNA Methylation; DNA-Directed RNA Polymerase; Development; Disease; Distant; Drops; Epigenetic Process; Estrogen receptor negative; Estrogen receptor positive; Excision; Extracellular Space; Functional RNA; Gene Expression; Genes; Genetic Polymorphism; Genetic Transcription; Goals; Health; Histones; IL6 gene; Immune response; Inflammation; Interleukin-6; Lead; Lipids; Liver; Liver diseases; Long-Term Effects; Lung; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of prostate; Measures; Mediating; MicroRNAs; Mouse Mammary Tumor Virus; Mus; Neoadjuvant Therapy; Normal tissue morphology; Nucleic Acid Regulatory Sequences; Operative Surgical Procedures; Organ; Patients; Pattern; Persons; Play; RNA; RNA Polymerase II; RNA Polymerase III; Recording of previous events; Rest; Role; Serum; Signal Pathway; Small RNA; Source; Structure of parenchyma of lung; Testing; Tumor Burden; Tumor-Derived; U6 small nuclear RNA; Xenograft procedure; arm; base; cancer cell; cytokine; healthy volunteer; histone modification; malignant breast neoplasm; minimally invasive; neoplastic cell; outcome forecast; promoter; regenerative; research study; tumor; tumor progression

DESCRIPTION (provided by applicant): The development of clinically applicable biomarkers for cancer, particularly minimally invasive biomarkers, has been an insurmountable challenge for a number of years. Recent discovery of circulating microRNAs provided some hope; these microRNAs have been suggested to be biomarkers of breast, lung, and prostate cancer. The source of these circulating microRNAs is unknown but is believed to be derived from cancer either through secreted exosomes or released by dying cancer cells. Intriguingly, the levels of several microRNAs are lower in the serum of cancer patients compared to healthy suggesting that cancer is either causing destruction of the adjoining normal tissues from which these microRNAs are normally secreted or it is influencing the expression and secretion of these microRNAs from distant organs. Our recent studies support the latter possibility; serum from breast cancer patients, who are clinically disease-free, displayed elevated levels of miR-451 and miR-101 but lower levels of miR-370, miR-574-3p, miR-342-3p, and miR-197. Additionally, serum from patients contained elevated levels of U6 and 5S, which are small RNAs transcribed by RNA polymerase III. The following hypotheses will be tested in this proposal: Chronic inflammation-like condition in cancer patients lead to permanent changes in microRNA/small RNA expression in distant organs, particularly in organs with regenerative capacity such as liver. Cancer or host-response to cancer causes elevated levels of circulating cytokines such as interleukin 6 (IL-6), which may mediate the long-term effects of cancer on microRNA expression in distant organs. Consequently, distinct microRNA and U6 RNA levels persist in the serum of cancer patients even after these patients are clinically free of the disease. Experiments described in two specific aims will test the above hypotheses: I) Determine whether genes corresponding to microRNAs/small RNA differentially expressed in the serum of cancer bearing animals show altered expression, histone modifications, and RNA polymerase occupancy in liver and lungs. The animal models of breast cancer including MMTV-PyMT, MMTV-neu and xenografts with ER1- positive and ER1-negative breast cancer cells along with appropriate controls will be used to measure serum microRNA profile and cancer-associated changes in microRNA, U6 and 5S expression in liver and lungs. Recruitment of modified histones and RNA polymerase II/III to regulatory regions of U6, 5S, and select microRNA genes in liver and lungs will be measured in control and cancer bearing animals. Serum will be analyzed for mouse IL-6. II) Investigate prospectively whether serum U6, 5S and select miRNA levels change during breast cancer treatment. Serum from breast cancer patients prior to starting neoadjuvant therapy, immediately after completing therapy but before surgery, and after surgery will be examined for U6, 5S RNA and select microRNAs. If cancer is the primary source of the above microRNA/ small RNAs in the serum, their levels should drop after neoadjuvant therapy and/or surgery. If specific microRNA is host -derived and changes in its expression in distant organ is permanent, therapy should not influence its expression. Long-term goal is to determine cancer-induced collateral damage to other organs and how this damage may influence overall health of cancer patients. PUBLIC HEALTH RELEVANCE: This proposal will examine whether microRNAs and small RNAs detected in the serum of breast cancer patients originate from cancer or from the distant organs. It is likely that the chronic inflammation-like condition in cancer patients may cause permanent gene expression changes in liver and lungs, which leads to altered levels of serum microRNAs and small RNAs. The circulating microRNAs may be the invisible long-arm of cancer that reaches distant organs as circulating microRNAs can fuse with heterotypic cells and influence gene expression.

描述(申请人提供)：临床可应用于癌症的生物标记物，特别是微创生物标记物的发展，多年来一直是一个不可逾越的挑战。最近发现的循环microRNAs提供了一些希望；这些microRNAs被认为是乳腺癌、肺癌和前列腺癌的生物标记物。这些循环中的microRNA的来源尚不清楚，但据信来自癌症，要么是通过分泌的外切体，要么是由濒临死亡的癌细胞释放的。有趣的是，与健康人相比，癌症患者血清中几种microRNAs的水平较低，这表明癌症要么破坏了正常分泌这些microRNAs的邻近正常组织，要么影响了来自远处器官的这些microRNAs的表达和分泌。我们最近的研究支持后一种可能性；临床上没有疾病的乳腺癌患者血清中miR-451和miR-101水平升高，而miR-370、miR-574-3p、miR-342-3p和miR-197水平降低。此外，患者的血清中U6和5S水平升高，这是由RNA聚合酶III转录的小RNA。在这项建议中，将检验以下假设：癌症患者的慢性炎症状态导致远处器官，特别是肝脏等具有再生能力的器官中microRNA/小RNA表达的永久性变化。癌症或宿主对癌症的反应导致循环细胞因子水平升高，如白细胞介素6(IL-6)，这可能介导癌症对远处器官microRNA表达的长期影响。因此，癌症患者的血清中存在明显的microRNA和U6RNA水平，即使这些患者在临床上没有患病。有两个特定目的的实验将验证上述假设：i)确定在荷癌动物血清中差异表达的microRNAs/Small RNA对应的基因是否显示出表达变化、组蛋白修饰以及RNA聚合酶在肝脏和肺中的占有率。包括MMTV-PYMT、MMTV-neu在内的乳腺癌动物模型和ER1阳性和ER1阴性乳腺癌细胞的异种移植模型以及适当的对照将被用来测量血清microRNA谱以及与癌症相关的肝脏和肺部microRNA、U6和5S表达的变化。在对照组和荷癌动物中，将测量修饰的组蛋白和RNA聚合酶II/III到U6、5S的调节区以及肝脏和肺中选定的microRNA基因的募集。将对小鼠血清进行IL-6分析。2)前瞻性研究乳腺癌治疗过程中血清U6、5S和部分miRNA水平是否发生变化。乳腺癌患者在开始新辅助治疗前、治疗结束后立即但手术前以及手术后的血清将检测U6、5S RNA和选定的microRNAs。如果癌症是血清中上述微RNA/小RNA的主要来源，则在新辅助治疗和/或手术后，它们的水平应该会下降。如果特定的microRNA是宿主来源的，并且它在远处器官中表达的变化是永久性的，治疗不应该影响它的表达。长期目标是确定癌症对其他器官的附带损害，以及这种损害可能如何影响癌症患者的整体健康。 与公共卫生相关：这项提案将检查在乳腺癌患者血清中检测到的微小RNA和小RNA是来自癌症还是来自远处的器官。癌症患者的慢性炎症样状态可能会导致肝脏和肺部的永久性基因表达变化，从而导致血清microRNAs和Small RNAs水平的变化。循环中的microRNA可能是癌症的看不见的长臂，可以到达远处的器官，因为循环中的microRNA可以与异型细胞融合，并影响基因表达。