Characterization of salivary microvesicles microRNA content

唾液微泡 microRNA 含量的表征

• 批准号: 8553354
• 负责人: Ilias Alevizos
• 金额: $ 63.1万
• 依托单位: NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH
• 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/8553354
• 关键词: Affect; Biological Markers; Biopsy; Cell Proliferation; Cell membrane; Cells; Characteristics; Diagnostic; Disease; Health; Inflammatory Response; Liquid substance; Messenger RNA; MicroRNAs; Multivesicular Body; Nucleic Acids; Oral; Pathogenesis; Pathology; Patients; Peripheral; Physiological; Plasma; Play; Process; Production; Proteins; Protocols documentation; Regulation; Research; Role; Saliva; Salivary; Salivary Gland Diseases; Salivary Gland Neoplasms; Salivary Glands; Side; Sjogren' s Syndrome; Small RNA; Source; Surface; Tissues; Urine; Vesicle; base; cancer cell; cell type; healthy volunteer; in vivo; insight; tool

Microvesicles are small, right-side out cell-secreted vesicles of about 30-100 nm, derived from fusion of multivesicular bodies to plasma membranes. Beyond their characteristic repertoire of surface markers, they feature a wide range of surface and internal proteins specific to their source, and recent studies found that they can also transport mRNA and microRNA (miRNA). We developed and optimized protocols for the routine isolation of microvesicles (most of them being exosomes, based on surface markers and electromicroscopy) from both whole and glandular saliva and we showed that microvesicles can be readily isolated from saliva, and that they contain miRNAs in quantities adequate for both qPCR and microarray hybridization. We also successfully hybridized microRNA microarrays with microRNAs isolated from salivary microvesicles from a healthy volunteer and a Sjogren's syndrome patient as a proof of concept. We are currently in the process of further and more extensively characterizing the small nucleic acid content of microvesicles since salivary microvesicular miRNAs may be valuable not only as a diagnostic tool but may also provide an insight in the role miRNAs play in the underlying pathophysiologic processes of various salivary gland diseases. Among others they may help understanding if specific miRNAs are involved abnormalities in saliva production or regulation of the peripheral inflammatory response in the salivary gland and oral characteristic of Sjogren's syndome. Exosomal miRNA analysis may also be valuable in understanding the pathogenesis of salivary gland tumors as a number of studies have identified miRNA dysregulation as a characteristic marker in cancer cell proliferation in vivo have found distinctive exosomal miRNA profiles in blood plasma, urine, and other fluids. In summary, isolation of salivary gland derived microvesicle miRNAs holds the promise of focused biomarker discovery for pathologies that directly or indirectly affect the salivary glands.

微泡是一种小的、右侧向外的细胞分泌的小囊泡，大小约为30-100 nm，来源于多囊泡体与质膜的融合。除了其特有的表面标记，它们还具有广泛的表面和内部蛋白质，特定于其来源，最近的研究发现，它们还可以运输mRNA和microRNA(MiRNA)。 我们开发和优化了从整个唾液和腺唾液中常规分离微囊(基于表面标记和电子显微镜，其中大多数是外体)的方案，我们表明可以很容易地从唾液中分离微泡，并且它们含有足够数量的miRNA，用于qPCR和微阵列杂交。我们还成功地将microRNA微阵列与从一名健康志愿者和一名干燥综合征患者的唾液微泡中分离出来的microRNA杂交，作为概念的证明。 我们目前正在进一步和更广泛地表征微泡的小核酸含量，因为唾液微泡miRNAs可能不仅作为一种诊断工具有价值，而且可能提供对miRNAs在各种唾液腺疾病的潜在病理生理过程中所起作用的洞察。在其他方面，它们可能有助于了解特定的miRNAs是否涉及唾液产生的异常或唾液腺外周炎症反应的调节以及干燥综合征的口腔特征。外体miRNA分析也可能对理解涎腺肿瘤的发病机制有价值，因为许多研究发现，体内癌细胞增殖的特征标志是miRNA异常调节，并在血浆、尿液和其他液体中发现了独特的外体miRNA图谱。 总之，分离唾液腺来源的微囊miRNAs有望为直接或间接影响唾液腺的病理发现有针对性的生物标记物。