Targeted proteomics and miRNomics to tackle biomarkers of infectious and neurodegenerative diseases and cancer

靶向蛋白质组学和 miRNA 组学来解决传染病、神经退行性疾病和癌症的生物标志物

• 批准号: 578469-2022
• 负责人: Berezovski, MaximMV
• 金额: $ 1.82万
• 依托单位: University of Ottawa
• 依托单位国家: 加拿大
• 项目类别: Alliance Grants
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=747305
• 关键词: Targeted; proteomics; miRNomics; tackle; biomarkers

Nowadays, research on molecular biomarkers is widely recognized as fundamental to improving the prevention, diagnosis, prognosis, monitoring and treatment of infectious and neurodegenerative diseases, and cancer. This project is focused on the development of innovative analytical methods for purification, preconcentration, separation, identification and characterization in biological fluids of proteins, glycoproteins and miRNA biomarkers, specifically related to COVID-19 (spike glycoprotein and nucleocapside protein), sepsis (procalcitonin protein), Parkinsons and Alzheimers diseases (alpha-synuclein protein and clusterin glycoprotein, respectively), chronic lymphocytic leukemia (miR21) and colorectal cancer (carcinoembryonic antigen glycoprotein, miR-421, miR130b-3p and miR27a-3p). Different strategies will be investigated, including the direct analysis of the biomarkers at the intact level or by bottom-up approaches, after their enzymatic digestion to obtain characteristic fragments (e.g. peptides, phosphopeptides, glycopeptides or glycans). Likewise, possible differences and alterations in the structure, composition or concentration of the potential biomarkers will be evaluated from a comparison of control and pathological samples. For these purposes, it will be primarily used high-resolution microscale separation techniques coupled to mass spectrometry (capillary liquid chromatography and capillary electrophoresis coupled to mass spectrometry, CapLC-MS and CE-MS) in full scan MS and tandem MS/MS modes, and online purification and preconcentration techniques, such as on-line solid-phase extraction capillary electrophoresis-mass spectrometry (on-line SPE-CE-MS). Finally, the application of advanced data analysis tools will provide a more systematic and comprehensive interpretation of the enormous amount of complex multidimensional data generated in the different comparative MS-based studies.

如今，对分子生物标志物的研究被广泛认为是改善感染性和神经退行性疾病以及癌症的预防、诊断、预后、监测和治疗的基础。该项目的重点是开发创新的分析方法，用于纯化，预浓缩，分离，鉴定和表征生物液体中的蛋白质，糖蛋白和miRNA生物标志物，特别是与COVID-19相关的生物标志物（刺突糖蛋白和核衣壳蛋白）、脓毒症（降钙素原蛋白），帕金森病和阿尔茨海默病（分别为α-突触核蛋白和丛生蛋白糖蛋白）、慢性淋巴细胞白血病（miR 21）和结肠直肠癌（癌胚抗原糖蛋白、miR-421、miR 130 b-3 p和miR 27 a-3 p）。将研究不同的策略，包括在酶消化后直接分析完整水平的生物标志物或通过自下而上的方法，以获得特征片段（例如肽、磷酸肽、糖肽或聚糖）。同样，将通过对照和病理学样本的比较，评价潜在生物标志物的结构、组成或浓度的可能差异和变化。为此，将主要使用高分辨率微量分离技术与质谱联用（毛细管液相色谱和毛细管电泳与质谱联用，CapLC-MS和CE-MS），采用全扫描MS和串联MS/MS模式，以及在线纯化和预浓缩技术，如在线固相萃取毛细管电泳-质谱（在线SPE-CE-MS）。最后，先进数据分析工具的应用将为不同的基于MS的比较研究中生成的大量复杂多维数据提供更系统、更全面的解释。