COVID-19 biomarker discovery

COVID-19 生物标志物的发现

基本信息

批准号：
10272260
负责人：
Aleksandra Nita-Lazar
金额：
$ 0.56万
依托单位：
NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
依托单位国家：
美国
项目类别：
财政年份：
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/10272260
关键词：
2019-nCoV Accounting Binding Biological Biological Assay Biological Markers COVID-19 Cell Surface Receptors Cell membrane Cells Centers for Disease Control and Prevention (U.S.)Cessation of life Clinical Code Coronavirus Detection Development Diagnosis Diagnostic Disease Disease Progression Evaluation Genes Genomics Glycoproteins Goals Human Immunodominant Epitopes Individual Infection Maps Mediating Membrane Proteins Monitor Nucleocapsid Nucleocapsid Proteins Patients Proteins Proteome Proteomics RNA Research Proposals Reverse Transcriptase Polymerase Chain Reaction Risk Role Saliva Sampling Serum Sputum Structural Protein Testing Time United States National Institutes of Health Urine Viral Viral Structural Proteins Virion Virus Work assay development base biomarker discovery comparative diagnostic assay diagnostic biomarker env Gene Products genomic data novel coronavirus novel therapeutics novel vaccines prognostic protein profiling receptor binding respiratory treatment response

项目摘要

The focus of the proteomic approach is to identify and quantify viral structural proteins, specifically spike (S) glycoprotein as well as envelope (E) protein, membrane (M) protein, and nucleocapsid (N) protein in patient samples. The S protein mediates attachment of the virus to the host cell surface receptors and subsequent fusion of the viral and host cell membranes to facilitate viral entry into the host cell. The S1 subunit of the ectodomain mediates binding of the virion to host cell-surface receptors through its receptor-binding domain and is the immunodominant epitope. Due to its critical role in cell entry, we anticipate high expression levels of the S protein during active infection making it an attractive diagnostic marker for SARS-CoV-2 infection. Additionally, levels of this protein are expected to change during the course of infection and can be used to monitor individual disease progression. Finally, our proposed proteomic based detection of the S protein could prove helpful in studies evaluating the efficacy of novel drugs and vaccines. Specific goals: 1) Development of proteomic based assay for detection and quantitation of SARS-CoV-2 proteome in biological samples. Work will focus on four structural proteins: spike (S) glycoprotein (primary target), envelope (E) protein, membrane (M) protein, and nucleocapsid (N) protein. The E and N gene (coding for the envelop and nucleocapsid protein) have been used for PCR based diagnostic assays. We therefore aim to explore using their encoded products as additional targets in our proteomics assay development. 2) Characterize and compare SARS-CoV-2 protein profiles among patients and at different times of the course of disease using nasopharyngeal and respiratory samples from patients admitted at the NIH previously tested with N1, N2 RT-PCR CDC RT-PCR assay and using genomic data from the research proposal Genomics of SARS-CoV-2 3) Comparison of protein profiles and correlation with RNA detection assays. This will provide protein level information regarding observed differences between N1 and N2 levels among patients as observed by RT-PCR assay. 4) Selection of viral proteomic biomarker(s) for further evaluation as diagnostic and prognostic target in different clinical samples (saliva, urine, serum)

蛋白质组学方法的重点是识别和定量患者样本中的病毒结构蛋白，特别是刺突 (S) 糖蛋白以及包膜 (E) 蛋白、膜 (M) 蛋白和核衣壳 (N) 蛋白。 S蛋白介导病毒与宿主细胞表面受体的附着以及随后病毒和宿主细胞膜的融合以促进病毒进入宿主细胞。胞外域的 S1 亚基通过其受体结合域介导病毒粒子与宿主细胞表面受体的结合，并且是免疫显性表位。由于其在细胞进入中的关键作用，我们预计 S 蛋白在主动感染期间会出现高表达水平，使其成为 SARS-CoV-2 感染的有吸引力的诊断标记物。此外，这种蛋白质的水平预计会在感染过程中发生变化，可用于监测个体疾病进展。最后，我们提出的基于蛋白质组学的 S 蛋白检测可能有助于评估新药和疫苗功效的研究。具体目标： 1) 开发基于蛋白质组学的检测和定量生物样品中 SARS-CoV-2 蛋白质组的方法。工作重点是四种结构蛋白：刺突 (S) 糖蛋白（主要目标）、包膜 (E) 蛋白、膜 (M) 蛋白和核衣壳 (N) 蛋白。 E 和 N 基因（编码包膜蛋白和核衣壳蛋白）已用于基于 PCR 的诊断测定。因此，我们的目标是探索使用它们的编码产物作为我们蛋白质组学检测开发中的附加靶标。 2) 使用先前使用 N1、N2 RT-PCR CDC RT-PCR 检测方法测试过的 NIH 入院患者的鼻咽和呼吸道样本，对患者和病程不同时间的 SARS-CoV-2 蛋白谱进行表征和比较SARS-CoV-2 基因组学研究提案中的基因组数据 3) 蛋白质谱比较以及与RNA检测分析的相关性。这将提供有关通过 RT-PCR 测定观察到的患者 N1 和 N2 水平之间观察到的差异的蛋白质水平信息。 4) 选择病毒蛋白质组生物标志物以进一步评估作为不同临床样本（唾液、尿液、血清）中的诊断和预后目标