Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences ICCBS), University of Karachi, Karachi, 75270, Pakistan.
Department of Biotechnology, University of Sargodha, Sargodha, Pakistan.
Sci Rep. 2021 Nov 23;11(1):22766. doi: 10.1038/s41598-021-02211-4.
In the current study, APOB (rs1052031) genotype-guided proteomic analysis was performed in a cohort of Pakistani population. A total of 700 study subjects, including Coronary Artery Disease (CAD) patients (n = 480) and healthy individuals (n = 220) as a control group were included in the study. Genotyping was carried out by using tetra primer-amplification refractory mutation system-based polymerase chain reaction (T-ARMS-PCR) whereas mass spectrometry (Orbitrap MS) was used for label free quantification of serum samples. Genotypic frequency of GG genotype was found to be 90.1%, while 6.4% was for GA genotype and 3.5% was for AA genotypes in CAD patients. In the control group, 87.2% healthy subjects were found to have GG genotype, 11.8% had GA genotype, and 0.9% were with AA genotypes. Significant (p = 0.007) difference was observed between genotypic frequencies in the patients and the control group. The rare allele AA was found to be strongly associated with the CAD [OR: 4 (1.9-16.7)], as compared to the control group in recessive genetic model (p = 0.04). Using label free proteomics, altered expression of 60 significant proteins was observed. Enrichment analysis of these protein showed higher number of up-regulated pathways, including phosphatidylcholine-sterol O-acyltransferase activator activity, cholesterol transfer activity, and sterol transfer activity in AA genotype of rs562338 (G>A) as compared to the wild type GG genotype. This study provides a deeper insight into CAD pathobiology with reference to proteogenomics, and proving this approach as a good platform for identifying the novel proteins and signaling pathways in relation to cardiovascular diseases.
在当前的研究中，对巴基斯坦人群进行了 APOB(rs1052031)基因型指导的蛋白质组分析。共纳入 700 例研究对象，包括冠心病（CAD）患者（n=480）和健康个体（n=220）作为对照组。采用四引物扩增受阻突变系统-聚合酶链反应（T-ARMS-PCR）进行基因分型，而采用无标记定量的质谱（Orbitrap MS）分析血清样本。在 CAD 患者中，GG 基因型的基因型频率为 90.1%，GA 基因型为 6.4%，AA 基因型为 3.5%。在对照组中，87.2%的健康受试者为 GG 基因型，11.8%为 GA 基因型，0.9%为 AA 基因型。患者和对照组之间的基因型频率存在显著差异（p=0.007）。与对照组相比，隐性遗传模型下，罕见等位基因 AA 与 CAD 强烈相关[OR：4（1.9-16.7）]，（p=0.04）。采用无标记蛋白质组学，观察到 60 个显著蛋白的表达发生改变。这些蛋白的富集分析显示，AA 基因型的 rs562338（G>A）中上调途径数量更高，包括磷酸胆碱-固醇 O-酰基转移酶激活活性、胆固醇转移活性和固醇转移活性。本研究通过蛋白质基因组学为 CAD 病理生理学提供了更深入的了解，并证明了这种方法是识别与心血管疾病相关的新型蛋白和信号通路的良好平台。
