Shoraka Shahrzad, Mohebbi Seyed Reza, Hosseini Seyed Masoud, Zali Mohammad Reza
Department of Microbiology and Microbial Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran.
Gastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Front Microbiol. 2023 Oct 6;14:1256042. doi: 10.3389/fmicb.2023.1256042. eCollection 2023.
Since the beginning of the COVID-19 pandemic, a wide clinical spectrum, from asymptomatic infection to mild or severe disease and death, have been reported in COVID-19 patients. Studies have suggested several possible factors, which may affect the clinical outcome of COVID-19. A pro-inflammatory state and impaired antiviral response have been suggested as major contributing factors in severe COVID-19. Considering that mitochondria have an important role in regulating the immune responses to pathogens, pro-inflammatory signaling, and cell death, it has received much attention in SARS-CoV-2 infection. Recent studies have demonstrated that high levels of cell-free mitochondrial DNA (-mtDNA) are associated with an increased risk of COVID-19 intensive care unit (ICU) admission and mortality. However, there have been few studies on -mtDNA in SARS-CoV-2 infection, mainly focusing on critically ill COVID-19 cases. In the present study, we investigated -mtDNA copy number in COVID-19 patients and compared between asymptomatic and symptomatic cases, and assessed the clinical values. We also determined the -nuclear DNA (-nDNA) copy number and mitochondrial transcription factor A (TFAM) mRNA level in the studied groups.
Plasma and buffy coat samples were collected from 37 COVID-19 patients and 33 controls. Briefly, after total DNA extraction, plasma -mtDNA, and -nDNA copy numbers were measured by absolute qPCR using a standard curve method. Furthermore, after total RNA extraction from buffy coat and cDNA synthesis, TFAM mRNA levels were evaluated by qPCR.
The results showed that -mtDNA levels in asymptomatic COVID-19 patients were statistically significantly higher than in symptomatic cases ( value = 0.01). However, -nDNA levels were higher in symptomatic patients than in asymptomatic cases ( value = 0.00). There was no significant difference between TFAM levels in the buffy coat of these two groups ( value > 0.05). Also, -mtDNA levels showed good diagnostic potential in COVID-19 subgroups.
-mtDNA is probably important in the outcome of SARS-CoV-2 infection due to its role in inflammation and immune response. It can also be a promising candidate biomarker for the diagnosis of COVID-19 subgroups. Further investigation will help understanding the COVID-19 pathophysiology and effective diagnostic and therapeutic strategies.
自新冠疫情开始以来,新冠患者出现了广泛的临床症状谱,从无症状感染到轻症或重症疾病乃至死亡。研究提出了几个可能影响新冠临床结局的因素。促炎状态和抗病毒反应受损被认为是重症新冠的主要促成因素。鉴于线粒体在调节对病原体的免疫反应、促炎信号传导和细胞死亡中起重要作用,它在新冠病毒感染中受到了广泛关注。最近的研究表明,高水平的游离线粒体DNA(-mtDNA)与新冠患者入住重症监护病房(ICU)及死亡风险增加有关。然而,关于新冠病毒感染中-mtDNA的研究较少,主要集中在重症新冠病例。在本研究中,我们调查了新冠患者的-mtDNA拷贝数,并比较了无症状和有症状病例之间的差异,评估了其临床价值。我们还测定了研究组中的-核DNA(-nDNA)拷贝数和线粒体转录因子A(TFAM)mRNA水平。
从37名新冠患者和33名对照中采集血浆和血沉棕黄层样本。简要地说,在提取总DNA后,使用标准曲线法通过绝对定量PCR测量血浆中的-mtDNA和-nDNA拷贝数。此外,从血沉棕黄层提取总RNA并进行cDNA合成后,通过定量PCR评估TFAM mRNA水平。
结果显示,无症状新冠患者的-mtDNA水平在统计学上显著高于有症状病例( 值 = 0.01)。然而,有症状患者的-nDNA水平高于无症状病例( 值 = 0.00)。这两组血沉棕黄层中TFAM水平之间无显著差异( 值 > 0.05)。此外,-mtDNA水平在新冠亚组中显示出良好的诊断潜力。
-mtDNA可能因其在炎症和免疫反应中的作用而对新冠病毒感染的结局很重要。它也可能是诊断新冠亚组的一个有前景的候选生物标志物。进一步的研究将有助于理解新冠的病理生理学以及有效的诊断和治疗策略。