Kawasaki Takahiro, Takeda Yoshito, Edahiro Ryuya, Shirai Yuya, Nogami-Itoh Mari, Matsuki Takanori, Kida Hiroshi, Enomoto Takatoshi, Hara Reina, Noda Yoshimi, Adachi Yuichi, Niitsu Takayuki, Amiya Saori, Yamaguchi Yuta, Murakami Teruaki, Kato Yasuhiro, Morita Takayoshi, Yoshimura Hanako, Yamamoto Makoto, Nakatsubo Daisuke, Miyake Kotaro, Shiroyama Takayuki, Hirata Haruhiko, Adachi Jun, Okada Yukinori, Kumanogoh Atsushi
Department of Respiratory Medicine and Clinical Immunology, Graduate School of Medicine, Osaka University, Osaka, 565-0871, Japan.
Laboratory of Immunopathology, World Premier International Immunology Frontier Research Center (WPI-IFReC), Osaka University, Osaka, 565-0871, Japan.
Inflamm Regen. 2022 Nov 30;42(1):53. doi: 10.1186/s41232-022-00243-5.
The coronavirus disease 2019 (COVID-19) pandemic is widespread; however, accurate predictors of refractory cases have not yet been established. Circulating extracellular vesicles, involved in many pathological processes, are ideal resources for biomarker exploration.
To identify potential serum biomarkers and examine the proteins associated with the pathogenesis of refractory COVID-19, we conducted high-coverage proteomics on serum extracellular vesicles collected from 12 patients with COVID-19 at different disease severity levels and 4 healthy controls. Furthermore, single-cell RNA sequencing of peripheral blood mononuclear cells collected from 10 patients with COVID-19 and 5 healthy controls was performed.
Among the 3046 extracellular vesicle proteins that were identified, expression of MACROH2A1 was significantly elevated in refractory cases compared to non-refractory cases; moreover, its expression was increased according to disease severity. In single-cell RNA sequencing of peripheral blood mononuclear cells, the expression of MACROH2A1 was localized to monocytes and elevated in critical cases. Consistently, single-nucleus RNA sequencing of lung tissues revealed that MACROH2A1 was highly expressed in monocytes and macrophages and was significantly elevated in fatal COVID-19. Moreover, molecular network analysis showed that pathways such as "estrogen signaling pathway," "p160 steroid receptor coactivator (SRC) signaling pathway," and "transcriptional regulation by STAT" were enriched in the transcriptome of monocytes in the peripheral blood mononuclear cells and lungs, and they were also commonly enriched in extracellular vesicle proteomics.
Our findings highlight that MACROH2A1 in extracellular vesicles is a potential biomarker of refractory COVID-19 and may reflect the pathogenesis of COVID-19 in monocytes.
2019年冠状病毒病（COVID-19）大流行广泛传播；然而，难治性病例的准确预测指标尚未确立。参与多种病理过程的循环细胞外囊泡是探索生物标志物的理想资源。
为了鉴定潜在的血清生物标志物并研究与难治性COVID-19发病机制相关的蛋白质，我们对从12例不同疾病严重程度的COVID-19患者和4例健康对照者收集的血清细胞外囊泡进行了高覆盖蛋白质组学分析。此外，还对从10例COVID-19患者和5例健康对照者收集的外周血单个核细胞进行了单细胞RNA测序。
在鉴定出的3046种细胞外囊泡蛋白中，与非难治性病例相比，MACROH2A1在难治性病例中的表达显著升高；此外，其表达随疾病严重程度增加。在外周血单个核细胞的单细胞RNA测序中，MACROH2A1的表达定位于单核细胞，在重症病例中升高。同样，肺组织的单核RNA测序显示，MACROH2A1在单核细胞和巨噬细胞中高表达，在致命性COVID-19中显著升高。此外，分子网络分析表明，“雌激素信号通路”、“p160类固醇受体共激活因子（SRC）信号通路”和“STAT转录调控”等通路在外周血单个核细胞和肺单核细胞的转录组中富集，它们也常见于细胞外囊泡蛋白质组学中。
我们的研究结果表明，细胞外囊泡中的MACROH2A1是难治性COVID-19的潜在生物标志物，可能反映了COVID-19在单核细胞中的发病机制。
