Next-generation proteomics of serum extracellular vesicles combined with single-cell RNA sequencing identifies MACROH2A1 associated with refractory COVID-19.

IF 5 3区医学 Q2 IMMUNOLOGY

Inflammation and Regeneration Pub Date : 2022-11-30 DOI:10.1186/s41232-022-00243-5

Takahiro Kawasaki, Yoshito Takeda, Ryuya Edahiro, Yuya Shirai, Mari Nogami-Itoh, Takanori Matsuki, Hiroshi Kida, Takatoshi Enomoto, Reina Hara, Yoshimi Noda, Yuichi Adachi, Takayuki Niitsu, Saori Amiya, Yuta Yamaguchi, Teruaki Murakami, Yasuhiro Kato, Takayoshi Morita, Hanako Yoshimura, Makoto Yamamoto, Daisuke Nakatsubo, Kotaro Miyake, Takayuki Shiroyama, Haruhiko Hirata, Jun Adachi, Yukinori Okada, Atsushi Kumanogoh

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引用次数: 0

Abstract

Background: 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.

Methods: 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.

Results: 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.

Conclusions: 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.

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新一代血清细胞外囊泡蛋白质组学结合单细胞RNA测序鉴定出与难治性COVID-19相关的MACROH2A1

背景:2019冠状病毒病(COVID-19)大流行广泛存在;然而，对难治性病例的准确预测尚未建立。循环细胞外囊泡参与了许多病理过程，是探索生物标志物的理想资源。方法:为了鉴定难治性COVID-19的潜在血清生物标志物并检测与发病机制相关的蛋白质，我们对12例不同疾病严重程度的COVID-19患者和4名健康对照者的血清细胞外囊泡进行了高覆盖率的蛋白质组学分析。此外，对10例COVID-19患者和5名健康对照者的外周血单个核细胞进行单细胞RNA测序。结果:在鉴定的3046个细胞外囊泡蛋白中，与非难治性病例相比，难治性病例中MACROH2A1的表达显著升高;其表达随疾病严重程度的增加而增加。在外周血单核细胞的单细胞RNA测序中，MACROH2A1的表达定位于单核细胞，在危重病例中表达升高。与此一致，肺组织单核RNA测序结果显示，MACROH2A1在单核细胞和巨噬细胞中高表达，在致死性COVID-19中显著升高。此外，分子网络分析显示，外周血单核细胞和肺的转录组中富集了“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，细胞外囊泡蛋白质组学中也普遍富集。结论: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。

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来源期刊

Inflammation and Regeneration Multiple-

CiteScore

11.10

自引率

1.20%

发文量

审稿时长

11 weeks

期刊介绍： Inflammation and Regeneration is the official journal of the Japanese Society of Inflammation and Regeneration (JSIR). This journal provides an open access forum which covers a wide range of scientific topics in the basic and clinical researches on inflammation and regenerative medicine. It also covers investigations of infectious diseases, including COVID-19 and other emerging infectious diseases, which involve the inflammatory responses. Inflammation and Regeneration publishes papers in the following categories: research article, note, rapid communication, case report, review and clinical drug evaluation.