Single-Cell RNA Sequencing Revealed CD14+ Monocytes Increased in Patients With Takayasu’s Arteritis Requiring Surgical Management

Abstract

Objectives: Takayasu Arteritis (TA) is a highly specific vascular inflammation and poses threat to patients’ health. Although some patients have accepted medical treatment, their culprit lesions require surgical management (TARSM). This study aimed at dissecting the transcriptomes of peripheral blood mononuclear cells (PBMCs) in these patients and to explore potential clinical markers for TA development and progression.Methods: Peripheral blood were collected from four TA patients requiring surgical management and four age-sex matched healthy donors. Single cell RNA sequencing (scRNA-seq) was adopted to explore the transcriptomic diversity and function of their PBMCs. ELISA, qPCR, and FACS were conducted to validate the results of the analysis.Results: A total of 29918 qualified cells were included for downstream analysis. Nine major cell types were confirmed, including CD14+ monocytes, CD8+ T cells, NK cells, CD4+ T cells, B cells, CD16+ monocytes, megakaryocytes, dendritic cells and plasmacytoid dendritic cells. CD14+ monocytes (50.0 vs. 39.3%, p < 0.05) increased in TA patients, as validated by FACS results. TXNIP, AREG, THBS1, and CD163 increased in TA patients. ILs like IL-6, IL-6STP1, IL-6ST, IL-15, and IL-15RA increased in TA group.Conclusion: Transcriptome heterogeneities of PBMCs in TA patients requiring surgical management were revealed in the present study. In the patients with TA, CD14+ monocytes and gene expressions involved in oxidative stress were increased, indicating a new treatment and research direction in this field.

Transcriptome profiling of abdominal aortic tissues reveals alterations in mRNAs of Takayasu arteritis

“…To the best of our knowledge, this study firstly performed bulk RNA-sequencing of TA vessel tissues to obtain the overall alterations of transcriptome in TA. Regarding the materials used for TA research, most studies detected RNA, cytokines, chemokines by using peripheral blood samples ( Matsumoto et al, 2021 ; Qing et al, 2021 ; Kabeerdoss et al, 2022 ; Kong et al, 2022 ). In this way, several susceptibility locus have been identified by genome-wide association studies to pinpoint potential clues for new therapeutic targets ( Renauer et al, 2015 ; Ortiz-Fernández et al, 2021 ).…”

“…The protocol for single-cell RNA sequencing followed the instructions of the GEXSCOPETM Single-Cell RNA Library Kit (Singleron Biotechnologies). Details have been described and can be referred to Gao et al ( Qing et al, 2021 ).…”

“…Human specimens were then collected and adopted for TA-related research but most published studies focused on peripheral blood including plasma, serum and blood cells. Our team has explored the cellular heterogeneities of peripheral blood and found that CD14 monocytes and gene expressions involving in oxidative stress were increased in TA patients by applying single-cell RNA sequencing (scRNA-seq) ( Qing et al, 2021 ). Profiled chemokines with peripheral blood, suggesting its important roles in TA ( Kong et al, 2022 ).…”

“…Previous studies have revealed critical roles of monocyte and fibroblast in TA ( Wei et al, 2015 ; Wu et al, 2021a ; Qing et al, 2021 ), and some studies have shown that interleukin (ILs) play a critical role in TA, especially IL-6 ( Kong et al, 2016 ; Yang et al, 2016 ; Choy et al, 2020 ; Ma et al, 2020 ; Kadoba et al, 2021 ). A previous CANTOS study has shown the IL-1β monoclonal antibody, Canakinumab, may play an important role in atherosclerosis ( Ridker et al, 2017 ).…”

Design and Protocol for Beijing Hospital Takayasu Arteritis (BeTA) Biobank

“…Patients who have suffered re-interventions because of re-stenosis or graft occlusion will also be admitted into this project. Another focus of the biobank is to provide unprecedented opportunities for tissue sequencing with state-of-the-art techniques like bulk RNA-seq [ 25 ], single-cell RNA sequencing [ 26 ] and multiomics. At the same time, using the sequencing data, we can also find more genes and polymorphisms related to TAK in Asian populations and compare them with Caucasian populations to find the genetic load of populations with different genetic backgrounds.…”