AUTHOR=Yu Leilin , Lin Wei , Shen Chanjuan , Meng Ting , Jin Peng , Ding Xiang , Eggenhuizen Peter J. , Ooi Joshua D. , Tang Rong , Nie Wannian , Li Xia , Xiao Xiangcheng , Zhong Yong 

TITLE=Intrarenal Single-Cell Sequencing of Hepatitis B Virus Associated Membranous Nephropathy

JOURNAL=Frontiers in Medicine

VOLUME=Volume 9 - 2022

YEAR=2022

URL=https://www.frontiersin.org/journals/medicine/articles/10.3389/fmed.2022.869284

DOI=10.3389/fmed.2022.869284

ISSN=2296-858X

ABSTRACT=Hepatitis B virus (HBV) infection has been associated with the development of various kidney diseases. Among them, membranous nephropathy (MN) is the most common kidney manifestation of HBV and usually develops in the chronic phase. To date, the pathogenesis of HBV-associated MN has been less elucidated. This study aimed to decipher the etiopathogenesis of HBV-associated MN by performing single-cell RNA sequencing (scRNA-seq) of kidney biopsy specimens from a patient with HBV-associated MN and two healthy individuals. Based on the scRNA-seq, we generated 4,114 intrarenal single-cell transcriptomes from the HBV-associated MN patient by scRNA-seq. We found that compared to healthy individuals, podocytes in the HBV-associated MN patient showed an increased expression of extracellular matrix formation-related genes, including HSPA5, CTGF, and EDIL3. Kidney endothelial cells (ECs) in the HBV-associated MN sample were enriched in inflammatory pathways, these include both innate and adaptive inflammation, in addition to inflammatory and physiologic responses including antigen processing and presentation, protein processing in endoplasmic reticulum, NF-kappa B signaling, IL-17 signaling, TNF signaling and NOD-like receptor signaling. Next, gene ontology (GO) functional enrichment analysis and Gene Set Variation Analysis (GSVA) further revealed that differentially expressed genes (DEGs) of ECs from the HBV-associated MN patients were enriched in apoptotic signaling pathway, response to cytokine, leukocyte cell-cell adhesion and response to interferon-gamma when compared with healthy subjects. The up-regulated DEGs in glomerular ECs of HBV-associated MN patients were involved in biological processes such as viral gene expression, and protein targeting to endoplasmic reticulum. Moreover, DEGs and enrichment analyses were compared between HBV-associated MN and anti-phospholipase A2 receptor (PLA2R)-positive idiopathic membranous nephropathy (IMN) using scRNA-seq of renal cells. The results verified that the overexpressed genes in ECs from HBV-associated MN were mainly enriched in regulation of protein targeting to endoplasmic reticulum, exocytosis, viral gene expression, IL-6 and IL-1 secretion when compared with anti-PLA2R positive IMN. The receptor-ligand crosstalk analysis revealed potential interactions between endothelial cells and other cells in HBV-associated-MN. These results offer new insight into the pathogenesis of HBV-associated MN and may identify new therapeutic targets for HBV-associated MN.