Transcriptomic Profiling of the Liver Sinusoidal Endothelium during Cirrhosis Reveals Stage-Specific Secretory Signature

Abstract

The poor prognosis of chronic liver disease (CLD) generates the need to investigate the evolving mechanisms of disease progression, thus disclosing therapeutic targets before development of clinical complications. Considering the central role of liver sinusoidal endothelial cells (LSECs) in pre-neoplastic advanced CLD, the present study aimed at investigating the progression of CLD from an endothelial holistic perspective. RNAseq defined the transcriptome of primary LSECs isolated from three pre-clinical models of advanced CLD, during the progression of the disease, and from fresh human cirrhotic tissue. At each stage of the disease, the effects of LSECs secretome on neighboring cells and proteomic analysis of LSECs-derived extracellular vesicles (EVs) were also determined. CLD was associated with deep common modifications in the transcriptome of LSECs in the pre-clinical models. Pathway enrichment analysis showed predominance of genes related with pro-oncogenic, cellular communication processes, and EVs biogenesis during CLD progression. Crosstalk experiments revealed endothelial EVs as potent angiocrine effectors. The proteome of LSECs EVs showed stage-specific signatures, including over-expression of tropomyosin-1. Proof-of-principle experiments treating cirrhotic HSCs with recombinant tropomyosin-1 suggested de-activating effects. Our data provide the basis for discovering novel biomarkers and therapeutic targets for new disease-modifying treatments for patients with advanced CLD.

Senescence-induced endothelial phenotypes underpin immune-mediated senescence surveillance

“…This mirrors potential signaling changes seen in independent transcriptional profiling data from human LSECs during the transition from health to chronic liver disease, where RELA motifs were also highly enriched in genes up-regulated in cirrhosis ( Supplemental Fig. S1E ; Manicardi et al 2021 ).…”

“…Enrichment results were plotted using the R package ggplot2 ( Wickham 2009 ). Other data sets used were as follows: Endothelial NF-κB target genes were obtained from the primary publication ( Kempe et al 2005 ), and regulated LSEC genes in health and chronic liver disease ( Manicardi et al 2021 ) were obtained from the Gene Expression Omnibus (GSE164799).…”

Gene Signatures Detect Damaged Liver Sinusoidal Endothelial Cells in Chronic Liver Diseases

“…After incubation with the antibodies, cells were washed and resuspended in a buffer solution without calcium and supplemented with DNase I (3:1, 10104159001, Roche, Switzerland) before cell isolation using FACS (FACS Aria IIu, BD Biosciences, Belgium). FACS was used to sort viable cells (negative selection based on propidium iodide) and LSECs were selected and sorted based on a positive signal for CD32 ( 27 – 29 ) and a negative signal for UV, F4/80, CD45. CD32b is expressed in all LSECs across the liver sinusoid ( Supplementary Figure 1A ) ( 30 ).…”

Cell Death in Hepatocellular Carcinoma: Pathogenesis and Therapeutic Opportunities

“…Phenotypic alterations in LSECs occur from the initial stages of liver injury to HCC development through a process known as capillarization ( Figure 4 B). In this scenario, LSECs lose their characteristic transmembrane pores (fenestrae) and acquire vasoconstrictor, proinflammatory, prothrombotic, and pro-tumorigenic properties [ 3 , 126 ]. At the molecular level, LSECs lose the expression of homeostatic proteins such as LYVE-1 and STAB1 and 2 [ 127 ].…”