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

Nicolò Manicardi,Martí Ortega-Ribera,Jaime Bosch,Felix Royo,Javier Vaquero,Agustín Albillos,Felix Elortza,Juan Carlos García-Pagán,David Sanfeliu-Redondo,Francisco Javier Cubero,Juan M Falcón-Pérez,Amelia J Hessheimer,Ana Martínez-Alcocer,Anabel Fernández-Iglesias,Constantino Fondevila,Jordi Gracia-Sancho,Mikel Azkargorta,Laia Abad-Jordà,Juan José Lozano

doi:10.3390/cancers13112688

Abstract

Simple SummaryWe define the transcriptome and secretome of primary LSECs during the progression of cirrhosis, revealing specific molecular signatures, novel biomarkers and therapeutic targets for new disease-modifying treatments for patients with advanced chronic liver disease.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.

Highlights

Cirrhosis represents the final pre-neoplastic stage of a group of chronic liver diseases (CLD) characterized by a complex ensemble of longstanding pathophysiological processes that alters liver function, architecture and hemodynamics
RNA sequencing of liver sinusoidal endothelial cells (LSECs) isolated from cirrhotic CCl4 (12–14 weeks), TAA and cBDL pre-clinical models of cirrhosis identified 7028 differentially expressed genes (DEG) in cirrhotic CCl4-LSECs (56.2% up-regulated and 43.8% down-regulated) (Table S1), 3382 DEG in cirrhotic TAA-LSECs (55.6% up-regulated and 44.4% down-regulated) (Table S2), and 4578 DEG in cBDL-LSECs (55.4% up-regulated and 44.6% down-regulated) (Table S3) compared with their corresponding healthy LSECs (Figure 1A)
The main molecular processes defined in cirrhotic LSECs de-regulation were cell communication and organization, pro-oncogenic processes, cell death and survival, immune response, lipid metabolism, and cellular growth and proliferation, among others (Figure 1B)

Summary

Introduction

Cirrhosis represents the final pre-neoplastic stage of a group of chronic liver diseases (CLD) characterized by a complex ensemble of longstanding pathophysiological processes that alters liver function, architecture and hemodynamics. Many studies on the underlying mechanisms of CLD and PH have been made only at the end-stage of the disease, ignoring the chronological changes that promote progression towards cirrhosis. In this context, animal models of liver disease represent an optimal choice when it comes to study the sequential mechanisms of these alterations [6]

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Conclusion