Ductular Reaction Cells Display an Inflammatory Profile and Recruit Neutrophils in Alcoholic Hepatitis

Abstract

Chronic liver diseases are characterized by the expansion of ductular reaction (DR) cells and the expression of liver progenitor cell (LPC) markers. In alcoholic hepatitis (AH), the degree of DR expansion correlates with disease progression and short‐term survival. However, little is known about the biological properties of DR cells, their impact on the pathogenesis of human liver disease, and their contribution to tissue repair. In this study, we have evaluated the transcriptomic profile of DR cells by laser capture microdissection in patients with AH and assessed its association with disease progression. The transcriptome analysis of cytokeratin 7‐positive (KRT7+) DR cells uncovered intrinsic gene pathways expressed in DR and genes associated with alcoholic liver disease progression. Importantly, DR presented a proinflammatory profile with expression of neutrophil recruiting C‐X‐C motif chemokine ligand (CXC) and C‐C motif chemokine ligand chemokines. Moreover, LPC markers correlated with liver expression and circulating levels of inflammatory mediators such as CXCL5. Histologically, DR was associated with neutrophil infiltration at the periportal area. In order to model the DR and to assess its functional role, we generated LPC organoids derived from patients with cirrhosis. Liver organoids mimicked the transcriptomic and proinflammatory profile of DR cells. Conditioned medium from organoids induced neutrophil migration and enhanced cytokine expression in neutrophils. Likewise, neutrophils promoted the proinflammatory profile and the expression of chemokines of liver organoids. Conclusion: Transcriptomic and functional analysis of KRT7+ cells indicate that DR has a proinflammatory profile and promote neutrophil recruitment. These results indicate that DR may be involved in the liver inflammatory response in AH, and suggest that therapeutic strategies targeting DR cells may be useful to mitigate the inflammatory cell recruitment in AH.

Canonical Wnt signalling is activated during BEC-to-hepatocyte conversionin vivoand modulates liver epithelial cell plasticity in hepatic organoids

“…BD organoid cells were also expressed various Wnt ligands with described canonical and non-canonical activity, with Wnt7b, Wnt7a and Wnt4 the most abundantly expressed (Figure 4, J) (Eubelen et al, 2018; Vallon et al, 2018; Yu et al, 2008) (Alok et al, 2017; Cho et al, 2017; Ferrari et al, 2018; Le Grand et al, 2009; Posokhova et al, 2015). To assess whether the panel of Wnts expressed by BD organoids might be relevant in a clinical setting, we examined the expression of Wnt ligands in the ductal reactions of human cirrhotic patients using the public data from laser capture microdissection (LCM) experiments (Aguilar Bravo et al, 2019). Wnt7b was the most abundantly expressed Wnt ligand in ductal reactions of cirrhotic human samples dissected based on CK7+ expression, suggesting that the biliary epithelium might be an important contributing source to the ductal reactions Wnt microenvironment of these patients (Figure 4, K).…”

“…Organoids primarily expressed Wnt4, Wnt7b and Wnt7a and Rspo1 HPC-induced signature was dependent on these endogenously produced Wnt ligands. Of the organoid expressed ligands, Wnt7b was strongly expressed in DR of human cirrhotic patients (Aguilar Bravo et al, 2019). A range of other studies also demonstrated expression of Wnt7a and Wnt7b ligands in BECs following liver injury (Okabe et al, 2016; Pepe-Mooney et al, 2019;Boulter et al, 2015).…”

“…The appearance of HPCs is thought to be associated with the creation of cellular niches in response to chronic damage and regeneration such as an unbalanced diet or viral infection. BEC-to-hepatocyte conversion in particular may be of special clinical relevance in liver pathological settings resulting from chronic and repeated injury such is long-term alcohol abuse (Aguilar Bravo et al, 2019).…”

Wnt/β‐catenin signalling controls bile duct regeneration by regulating differentiation of ductular reaction cells

“…8 Related studies have shown that DRCs are a cell group different from liver progenitor cells (HPCs) and have their unique repair function. 36 An essential prerequisite for biliary repair is that DRCs can reconstruct the biliary structure. 37 In this study, In DR studies, EpCAM and SOX9 are often identified as stem cell markers.…”

Neutrophils interact with cholangiocytes to cause cholestatic changes in alcoholic hepatitis

“…14 Similarly, a role for neutrophils in the pathogenesis of AH has always been presumed, and in fact their presence in the hepatic lobule is part of the criteria for histological diagnosis. 12 However, to our knowledge, it has not previously been considered that neutrophils interact with cholangiocytes in this disease (except for with reactive bile ducts), 52 nor that their interaction must include direct contact. By providing evidence that neutrophils bind to cholangiocytes via ITGB1 in AH and that the degree of interaction reflects the degree of jaundice (figures 1 and 6), the current work provides unexpected new cell and molecular targets for therapy.…”