EREG-secreting THBS1+ tissue monocytes are recruited by C5a to promote rapid liver regeneration in patients and mice during the ALPPS procedure.

An ongoing debate surrounds the impact of chemotherapy on post-hepatectomy liver regeneration in patients with colorectal cancer liver metastases (CRLM), with unclear regulatory mechanisms. This study sought to delve into liver regeneration post-resection in CRLM patients, specifically examining the roles of hepatocyte growth factor (HGF) and transforming growth factor β1 (TGF-β1). In this longitudinal observational study, 17 patients undergoing major liver resection for CRLM and 17 with benign indications as controls were enrolled. Liver regeneration within 30 postoperative days was assessed via CT, considering clinicopathological characteristics, liver enzymes, liver stiffness by elastography, and the impact of HGF and TGF-β1 on liver regeneration. The results revealed that the control group exhibited significantly higher mean liver regeneration volume (200 ± 180 mL) within 30 days postoperatively compared to the CRLM group (72 ± 154 mL); p = 0.03. Baseline alkaline phosphatase (AP) and TGF-β1 blood levels were notably higher in the CRLM group. Immunohistochemical analysis indicated a higher proportion of CRLM patients with high TGF-β1 expression in liver tissues compared to the control group (p = 0.034). Correlation analysis showed that resected liver volume, baseline plasma HGF, AP, and albumin levels significantly correlated with liver regeneration volume. However, in multivariable analysis, only resected liver volume (β: 0.31; 95% CI: 0.14–0.47, p = 0.01) remained significant. In conclusion, this study highlights compromised liver regeneration in CRLM patients post-chemotherapy. Additionally, these patients exhibited lower serum TGF-β1 levels and reduced TGF-β1 expression in liver tissue, suggesting TGF-β1 involvement in mechanisms hindering liver regeneration capacity following major resection after chemotherapy.

Rapid liver regeneration is required after living-donor liver transplantation and oncologic liver resections to warrant sufficient liver function and prevent small-for-size syndrome. Recent evidence highlights the therapeutic potential of mesenchymal stem cells (MSC) for treatment of toxic liver injury, but whether MSC and their secreted factors stimulate liver regeneration after surgical injury remains unknown. Therefore, the aim of this study is to investigate the effect of human liver-derived MSC-secreted factors in an experimental liver resection model. C57BL/6 mice were subjected to a 70% partial hepatectomy and treated with either concentrated MSC-conditioned culture medium (MSC-CM) or vehicle control. Animals were analyzed for liver and body weight, hepatocyte proliferation, and hepatic gene expression. Effects of MSC-CM on gene expression in a human hepatocyte-like cell line (Huh7 cells) were analyzed using genome-wide gene expression arrays. Liver regeneration was significantly stimulated by MSC-CM as shown by an increase in liver to body weight ratio and hepatocyte proliferation. MSC-CM upregulated hepatic gene expression of cytokines and growth factors relevant for cell proliferation, angiogenesis, and anti-inflammatory responses. In vitro, treatment of Huh7 cells with MSC-CM significantly altered expression levels of ~3,000 genes. Functional analysis revealed strong effects on networks associated with protein synthesis, cell survival, and cell proliferation. This study shows that treatment with MSC-derived factors can promote hepatocyte proliferation and regenerative responses in the early phase after surgical resection. MSC-CM may represent a feasible new strategy to promote liver regeneration in patients undergoing extensive liver resection or after transplantation of small liver grafts.

Background: There is limited knowledge about the mechanisms behind the unparalleled growth of the future liver remnant (FLR) linked to associating liver partition and portal vein ligation for staged hepatectomy (ALPPS). In this study, liver regenerative markers were examined in patients subjected to ALPPS. Methods: Ten patients with colorectal liver metastases treated with neoadjuvant chemotherapy and ALPPS were included. Plasma was sampled at 6 time points and biopsies from both liver lobes were collected at both stages of ALPPS. The levels of interleukin (IL)-6, hepatocyte growth factor (HGF), tumor necrosis factor-α, epidermal growth factor, and vascular endothelial growth factor in plasma were measured at each time point. Expression of mRNA for markers of proliferation and apoptosis was studied in the biopsies from both liver lobes taken at both stages. Results: ALPPS resulted in a peak of IL-6 after stage 1 (p = 0.004), which decreased rapidly and did not increase again after stage 2. HGF also increased after stage 1 (p = 0.048), and the HGF levels correlated significantly with the degree of growth of the FLR before stage 2 (p = 0.02, r² = 0.47). There was a correlation between peak levels of IL-6 and HGF (p = 0.03, r² = 0.84). Conclusions: IL-6 and HGF seem to be early mediators of hypertrophy after stage 1 in the ALPPS procedure. The peak HGF plasma level correlates with the degree of FLR growth in patients subjected to ALPPS.

Portal vein embolization (PVE) can decrease the resection ratio for major hepatectomy. (99m)Tc-galactosyl human serum albumin (GSA) scintigraphy is useful for evaluating quantitative functional liver volume. Branched chain amino acids (BCAAs) modulate liver function and regeneration. We analyzed the effects of BCAAs, in terms of liver function and regeneration after PVE, in combination with major hepatectomy. This randomized controlled trial was conducted for patients receiving PVE through to complete hepatectomy from September, 2011 to June, 2013. BCAA granules were added two times a day to a conventional diet in the BCAA administration group (BCAA group). The primary end point was functional liver regeneration of the future remnant liver after PVE followed by hepatic resection. Functional liver regeneration was assessed by the liver uptake value obtained from (99m)Tc-GSA scintigraphy single-photon-emission computed tomography/computed tomography fusion images. The secondary end points were volumetric liver regeneration and changes in liver function and laboratory data. A BCAA group (n = 13) and a non-BCAA group (control group; n = 15) were included. The primary end point was partially met: the liver uptake value significantly increased in the BCAA group compared with the control group 6 months after hepatic resection (266.7% vs 77.6%, P = 0.04) and marginally increased after PVE (43.8% vs 17.4%, P = 0.079). Following PVE, the increment of the uptake ratio of the liver to the liver plus heart at 15 min was significantly less in the BCAA group than in the control group (0.0 and 0.01, P = 0.023). BCAA supplementation improved functional liver regeneration and function in patients undergoing PVE followed by major hepatic resection.

Integrating CyTOF with scRNA-seq reveals that Cyclovirobuxine D inhibits HCC progression through hepatic immune microenvironment remodeling.

BackgroundThe influence of pyroptosis on tumors is complex and diverse. However, its specific impact on hepatocellular carcinoma (HCC) is still not well understood. Therefore, the objective of this study was to develop a prognostic signature for HCC based on pyroptosis-related genes.MethodsThe single-cell RNA sequencing (scRNA-seq) data, mRNA expression files and corresponding clinical information of HCC were obtained from the The Cancer Genome Atlas and Gene Expression Omnibus databases. Python was used to process scRNA-seq data and calculated the enrichment score of pyroptosis-related genes (PRGs). Weight Co-Expression Network Analysis was used to identify pyroptosis-related hub genes. By overlapping the PRGs from scRNA-seq analysis and bulk RNA-seq analysis, respectively. Then, Univariate cox and LASSO regression were used to construct the pyroptosis prognostic model. Multivariate cox was used to identify independent factors for HCC and then developed a nomogram. The biological functions, survival analysis, immune characteristics, therapy response, and m6A modification status were analyzed.ResultsBased on the scRNA-seq analysis and bulk RNA-seq analysis, hub PRGs were identified in HCC. Of those genes, five PRGs (ADGRE2, FCER1G, SLC9A9, CYBB, SLAMF6) were selected as a prognostic signature. The risk score established from the prognostic signature was an independent prognostic factor for HCC. The high-risk score group is associated with a poor prognosis, characterized by immunosuppressive features.ConclusionThis study uniquely integrates single-cell and bulk transcriptomic data to systematically identify pyroptosis-related prognostic biomarkers, pinpointing their cellular origin within the tumor microenvironment.

Liver cirrhosis is prognostically associated with poor life expectancy owing to subsequent liver failure. Thus, understanding liver regeneration processes during cirrhotic injury is highly important. This study explored the role of macrophage heterogeneity in liver regeneration following splenectomy. We collected detailed clinical information from 54 patients with decompensated cirrhosis before and after splenectomy. Obvious liver regeneration was observed after splenectomy in cirrhotic patients. Single-cell RNA sequencing (scRNA-seq) was performed on three paired liver tissues from patients before and after surgery to explore the immune microenvironment map and the characteristics of liver regeneration-associated macrophages (RAMs). scRNA-seq analysis revealed that the composition of hepatic immune cells changed after splenectomy; among these changes, the proportion of CD300E+ RAMs significantly increased after surgery, and high expression levels of functional genes associated with cell proliferation promoted liver regeneration. Moreover, a mouse model of carbon tetrachloride-induced cirrhosis and a coculture system consisting of primary bone marrow-derived macrophages and hepatocytes were established for validation. We observed a similar phenomenon of liver regeneration in cirrhotic mice and further confirmed that CD300E+ monocyte-derived macrophages facilitated hepatocyte NAD+ synthesis via the secretion of NAMPT, which subsequently promoted hepatocyte proliferation. This study characterized the hepatic immune microenvironment in patients with cirrhosis following splenectomy. Our findings demonstrated that CD300E+ macrophages play a crucial role in remodeling the hepatic immune microenvironment after splenectomy, thereby promoting liver regeneration in patients with decompensated cirrhosis. CD300E+ macrophages are anticipated to emerge as a novel therapeutic strategy for the treatment of liver cirrhosis.

The Zinc Transporter Zip14 Influences c-Met Phosphorylation and Hepatocyte Proliferation During Liver Regeneration in Mice

PurposeLiver resection is currently considered the most effective treatment for patients with liver cancer. To the best of our knowledge, no study has investigated the association between gadoxetic acid-enhanced magnetic resonance imaging (MRI) findings and liver regeneration in patients who underwent hemihepatectomy. We aimed to clarify the relationship between the signal intensity (SI) of the liver parenchyma on gadoxetic acid-enhanced MRI and the degree of liver regeneration in patients who underwent hemihepatectomy.Materials and methodsForty-one patients who underwent gadoxetic acid-enhanced MRI before hemihepatectomy were enrolled. We calculated the liver-to-erector spinae muscle SI ratio (LMR) in the hepatobiliary phase and the precontrast images. ΔLMR was calculated using the following equation: ΔLMR = (LMR in the hepatobiliary phase−LMR in the precontrast image)/LMR in the precontrast image. The preoperative and postoperative remnant liver volumes (LVs) were calculated using CT volumetry. We calculated the resection rate (RR) and liver regeneration index (LRI) using the following formulas: RR = Resected LV/Total LV × 100 and LRI = (postoperative remnant LV−preoperative remnant LV)/preoperative remnant LV × 100. The relationships among LRI, imaging, and clinicopathological factors were analyzed.ResultsUnivariate analysis showed RR and ΔLMR showed a positive correlation with LRI (ρ = 0.4133, p = 0.0072 and ρ = 0.7773, p < 0.001, respectively). Spleen volume showed a negative correlation with LRI (ρ = −0.3138, p = 0.0486). Stepwise multiple regression analysis showed ΔLMR and RR were independently correlated with LRI (β coefficient = 44.8771, p = 0.0198 and β coefficient = 1.9653, p < 0.001, respectively).ConclusionΔLMR may serve as a preoperative predictor of liver regeneration in patients undergoing hemihepatectomy.

To explore whether intravoxel incoherent motion (IVIM) parameters could evaluate liver regeneration preoperatively. A total of 175 HCC patients were initially recruited. The apparent diffusion coefficient, true diffusion coefficient (D), pseudodiffusion coefficient (D*), pseudodiffusion fraction (f), diffusion distribution coefficient, and diffusion heterogeneity index (Alpha) were measured by two independent radiologists. Spearman's correlation test was used to assess correlations between IVIM parameters and the regeneration index (RI), calculated as 100% × (the volume of the postoperative remnant liver - the volume of the preoperative remnant liver) / the volume of the preoperative remnant liver. Multivariate linear regression analyses were used to identify the factors for RI. Finally, 54 HCC patients (45 men and 9 women, mean age 51.26 ± 10.41 years) were retrospectively analyzed. The intraclass correlation coefficient ranged from 0.842 to 0.918. In all patients, fibrosis stage was reclassified as F0-1 (n = 10), F2-3 (n = 26), and F4 (n = 18) using the METAVIR system. Spearman correlation test showed D* (r = 0.303, p = 0.026) was associated with RI; however, multivariate analysis showed that only D value was a significant predictor (p< 0.05) of RI. D and D*showed moderate correlations with fibrosis stage (r = -0.361, p= 0.007; r = -0.457, p = 0.001). Fibrosis stage showed a negative correlation with RI (r = -0.263, p= 0.015). In the 29 patients who underwent minor hepatectomy, only the D value showed a positive association (p< 0.05) with RI, and a negative correlation with fibrosis stage (r = -0.360, p= 0.018). However, in the 25 patients who underwent major hepatectomy, no IVIM parameters were associated with RI (p > 0.05). The D and D* values, especially the D value, may be reliable preoperative predictors of liver regeneration. • The D and D* values, especially the D value, derived from IVIM diffusion-weighted imaging may be useful markers for the preoperative prediction of liver regeneration in patients with HCC. • The D and D* values derived from IVIM diffusion-weighted imaging show significant negative correlations with fibrosis, an important predictor of liver regeneration. • No IVIM parameters were associated with liver regeneration in patients who underwent major hepatectomy, but the D value was a significant predictor of liver regeneration in patients who underwent minor hepatectomy.

Whether keratoconus (KC) is an inflammatory disease is currently debated. Hence, we aimed to investigate the immune-related features of KC based on single-cell RNA sequencing (scRNA-seq) and bulk RNA sequencing (bulk RNA-seq) data. scRNA-seq data were obtained from the Genome Sequence Archive (GSA), bulk RNA-seq data were obtained from the Gene Expression Omnibus (GEO), and immune-associated genes(IAGs) were obtained from the ImmPort database. Cell clusters of KC were annotated, and different cell clusters were then selected. The IAG score of each cell was calculated using the AUCell package. Three bulk RNA-seq datasets were merged and used to identify the differentially expressed genes (DEGs), biological functions, and immune characteristics. Weighted gene coexpression network analysis (WGCNA) was used to select the IAG score-related hub genes. Based on scRNA-seq and bulk RNA-seq analyses, three machine learning algorithms, including random forest (RF), support vector machine (SVM), and least absolute shrinkage and selection operator (LASSO) regression analysis, were used to identify potential prognostic markers for KC. A predictive nomogram was developed based on prognostic markers. Six cell clusters were identified in KC, and decreased corneal stromal cell-5 (CSC-5) and increased CSC-6 were found in KC. CSC and immune cell clusters had the highest IAG scores. The bulk RNA-seq analysis identified 1362 DEGs (553 upregulated and 809 downregulated) in KC. We found different immune cell populations and differentially expressed cytokines in KC. More than three key IAG score-related modules and 367 genes were identified. By integrating the scRNA-seq and bulk RNA-seq analyses, 250 IAGs were selected and then incorporated into three machine learning models, and 10 IAGs (CEP112, FYN, IFITM1, IGFBP5, LPIN2, MAP1B, RNASE1, RUNX3, SMIM10, and SRGN) were identified as potential prognostic genes that were significantly associated with cytokine and matrix metalloproteinase(MMP)1-14 expression. Finally, a predictive nomogram was constructed and validated. Taken together, our results identified CSCs and immune cell clusters that may play a key role during KC progression by regulating immunological features and maintaining cell stability.

An innovative approach, named associating liver partition and portal vein ligation for staged hepatectomy (ALPPS), has made possible a marked increase in future liver remnant (FLR) over a short period of time, and has challenged the concept of unresectability. To date, ALPPS has been employed for a variety of primary and metastatic liver tumors from many centers across the world. Many technical variations of ALPPS have been described. However, much controversy has surrounded this procedure, and many problems remain to be solved, such as the reasons for rapid hypertrophy, the technical feasibility, safety and long-term oncologic outcomes, and the result compared with the conventional two-stage hepatectomy. These aspects are needed to be evaluated further in large-scale studies. We believe that the ALPPS procedure will has much room for further development. Key words: Liver neoplasms; Associating liver partition and portal vein ligation for staged hepatectomy

BackgroundThe liver has the remarkable capacity to regenerate in order to compensate for lost or damaged hepatic tissue. However, pre-existing pathological abnormalities, such as hepatic steatosis (HS), inhibits the endogenous regenerative process, becoming an obstacle for liver surgery and living donor transplantation.Recent evidence indicates that multipotent mesenchymal stromal cells (MSCs) administration can improve hepatic function and increase the potential for liver regeneration in patients with liver damage. Since HS is the most common form of chronic hepatic illness, in this study we evaluated the role of MSCs in liver regeneration in an animal model of severe HS with impaired liver regeneration.MethodsC57BL/6 mice were fed with a regular diet (normal mice) or with a high-fat diet (obese mice) to induce HS. After 30 weeks of diet exposure, 70% hepatectomy (Hpx) was performed and normal and obese mice were divided into two groups that received 5 × 105 MSCs or vehicle via the tail vein immediately after Hpx.ResultsWe confirmed a significant inhibition of hepatic regeneration when liver steatosis was present, while the hepatic regenerative response was promoted by infusion of MSCs. Specifically, MSC administration improved the hepatocyte proliferative response, PCNA-labeling index, DNA synthesis, liver function, and also reduced the number of apoptotic hepatocytes.These effects may be associated to the paracrine secretion of trophic factors by MSCs and the hepatic upregulation of key cytokines and growth factors relevant for cell proliferation, which ultimately improves the survival rate of the mice.ConclusionsMSCs represent a promising therapeutic strategy to improve liver regeneration in patients with HS as well as for increasing the number of donor organs available for transplantation.

The liver is the only organ that can regenerate and regain its original tissue-to-body weight ratio within a short period of time after tissue loss. Insufficient liver regeneration in patients after partial hepatectomy or liver transplantation with partial liver grafts often leads to post-hepatectomy liver failure or small-for-size syndrome, respectively. Enhancing liver regeneration after liver injury might improve outcomes and increase patient survival. Liver regeneration comprises hepatocyte proliferation, and hepatic progenitor cell expansion and differentiation into hepatocytes. The immune system is intensively involved in liver regeneration. The current review provides a comprehensive overview of the diverse roles played by immune cells in liver regeneration. Macrophages, neutrophils, eosinophils, basophils, mast cells, platelets, dendritic cells, type 1 innate lymphoid cells, B cells, and T cells are implicated in promoting liver regeneration, while natural killer cells and overactivated natural killer T cells are supposed to inhibit hepatocyte proliferation. We also highlight the predominant underlying mechanisms mediated by immune cells, which may contribute to the development of novel strategies for promoting liver regeneration in patients with liver diseases.

Hepatocellular carcinoma (HCC) is an aggressive malignancy associated with an immunosuppressive tumor microenvironment (TME), which contributes to resistance to therapy. Lycorine, a natural alkaloid, has demonstrated anticancer potential, but its effects on HCC remain insufficiently explored. This study aimed to systematically assess the impact of lycorine on HCC and elucidate its underlying mechanisms of action. In vivo orthotopic HCC mouse models were employed, incorporating high-dimensional single-cell techniques to comprehensively analyze the effects of lycorine on the TME. Mice bearing orthotopic HCC tumors were treated with lycorine at doses of 10 or 20 mg/kg for 14 days. Tumor progression was assessed by measuring tumor weight. The immune landscape was examined using cytometry by time of flight (CyTOF), and cellular heterogeneity was evaluated via single-cell RNA sequencing (scRNA-seq). Cell-cell communication networks were reconstructed through CellChat analysis. Lycorine treatment significantly reduced tumor burden and modulated cytokine expression profiles. CyTOF analysis revealed that lycorine reprogrammed the immune landscape, enhancing macrophage and neutrophil infiltration while decreasing PD-L1 expression on neutrophils. scRNA-seq analysis indicated that lycorine regulated specific cell subpopulations, including malignant epithelial cells, Notch2+ neutrophils, and Spp1+ macrophages. Cell-cell communication analysis identified the regulation of the Mif-(Cd74+Cd44/Cxcr4) signaling axis as a key underlying mechanism. Lycorine remodels the HCC TME through coordinated actions on malignant epithelial cells and immune populations, notably neutrophil and macrophage subpopulations. These findings support the potential development of lycorine-based combination therapies for HCC.