Letter to the Editor: Re-evaluating the clinical relevance of the RIPPLY1-TBX19 axis in CTNNB1-mutant hepatocellular carcinoma.

Met-thodology

Simple SummaryLiver cancer is one of the most commonly diagnosed cancers worldwide and the fourth leading cause of cancer-related deaths. Hepatocellular carcinoma (HCC) accounts for at least 80% of all malignant liver primary tumors. A better characterization of molecular mechanisms underlying HCC onset and progression may lead to discover new therapeutic targets and biomarkers. In this study, we performed an integrative transcriptomics analysis to evaluate the clinical relevance of genes associated with hepatocyte differentiation in human HCC. The HepaRG cell line model was used to define a gene expression signature reflecting the status of tumor hepatocyte differentiation. This signature was able to stratify HCC patients into clinically relevant molecular subtypes. Then, a minimal subset of seven differentiation-associated genes was identified to predict a poor prognosis in several cancer datasets.Hepatocellular carcinoma (HCC) is a deadly cancer worldwide as a result of a frequent late diagnosis which limits the therapeutic options. Tumor progression in HCC is closely correlated with the dedifferentiation of hepatocytes, the main parenchymal cells in the liver. Here, we hypothesized that the expression level of genes reflecting the differentiation status of tumor hepatocytes could be clinically relevant in defining subsets of patients with different clinical outcomes. To test this hypothesis, an integrative transcriptomics approach was used to stratify a cohort of 139 HCC patients based on a gene expression signature established in vitro in the HepaRG cell line using well-controlled culture conditions recapitulating tumor hepatocyte differentiation. The HepaRG model was first validated by identifying a robust gene expression signature associated with hepatocyte differentiation and liver metabolism. In addition, the signature was able to distinguish specific developmental stages in mice. More importantly, the signature identified a subset of human HCC associated with a poor prognosis and cancer stem cell features. By using an independent HCC dataset (TCGA consortium), a minimal subset of seven differentiation-related genes was shown to predict a reduced overall survival, not only in patients with HCC but also in other types of cancers (e.g., kidney, pancreas, skin). In conclusion, the study identified a minimal subset of seven genes reflecting the differentiation status of tumor hepatocytes and clinically relevant for predicting the prognosis of HCC patients.

Background and Objective: Hepatocellular carcinoma (HCC) is the dominant form of primary liver tumor and the second most lethal malignancy in worldwide. From transcriptome sequencing analysis, we observed that S100A10 was overexpressed in our cohort of 16 HCC patients. S100A10 is a calcium binding protein and accumulating studies have indicated that annexin A2- S100A10 heterotetramer on the cell surface is important for the conversion of plasminogen to plasmin, which promotes activation of matrix metalloproteinases and degradation of extracellular matrix. This enhances cancer invasion and migration. While annexin A2 overexpression has been documented in various malignancies and shows significant association with local invasion and metastasis, information regarding the expression of S100A10 in human cancers is scarce. Furthermore, how S100A10 was regulated remains unexplored. Hypoxia is a common phenomenon in HCC microenvironment and stabilizies transcription factors, hypoxia inducible factor (HIF)-1α and -2α, to initiate transcription of a wide repertoire of genes for cancer progression. The major aim of this study is to evaluate the possible clinical relevance and diagnostic value of S100A10 in HCC and study the molecular mechanism that drives S100A10 overexpression under the influence of hypoxia in HCC. Experimental Procedures: We evaluated S100A10 expression in human HCC and the corresponding non-tumorous liver (NT) tissues in a cohort of 67 HCC patients by RT-PCR. S100A10 expression was correlated with various HCC clinico-pathological features by Student's t-test and survival outcome of HCC patients by Kaplan-Meier plot followed by log rank test. Promoter of S100A10 was analyzed for potential hypoxia responsive elements -A/GCGTG- and confirmed by ChIP assay with HIF antibodies. To evaluate the impact of HIF on S100A10 expression, we knocked-down (shRNA) and knocked-out (TALEN) HIF-1/2α in a HCC cell line, MHCC97L and evaluated S100A10 mRNA and protein expression. Results: S100A10 was significantly upregulated (P = 0.001, Wilcoxon signed rank test) in HCC as compared to NT tissues. Overexpression of S100A10 (≥ 2 fold) was found in 50.746% (34/67) of HCC patients and was closely correlated with direct liver invasion (P = 0.038), tumor microsatellite formation (P = 0.050) and absence of tumor encapsulation (P = 0.048). More importantly, S100A10 overexpression was associated with poorer 5-year survival rate in HCC patients (P = 0.062). ROC analysis suggested that S100A10 could be a potential diagnostic marker for HCC detection (Area Under Curve = 0.685, P = 0.001). Hypoxia induced S100A10 mRNA and protein expression levels in multiple HCC cell lines and this was abolished in HIF-1/2α knockdown or knockout HCC cells. Conclusion: Our current data have presented evidence that S100A10 can be a potential diagnostic and prognostic marker in HCC and S100A10 is a transcriptional target of HIF. Citation Format: David Kung-Chun Chiu, Carmen Chak-Lui Wong, Irene Oi-Lin Ng, Aki Pui-Wah Tse. S100A10 as a novel biomarker in hepatocellular carcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 533. doi:10.1158/1538-7445.AM2015-533

SCYL3, as a novel binding partner and regulator of ROCK2, promotes hepatocellular carcinoma progression

Mevalonate pathway promotes liver cancer by suppressing ferroptosis through CoQ10 production and selenocysteine-tRNA modification.

The combination of ALBI and AFP response: A small step forward in HCC.

Deciphering Hepatocellular Carcinoma: From Bench to Bedside and Back

Simple SummaryNon-alcoholic fatty liver disease (NAFLD) is a major health problem globally linked with the growing prevalence of metabolic syndrome. A subset of patients with NAFLD progress to non-alcoholic steatohepatitis (NASH), which increases the risk of hepatocellular carcinoma (HCC). However, the mechanisms responsible for the progression to HCC are unclear, and no preventative modalities have been developed. To address this issue, the present study used the natural compound honokiol to clarify the mechanism of this process. The results illustrated that epidermal growth factor receptor (EGFR) was upregulated in mice with NASH, and treatment with honokiol inhibited EGFR and the progression to HCC. Further analysis illustrated that honokiol increased glucocorticoid receptor (GR) nuclear translocation and mitogen-inducible gene 6 (MIG6)/ERBB receptor feedback inhibitor 1 (ERRFI1) expression, thereby promoting EGFR degradation. These findings were confirmed in tissues from patients with NASH and HCC.Non-alcoholic steatohepatitis (NASH) has become a serious public health problem associated with metabolic syndrome. The mechanisms by which NASH induces hepatocellular carcinoma (HCC) remain unknown. There are no approved drugs for treating NASH or preventing NASH-induced HCC. We used a genetic mouse model in which HCC was induced via high-fat diet feeding. This mouse model strongly resembles human NASH-induced HCC. The natural product honokiol (HNK) was tested for its preventative effects against NASH progression to HCC. Then, to clarify the mechanisms underlying HCC development, human HCC cells were treated with HNK. Human clinical specimens were also analyzed to explore this study’s clinical relevance. We found that epidermal growth factor receptor (EGFR) signaling was hyperactivated in the livers of mice with NASH and human HCC specimens. Inhibition of EGFR signaling by HNK drastically attenuated HCC development in the mouse model. Mechanistically, HNK accelerated the nuclear translocation of glucocorticoid receptor (GR) and promoted mitogen-inducible gene 6 (MIG6)/ERBB receptor feedback inhibitor 1 (ERRFI1) expression, leading to EGFR degradation and thereby resulting in robust tumor suppression. In human samples, EGFR-positive HCC tissues and their corresponding non-tumor tissues exhibited decreased ERRFI1 mRNA expression. Additionally, GR-positive non-tumor liver tissues displayed lower EGFR expression. Livers from patients with advanced NASH exhibited decreased ERRFI1 expression. EGFR degradation or inactivation represents a novel approach for NASH–HCC treatment and prevention, and the GR–MIG6 axis is a newly defined target that can be activated by HNK and related compounds.

Background & Aims: The molecular pathogenesis of primary liver cancer is extremely complex and heterogeneous, which was further complicated by multiple carcinogenesis. We aimed to elucidate the pathogenesis of multifocal primary liver cancer by genomic profiling. Methods: A patient with synchronous hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (ICC) who underwent resection and experienced post-operative intrahepatic metastases (IMs) was enrolled. Multiregion whole-exome sequencing was applied on the three primary tumors and two IMs to infer tumor clonality and evolution. The mutational and expression profiles, clinical relevance (n = 236) and functional implications of the identified potential driver gene FAT4 in HCC were investigated. Results: In total, we identified 12, 184, 126, and 54 of protein coding mutation in the cirrhotic liver, two HCCs (designated as HCC-A and HCC-B) and one ICC respectively. The mutational data of the two HCCs and one ICC showed almost no overlaps, suggesting that they developed through an accumulation of complete different sets of genetic alterations. For each tumor, multiregion sequencing data showed varied intratumoral heterogeneity (18.1% in HCC-A, 15.7% in HCC-B, 45.6% in ICC). The mutational profile of two IMs showed obvious similarity with HCC-A (87.6% and 87.7% shared with HCC-A respectively), rather than HCC-B and ICC tumors, indicating that they originated from HCC-A. Notably, FAT4 was the only gene that mutated in both HCCs and the two IMs, but in different locations (HCC-A and two IMs have the same G to A mutation at the coding position 2530, while HCC-B genomes have the A to C mutation at the coding position 14804). Prevalence screen revealed that FAT4 was mutated in over 15% of HCC patients and also several HCC cell lines. Significant down-regulation of FAT4 mRNA and protein expression was found in HCC, and loss of FAT4 independently correlated with early recurrence in HCC patients. In HCC cell lines, knockdown of FAT4 promoted and re-expression of FAT4 reduced cell growth and invasion. Conclusions: Spatial and temporal dissection of genomic alterations during the progression of liver cancer may help elucidate the basis for its dismal prognosis. FAT4 acts as a putative tumor suppressor that is frequently inactivated in human HCC. Citation Format: Qiang Gao, Zhi-Chao Wang, Rui-Bin Xi, Jia Fan. Inferring liver cancer evolution from spatial and temporal genomic heterogeneity. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4814. doi:10.1158/1538-7445.AM2015-4814

Hepatocellular carcinoma (HCC) is one of the most prevalent and aggressive forms of liver cancer, with high morbidity and poor prognosis due to late diagnosis and limited treatment options. Despite advances in understanding its molecular mechanisms, effective biomarkers for early detection and targeted therapy remain scarce. Zinc finger protein 71 (ZNF71), a zinc-finger protein, has been implicated in various cancers, yet its role in HCC remains largely unexplored. This gap in knowledge underscores the need for further investigation into the ZNF71 of potential as a diagnostic or therapeutic target in HCC. To explore the expression levels, clinical relevance, and molecular mechanisms of ZNF71 in the progression of HCC. The study evaluated ZNF71 expression in 235 HCC specimens and 13 noncancerous liver tissue samples using immunohistochemistry. High-throughput datasets were employed to assess the differential expression of ZNF71 in HCC and its association with clinical and pathological features. The impact of ZNF71 on HCC cell line growth was examined through clustered regularly interspaced short palindromic repeat knockout screens. Co-expressed genes were identified and analyzed for enrichment using LinkedOmics and Sangerbox 3.0, focusing on significant correlations (P < 0.01, correlation coefficient ≥ 0.3). Furthermore, the relationship between ZNF71 expression and immune cell infiltration was quantified using TIMER2.0. ZNF71 showed higher expression in HCC tissues vs non-tumorous tissues, with a significant statistical difference (P < 0.05). Data from the UALCAN platform indicated increased ZNF71 levels across early to mid-stage HCC, correlating with disease severity (P < 0.05). High-throughput analysis presented a standardized mean difference in ZNF71 expression of 0.55 (95% confidence interval [CI]: 0.34-0.75). The efficiency of ZNF71 mRNA was evaluated, yielding an area under the curve of 0.78 (95%CI: 0.75-0.82), a sensitivity of 0.63 (95%CI: 0.53-0.72), and a specificity of 0.82 (95%CI: 0.73-0.89). Diagnostic likelihood ratios were positive at 3.61 (95%CI: 2.41-5.41) and negative at 0.45 (95%CI: 0.36-0.56). LinkedOmics analysis identified strong positive correlations of ZNF71 with genes such as ZNF470, ZNF256, and ZNF285. Pathway enrichment analyses highlighted associations with herpes simplex virus type 1 infection, the cell cycle, and DNA replication. Negative correlations involved metabolic pathways, peroxisomes, and fatty acid degradation. TIMER2.0 analysis demonstrated positive correlations of high ZNF71 expression with various immune cell types, including CD4+ T cells, B cells, regulatory T cells, monocytes, macrophages, and myeloid dendritic cells. ZNF71 is significantly upregulated in HCC, correlating with the disease's clinical and pathological stages. It appears to promote HCC progression through mechanisms involving the cell cycle and metabolism and is associated with immune cell infiltration. These findings suggest that ZNF71 could be a novel target for diagnosing and treating HCC.

Simple SummaryPrimary liver cancers (PLCs) mainly comprise hepatocellular carcinoma (HCC), intrahepatic cholangiocarcinoma (iCCA), and combined (c)HCC-CCA. Both small duct types iCCA (a subtype pf iCCA) and cHCC-CCA are known to be tumors with histological heterogeneity. Understanding key tumor heterogeneity is crucial as it reflects tumor aggressiveness, patient outcome, treatment choice, and is predictive of treatment efficacy. In addition, PLCs often present with multiple liver tumors, which can be a combination of different types of PLCs or HCCs (intrahepatic metastasis or multicentric occurrence), and the pathological interpretation plays an important role in these cases. The aim of this review is to clarify the pathological features of HCC, iCCA, and cHCC-CCA, including their diagnostic pitfalls, molecular profiles, and the correlation between tumor subtypes and treatment choice.Primary liver cancers (PLCs) mainly comprise hepatocellular carcinoma (HCC), intrahepatic cholangiocarcinoma (iCCA), and cHCC-CCA. Combined HCC-CCA and small duct type iCCA show similar clinical presentations, and their histological features are more complex than seen in HCC. Therefore, while their treatment strategy differs, it is difficult to properly diagnose these tumors. Currently, HCC is the only tumor that can be treated by liver transplantation. In addition, small duct type iCCA harbors IDH1/2 mutations and FGFR2 fusions, which can be used for targeted therapy. Thus, improving diagnostic accuracy is crucial. A further point to note is that PLCs often present as multiple liver tumors, and they can be a combination of different types of PLCs or HCCs. In the case of HCCs, two different scenarios are possible, namely intrahepatic metastasis, or multicentric occurrence. Therefore, it is essential to characterize the type of multiple liver tumors. This review aims to clarify the pathological features of HCC, iCCA and cHCC-CCA, including their diagnostic pitfalls and clinical relevance. It is designed to be of use to clinicians who are dealing with PLCs, to provide a better understanding of the pathology of these tumors, and to enable a more accurate diagnosis and optimal treatment choice.

Purpose: Traditional clinical evaluations based on pathological classification have shown limited effectiveness in predicting prognosis and guiding treatment for patients with hepatocellular carcinoma (HCC). This study aims to identify a robust molecular biomarker for improving prognosis and therapy in HCC. Methods: The International Cancer Genome Consortium (ICGC), Gene Expression Omnibus (GEO), and The Cancer Genome Atlas (TCGA) provided expression data and clinicopathological information for several cohorts. . First, Cox regression analysis and differentially expressed analysis were performed to identify robust prognostic genes. Next, machine learning algorithms, including 101 statistical models, were employed to pinpoint key genes in HCC. Single-cell sequencing analysis was conducted to explore the potential subcellular functions of each key gene. Based on these findings, an HCC Prognosis- Related Index (HPRI) was developed from the identified key genes, and HPRIbased nomogram models were validated across multiple cohorts. Additionally, tumor microenvironment analysis and drug sensitivity analysis were performed further to assess the clinical significance of the HPRI in HCC. Results: A total of 36 robust prognostic genes with differential expression in HCC were identified, from which seven key genes-DCAF13, EEF1E1, GMPS, OLA1, PLOD2, PABPC1, and PPARGC1A-were filtered using machine learning algorithms. Except for PPARGC1A, all these genes were highly expressed in malignant cells, followed by fibroblasts. Notably, we developed the HPRI based on the key genes and validated its clinical relevance. Results demonstrated that the HPRI and HPRI-derived nomogram models had good predictive performance across multiple cohorts. Following tumor microenvironment analysis revealed that a high HPRI was linked to a higher likelihood of immune evasion. Drug sensitivity analysis suggested that patients with a high HPRI might benefit from chemotherapeutic agents like sorafenib, as well as novel compounds such as ML323 and MK-1775. Conclusion: Our study demonstrates a well-rounded approach by integrating gene expression, machine learning, tumor microenvironment analysis, and drug sensitivity profiling. HPRI may serve as a promising predictor for guiding prognosis and personalized treatment in HCC.

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ABSTRACTObjectives: miRNAs are frequently deregulated in cancer and have shown promise as tissue-based markers for cancer classification and prognostication. Emerging evidence indicates that miR-17-5p plays an important role in carcinogenesis. However, the expression of miR-17-5p in HCC tissues and its clinical relevance has not been systematically studied yet, and whether miR-17-5p expression has influence on prognosis of HCC is still unknown. In this study, we investigate the expression and clinical significance of miR-17-5p in human HCC. Methods: The expression levels of miR-17-5p were measured in 120 paired hepatocellular carcinoma (HCC) and paracarcinomatous liver tissues (PCLTs) derived from patients who underwent hepatic resection by qRT-PCR. Furthermore, the correlation of miR-17-5p levels with clinicopathologic variables and prognosis was analyzed. Results: miR-17-5p was significantly upregulated in HCCs (p < .001). Furthermore, HCC with metastasis had higher miR-17-5p levels than that without metastasis (p < .001). Importantly, the elevated expression of miR-17-5p correlated with multiple tumor nodules (p = .046), worse Edmondson–Steiner grade (p = .024), vein invasion (p = .001), shortened overall survival (p = .012), and disease-free survival (p = .011) of HCC. Multivariable Cox regression analysis revealed that miR-17-5p was an independent risk factor for overall survival and disease-free survival (p = .002 and p = .042, respectively). Conclusion: miR-17-5p is highly elevated in HCC, especially in HCC with metastasis. miR-17-5p can serve as a novel prognostic marker for HCC.

Background: Hepatocellular carcinoma (HCC) is one of the most common malignancies. The treatment of choice for HCC is liver resection and in certain cases liver transplantation. Tumour recurrence is, however, frequent and may be explained by the existence of undetectable tumour cells. The reverse transcriptase polymerase chain reaction (RT-PCR) can detect even small numbers of tumour cells in peripheral blood. Methods: Peripheral blood from 47 patients (group A: 19 patients with newly diagnosed HCC; group B: 20 control patients with non-HCC liver lesions; group C: eight HCC aftercare patients) was tested for the presence of circulating HCC cells by demonstration of α-fetoprotein (AFP) mRNA with RT-PCR technique. Results: Six patients with newly diagnosed HCC tested positive for circulating HCC cells. No patient in the control group or in the aftercare group was positive for AFP mRNA. Conclusions: The specificity (100%) of AFP mRNA as a marker for HCC cells in circulating blood was satisfactory, but the sensitivity (31.6%) was disappointing. This method of detecting circulating HCC cells did not provide any clinically relevant information that was not available from routine imaging or laboratory tests.