Aggressive Hepatocellular Carcinoma Research Articles

Prognostic Role of 18F-FDG Positron Emission Tomography in Patients with Resectable Hepatocellular Carcinoma

Introduction: 18F-fluorodeoxyglucose positron emission tomography(18F-FDG PET) could predict tumour biology and aggressive of hepatocellular carcinoma(HCC). We hypothesized that 18F-FDG PET predicted survival of patients with HCC who underwent hepatectomy. Method: This was a retrospective study at a university center in Hong Kong. Data was retrieved from a prospectively entered database. Maximum standard uptake value(SUVmax) of tumor/ normal liver uptake(T/L ratio) was recorded. The PET positivity for overall survival(OS) was determined using T/L ratio by c-statistics using receiver operating characteristic curve. Multivariate analyses with preoperative, pathological variables and PET positivity were performed to determine the prognostic significance of post-hepatectomy OS and recurrence. Results: From 2004 to 2019, 267 patients who had 18F-FDG PET before hepatectomy. The 5-year OS and disease free survival(DFS) were 64.8% and 40.5% respectively. The median follow-up was 45.3 months. Table 1 showed the baseline demographics, operative and pathological characteristics of all patients. The optimal cut-off of T/L ratio was 1.526 based on ROC(AUC 0.674, 95% CI 0.608-0.741, P<0.001). Using a T/L ratio cutoff at 1.526, it was found to be an independent predictor of OS [hazard ratio(HR) 1.762 (1.120-2.773), p=0.014] (Table2) and was associated with a better OS.(Figure1) PET positive HCC were more likely to recur (with 5-year DFS 45.8% vs. 33.1%, p=0.001) and had a shorter DFS (median survival 8.5 vs 50 months, p=0.001).(Figure 2) Conclusion: PET positivity using T/L ratio was an independent factor for OS in HCC patients who underwent hepatectomy. 18F-FDG PET could be explored as a potential radiological biomarkers for HCC aggressiveness.

GDI2 is a novel diagnostic and prognostic biomarker in hepatocellular carcinoma.

Background: GDP Dissociation inhibitor 2 (GDI2) gene has been correlated with some important biological processes in a variety of cancers, whereas the role of GDI2 in hepatocellular carcinoma (HCC) is ill-defined. We aimed to demonstrate the relationship between GDI2 and HCC based on The Cancer Genome Atlas (TCGA) data mining.Methods: The expression of GDI2 was compared between cancer and normal tissues of 371 HCC patients collected from TCGA-LIHC, and verified in HCC cell lines. Gene set enrichment analysis (GSEA) was applied to annotate biological function of GDI2. Furthermore, Wilcoxon rank sum test, Logistics regression, as well as Cox regression and Kaplan-Meier survival analysis, were employed to evaluate the association of GDI2 expression with clinicopathological characteristics, and survival status of HCC patients, respectively.Results: It showed that the expression of GDI2 was much higher in tumor tissues than in normal tissues (P < 0.001) of HCC patients. And the elevated expression of GDI2 was correlated with more aggressive HCC tumor status, including severe primary tumor extent, advanced pathological stage, serious histologic grade, and mutated TP53 status (P < 0.05). Moreover, high GDI2 expression was strongly associated with a poor survival rate (P < 0.001). Both enrichment and immune infiltration analyses implied that GDI2-associated signaling mainly involve lipid metabolism and extracellular matrix (ECM) constructing pathways related to tumor microenvironment (TME) (P < 0.05).Conclusions: The elevated expression of GDI2 predicts poor prognosis in HCC patients, indicating that GDI2 could be applied as a predictive biomarker for diagnosis and prognosis of HCC.

SQSTM1 Expression in Hepatocellular Carcinoma and Relation to Tumor Recurrence After Radiofrequency Ablation

Autophagy is a process that allows the degradation of detrimental components through the lysosome to maintain cellular homeostasis under variable stimuli. SQSTM1 is a key molecule involved in functional autophagy and is linked to different signaling pathways, oxidative responses, and inflammation. Dysregulation of autophagy is reported in a broad spectrum of diseases. Accumulation of SQSTM1 reflects impaired autophagy, which is related to carcinogenesis and progression of various tumors, including hepatocellular carcinoma (HCC). This study investigated SQSTM1 protein expression in HCC and its relation to the clinicopathological features and the likelihood of tumor recurrence after radiofrequency ablation (RFA). This study included 50 patients with cirrhotic HCC of Barcelona Clinic Liver Cancer stages 0/A-B eligible for RFA. Tumor and peritumor biopsies were obtained just prior to local ablation and assessed for tumor pathological grade and SQSTM1 expression by immunohistochemistry. Patients were followed for one year after achieving complete ablation to detect any tumor recurrence. Serum alpha-fetoprotein level (U=149.50, P=0.027∗) and pathological grade of the tumor (χ2=12.702, P=0.002∗) associated significantly with the tumor response to RFA. SQSTM1 expression level was significantly increased in HCC compared to the adjacent peritumor cirrhotic liver tissues (Z=5.927, P<0.001∗). Significant direct relation was found between SQSTM1 expression level in HCC and the pathological grade of the tumor (H=33.789, P<0.001∗). On follow-up, tumor and peritumor SQSTM1 expression levels performed significantly as a potential predictor of the overall survival, but not the disease recurrence. SQSTM1 expression could determine aggressive HCC, even with reasonable tumor size and number, and identify the subset of HCC patients with short overall survival and unfavorable prognosis. SQSTM1 expression could not predict post-RFA intrahepatic HCC recurrence. SQSTM1 may be a potential biomarker and target for the selection of HCC patients for future therapies.

Non-alcoholic fatty liver disease and diabetes mellitus: bidirectional relationship

To date, nonalcoholic fatty liver disease (NAFLD) is the most common cause of chronic diffuse liver disease. Under adverse conditions, its natural course involves progression from simple steatosis and nonalcoholic steatohepatitis (NASH) to the development of liver cirrhosis and hepatocellular carcinoma (HCC). In recent years, there has been much convincing evidence that NAFLD is a multisystem disease that contributes to damage to extrahepatic organs and systems, primarily increasing the risk of cardiovascular diseases, type 2 diabetes, chronic kidney disease and other diseases. In particular, numerous studies in recent years indicate that NAFLD increased the risk of diabetes by at least twice. Currently, the complex and bidirectional relationship between NAFLD and type 2 diabetes are well studied. NAFLD, hepatic and systemic insulin resistance, gut dysbiosis and lipotoxicity are the main factors determining the development of diabetes mellitus in predisposed individuals. Once type 2 diabetes manifests clinically, the likelihood of progressive liver damage increases. Non-alcoholic fatty liver disease, which is associated with type 2 diabetes, is thought to be a sign of a severe clinical course with serious clinical consequences in the form of NASH, liver cirrhosis and HCC. This combination requires a more aggressive therapeutic strategy, HCC screening, and long-term follow-up. A vicious circle is formed, which leads to adverse clinical consequences, worsens the prognosis and requires an active diagnostic and treatment strategy.&#x0D;

Circulating Tumor Cell-Based Messenger RNA Scoring System for Prognostication of Hepatocellular Carcinoma: Translating Tissue-Based Messenger RNA Profiling Into a Noninvasive Setting.

Numerous studies in hepatocellular carcinoma (HCC) have proposed tissue-based gene signatures for individualized prognostic assessments. Here, we develop a novel circulating tumor cell (CTC)-based transcriptomic profiling assay to translate tissue-based messenger RNA (mRNA) signatures into a liquid biopsy setting for noninvasive HCC prognostication. The HCC-CTC mRNA scoring system combines the NanoVelcro CTC Assay for enriching HCC CTCs and the NanoString nCounter platform for quantifying the HCC-CTC Risk Score (RS) panel in enriched HCC CTCs. The prognostic role of the HCC-CTC RS was assessed in The Cancer Genome Atlas (TCGA) HCC cohort (n=362) and validated in an independent clinical CTC cohort (n=40). The HCC-CTC RS panel was developed through our integrated data analysis framework of 8 HCC tissue-based gene signatures and identified the top 10 prognostic genes (discoidin domain receptor tyrosine kinase 1 [DDR1], enoyl-CoA hydratase and 3-hydroxyacyl CoA dehydrogenase [EHHADH], androgen receptor [AR], lumican [LUM], hydroxysteroid 17-beta dehydrogenase 6[HSD17B6], prostate transmembrane protein, androgen induced 1 [PMEPA1], tsukushi, small leucine rich proteoglycan [TSKU], N-terminal EF-hand calcium binding protein 2 [NECAB2], ladinin 1 [LAD1], solute carrier family 27 member 5 [SLC27A5]) highly expressed in HCC with low expressions in white blood cells. The panel accurately discriminated overall survival in TCGA HCC cohort (hazard ratio [HR], 2.0; 95% confidence interval [CI], 1.4-2.9). The combined use of the scoring system and HCC-CTC RS panel successfully distinguished artificial blood samples spiked with an aggressive HCC cell type, SNU-387, from those spiked with PLC/PRF/5 cells (P=0.02). In the CTC validation cohort (n=40), HCC-CTC RS remained an independent predictor of survival (HR, 5.7; 95% CI, 1.5-21.3; P=0.009) after controlling for Model for End-Stage Liver Disease score, Barcelona Clinic Liver Cancer stage, and CTC enumeration count. Our study demonstrates a novel interdisciplinary approach to translate tissue-based gene signatures into a liquid biopsy setting. This noninvasive approach will allow real-time disease profiling and dynamic prognostication of HCC.

MRPS31 loss is a key driver of mitochondrial deregulation and hepatocellular carcinoma aggressiveness

Deregulated mitochondrial energetics is a metabolic hallmark of cancer cells. However, the causative mechanism of the bioenergetic deregulation is not clear. In this study, we show that somatic copy number alteration (SCNA) of mitoribosomal protein (MRP) genes is a key mechanism of bioenergetic deregulation in hepatocellular carcinoma (HCC). Association analysis between the genomic and transcriptomic profiles of 82 MRPs using The Cancer Genome Atlas-Liver HCC database identified eight key SCNA-dependent MRPs: MRPS31, MRPL10, MRPL21, MRPL15, MRPL13, MRPL55, and DAP3. MRPS31 was the only downregulated MRP harboring a DNA copy number (DCN) loss. MRPS31 loss was associated specifically with the DCN losses of many genes on chromosome 13q. Survival analysis revealed a unique dependency of HCC on the MRPS31 deficiency, showing poor clinical outcome. Subclass prediction analysis using several public classifiers indicated that MRPS31 loss is linked to aggressive HCC phenotypes. By employing hepatoma cell lines with SCNA-dependent MRPS31 expression (JHH5, HepG2, Hep3B, and SNU449), we demonstrated that MRPS31 deficiency is the key mechanism, disturbing the whole mitoribosome assembly. MRPS31 suppression enhanced hepatoma cell invasiveness by augmenting MMP7 and COL1A1 expression. Unlike the action of MMP7 on extracellular matrix destruction, COL1A1 modulated invasiveness via the ZEB1-mediated epithelial-to-mesenchymal transition. Finally, MRPS31 expression further stratified the high COL1A1/DDR1-expressing HCC groups into high and low overall survival, indicating that MRPS31 loss is a promising prognostic marker.SignificanceOur results provide new mechanistic insight for mitochondrial deregulation in HCC and present MRPS31 as a novel biomarker of HCC malignancy.

AnnexinA7 promotes epithelial\u2013mesenchymal transition by interacting with Sorcin and contributes to aggressiveness in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide, and metastasis is the major cause of the high mortality of HCC. In this study, we identified that AnnexinA7 (ANXA7) and Sorcin (SRI) are overexpressed and interacting proteins in HCC tissues and cells. In vitro functional investigations revealed that the interaction between ANXA7 and SRI regulated epithelial–mesenchymal transition (EMT), and then affected migration, invasion, and proliferation in HCC cells. Furthermore overexpression/knockdown of ANXA7 was remarkably effective in promoting/inhibiting tumorigenicity and EMT in vivo. Altogether, our study unveiled a mechanism that ANXA7 promotes EMT by interacting with SRI and further contributes to the aggressiveness in HCC, which provides a novel potential therapeutic target for preventing recurrence and metastasis in HCC.

The Presence of Circulating Tumor Cell Cluster Characterizes an Aggressive Hepatocellular Carcinoma Subtype.

BackgroundGrowing evidence suggests that circulating tumor cell (CTC) clusters may be an important factor in the metastatic process, but their role in hepatocellular carcinoma (HCC) remains unclear. This study aimed to characterize the molecular and clinical features of CTC cluster-positive human HCC and to assess its prognostic value in HCC patients.MethodsThe CTCs and CTC clusters were evaluated in 204 HCC patients using CellSearch™ System. The counts of CTCs and CTC clusters were correlated with different clinical features, while their associations with progression-free survival (PFS) and overall survival (OS) were evaluated integrally and hierarchically by Kaplan–Meier estimates or Cox proportional regression analysis. Five cases each of CTC cluster-negative and cluster-positive patients were selected for RNA-sequencing analysis. The results of gene enrichment analysis were further verified using tissue microarray (TMA) by immunohistochemistry (IHC).ResultsCTCs and CTC clusters were detected in 76 (37.3%) and 19 (9.3%) of 204 preoperative samples, respectively. CTC cluster-positive HCC represented an aggressive HCC phenotype with larger tumor size, more frequent microvascular invasion, and higher tumor stages. The survival of HCC patients utilizing CTCs and CTC clusters individually showed prognostic significance, while joint analysis revealed patients in Group III (CTC ≥ 2 and CTC cluster > 0) had the worst outcome. Stratified analysis of outcomes in Barcelona Clinic Liver Cancer (BCLC) and tumor–node–metastasis (TNM) stages indicated that patients with CTC clusters had significantly poorer prognosis in each stage than those without CTC clusters. Moreover, the RNA sequencing and TMA staining results showed that CTC cluster-positive HCCs were usually associated with Wnt/β-catenin signaling activation.ConclusionThe presence of CTC clusters characterizes an aggressive HCC subtype. CTC clusters may be used as a biomarker in predicting the prognosis on each stage of malignancy in HCC, which provides evidence for formulating therapeutic strategies for more precise treatment.

Predictors of Macrovascular Invasion and Extrahepatic Metastasis in Treatment Naive Hepatocellular Carcinoma: When Is [18F] FDG PET/CT Relevant?

Hypermetabolic macrovascular invasion (MVI) and extrahepatic metastasis (EHM) occur in aggressive hepatocellular carcinoma (HCC) and carry unfavorable prognosis. [18F] FDG PET/CT, despite having low sensitivity in primary HCC, is valuable in patients with aggressive HCC for detection of hypermetabolic MVI and EHM. The study aimed at identifying the parameters that could predict hypermetabolic MVI and/or EHM in treatment naive HCC patients for tailored approach to utilize [18F] FDG PET/CT. Data of 131 treatment naive HCC patients (median age, 60years; range, 21-80years; 90.8% males) who underwent [18F] FDG PET/CT were retrospectively analyzed to determine the proportion of patients with hypermetabolic MVI and/or EHM. Logistic regression analysis was performed to define independent predictors of hypermetabolic MVI and/or EHM. 78/131 (59.5%) patients had hypermetabolic MVI and/or EHM. 52/131 (39.7%) patients had EHM. 56/131 (42.7%) patients had hypermetabolic MVI of which, 30 had concomitant EHM with majority (90%; 27/30) having distant metastasis. 26/131 (19.8%) patients had hypermetabolic MVI without EHM while 22/131 (16.8%) patients had EHM without hypermetabolic MVI of which, majority (95.5%; 21/22) had distant metastasis. Hypermetabolic MVI was associated with EHM (χ2 = 7.868; p value = 0.007). AFP > 93.7ng/ml, SUVmax > 3.5, and maximum tumor size > 5.0cm were the independent predictors of hypermetabolic MVI and/or EHM. In treatment naive HCC patients with AFP > 93.7ng/ml or maximum tumor size > 5.0cm, [18F] FDG PET/CT can be valuable.

CCL23 in Balancing the Act of Endoplasmic Reticulum Stress and Antitumor Immunity in Hepatocellular Carcinoma.

Endoplasmic reticulum (ER) stress is a cellular process in response to stress stimuli in protecting functional activities. However, sustained hyperactive ER stress influences tumor growth and development. Hepatocytes are enriched with ER and highly susceptible to ER perturbations and stress, which contribute to immunosuppression and the development of aggressive and drug-resistant hepatocellular carcinoma (HCC). ER stress-induced inflammation and tumor-derived chemokines influence the immune cell composition at the tumor site. Consequently, a decrease in the CCL23 chemokine in hepatic tumors is associated with poor survival of HCC patients and could be a mechanism hepatic tumor cells use to evade the immune system. This article describes the prospective role of CCL23 in alleviating ER stress and its impact on the HCC tumor microenvironment in promoting antitumor immunity. Moreover, approaches to reactivate CCL23 combined with immune checkpoint blockade or chemotherapy drugs may provide novel opportunities to target hepatocellular carcinoma.

Characteristics of De novo Hepatocellular carcinoma (HCC) in Chronic HCV Egyptian patients received Direct Acting Antivirals (DAAs) therapy: A pilot study

Abstract Background Direct-acting antiviral agents for chronic hepatitis C have initiated a revolution in the management and control of this important liver disease with cure rates over 90%. However, it has been suggested that HCC may occur or recur in patients with chronic HCV infection who received DAAs therapy. Aim To compare characteristics and behavior of de novo Hepatocellular carcinoma (HCC) in chronic HCV patients who received direct acting antiviral treatment with those who didn't receive DAAs . Patients and methods The current study was conducted on patients with de novo HCC presented at HCC clinic, Tropical medicine department Ain Shams University Hospitals between December 2017 and December 2018, aged (18-70 years old) . Total number of HCC cases visited the clinic during the study period was 416 patients. 160 patients with de novo HCC were enrolled in the study fullfiling the inclusion criteria and divided to two groups, 80 patients with HCC who received Direct antiviral treatment, the other 80 patients who didn't receive Direct antiviral treatment. Results Unlike some previous concerns about increased biologic aggressiveness of HCC after DAA treatment, resulting in infiltrative pattern and multinodular HCC outside Milan criteria, our results on de novo HCC after DAAs were mostly single, small, and diagnosed in early stages, resulting in BCLC stage 0-B in the majority of patients. There were no statistical differences regarding total size of tumor,or vascular invasion or AFP, but there’s a statistical difference in largest size of HFLs that was more significant in group (A) P = 0.04 The mean Child score was 5.75 in group A, with 61 patients (76.3%) were Child class A,19 patients (23.8%) were Child class B and 0 patients were child C. For the patients in group (B) the mean Child score was 6.15, with 50 patients (62.5%) were Child class A, 26 patients (32.5%) were Child class B and 4 patients (5%) were child C. Conclusion The current study shows no differences between the 2 studied groups as regard tumor aggressiveness, but data on risk of HCC and behaviour after DAAs is still conflicting and further studies are needed

Targeting ER stress in the hepatic tumor microenvironment.

Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer. It currently ranks as one of the most aggressive and deadly cancers worldwide, with an increasing mortality rate and limited treatment options. An important hallmark of liver pathologies, such as liver fibrosis and HCC, is the accumulation of misfolded and unfolded proteins in the lumen of the endoplasmic reticulum (ER), which induces ER stress and leads to the activation of the unfolded protein response (UPR). Upon accumulation of misfolded proteins, ER stress is sensed through three transmembrane proteins, IRE1α, PERK, and ATF6, which trigger the UPR to either alleviate ER stress or induce apoptosis. Increased expression of ER stress markers has been widely shown to correlate with fibrosis, inflammation, drug resistance, and overall HCC aggressiveness, as well as poor patient prognosis. While preclinical in vivo cancer models and in vitro approaches have shown promising results by pharmacologically targeting ER stress mediators, the major challenge of this therapeutic strategy lies in specifically and effectively targeting ER stress in HCC. Furthermore, both ER stress inducers and inhibitors have been shown to ameliorate HCC progression, adding to the complexity of targeting ER stress players as an anticancer strategy. More studies are needed to better understand the dual role and molecular background of ER stress in HCC, as well as its therapeutic potential for patients with liver cancer.

International study on the outcome of locoregional therapy for liver transplant in hepatocellular carcinoma beyond Milan criteria

Background & AimsGood outcomes after liver transplantation (LT) have been reported after successfully downstaging to Milan criteria in more advanced hepatocellular carcinoma (HCC). We aimed to compare post-LT outcomes in patients receiving locoregional therapies (LRT) before LT according to Milan criteria and University of California San Francisco downstaging (UCSF-DS) protocol and ‘all-comers’. MethodsThis multicentre cohort study included patients who received any LRT before LT from Europe and Latin America (2000–2018). We excluded patients with alpha-foetoprotein (AFP) above 1,000 ng/ml. Competing risk regression analysis for HCC recurrence was conducted, estimating subdistribution hazard ratios (SHRs) and corresponding 95% CIs. ResultsFrom 2,441 LT patients, 70.1% received LRT before LT (n = 1,711). Of these, 80.6% were within Milan, 12.0% within UCSF-DS, and 7.4% all-comers. Successful downstaging was achieved in 45.2% (CI 34.8–55.8) and 38.2% (CI 25.4–52.3) of the UCSF-DS group and all-comers, respectively. The risk of recurrence was higher for all-comers (SHR 6.01 [p <0.0001]) and not significantly higher for the UCSF-DS group (SHR 1.60 [p = 0.32]), compared with patients remaining within Milan. The all-comers presented more frequent features of aggressive HCC and higher tumour burden at explant. Among the UCSF-DS group, an AFP value of ≤20 ng/ml at listing was associated with lower recurrence (SHR 2.01 [p = 0.006]) and better survival. However, recurrence was still significantly high irrespective of AFP ≤20 ng/ml in all-comers. ConclusionsPatients within the UCSF-DS protocol at listing have similar post-transplant outcomes compared with those within Milan when successfully downstaged. Meanwhile, all-comers have a higher recurrence and inferior survival irrespective of response to LRT. Additionally, in the UCSF-DS group, an ALP of ≤20 ng/ml might be a novel tool to optimise selection of candidates for LT. Clinical trial numberThis study was registered as part of an open public registry (NCT03775863). Lay summaryPatients with more extended HCC (within the UCSF-DS protocol) successfully downstaged to the conventional Milan criteria do not have a higher recurrence rate after LT compared with the group remaining in the Milan criteria from listing to transplantation. Moreover, in the UCSF-DS patient group, an ALP value equal to or below 20 ng/ml at listing might be a novel tool to further optimise selection of candidates for LT.

Abstract 50: Factors Affecting the Aggressiveness of Hepatocellular Carcinoma among Hepatitis C Related Cirrhosis Patients

Abstract Purpose: The study was conducted to assess possible epidemiological, clinical and laboratory factors predicting the aggressiveness of Hepatocellular carcinoma (HCC) in HCV-related liver cirrhosis patients. Methods: A comparative hospital-based cross-sectional study was conducted. Exclusion criteria were HBsAg positive patients, and patients undergoing any treatment modalities for HCC. Complete history was taken focusing on signs and symptoms of the chronic liver disease, and clinical examination was done. Results: Among 173 patients, only 14.4% of the patients were classified as Child A. In near percentages, patients with Child B represented 43.4%, and patients with Child C were 42.2% of the studied patients. Most of the patients were males (75.7%), while the rest were females (24.3%). Patients were divided to 2 groups: group (I) included 36 patients with non-aggressive HCC (score = 4), and group (II) included 137 patients with aggressive HCC (score &amp;gt; 4) according to the aggressiveness index (AgI) described by Carr et al that takes in count AFP, Maximum Tumor Diameter, number of nodules and presence of portal vein thrombosis. HCC patients in group (II) were 1.9 times more likely to have positive HBc Ab as compared to patients in group (I) where (OR = 1.9, CI = 1.3 - 3.6). Multiple logistic regression analysis for predicting the aggressiveness of HCC explains that 86.7% of factors (age, Child Pugh score, BCLC staging, Gamma glutamyl transpeptidase (GGT), alkaline phosphatase (ALP), Platelets, HBcore ab and AFP) were contributing to aggressiveness among HCC-Hepatitis C related cirrhosis patients (R 2 = 0.867) with overall significant where chi-square = 21.876, p = 0.001. Conclusion: In conclusion, age, platelets, Child Pugh score, BCLC staging, HBcore ab, ALP, GGT and AFP are significant predictors for the aggressiveness of HCC. Citation Format: Randa Salah Abd Elmoneim, Fathalla Sedki, Aida Mohey, Ehab Hassouna, Eman Tayae. Factors Affecting the Aggressiveness of Hepatocellular Carcinoma among Hepatitis C Related Cirrhosis Patients [abstract]. In: Proceedings of the 9th Annual Symposium on Global Cancer Research; Global Cancer Research and Control: Looking Back and Charting a Path Forward; 2021 Mar 10-11. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2021;30(7 Suppl):Abstract nr 50.

Abstract 764: Circulating tumor cell-based mRNA scoring system for prognostication of hepatocellular carcinoma - Translating HCC tissue-based mRNA profiling into a non-invasive setting

Abstract Background and Aims Numerous studies in hepatocellular carcinoma (HCC) have proposed tissue-based gene signatures for individualized prognostic assessments. In this study, we aim to develop the HCC circulating tumor cell (CTC) mRNA Scoring System and the HCC-CTC Risk Score (RS) panel to translate tissue-based mRNA signatures into a liquid biopsy setting for non-invasive HCC prognostication. Methods We developed the HCC CTC mRNA Scoring System by combining the NanoVelcro CTC Assay for enriching HCC CTCs and the NanoString nCounter platform for quantifying the HCC-CTC RS panel in enriched HCC CTCs. The prognostic role of HCC-CTC RS was assessed in the Cancer Genome Atlas (TCGA) HCC cohort (n=362) and validated in an independent CTC cohort (n=40). Results The HCC-CTC RS panel was developed from 8 HCC tissue-based prognostic gene signatures. Through our integrated data analysis framework, the top 10 prognostic genes highly expressed in HCC cells and lowly expressed in white blood cells were selected as the HCC-CTC RS panel. The panel accurately discriminated overall survival in TCGA HCC cohort (hazard ratio [HR]: 2.01, 95% confidence intervals [CI]: 1.39-2.91). In addition, the combined use of Scoring System and panel successfully distinguished artificial blood samples spiked with an aggressive HCC cell type, SNU-387, from those spiked with PLC/PRF/5 cells (P=0.02). Finally, the prognostic value of HCC-CTC RS was further validated in the CTC cohort (HR: 8.48, 95% CI: 1.89-38.03) by multivariable analysis. Conclusions Our pilot study demonstrates a novel interdisciplinary approach to translate tissue-based gene signatures into a liquid biopsy setting. This non-invasive approach will allow real-time disease profiling and dynamical prognostication of HCC. Citation Format: Yi-Te Lee, Na Sun, Minhyung Kim, Jasmine J. Wang, Benjamin V. Tran, Ryan Y. Zhang, Ceng Zhang, Saeed Sadeghi, Richard S. Finn, Sammy Saab, Ronald W. Busuttil, Yazhen Zhu, Hsian-Rong Tseng, Sungyong You, Ju Dong Yang, Vatche G. Agopian. Circulating tumor cell-based mRNA scoring system for prognostication of hepatocellular carcinoma - Translating HCC tissue-based mRNA profiling into a non-invasive setting [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 764.

Abstract LB232: Imbalance of TGF-β1/BMP-7 pathway induced by M2-polarized macrophages promotes hepatocellular carcinoma aggressiveness

Abstract The transforming growth factor-beta (TGF-β) signaling represented the predominant cytokine signaling involved in the development and progression of hepatocellular carcinoma (HCC). Bone morphogenetic protein (BMP), another member of TGF-β superfamily, was frequently found to have cross-talk with TGF-β pathway. However, the complex interaction between TGF-β and BMP pathways has not been fully elucidated in HCC. In this study, we found that the imbalance of TGF-β1/BMP-7 pathway was associated with aggressive pathological features and poor clinical outcome in HCC, and it could significantly promote HCC cells invasion and stemness by increasing inhibitor of differentiation 1 (ID1). Next, we indicated that the miR-17-92 cluster could disrupted the balance of TGF-β1/BMP-7 pathway in HCC cells by inducing TGFBR2 post-transcriptional silence and inhibiting ACVR1 post-translational ubiquitylation via targeting Smurf1. Furthermore, we demonstrated that miR-17-92 cluster was originated from the extracellular vesicles (EVs) of M2-polarized tumor associated macrophages (M2-TAMs) infiltrated in HCC microenvironment and induced the imbalance of TGF-β1/BMP-7 pathway. Finally, we found that sh-MIR17HG lentivirus, TGFBR2 lentivirus and ACVR1 inhibitor could profoundly inhibit M2-TAMs-stimulated the growth and metastasis of HCC xenografts in vivo. We propose that imbalance of TGF-β1/BMP-7 pathway may serve as a prognostic biomarker, M2-TAMs and miR-17-92 cluster could be a therapeutic target for this subtype of HCC. Citation Format: Junya Ning, Yingnan Ye, Jinpu Yu. Imbalance of TGF-β1/BMP-7 pathway induced by M2-polarized macrophages promotes hepatocellular carcinoma aggressiveness [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr LB232.

ADV score is a quantifiable prognostic prediction model for surgical resection of hepatocellular carcinoma: A Korea-Japan collaborative validation study with 10,606 patients

We previously demonstrated that multiplication of α-fetoprotein (AFP), des-γ-carboxy prothrombin (DCP) and tumor volume (TV) (ADV score, expressed in log10) is an integrated surrogate marker of post-resection prognosis for hepatocellular carcinoma (HCC). This study aimed to validate the ADV score as a quantifiable biomarker for oncological aggressiveness of HCC on its prognostic impact following resection of HCC. A multicenter validation cohort with 10,606 patients (4,900 [46.2%] from Korea and 5,706 [53.8%] from Japan) was established through the Korea-Japan collaboration study groups managed by the Korea and Japan Society of Hepatobiliary Pancreatic Surgery. In the Korean cohort, disease-free survival (DFS) and overall survival (OS) rate at 5 years were 66.7% and 93.0% in ADV score < 2log; 56.0% and 84.1% in ADV score 2.0-2.9log; 51.0% and 79.7% in ADV score 3.0-3.9log; 45.7% and 75.3% in ADV score 4.0-4.9log; 44.4% and 69.4% in ADV score 5.0-5.9log; 38.4% and 64.3% in ADV score 6.0-6.9log; 32.6% and 62.9% in ADV score 7.0-7.9log; 28.7% and 53.2% in ADV score 8.0-8.9log; 30.4% and 46.5% in ADV score 9.0-9.9log; 30.5% and 47.7% in ADV score 10.0-10.9log; 22.5% and 35.7% in ADV score 11.0-11.9log; and 18.1% and 33.8% in ADV score ≥ 12log, respectively (both p < 0.001). The ADV-score dependent DFS and OS rates in the Japanese cohort were very similar with those of the Korean cohort. Consequently, the overall study cohort also revealed similar ADV-score dependent DFS and OS rates. Multiplication of AFP and DCP (AD score) and TV were independent risk factors for DSF and OS. The 5-year DFS and OS rates were visualized according to AD score and TV as like weather prediction diagrams. This high-volume international validation study revealed that the preoperative ADV score is a reliable surrogate biomarker for quantifiable prediction of post-resection prognosis in the Korean and Japanese patients with HCC.

Loss of TARBP2 Drives the Progression of Hepatocellular Carcinoma via miR-145-SERPINE1 Axis.

The clinical outcomes of hepatocellular carcinoma (HCC) remain dismal. Elucidating the molecular mechanisms for the progression of aggressive HCC holds the promise for developing novel intervention strategies. The transactivation response element RNA-binding protein (TRBP/TARBP2), a key component of microRNA (miRNA) processing and maturation machinery has been shown to play conflicting roles in tumor development and progression. We sought to investigate the expression of TARBP2 in HCC using well-characterized HCC cell lines, patient-derived tissues and blood samples. Additionally, the potential prognostic and diagnostic value of TARBP2 in HCC were analyzed using Kaplan-Meier plots and ROC curve. Cell counting kit‐8 (CCK‐8), wound healing and transwell assays examined the ability of TARBP2 to induce cell proliferation, migration, and invasion in HCC cell lines. RNA sequencing was applied to identify the downstream elements of TARBP2. The interaction of potential targets of TARBP2, miR‐145 and serpin family E member 1 (SERPINE1), was assessed using luciferase reporter assay. TARBP2 expression was down-regulated in HCC cell lines relative to normal hepatocyte cells, with a similar pattern further confirmed in tissue and blood samples. Notably, the loss of TARBP2 was demonstrated to promote proliferation, migration, and invasion in HCC cell lines. Interestingly, the reduction of TARBP2 was shown to result in the upregulation of SERPINE1, also known as plasminogen activator inhibitor (PAI-1), which is a vital gene of the HIF-1 signaling pathway. Knockdown of SERPINE1 rescued the TARBP2-lost phenotype. Moreover, TARBP2 depletion induced the upregulation of SERPINE1 through reducing the processing of miR-145, which directly targets SERPINE1. Finally, overexpression of miR-145 repressed SERPINE1 and rescued the functions in sh-TARBP2 HCC cells. Our findings underscore a linear TARBP2-miR-145-SERPINE1 pathway that drives HCC progression, with the potential as a novel intervention target for aggressive HCC.

Induced Mitochondrial Alteration and DNA Damage via IFNGR-JAK2-STAT1-PARP1 Pathway Facilitates Viral Hepatitis Associated Hepatocellular Carcinoma Aggressiveness and Stemness

Hepatitis virus is a major risk factor for liver cancer. The mitochondrial dysfunction IFN gamma-related pathways are activated after virus infection. Jak family-related protein is involved in the downstream of IFN gamma-related pathways. However, the effect of the IFNGR-JAK-STAT pathway acting as functional regulators of their related protein expression on virus infection and hepatocellular carcinoma (HCC) remains unclear. Interestingly, the role of the DNA repair gene (PARP1) in therapy resistant cancers also has not been studied and explored well. In this study, we hypothesized that momelotinib could suppress the progression of HCC by targeting Jak family related and PARP1 DNA repair protein. Based on this observation, we link the relevant targets of the JAK family and the potential applications of targeted therapy inhibitors. We analyzed possible synergism between momelotinib and sorafenib in hepatitis virus-associated liver cancer. Immunostaining, colony formation assay, cell invasion, migration, and tumorsphere-formation assay were used for drug cytotoxicity, cell viability, and possible molecular mechanism. We first demonstrated that the expression of Jak1 and 2 is significantly upregulated in vHCC than in nvHCC/normal liver tissues. In addition, the gene expression of IFN gamma-related pathways is activated after virus infection. Additionally, we found that momelotinib significantly inhibited the growth of HCC cells and reduces the expression of Jak2, which showed the importance of momelotinib in targeting Jak2 and reducing tumorigenesis in HCC. Meanwhile, momelotinib effectively inhibited the IFNGR-JAK-STAT pathway and reduced the migratory/invasive ability of vHCC cells through down-regulating EMT biomarkers (E-cadherin and vimentin), transcription factor (Slug), and significantly inhibits the DNA damage repair enzyme PARP1. It also induced cell apoptosis of vHCC cells. Furthermore, the combined effect of momelotinib and sorafenib both at in vitro and in vivo synergistically suppresses the proliferation of vHCC cells and effectively reduces the tumor burden. Our results showed that momelotinib effectively suppressed the expression of the IFNGR-JAK-STAT-PARP1 pathway, which results in the downregulation of cancer stem cell genes and enhances the antitumor efficacy of sorafenib by initiating the expression of apoptosis-related genes and inhibiting the DNA repair gene in vHCC cells, thus maximizing its therapeutic potential for patients with HCC.

Prognostic Values of Alpha-Fetoprotein and Des-Gamma-Carboxyprothrombin in Hepatocellular Carcinoma in China: An Analysis of 4792 Patients.

BackgroundThe importance of alpha-fetoprotein (AFP) and des-gamma-carboxyprothrombin (DCP) in hepatocellular carcinoma (HCC) has been studied extensively in Japan, where hepatitis C virus is the predominant aetiology of HCC. The clinical profiles of HCC regarding the state of AFP and DCP in a hepatitis B virus epidemic area have not been comprehensively investigated, and the value of these tumour markers in evaluating the response to treatment and the detection of recurrence has yet to be determined.Patients and MethodsA total of 4792 patients treated in our centre were continuously analysed regarding accessible AFP and DCP data pre- and posttreatment. Baseline characteristics were summarized, and comparisons of progression-free survival (PFS) and overall survival (OS) rates were made independently. The prognostic significance of each factor was tested with the Cox proportional hazards model. Patients who had AFP and DCP data pretreatment, pre- and posttreatment, and those who were continuously monitored more than twice were analysed separately.ResultsA total of 2600 patients (53.4%) were positive for AFP and DCP; 362 (7.6%) and 1211 (25.3%) patients were AFP- or DCP-positive, respectively, and 619 patients (12.9%) were negative for both AFP and DCP. Patients in the AFP single-positive or double-negative groups had the best OS (P<0.001). Patients with less than 50% responses in AFP and DCP after treatments suffered from worse prognostic survival (P<0.001). In the multivariate analysis, elevated AFP and DCP were identified as independent prognostic factors of PFS and OS. In addition, different tumour markers were related to different clinical and pathological traits.ConclusionThe present study comprehensively explored the clinical value of classical tumour markers for HCC using the “point-to-line” method. Positivity of pretreatment AFP and DCP or less than 50% treatment response rates exhibited more aggressive HCC, resulting in poor PFS and OS in HCC patients.