HCC Cases Research Articles

Role of CD34, SMA & Ki-67 Immunohistochemistry– An Important Diagnostic Tool in Differentiating Early Well Differentiated Hepatocellular Carcinoma from Benign Hepatic Mimickers: A Retrospective Study from North East India

North East India has higher incidence of cancers than rest of the country& with advent of early screening, Hepatocellular carcinoma is getting detected at an early stage. Core needle biopsies have become indispensible tool for diagnosis of HCC and it is difficult to identify cell thickness in early well-differentiated HCC from benign mimickers on histomorphology. To overcome this difficulty, present study was designed to see the utility of CD 34, SMA & Ki-67 in this regard and also to see role of these markers in differentiating Early well differentiated hepatocellular carcinoma from benign hepatic mimickers. 48 FFPE blocks of various lesions of liver were retrieved from Oncopathology Department, BBCI Guwahati in last one year. Cases were divided into Group A (n=28-HCC) Group B (n=20-benign mimickers). IHC CD34, SMA & Ki-67 were done on all cases. Demographic &clinical details with histomorphological findings were correlated. In Group A- 26 out of 28 HCC cases (92.8%), CD34 was strongly, completely & diffusely expressed by sinusoidal vessels with more than 3 cell plate thickness. Group B- In normal liver (10 out of 20), hepatic sinusoids are negative to weak positive for CD34.In 7 out of 20 (35.1%) benign conditions, it is sparsely expressed in capillarized sinusoids at periportal & perinodular area& high-grade dysplastic nodules showed peripheral and focal staining for CD34. Stromal cells are strongly Positive for SMA in 85.7% (group A 24 out of 28). Ki-67 was slightly increased in Group A (6-10%) than in Group B (1-2 %). Our data demonstrates that CD34, SMA & Ki-67 has significant increased expression in early WD HCC as compared to benign mimickers. Hence, CD34, SMA & ki-67 are significant helpful IHC tool for reaching to a definite diagnosis of Early WD HCC when disease is yet at early stage & and ideal for setup where specific markers of malignant hepatic tumors are not available. Thus, immunohistochemical markers essentially expressed in HCC in a specific ...

Clinicopathologic features and prognosis of steatohepatitic hepatocellular carcinoma based on varying cutoffs of tumoral steatohepatitic changes.

Steatohepatitic hepatocellular carcinoma (SH-HCC) is currently recognized as a distinct histologic subtype of HCC. The prognosis and specific criteria for determining the amount of steatohepatitis required to define SH-HCC are still unclear. After excluding all recognized HCC subtypes from 505 HCC cases (2010-2019), the remaining cases were categorized as conventional HCC (CV-HCC) (n = 223). The cases classified as SH-HCC (n = 171) were further divided into groups based on the percentage of steatohepatitis: 5% or more, 30% or more, and 50% or more. Hepatitis C virus infection was the predominant underlying liver disease in both the CV-HCC and SH-HCC groups. Metabolic dysfunction-associated steatotic liver disease (formerly nonalcoholic fatty liver disease) was more prevalent in all cases of SH-HCC with different steatohepatitic cutoffs than in cases of CV-HCC. There were no differences in the stage of fibrosis of the background liver between the CV-HCC and SH-HCC groups. SH-HCC with different cutoffs exhibited a notable increase in the presence of glycogenated nuclei, Mallory-Denk bodies, and hyaline globules in tumor cells. Survival analysis did not reveal substantial differences in overall survival between the CV-HCC and SH-HCC groups and among patients with SH-HCC with different steatohepatitis cutoffs. The degree of intratumoral steatohepatitis in patients with SH-HCC does not appear to be a notable prognostic factor. The presence of steatohepatitis in the tumor is better recognized as 1 of the histopathologic patterns of HCC.

A combined model of six serum microRNAs as diagnostic markers for hepatocellular carcinoma

BackgroundHepatocellular carcinoma (HCC) is associated with high morbidity and mortality, and its poor prognosis is mainly due to the lack of an effective means of early diagnosis. This study aimed to identify a group of serum microRNAs (miRNAs) as potential biomarkers for the diagnosis of HCC. MethodsWe collected 190 HCC cases, 109 benign lesions of the liver, 40 cases of non-HCC tumors, and 130 healthy controls. The 469 participants were divided into training and validation sets. A literature search revealed 12 miRNAs closely associated with HCC. In the training set, significantly differentially expressed miRNAs (DEmiRNAs) were screened using real-time quantitative PCR, and a diagnostic model of HCC was constructed using logistic regression analysis. An independent validation was performed using a validation set. The identified DE miRNAs were subjected to target gene prediction and functional analyses. ResultsCompared to the controls, the levels of miR-21, miR-221, miR-801, and miR-1246 significantly decreased in HCC (P < 0.05), while the levels of miR-26a and miR-122 significantly increased (P < 0.05). A diagnostic model based on the six DE miRNAs was successfully constructed, with AUC values of 0.953 for the training set and 0.952 for the verification set. Finally, 100 target genes of the DE miRNAs were predicted and were significantly enriched in the B cell receptor, neurotrophin, ferroptosis, and EGFR tyrosine kinase inhibitor resistance signaling pathways. ConclusionsThe constructed diagnostic model based on six DE miRNA combinations has important clinical value for the early diagnosis of HCC

Disparities in hepatocellular carcinoma incidence among Hispanic and non-Hispanic adults in Arizona: Trends between 2009-2017

BackgroundHepatocellular carcinoma (HCC) is a highly lethal cancer with few treatment options available to patients. Most HCC cases in Arizona, a state with a high proportion of Hispanic adults, have not been included in recent reports of HCC incidence. This study describes trends in HCC incidence and stage at diagnosis among Arizona residents between 2009–2017 and reports on racial and ethnic disparities for these outcomes. MethodsThe Arizona Cancer Registry was used to identify Arizonans aged 19 or older diagnosed with liver cell carcinoma diagnosed between 2009–2017. A total of 5043 cases were examined. Adjusted annual and 3-year HCC incidence rates (per 100,000) were examined for non-Hispanic White (NHW) and Hispanic adults. ResultsThe total age-adjusted HCC incidence rate increased significantly between 2009–2012 and then declined significantly between 2012–2017. Across nearly all years, age-adjusted HCC incidence in Hispanic adults was twice that of NHW adults. Hispanic adults were more likely to be diagnosed at a later stage across all time periods. The disparity in 3-year age-adjusted HCC incidence rate between NHW and Hispanic adults decreased between 2009–2017. ConclusionWhe total age-adjusted HCC incidence rate increased significantly between 2009–2012 and then declined significantly between 2012–2017. Across nearly all years, age-adjusted HCC incidence in Hispanic adults was twice that of NHW adults. Hispanic adults were more likely to be diagnosed at a later stage across all time periods. The disparity in 3-year age-adjusted HCC incidence rate between NHW and Hispanic adults decreased between 2009–2017.

Diet, lifestyle factors, comorbidities, and hepatocellular carcinoma risk in a middle eastern country: a case-control study

BackgroundHepatocellular Carcinoma (HCC) can be classified as one of the most common malignancies worldwide. There is scarcity of the published data on the risk factors for HCC in the Gulf Cooperation Council countries specifically Kuwait. Therefore, this case-control study sought to examine the risk factors associated with HCC in Kuwait.MethodsFifty-three histopathologically confirmed HCC cases were recruited from the Kuwait Cancer Control Center Registry. One hundred ninety-six controls (1:4 ratio) were selected from medical and/ or surgical outpatient’s clinics at all six public hospitals of Kuwait. A structured questionnaire was used to collect the data both from cases and controls through face-to-face interviews. A multivariable logistic regression model was fitted to the case-control data. Adjusted odds ratios (ORadj) and their 95% confidence intervals (CI) were computed using the parameters’ estimates of the final model and used for interpretation of the model.ResultsThe HCC cases compared with the controls were 41.6 times more likely to have had the history of non-alcoholic fatty liver disease (NAFLD) (ORadj = 41.6; 95% CI: 8.9–193.5; p < 0.001). The cases compared with the controls were more likely to have reported the history of heavy alcohol drinking (ORadj = 14.2; 95% CI: 1.2–173.4; p = 0.038). Furthermore, compared with the controls, the HCC cases tended to frequently consume milk and/or milk substitutes (≥ 3 glass/ week) (ORadj = 7.2; 95% CI: 1.2–43.4). Conversely however, there was a significant protective effect if the participants reportedly have had regularly used olive oil in their routine diet as a source of fat (ORadj = 0.17; 95% CI: 0.04–0.80) or regularly used non-steroid anti-inflammatory drugs (NSAIDs) (ORadj = 0.20; 95% CI: 0.05–0.71).ConclusionsThis study showed that heavy alcohol consumption, NAFLD history, and excessive consumption of milk/ milk substitutes were associated with a significantly increased HCC risk. Conversely however, regular use of olive oil in the diet as a source of fat or regular use of NSAIDs had a significantly protective effect against HCC risk. Adapting healthy dietary habits and preventing/ treating NAFLD may minimize the HCC risk. Future research with a larger sample size may contemplate validating the results of this study and unraveling additional risk factors contributing to HCC risk. The resultant data may help design and implement evidence-based educational programs for the prevention of HCC in this and other similar settings.

Hepatocellular carcinoma in hepatitis C virus patients treated with direct acting antivirals (DAAs) and patients not exposed to DAAs: a large center comparative study

BackgroundHepatocellular carcinoma (HCC) is the first cause of cancer in Egypt. Recently, HCC developed post direct-acting antivirals (DAAs) differ in some characteristics from those developed without DAAs exposure regarding the biological features and behavior of HCC. We aimed to assess the epidemiological, clinical, laboratory, and radiological findings besides the biological behavior of HCC patients post DAAs in comparison to HCC not exposed to DAAs. An analytic cross-sectional research was performed at the National Liver Institute which is a tertiary multidisciplinary HCC center. Subjects included hepatitis C virus patients and were allocated into two groups: group I included 2036 HCC cases post-DAA treatment and group II included 6338 HCC cases who did not receive DAAs. Subjects were examined to evaluate clinical, laboratory, and radiological findings. Tumor staging was done using the BCLC staging system.ResultsGroup II showed a more advanced Child–Pugh score, FIB-4 index, and MELD score than Group I (P = 0.001). The multiplicity of hepatic focal lesions was elevated in group I than in group II (P = 0.033). AFP level was significantly elevated in group I than in group II (p = 0.012). Portal vein invasion was significantly elevated in group I than in group II patients (P = 0.001). Extrahepatic spread of HCC was significantly elevated in group I than in group II (P = 0.001). Infiltrative lesions were significantly elevated in group I than in group II (P = 0.002).ConclusionOur study detected that the behavior in HCC post DAAs treatment is more aggressive in respect of the number of lesions, PV invasion; local and distant metastasis, and serum AFP level than in patients unexposed to DAAs. Strict surveillance in cirrhotic patients treated with DAA should be followed according to the international guidelines for early diagnosis and treatment of HCC.

Abstract 3974: Macronutrient composition dictates MASH-associated HCC immunosurveillance through microbiota alteration and metabolic adaptation

Abstract Hepatocellular carcinoma (HCC) is the most common form of liver cancer, with an estimated incidence of over one million cases by 2025. The advances in vaccines against HBV and HCV have led to a decrease in the number of virus-related cases; however, the obesity epidemic has contributed to a drastic increase in metabolic dysfunction-associated steatohepatitis (MASH) related HCC cases (e.g., obesity and type 2 diabetes induced HCC). Considering this trend, we aim to understand how lifestyle choices, like food composition, in combination with alcohol consumption at a “social drinking level” impacts host metabolism, and subsequently the development of MASH-induced HCC. We hypothesize that the impact of dietary composition on dysbiosis and immune cells, particularly adaptive immune cell metabolic status, will drive the phenotype. We have developed a MASH-induced HCC murine model, with long-term dietary intervention including seven different customized diets: normal chow (NC), ketogenic, solid high fat diet (sHFD), or isocaloric liquid Lieber DeCarli Diets (LDC): high fat/low carb (HF), high fat/low carb with ethanol (HF-EtOH), low fat/high carb (LF), and low fat/high carb with ethanol (LF-EtOH). Mice began special diet in late adolescences and disease progression was monitored over the course of a year, where mice developed HCC. Our model allowed for a comprehensive understanding of MASH-induced HCC where disease initiation could be studied beyond known contributors like obesity or body mass index (BMI). The animal model was analyzed comprehensively with high-parameter flow cytometry (FC), 16S sequencing, bulk RNA-seq, scRNA/ATAC-seq, and WGS. Long-term diet intervention led to macronutrient specific microbiota dysregulation, alterations in systemic metabolism and suppression of adaptive immune response, consequently impacting the progression of HCC. Customized dietary interventions showed a specific phenotype regarding HCC development and progression with distinct mutational genotype and particular response to immunotherapy. T and B cell subpopulation deficiency had a significant impact on MASH-induced HCC development in a macronutrient composition dependent manner. In summary, each dietary intervention showed a distinct pattern and effect on dysbiosis, systemic metabolic reprograming, cellular transcriptomic and epigenetic alteration, and particularly immune cell effector function. Additionally, some compositions supported the immuno-escape mechanism of malignant cells through regulation of their MHC machinery. Collectively, these accumulated data construct an atlas for understanding the underlying molecular mechanisms contributing to lifestyle-induced HCC providing the knowledge needed to advance effective strategies for treatment and prevention. Citation Format: Nicolas T. Ryujin, Jian Huang, Albert Nguyen, Jared Edwards, Yuhe Cheng, Jessica Wen, Ashish Damania, Nadim Ajami, Spencer Rosario, Mark Long, Ludmil Alexandrov, Shabnam Shalapour. Macronutrient composition dictates MASH-associated HCC immunosurveillance through microbiota alteration and metabolic adaptation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3974.

Abstract 6871: Tumor-infiltrating mregDC restrains anti-tumor immunity to promote cancer relapse in HCC

Abstract Background and aims: Cancer relapse is one of the major causes of mortality in patients after curative resection of primary tumors. Furthermore, significant proportion of patients with recurrent tumor lack of effective treatments, thus novel therapeutic strategies are urgently needed. Although there have been several single-cell studies of primary tumor, functional roles and mechanism of tumor-infiltrating immune cells, especially those of myeloid cells in tumor relapse have remained to be elucidated. Methods: We analyzed the RNA profile of 16498 cells from 12 primary and 6 early-relapse HCC cases. Digital pathology, multiplex immunofluorescent staining and digital spatial profiling (DSP) of paired primary and relapsed tumors from 41 HCC patients were also performed as our validation set. The molecular mechanisms associated with DCs was further tested through ex vivo and in vivo experiments. Results: We performed single-cell clustering and visualized 3 DC clusters (cDC1, cDC2, and DC3). Compared with the other two subsets, the enrichment of DC3 in relapsed HCC was correlated with a higher second recurrence rate. Deeper inspections of the signature genes indicated this subset as ‘mregDC’, a DC subset enriched in maturation and regulatory molecules. Our results demonstrated that mregDC recruited CD161+CD8+ T cells that had an innate-like, low-cytotoxic, and low-clonal-expansion phenotype. CD161+CD8+ T cells secreted large amounts of TNF to activate non-canonical NF-κB pathway via tumor necrosis factor receptor 2 (TNFR2) and promoted the differentiation of immature DC into mregDC. This positive feedback loop between mregDC and CD161+CD8+ T cells shaped a dysfunctional anti-tumor immunity, facilitating cancer relapse in HCC. In vivo assay further indicated that therapy combining anti-TNFR2 and PD-L1 antibodies significantly inhibited HCC tumor growth via impeding the infiltration of mregDC compared with anti-PD-L1 monotherapy. We also found mregDC recruited FCN1+ monocyte, a MDSC-like cell population to create an immunosuppressive niche of relapsed HCC. Conclusions: We identified this population of mregDC with immunomodulatory function in recurrent HCC and demonstrated the role of these special DCs in shaping the immunosuppressive microenvironment of recurrent tumors. Our study suggests that mregDC may be a new target for the treatment of recurrent HCC in the future. Citation Format: Zefan Zhang, Jian Zhou, Jia Fan, Yu Zhong, Liang Wu, Yunfan Sun. Tumor-infiltrating mregDC restrains anti-tumor immunity to promote cancer relapse in HCC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6871.

Routes to diagnosis for hepatocellular carcinoma patients: predictors and associations with treatment and mortality

BackgroundHepatocellular carcinoma (HCC) incidence has increased rapidly, and prognosis remains poor. We aimed to explore predictors of routes to diagnosis (RtD), and outcomes, in HCC cases.MethodsHCC cases diagnosed 2006–2017 were identified from the National Cancer Registration Dataset and linked to Hospital Episode Statistics and the RtD metric. Multivariable logistic regression was used to explore associations between RtD, diagnosis year, 365-day mortality and receipt of potentially curative treatment.Results23,555 HCC cases were identified; 36.1% via emergency presentation (EP), 30.2% GP referral (GP), 17.1% outpatient referral, 11.0% two-week wait and 4.6% other/unknown routes. Odds of 365-day mortality was >70% lower via GP or OP routes than EP, and odds of curative treatment 3–4 times higher. Further adjustment for cancer/cirrhosis stage attenuated the associations with curative treatment. People who were older, female, had alcohol-related liver disease, or were more deprived, were at increased risk of an EP. Over time, diagnoses via EP decreased, and via GP increased.ConclusionsHCC RtD is an important predictor of outcomes. Continuing to reduce EP and increase GP and OP presentations, for example by identifying and regularly monitoring patients at higher risk of HCC, may improve stage at diagnosis and survival.

Enhancement Pattern Mapping for Early Detection of Hepatocellular Carcinoma in Patients with Cirrhosis.

Limited methods exist to accurately characterize the risk of malignant progression of liver lesions. Enhancement pattern mapping (EPM) measures voxel-based root mean square deviation (RMSD) of parenchyma and the contrast-to-noise (CNR) ratio enhances in malignant lesions. This study investigates the utilization of EPM to differentiate between HCC versus cirrhotic parenchyma with and without benign lesions. Patients with cirrhosis undergoing MRI surveillance were studied prospectively. Cases (n=48) were defined as patients with LI-RADS 3 and 4 lesions who developed HCC during surveillance. Controls (n=99) were patients with and without LI-RADS 3 and 4 lesions who did not develop HCC. Manual and automated EPM signals of liver parenchyma between cases and controls were quantitatively validated on an independent patient set using cross validation with manual methods avoiding parenchyma with artifacts or blood vessels. With manual EPM, RMSD of 0.37 was identified as a cutoff for distinguishing lesions that progress to HCC from background parenchyma with and without lesions on pre-diagnostic scans (median time interval 6.8 months) with an area under the curve (AUC) of 0.83 (CI: 0.73-0.94) and a sensitivity, specificity, and accuracy of 0.65, 0.97, and 0.89, respectively. At the time of diagnostic scans, a sensitivity, specificity, and accuracy of 0.79, 0.93, and 0.88 were achieved with manual EPM with an AUC of 0.89 (CI: 0.82-0.96). EPM RMSD signals of background parenchyma that did not progress to HCC in cases and controls were similar (case EPM: 0.22 ± 0.08, control EPM: 0.22 ± 0.09, p=0.8). Automated EPM produced similar quantitative results and performance. With manual EPM, a cutoff of 0.37 identifies quantifiable differences between HCC cases and controls approximately six months prior to diagnosis of HCC with an accuracy of 89%.

Genome-wide association study identifies high-impact susceptibility loci for HCC in North America.

Despite the substantial impact of environmental factors, individuals with a family history of liver cancer have an increased risk for HCC. However, genetic factors have not been studied systematically by genome-wide approaches in large numbers of individuals from European descent populations (EDP). We conducted a 2-stage genome-wide association study (GWAS) on HCC not affected by HBV infections. A total of 1872 HCC cases and 2907 controls were included in the discovery stage, and 1200 HCC cases and 1832 controls in the validation. We analyzed the discovery and validation samples separately and then conducted a meta-analysis. All analyses were conducted in the presence and absence of HCV. The liability-scale heritability was 24.4% for overall HCC. Five regions with significant ORs (95% CI) were identified for nonviral HCC: 3p22.1, MOBP , rs9842969, (0.51, [0.40-0.65]); 5p15.33, TERT , rs2242652, (0.70, (0.62-0.79]); 19q13.11, TM6SF2 , rs58542926, (1.49, [1.29-1.72]); 19p13.11 MAU2 , rs58489806, (1.53, (1.33-1.75]); and 22q13.31, PNPLA3 , rs738409, (1.66, [1.51-1.83]). One region was identified for HCV-induced HCC: 6p21.31, human leukocyte antigen DQ beta 1, rs9275224, (0.79, [0.74-0.84]). A combination of homozygous variants of PNPLA3 and TERT showing a 6.5-fold higher risk for nonviral-related HCC compared to individuals lacking these genotypes. This observation suggests that gene-gene interactions may identify individuals at elevated risk for developing HCC. Our GWAS highlights novel genetic susceptibility of nonviral HCC among European descent populations from North America with substantial heritability. Selected genetic influences were observed for HCV-positive HCC. Our findings indicate the importance of genetic susceptibility to HCC development.

Is it time for China to prioritize pan-genotypic regimens for treating patients with hepatitis C?

IntroductionThe treatment of hepatitis C has entered the pan-genotypic era, but the effectiveness is not good for the genotype 3b patients who have a large proportion in China. The guidelines for hepatitis C recommend the use of gene-specific regimens when the regional 3b prevalence rate greater than 5%. This study is to explore rationality of this proportion and the cost-effectiveness to implement pan-genotypic regimens in China.MethodsA decision Markov model was developed from the health system perspective to evaluate the effectiveness and cost-effectiveness between pan-genotypic and gene-specific treatment regimens for hepatitis C patients. Additionally, we set a regional genotype 3b patient proportion of 0–100% to explore at which proportion it is necessary to perform genotype identification and typing therapy on patients. Model parameters were derived from published literature and public databases. Effectiveness was measured by cured patient numbers, newly diagnosed cases of decompensated cirrhosis, hepatocellular carcinoma, need for liver transplantation, and quality-adjusted life years (QALYs). Cost-effectiveness outcomes included costs and the incremental cost-effectiveness ratio (ICER). The 1–3 times 2022 Chinese per capita gross domestic product was used as the willingness-to-pay threshold. One-way and probabilistic sensitivity analyses were performed to assess the uncertainty of the model parameters.ResultsCompared with gene-specific regimens, pan-genotypic regimens resulted in an additional 0.13 QALYs and an incremental cost of $165, the ICER was $1,268/QALY. From the view of efficacy, the pan-genotypic regimens cured 5,868 more people per 100,000 patients than gene-specific regimens, avoiding 86.5% of DC cases, 64.6% of HCC cases, and 78.2% of liver transplant needs. Identifying 3b patients before treatment was definitely cost-effectiveness when their prevalence was 12% or higher. The results remained robust in sensitivity analyses.ConclusionsIn China, the prioritized recommendation of pan-genotypic therapeutics proves to be both cost-effective and efficacious. But, in regions where the prevalence of genotype 3b exceeds 12%, it is necessary to identify them to provision of more suitable therapies.

The dynamic variation position and predominant quasispecies of hepatitis B virus: Novel predictors of early hepatocarcinoma

To find the predictors of early HCC based on the dynamic changes of HBV quasispecies, this study utilizing the second-generation sequencing (NGS) and high-order multiplex droplet digital PCR (ddPCR) technology to examine the HBV quasispecies in serum of total 247 subjects recruited from high-incidence area of HCC. In the discovery stage, 15 non-synonymous Single Nucleotide Polymorphisms (SNPs) with higher variant proportion in HCC case group were founded (all P<0.05). Furthermore, the variant proportions in some of these SNPs were observed changing regularly within 5 years before the onset of HCC, and 5 of them located in HBX, 2 in HBS and 2 in HBC. The HBV predominant quasispecies and their consensus sequences were identified by genetic evolution analysis, in which the high HBS and HBC quasispecies heterogeneity were found associated with the forming of multifarious quasispecies clones, and the HBX gene had the highest proportion of predominant quasispecies (46.7 % in HBX vs 12.7 % and 13.8 % in HBS and HBC respectively) with the key variations (G1512A, A1630G, T1753C/G/A, A1762T and G1764A) determined. In the validation stage, we confirmed that the combined double mutations of G1512A+A1630G, A1762T+G1764A, and the combined triple mutations of T1753C/G/A + A1762T+G1764A, all expressed higher in early HCC cases when comparing with control group (all P<0.05). We also demonstrated the advantages of ddPCR using in multi-variations detection in large-sample for early HCC surveillance and screening. So we think that the dynamic of key HBV variation positions and their different combinations determined by quasispecies anlysis in this study can act as the novel predictors of early hepatocarcinoma and suitable to popularize and apply in HCC screening.

Mean platelet volume and platelet distribution width as predicting biomarkers of hepatitis C related hepatocellular carcinoma

Background and Aims: Besides their role in hemostasis and thrombosis; platelets have roles in innate and adaptive immune responses, atherosclerosis, lymphatic vessel development, angiogenesis and carcinogenesis .The relation between platelets and cancer is well-known and thrombocytosis may occur as a paraneoplastic syndrome. Several studies reported a correlation of high platelet count with poor prognosis in different malignancies. The aim of this study was to explore the predictive values of platelet count (PLC), mean platelet volume (MPV) &amp; platelet distribution width (PDW) in HCV related HCC, as well as their association with tumor burden. Method: The study was conducted on two groups of patients, group 1; included patients with naive HCV related HCC and group 2; included patients with chronic HCV cirrhosis. Liver biochemical tests, AFP, serum creatinine, multi-phasic contrast enhanced abdominal CT and or MRI were done at baseline for all cases. In Group 1, non-invasive diagnosis of HCC in cirrhotic patients according to the EASL guidelines &amp; Barcelona Clinic Liver Cancer (BCLC) staging was done. In Group 2, liver cirrhosis was diagnosed through liver histology or a combination of radiographic, clinical, and laboratory findings. CBC was done for all participating patients using automated hematology analyzer with reporting of PLC, MPV&amp; PDW. Results: Group 1 included 144 patients, while group 2 included 116 patients. In group 1, the age was significantly older &amp; the number of males was significantly higher. PLC was significantly lower in group 1 than group 2. In group 1, PLC was &lt; 150× 109/L (thrombocytopenia) in 71.6% of the cases, while it was normal in 28.4% of them. The median HCC diameter was significantly larger in patients with normal PLC than thrombocytopenic patients. There was significant positive correlation of PLC with BCLC stage. PLC was significantly higher in late than early BCLC stage. In group 1; PLC at cut off value of 134.5×109/L predicted late BCLC stages; with sensitivity of 44.1%, specificity of 82.5%, AUC of 0.65 &amp; p = 0.006. There was insignificant difference between both groups regarding PDW. There was insignificant difference between both groups regarding PDW. There was insignificant difference between both groups regarding the MPV. In group 1; PDW was ≥14.5% in 79% &amp; &lt;14.5% in 21% of the patients while MPV was ≥10.4 fl in 52.1% of the patients &amp; &lt;10.4 fl in 47.9% of them. There was insignificant correlation of MPV &amp; PDW with BCLC stage. PDW at cut off value of 15.05% predicted HCV related HCC with sensitivity = 62, 2%, specificity = 58, 1 %, AUC = 57, 8 %, p = 0.043. There was significant negative correlation of the MPV with serum AFP level. MPV was significantly smaller in late than early BCLC stage. On multivariate logistic regression analysis, gender, patient status of performance (PS), Alpha fetoprotein (AFP), PLC, Albumin -Bilirubin (ALBI) grade and Model of End stage Liver Disease (MELD) score, were significant predictors of HCV related HCC, while age, PDW, MPV and Child -Turcotte -Pugh (CTP) score were not significant predictors. Conclusion: PLC was significant predictors of HCV related HCC. HCV related HCC patients with normal PLC had larger tumor diameter. PLC positively correlated with BCLC stage. In HCV related HCC cases, PLC at cut off value of 134.5×109/L may predict late BCLC stages. PDW at cut off 15.05% may predict HCV related HCC. MPV was significantly smaller in late than early BCLC stages &amp; it negatively correlated with serum AFP level. MPV was not significant predictors of HCV related HCC.

Development and clinical validation of a novel algorithmic score (GAAD) for detecting HCC in prospective cohort studies.

Alpha-fetoprotein (AFP) and des-gamma carboxyprothrombin (DCP), also known as protein induced by vitamin K absence-II (PIVKA-II [DCP]) are biomarkers for HCC with limited diagnostic value when used in isolation. The novel GAAD algorithm is an in vitro diagnostic combining PIVKA-II (DCP) and AFP measurements, age, and gender (biological sex) to generate a semi-quantitative result. We conducted prospective studies to develop, implement, and clinically validate the GAAD algorithm for differentiating HCC (early and all-stage) and benign chronic liver disease (CLD), across disease stages and etiologies. Patients aged ≥18 years with HCC or CLD were prospectively enrolled internationally into algorithm development [n = 1084; 309 HCC cases (40.7% early-stage) and 736 controls] and clinical validation studies [n = 877; 366 HCC cases (47.6% early-stage) and 303 controls]. Serum samples were analyzed on a cobas® e 601 analyzer. Performance was assessed using receiver operating characteristic curve analyses to calculate AUC. For algorithm development, AUC for differentiation between early-stage HCC and CLD was 90.7%, 84.4%, and 77.2% for GAAD, AFP, and PIVKA-II, respectively. The sensitivity of GAAD for the detection of early-stage HCC was 71.8% with 90.0% specificity. Similar results were shown in the clinical validation study; AUC for differentiation between early-stage HCC and CLD was 91.4% with 70.1% sensitivity and 93.7% specificity. GAAD also showed strong specificity, with a lower rate of false positives regardless of disease stage, etiology, or region. The GAAD algorithm significantly improves early-stage HCC detection for patients with CLD undergoing HCC surveillance. Further phase III and IV studies are warranted to assess the utility of incorporating the algorithm into clinical practice.

Cabozantinib inhibits HBV-RNA transcription by decreasing STAT3 binding to the enhancer region of cccDNA.

Precision medicine and customized therapeutics based on the features of each patient are important for maximizing therapeutic effects. Because most cases of HCC occur in the damaged liver through various etiologies, such as hepatitis virus infection, steatohepatitis, and autoimmune hepatitis, there should be a rationale for the choice of therapeutic options based on these etiologies. Although cabozantinib, an oral multikinase inhibitor, has demonstrated clinical effectiveness in advanced HCC, subgroup analyses showed a lower HR for death in HBV-related HCC. This study aimed to determine the therapeutic effects of cabozantinib in HBV-related HCC. Using HBV infection models and gene knockout cells, we determined the crucial signaling axis responsible for the effects of cabozantinib on HBV. A chromatin immunoprecipitation assay was performed to determine the interaction between the signaling molecules and HBV DNA. Agonists and inhibitors were used for confirmation. Cabozantinib inhibited HBV replication through the HGF-mesenchymal-epithelial transition factor-signal transducer and activator of transcription 3 (MET-STAT3) signaling axis. The importance of STAT3 in viral replication has been confirmed using gene-edited STAT3 knockout cells. The chromatin immunoprecipitation assay revealed that the binding levels of phosphorylated STAT3 to enhancer region 1 of HBV covalently closed circular DNA were significantly increased by HGF stimulation. Cabozantinib has favorable therapeutic effects on HBV-related HCC because it inhibits HCC not only directly but also indirectly by means of inhibitory effects on HBV.

Circulating D-dimer level may predict tumor burden in chronic hepatitis C related hepatocellular carcinoma

Introduction: Coagulation and fibrinolysis activation has been implicated in angiogenesis, tumor cell invasion, tumor progression and metastatic spread. D-dimer is a biomarker that globally indicates the activation of hemostasis and fibrinolysis. Aim: To study the correlation of circulating D-dimer level with the tumor grade in HCV related HCC. Methods: HCC was diagnosed by non- invasive method in cirrhotic patients according to EASL criteria using contrast enhanced abdominal CT and or MRI. After metastatic work up, HCC grade was described as grade one if it was within Milan criteria (one nodule &lt;5cm or up to 3 nodules; the largest is &lt;3cm without vascular invasion or extra- hepatic involvement); grade two if it was beyond Milan criteria but non –metastatic &amp; grade three if it was metastatic (vascular invasion, lymph node metastasis, distant metastasis). BCLC staging depending on HCC burden, Child -Turcotte-Pugh (CTP) class and ECOG performance status (PST), was done for all studied cases. Chromic HCV was diagnosed by 4th generation ELIZA for HCV antibody and confirmed by quantitative PCR for HCV RNA. Circulating D dimer level, serum AFP level, liver biochemical tests, serum creatinine &amp; complete blood count were measured for all studied cases. Patients with history of previous treatment for HCC; recent benign thrombo-embolic disorder or renal impairment were excluded. Results: Fifty chronic HCV related HCC cases; 43 males and 7 females with age 62.2±7.4 years were enrolled. Circulating D-dimer level was significantly higher in HCC grade 2 and 3 than HCC grade 1 [(400-800) versus (0-400 ng/ml), p=0.001] &amp; in BCLC stage D than BCLC stage C HCC [(400-800) versus (0-700 ng/ml), p=0.021].There was significant positive correlation between circulating D-dimer level and HCC size and HCC grade (r =0.35, p = 0.012, r =0.33, p = 0.016 respectively). There was also significant positive correlation between circulating D-dimer level and both serum AFP level (r =0.45, p=0.001) and ECOG performance status (r =0.4, p = 0.002). Serum level of circulating D-dimer at cut off level ≥300 ng/ml can predict HCC grade beyond Milan criteria (G2&amp; 3) with PPV of 77.1%, NPV of 73.3%, AUC= 0.8 and p &lt;0.0005.Also, serum AFP level at cut off ≥394.5 can predict HCC grades beyond Milan criteria with PPV of 89.3%, NPV of 72.7%, AUC= 0.81 and P&lt;0.0005. Conclusion: In HCV related HCC, circulating D-dimer level was positively correlated with HCC grade and largest tumor size with higher level in HCC grades 2&amp;3 as well as BCLC stage D. Cut off level of circulating D-dimer level of ≥300 ng/ml can predict HCC grades beyond Milan criteria.

Study of some potential biomarkers in Egyptian hepatitis C virus patients in relation to liver disease progression and HCC

BackgroundEgypt has the greatest prevalence of hepatitis C worldwide according to the WHO reports, accounting for 13% of the global HCV infections. HCV is a substantial precursor for fibrosis, cirrhosis, and hepatocellular carcinoma. This study aimed to investigate the potential relevance of some cytokines, miR-122 and miR-221 for the diagnosis of liver disease progression associated to HCV infection.MethodsOne hundred and twenty blood samples were collected from patients with chronic liver disease, HCC, and healthy individuals. Total bilirubin, alanine aminotransferase, aspartate aminotransferase, platelet count, albumin, and creatinine were measured. Serum level of selected cytokines was conducted by ELISA. Serum miRNA expression was detected by RT-PCR.ResultsIL-2R was higher among HCC patients and the mean concentration of both TNF-αRII and IL-6R was higher among cirrhotic patients. The expression of miRNA-122 showed a little fold decrease in all studied groups; the highest level was observed in HCC patients. The expression of miRNA-221 showed a significant fold increase in HCC and cirrhotic groups.ConclusionsThis study revealed that there is no difference in liver disease progression in patients regarding sex and age. Routine liver function tests performed poorly in terms of early diagnosis of liver disease progression; however, serum total bilirubin gave somewhat useful guide for discrimination between fibrotic, cirrhotic and HCC cases. IL-2R showed a significant consistent increase in its level with disease progression. The miR-221 serum level showed significant fold increase with liver disease progression. Therefore, making miR-221 a potential non-invasive biomarker for liver disease progression in the diagnostic setting is recommended.

Long-Term Clinical Outcome of Carbon Ion Radio Therapy for Hepatocellular Carcinoma in the Caudate Lobe

Surgical resection is the first-line treatment for hepatocellular carcinoma in the caudate lobe (caudate HCC), but it is often difficult due to the tumor's location. In addition, radiofrequency ablation and transcatheter arterial chemoembolization are also difficult for the same reason. This study aimed to evaluate the safety and efficacy of carbon-ion radiation therapy (C-ion RT) for caudate HCC. We performed a retrospective cohort study of patients with hepatocellular carcinoma treated by C-ion RT between April 2000 and March 2020 in our institution. The eligibility criteria for this study were: (1) located mainly in the caudate lobe (2) the treatment protocols of 45.0-48.0 Gy/2 fractions or 52.8-60.0 Gy/4 fractions, which proved the safety and efficacy in the past clinical trials; (3) N0M0 status; (4) an Eastern Cooperative Oncology Group performance status (PS) of 0 to 2; (5) controllable ascites. The prescribed dose (Gy) used in this study is relative biological effectiveness (RBE) weighted dose. Overall survival (OS), progression-free survival (PFS), and local control rate (LC) were calculated by the Kaplan-Meier method. Adverse events were evaluated by NCI-CTCAE ver. 5.0. SPSS software version 27.0 (IBM Inc.) was used for all analyses. We defined p-value < 0.05 as statistically significant. A total of 25 patients met the criteria and were evaluated. The median patient age was 73 years (range 58-89), and 21 of 25 patients were male. The number of patients with PS 0 was 22, PS 1 was 1, and PS 2 was 2. The number of HBV-related HCC cases was in 8, HCV-related HCC cases was in 11, and non-B and non-C cases was in 6. The median maximum tumor diameter was 3.0 cm (1.1-4.8 cm). In 6 patients, identification of vascular invasion to the main trunk of the portal vein and/or major hepatic vein was confirmed. The Child-Pugh (CP) grade was A in 21 patients and B in 4. The modified albumin-bilirubin (mALBI) grade 1 is in 17 patients, 2a in 4, 2b in 4. Prescribed doses were 45 Gy / 2 fr in 3 cases, 48 Gy / 2 fr in 12 cases, 52.8 Gy / 4 fr in 7 cases, and 60 Gy / 4 fr in 3 cases. With a median follow-up period of 43.6 months (range 0.3-85.0), 3-year OS, PFS, and LC were 74% (95% confidence interval [CI], 54.8-93.8%), 32% (95% CI, 11.8-51.4%), and 93% (95% CI, 79.4-106%), respectively. All patients had no Grade 2 or higher adverse events during the observation period. The safety and efficacy of C-ion RT for caudate HCC were demonstrated. These results suggested that C-ion RT may be a promising treatment option for patients with caudate HCC.

Hypermethylation of DNA Methylation Markers in Non-Cirrhotic Hepatocellular Carcinoma.

Aberrant DNA methylation changes have been reported to be associated with carcinogenesis in cirrhotic HCC, but DNA methylation patterns for these non-cirrhotic HCC cases were not examined. Therefore, we sought to investigate DNA methylation changes on non-cirrhotic HCC using reported promising DNA methylation markers (DMMs), including HOXA1, CLEC11A, AK055957, and TSPYL5, on 146 liver tissues using quantitative methylation-specific PCR and methylated DNA sequencing. We observed a high frequency of aberrant methylation changes in the four DMMs through both techniques in non-cirrhotic HCC compared to cirrhosis, hepatitis, and benign lesions (p < 0.05), suggesting that hypermethylation of these DMMs is specific to non-cirrhotic HCC development. Also, the combination of the four DMMs exhibited 78% sensitivity at 80% specificity with an AUC of 0.85 in discriminating non-cirrhotic HCC from hepatitis and benign lesions. In addition, HOXA1 showed a higher aberrant methylation percentage in non-cirrhotic HCC compared to cirrhotic HCC (43.3% versus 13.3%, p = 0.039), which was confirmed using multivariate linear regression (p < 0.05). In summary, we identified aberrant hypermethylation changes in HOXA1, CLEC11A, AK055957, and TSPYL5 in non-cirrhotic HCC tissues compared to cirrhosis, hepatitis, and benign lesions, providing information that could be used as potentially detectable biomarkers for these unusual HCC cases in clinical practice.