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Background/Aim: Alteration of F-box and leucine-rich repeat protein 5 (FBXL5), an iron-sensing ubiquitin ligase, might be related with carcinogenesis of hepatocellular carcinoma (HCC), by disturbing cellular iron homeostasis. However, the clinical implications of FBXL5 expression using patient samples need to be elucidated. Patients and Methods: We collected HCC tissue samples from two institutes: Samsung Medical Center (n=259) and Hallym University Sacred Heart Hospital (n=115) and evaluated FBXL5 expression using immunohistochemistry. Using cut-off values determined by X-tile software, association between FBXL5 expression and several clinicopathological parameters was investigated. For external validation, the Cancer Genome Atlas (TCGA) cohort was used. Results: The best cutoff value for FBXL5 IHC expression associated with recurrence-free survival (RFS) was 5%. Low FBXL5 expression was found in 18.7% of the total 374 HCCs and was associated with non-viral etiology (p=0.019). Low FBXL5 expression was related with inferior disease-specific survival (DSS, p=0.002) and RFS (p=0.001) and also was an independent prognostic factor for DSS and RFS. In addition, cases with low FBLX5 mRNA levels showed inferior DSS and RFS (p

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The roles of chromatin remodelers and their underlying mechanisms of action in cancer remain unclear. In this study, SMARCB1, known initially as a bona fide tumor suppressor gene, was investigated in liver cancer. SMARCB1 was highly upregulated in patients with liver cancer and was associated with poor prognosis. Loss- and gain-of-function studies in liver cells revealed that SMARCB1 loss led to reduced cell proliferation, wound healing capacity, and tumor growth in vivo. Although upregulated SMARCB1 appeared to contribute to switch/sucrose nonfermentable (SWI/SNF) complex stability and integrity, it did not act using its known pathways antagonism with EZH2 or association between TP53 or AMPK. SMARCB1 knockdown induced a mild reduction in global H3K27 acetylation, and chromatin immunoprecipitation sequencing of SMARCB1 and acetylated histone H3K27 antibodies before and after SMARCB1 loss identified Nucleoporin210 (NUP210) as a critical target of SMARCB1, which bound its enhancer and changed H3K27Ac enrichment and downstream gene expression, particularly cholesterol homeostasis and xenobiotic metabolism. Notably, NUP210 was not only a putative tumor supporter involved in liver cancer but also acted as a key scaffold for SMARCB1 and P300 to chromatin. Furthermore, SMARCB1 deficiency conferred sensitivity to doxorubicin and P300 inhibitor in liver cancer cells. These findings provide insights into mechanisms underlying dysregulation of chromatin remodelers and show novel associations between nucleoporins and chromatin remodelers in cancer.Significance:This study reveals a novel protumorigenic role for SMARCB1 and describes valuable links between nucleoporins and chromatin remodelers in cancer by identifying NUP210 as a critical coregulator of SMARCB1 chromatin remodeling activity.

Background The treatment goal of ulcerative colitis (UC) has changed from the control of symptoms to mucosal healing, previously evaluated mainly by endoscopy. Recently, the importance of histologic activity has emerged. Therefore, this study aimed to investigate the risk of clinical relapse according to histologic activity in UC with a Mayo endoscopic subsccore (MES) of 0 or 1. Methods In a retrospective cohort after our center’s biopsy guideline for UC was instituted, 492 UC patients with an MES of 0 or 1 were enrolled and analyzed. The primary outcome was the development of a clinical relapse including changes in medication, hospitalization, colectomy, and the development of colorectal cancer during the follow-up period. Results During the median 549 days of follow-up, 92 (18.7%) patients had a clinical relapse. All the patients changed their medication, including 4 hospitalized patients. Histologic activity defined by a Geboes score of ≧3.1 (hazard ratio [HR], 1.732; P = .035) and steroid use history (HR, 1.762; P = .008) were independent factors associated with clinical relapse. When stratified, the 1- and 2-year incidence rates of clinical relapse were 4.1% and 10.6%, respectively, for patients with histologic improvement and no steroid use history, whereas the rates were 23.9% and 39.4% for patients with histologic activity and steroid use history. Conclusions In UC with an MES of 0 or 1, histologic activity and steroid use history can be used to stratify the risk of clinical relapse.

Background/Aim: Transient receptor potential vanilloid 6 (TRPV6), an endothelial Ca(2+)-selective entry channel, is expressed in various cancer types, and a selective TRPV6 inhibitor is currently being investigated in a clinical trial. However, TRPV6 expression in hepatocellular carcinoma (HCC) has not been reported. Materials and Methods: We evaluated TRPV6 expression in 219 cases of HCC and analyzed its association with clinicopathological parameters and prognostic significance. TRPV6 mRNA expression was compared between HCC and non-tumor liver tissues using various public datasets, and its prognostic effect was examined in The Cancer Genome Atlas (TCGA) cohort. Results: Low TRPV6 expression was found in 37.4% of patients, which was significantly associated with adverse histologic features, and patients with low TRPV6 expression had shorter recurrence-free and disease-free survival. TRPV6 mRNA expression was consistently lower in HCC compared to non-tumor liver samples in public datasets, at the whole tissue level as well as single-cell level. Patients with low TRPV6 expression in the TCGA cohort had shorter progression-free survival. Conclusion: TRPV6 expression is down-regulated in HCCs and associated with a poor prognosis. TRPV6 may be a prognostic biomarker in HCC.

Purpose: The aim of this study was to develop and validate a fully-automatic quantification of the hepatorenal index (HRI) calculated by a deep convolutional neural network (DCNN) comparable to the interpretations of radiologists experienced in ultrasound (US) imaging.Methods: In this retrospective analysis, DCNN-based organ segmentation with Gaussian mixture modeling for automated quantification of the HRI was developed using abdominal US images from a previous study. For validation, 294 patients who underwent abdominal US examination before living-donor liver transplantation were selected. Interobserver agreement for the measured brightness of the liver and kidney and the calculated HRI were analyzed between two board-certified radiologists and DCNN using intraclass correlation coefficients (ICCs).Results: Most patients had normal (n=95) or mild (n=198) fatty liver. The ICCs of hepatic and renal brightness measurements and the calculated HRI between the two radiologists were 0.892 (95% confidence interval [CI], 0.866 to 0.913), 0.898 (95% CI, 0.873 to 0.918), and 0.681 (95% CI, 0.615 to 0.738) for the first session and 0.920 (95% CI, 0.901 to 0.936), 0.874 (95% CI, 0.844 to 0.898), and 0.579 (95% CI, 0.497 to 0.650) for the second session, respectively; the results ranged from moderate to excellent agreement. Using the same task, the ICCs of the hepatic and renal measurements and the calculated HRI between the average values of the two radiologists and DCNN were 0.919 (95% CI, 0.899 to 0.935), 0.916 (95% CI, 0.895 to 0.932), and 0.734 (95% CI, 0.676 to 0.782), respectively, showing high to excellent agreement.Conclusion: Automated quantification of HRI using DCNN can yield HRI measurements similar to those obtained by experienced radiologists in patients with normal or mild fatty liver.

ZMYND8 (Zinc finger MYND (Myeloid, Nervy and DEAF-1)-type containing 8) has been known to play an important role in tumor regulation in various types of cancer. However, the results of ZMYND8 expression and their clinical significance in hepatocellular carcinoma (HCC) have not yet been published. In the present study, we investigate the expression of ZMYND8 protein and mRNA in HCC and elucidate its prognostic significance. ZMYND8 protein and mRNA expression in 283 and 234 HCCs were investigated using immunohistochemistry and microarray gene expression profiling data. The relationships between ZMYND8 expression with clinicopathologic features and prognosis of HCC patients were evaluated. Furthermore, we performed the invasion, migration, apoptosis, soft agar formation assay and sphere formation assay in HCC cell lines, and evaluated tumorigenicity in a nude mouse model, after ZMYND8 knockdown. Overexpression of ZMYND8 protein and mRNA was observed in 20.5% and 26.9% of HCC cases, respectively. High ZMYND8 expression showed significant correlations with microvascular invasion, high Edmondson grade, advanced American Joint Committee on Cancer, and increased alpha-fetoprotein level. ZMYND8 mRNA overexpression was an independent prognostic factor for predicting early recurrence as well as short recurrence-free survival (RFS). Downregulation of ZMYND8 reduced migration and invasion of HCC cells, and promoted apoptosis of HCC cells in an in vitro model. In a xenograft nude mouse model, knockdown of ZMYND8 significantly reduced tumor growth. ZMYND8 mRNA overexpression could be a prognostic marker of shorter RFS in HCC patients after curative resection. ZMYND8 might play an important role in the proliferation and progression of HCC and could be a promising candidate for targeted therapy.

Tumor budding is associated with adverse histology. It is a predictor of poor oncologic outcomes in colorectal cancer. However, it remains unclear whether tumor budding is a predictor of poor prognosis for rectal cancer patients regardless of neoadjuvant chemoradiotherapy (nCRT). This study analyzed 2888 rectal cancer patients who underwent radical surgery from 2007 to 2014. Among these patients, 939 underwent nCRT while 1949 did not receive nCRT. Tumor budding was defined as positive if the number of isolated tumor cells or small clusters of up to five tumor cells at the invasive front of the tumor was five or more. If the number was less than five, it was defined as negative. Patients were categorized according to tumor budding status. We used 1:1 propensity score matching to adjust for potential baseline confounders between the two groups. Among 2888 patients, 939 received nCRT while 1949 did not receive nCRT. A total of 418 patients who received nCRT were matched (209 in each group). A total of 1024 patients without nCRT were also matched (512 in each group). In matched patients, 5-year overall survival (OS) and 5-year disease-free survival (DFS) rates for the positive budding group were significantly lower than those in the negative budding group regardless of nCRT. On multivariate analysis of prognostic factors, positive budding was associated with poorer disease-free survival independent of nCRT. Tumor budding positivity is a prognostic indicator of poor outcomes in rectal cancer patients regardless of neoadjuvant chemoradiotherapy.

Hepatocellular carcinoma (HCC) has one of the highest mortality rates among solid cancers. Late diagnosis and a lack of efficacious treatment options contribute to the dismal prognosis of HCC. Immune checkpoint inhibitor (ICI)-based immunotherapy has presented a new milestone in the treatment of cancer. Immunotherapy has yielded remarkable treatment responses in a range of cancer types including HCC. Based on the therapeutic effect of ICI alone (programmed cell death (PD)-1/programmed death-ligand1 (PD-L)1 antibody), investigators have developed combined ICI therapies including ICI + ICI, ICI + tyrosine kinase inhibitor (TKI), and ICI + locoregional treatment or novel immunotherapy. Although these regimens have demonstrated increasing treatment efficacy with the addition of novel drugs, the development of biomarkers to predict toxicity and treatment response in patients receiving ICI is in urgent need. PD-L1 expression in tumor cells received the most attention in early studies among various predictive biomarkers. However, PD-L1 expression alone has limited utility as a predictive biomarker in HCC. Accordingly, subsequent studies have evaluated the utility of tumor mutational burden (TMB), gene signatures, and multiplex immunohistochemistry (IHC) as predictive biomarkers. In this review, we aim to discuss the current state of immunotherapy for HCC, the results of the predictive biomarker studies, and future direction.

Hepatic estrogen signaling, which is important in liver injury/recovery, is determined by the level of systemic estrogen and hepatic estrogen receptor. We aimed to evaluate whether females' advantage in the tolerance of hepatic ischemia-reperfusion injury decreases according to the age of 40 y (systemic estrogen decrease) and macrosteatosis (hepatic estrogen receptor decrease).We included 358 living liver donors (128 female and 230 male individuals). The tolerance of hepatic ischemia-reperfusion injury was determined by the slope of the linear regression line modeling the relationship between the duration of intraoperative hepatic ischemia and the peak postoperative transaminase level. Estrogen receptor content was measured in the biopsied liver samples using immunohistochemistry.In the whole cohort, the regression slope for aspartate transaminase was comparable between female and male individuals (P = 0.940). Within the subgroup of donors aged ≤40 y, the regression slope was significantly smaller in female individuals (P = 0.031), whereas it was comparable within donors aged40 y (P = 0.867). Within the subgroup of nonmacrosteatotic donors aged ≤40 y, the regression slope was significantly smaller in female individuals in univariable (P = 0.002) and multivariable analysis (P = 0.006), whereas the sex difference was not found within macrosteatotic donors aged ≤40 y (P = 0.685). Estrogen receptor content was significantly greater in female individuals within nonmacrosteatotic donors aged ≤40 y (P = 0.021), whereas it was not different in others of age40 y or with macrosteatosis (P = 0.450).The tolerance of hepatic ischemia-reperfusion injury was greater in female individuals than in male individuals only when they were40 y and without macrosteatosis. The results were in agreement with the hepatic estrogen receptor immunohistochemistry study.

Objective To evaluate the performance of dual internally cooled wet tip (ICWT) radiofrequency electrodes in comparison to dual internally cooled tip (ICT) electrodes. Methods Twenty ablation zones were created for each type of electrodes. Planned procedure time was 6 min. Diameters of the ablation zone along the x-, y-, and z-axes (Dx, Dy, and Dz), ablation zone sphericity, quantitative sphericity measurement, and ablation volume were measured and compared between the two electrode types. Circularity of the ablation zone on the surface with x- and z- axes (zx plane) and amount of energy applied were also compared. Results Dx and Dz were significantly longer with ICWT than those with ICT (Dx: 3.0 vs. 2.8 cm, p = .018; and Dz: 2.7 vs. 2.3 cm, p

Risk factors, including lymphatic/vascular/perineural invasion, are considered indication for adjuvant treatment in stage II colon cancer. However, tumor budding is not included in the above risk factors.The object of this study was to assess the value of tumor budding as a prognostic factor in stage II colon cancer.This is a retrospective cohort study.This study was conducted in a tertiary referral center.This study analyzed 1,390 patients with stage II colon cancer who received curative resection from 2007 to 2013 at an institution.These patients were classified according to tumor budding status: low-grade tumor budding (10 buds) and high-grade tumor budding (≥ 10 buds). Differences between the two groups were corrected by the propensity score matching.Disease free survival and overall survival were the primary end points.Among 1,390 patients, 146 (10.5%) had high-grade tumor budding. The high- grade tumor budding group showed adverse histological characteristics such as advanced T stage, histological grade of differentiation, and presence of lymphatic/perineural invasion. After matching, the 5-year disease-free survival rate for the high-grade tumor budding group was significantly lower than for the low-grade group. We also compared survival outcomes according to tumor budding grade for patients who did not have risk factors who and did not receive adjuvant treatment. The 5-year overall survival was similar between the two groups. However, the 5-year disease-free survival decreased significantly in the high-grade tumor budding group than the low-grade tumor budding group.This was a retrospective study with a single center design.High-grade tumor budding is a poor prognostic factor in stage II colon cancer and is considered as one of the risk factors for adjuvant treatment. See Video Abstract at http://links.lww.com/DCR/B962.

To evaluate the postoperative prognostic value of the Liver Imaging Reporting and Data System (LI-RADS) category on gadoxetic acid-enhanced MRI and 18F-fluorodeoxyglucose PET-CT in patients with primary liver carcinomas (PLCs).A total of 189 patients with chronic liver disease and surgically proven single PLC (42 intrahepatic cholangiocarcinomas and 21 combined hepatocellular-cholangiocarcinomas and 126 hepatocellular carcinomas [2:1 matching to non-HCC malignancies]) were retrospectively evaluated with gadoxetic acid-enhanced MRI and PET-CT. Two independent reviewers assigned an LI-RADS category for each observation. The tumor-to-liver standardized uptake value ratio (TLR) was calculated. The overall survival (OS), recurrence-free survival (RFS), and the associated factors were evaluated.In multivariable analysis, LI-RADS category (LR-4 or LR-5 [LR-4/5] vs. LR-M; OS, hazard ratio [HR] 2.24, p = 0.006; RFS, HR 1.61, p = 0.028) and TLR (low, 2.3 vs. high, ≥ 2.3; OS, HR 2.09, p = 0.014; RFS, HR 2.17, p 0.001) were the independent factors for OS and RFS. For the LR-M group, the high TLR group showed lower OS and RFS rates than the low TLR group (OS, p = 0.008; RFS, p 0.001). For the LR-4/5 group, the OS and RFS rates were not significantly different between the high TLR and low TLR groups (both p 0.05).Both LI-RADS category on MRI and TLR on PET-CT are associated with the postoperative prognosis of PLCs. The prognosis of PLCs classified as LR-M can be further stratified according to the TLR group, but not for the PLCs classified as LR-4/5.• The LI-RADS category (LR-4/5 vs. LR-M) and tumor-to-liver standardized uptake value ratio (TLR, low vs. high) were independent factors for postoperative prognosis of primary liver carcinomas (PLCs). • For PLCs classified as LR-M, the TLR group helps stratify the postoperative prognosis of PLCs, with the high TLR group having a poor prognosis and the low TLR group having a better prognosis (p = 0.008 for OS and p 0.001 for RFS). • For PLCs classified as LR-4/5, the OS and RFS rates were not significantly different between the high TLR and low TLR groups (both p 0.05).

To compare the presence of washout and the diagnostic performance of computed tomography (CT) and magnetic resonance imaging (MRI) for hepatocellular carcinoma (HCC) according to the presence of hepatic steatosis. This retrospective study included 566 patients with chronic liver disease who had undergone hepatic resection for hepatic tumors (482 HCCs and 84 non-HCCs) between January 2016 and June 2018 and had available multiphasic CT and MR images. Patients were allocated in the fatty liver (n = 141) or non-fatty liver (n = 425) group according to the presence of hepatic steatosis, defined as lipid droplets in at least 5% of hepatocytes on pathological examination. The presence of HCC washout and the diagnostic performance of CT and MRI for HCC were compared between the groups. HCC washout was less frequently seen in the fatty liver group than in the non-fatty liver group on CT (61.5% vs. 88.9%, p 0.05). Hepatic steatosis significantly decreased the performance of CT for the diagnosis of HCC, whereas it did not significantly alter the performance of MRI. • Unlike MRI, there is vanishing HCC washout on CT caused by the background hepatic steatosis. • The diagnostic performance of CT for the diagnosis of HCC was significantly altered by hepatic steatosis. • The optimal cutoff HU value of the liver parenchyma for the vanishing washout of HCC was

Background Hepatobiliary phase (HBP) hypointense nodules without arterial phase hyperenhancement (APHE) at gadoxetic acid-enhanced MRI may indicate hepatocellular carcinoma (HCC) or nonmalignant cirrhosis-associated nodules. Purpose To assess the distribution of pathologic diagnoses of HBP hypointense nodules without APHE at gadoxetic acid-enhanced MRI and to evaluate clinical and imaging features in differentiating their histologic grades. Materials and Methods This retrospective multicenter study included pathologic analysis-confirmed HBP hypointense nodules without APHE (≤30 mm) in patients with chronic liver disease or cirrhosis screened between January 2008 and June 2016. Central pathologic review by 10 pathologists determined final histologic grades as progressed HCC, early HCC, high-grade dysplastic nodule (DN), and low-grade DN or regenerative nodule. Gadoxetic acid-enhanced MRI features were analyzed by three radiologists. Multivariable logistic regression analyses with elastic net regularization were performed to identify clinical and imaging features for differentiating histologic grades. Results There were 298 patients (mean age, 59 years ± 10; 226 men) with 334 nodules evaluated, and progressed HCCs were diagnosed in 44.0% (147 of 334), early HCCs in 20.4% (68 of 334), high-grade DNs in 27.5% (92 of 334), and low-grade DNs or regenerative nodules in 8.1% (27 of 334). Serum α-fetoprotein level 100 ng/mL or greater (odds ratio, 2.7; P = .01) and MRI features including well-defined margin (odds ratio, 5.5; P = .003), hypointensity at precontrast T1-weighted imaging (odds ratio, 3.2; P < .001), intermediate hyperintensity at T2-weighted imaging (odds ratio, 3.4; P < .001), and restricted diffusion (odds ratio, 1.9; P = .04) were independent predictors for progressed HCC at multivariable analysis. Conclusion In patients at high risk for hepatocellular carcinoma (HCC), hepatobiliary phase hypointense nodules without arterial phase hyperenhancement at gadoxetic acid-enhanced MRI corresponded mainly to progressed HCCs, early HCCs, and high-grade dysplastic nodules. High α-fetoprotein level and some imaging features at MRI helped to differentiate progressed HCC from lower grade nodules. © RSNA, 2020 Online supplemental material is available for this article. See also the editorial by Motosugi in this issue.

PurposeThe purpose of this study was to investigate the role of carcinoembryonic antigen (CEA) levels in improving the performance of magnetic resonance imaging (MRI) for the prediction of pathologic response after the neoadjuvant chemoradiation (NCRT) for patients with rectalcancer.Materials and MethodsWe retrospectively reviewed the medical records of 524 rectal cancer patients who underwent NCRT and total mesorectal excision between January 2009 and December 2014. The performances of MRI with or without CEA parameters (initial CEA and CEA dynamics) for prediction of pathologic tumor response grade (pTRG) were compared by receiver-operating characteristic analysis with DeLong’s method. Cox regression was used to identify the independent factors associated to pTRG and disease-free survival (DFS) after NCRT.ResultsThe median follow-up was 64.0 months (range, 3.0 to 113.0 months). On multivariate analysis, poor tumor regression grade on MRI (mrTRG; p < 0.001), initial CEA (p < 0.001) and the mesorectal fascia involvement on MRI before NCRT (mrMFI; p=0.054) showed association with poor pTRG. The mrTRG plus CEA parameters showed significantly improved performances in the prediction of pTRG than mrTRG alone. All of mrTRG, mrMFI, and initial CEA were also identified as independent factors associated with DFS. The initial CEA further discriminated DFS in the subgroups with good mrTRG or that without mrMFI.ConclusionThe CEA parameters significantly improved the performance of MRI in the prediction of pTRG after NCRT for patients with rectal cancer. The DFS was further discriminated by initial CEA level in the groups with favorable MRI parameters.

This study analyzed factors related to allograft fibrosis in clinically stable pediatric liver transplantation patients. Pediatric patients who underwent liver transplantation from January 1997 to January 2008 and further underwent 10-year protocol biopsies were examined. Grades of inflammation and fibrosis were classified based on Banff criteria and the Liver Allograft Scoring (LAF) system, respectively. Risk factors for fibrosis were analyzed using logistic regression. Sixty-six patients with no clinical signs of chronic liver disease were included. Forty-one patients out of 66 (62.1%) had certain stage of allograft fibrosis. More than five events with aminotransferase >50 U/L was a risk factor for a LAF score 1–2 portal fibrosis (OR = 3.156, CI 1.059–9.410, P = 0.039). More than five events with aminotransferase >100 U/L was a risk factor for LAF score 2 portal fibrosis (OR = 13.978, CI 2.025–97.460, P = 0.007) and LAF score 1–2 sinusoidal fibrosis (OR = 4.897, CI 1.167–20.548, P = 0.030). Positive autoantibody (OR = 3.298, CI 1.039–10.473, P = 0.043) and gamma-glutamyl transferase 60 U/L (OR = 6.201, CI 1.096–35.097, P = 0.039) were related to sinusoidal fibrosis with LAF score of 1–2 and 2, respectively. Experience of post-transplantation lymphoproliferative disease was related to LAF score 1–2 portal fibrosis (OR = 7.371, CI 1.320–41,170, P = 0.023) and LAF score 1–2 centrolobular fibrosis (OR = 8.822, CI = 1.378–56.455, P = 0.022). Our results indicate that liver fibrosis is common in patients with no clinical signs of graft deterioration and repeated elevation of aminotransferases, positive autoantibodies, elevated gamma-glutamyl transferase and experience of post-transplantation lymphoproliferative disease are suspicious signs for fibrosis.

Background/aim Primary hepatic angiosarcoma (PHA) is a rare disease entity with variable morphologic features. Recent findings regarding ROS1 gene rearrangements in PHA may lead to new targeted therapies. Patients and methods Thirteen cases (4 resected specimens and 9 biopsy samples) underwent histologic review and morphologic patterns were classified according to a previous study as 1) sinusoidal, 2) peliotic, 3) vasoformative, and 4) solid (epithelioid/spindled). ROS1 immunohistochemistry and investigation of the presence of a ROS1 fusion gene by reverse transcription-polymerase chain reaction were performed in available cases. Results Eight of 13 cases (62%) showed vasoformative patterns. Three cases (23%) were classified as sinusoidal and two (15%) as solid patterns. Mortality rate was 90% (9/10) except for three patients lost in follow up. Only one patient is still alive and has survived for 8 months with the disease. All cases tested did not have ROS1 expression (0/9) or a ROS1 fusion gene (0/4). Conclusion We report 13 cases of PHA with 90% mortality. Vasoformative PHA is the most common histologic type. New findings on ROS1 fusion gene rearrangements could lead to the development of novel targeted therapeutics for PHA patients with dismal prognosis.

Background/aim Pregnancy up-regulated non-ubiquitous calmodulin kinase (PNCK) is a member of calmodulin kinase, and overexpression of PNCK with involvement in carcinogenesis have been reported in HER-2 amplified breast cancer, clear cell renal cell carcinoma and nasopharygeal carcinoma. However, the expression of PNCK and its clinical implication have not been elucidated in hepatocellular carcinoma (HCC). Materials and methods We investigated PNCK expression at both the protein and mRNA level using immunohistochemistry (IHC) and microarray gene expression profiling in HCC tissue samples, and evaluated its association with clinicopathological parameters and their potential prognostic significance. Results High PNCK protein expression and high PNCK mRNA level was observed in 61.7% and 34.7% of total HCC cases, respectively. PNCK mRNA level was higher in tumor tissues than in background non-tumor tissues, and significantly correlated with protein expression by IHC. High PNCK expression was associated with higher Edmondson grade, intrahepatic metastasis, microvascular invasion and higher AFP levels. Patients with high PNCK expression showed shorter recurrence-free survival and disease-specific survival, and high mRNA expression of PNCK was an independent prognostic factor in disease-specific survival. Conclusion Up-regulation of PNCK expression as well as its association with poor prognosis was demonstrated in HCC. PNCK might be a prognostic biomarker of HCC, and could be a potential candidate therapeutic target.

Checkpoint inhibitor approval for microsatellite instability-high (MSI-H) tumours has made MSI as a therapeutically important biomarker. Next-generation sequencing (NGS)-based MSI detection is being widely used for assessing MSI. However, MSI tumours detected using NGS and their relevance to MSI-polymerase chain reaction (PCR) and mismatch repair deficiency (dMMR) are unclear. In 1942 solid cancer cases tested using NGS-based comprehensive cancer panel with 523 genes (1.94 mb), the MSI score, tumour mutation burden (TMB; ≥ 10 mutations/mb), and frameshift mutations were analysed. GeneScan analyses of five mononucleotide markers (MSI-PCR) and MMR protein immunohistochemistry (IHC) were compared with the NGS-MSI results. With a ≥ 12% MSI score as a cut-off for MSI-H, two MSS cases were classified as MSI-H. With a ≥ 20% cut-off, 10 cases categorised as MSS by NGS were MSI-H/dMMR by MSI-PCR and MMR IHC. To avoid discrepant cases, we adopted a high MSI cut-off and a borderline MSI category. Finally, MSI-H (≥ 20%), borderline MSI (≥ 7% and 20%), and MSS ( 7%) were found in 35 (1.8%), 24 (1.2%), and 1883 (97%) cases, respectively. All MSI-H cases by NGS were MSI-H/dMMR by MSI-PCR and MMR IHC. Of the 24 borderline MSI cases by NGS, MSI-H/dMMR was 9 (37.5%) cases, MSS/dMMR was 1 (4.2%) case, and 11 (45.8%) of them had high TMB. All MSS cases by NGS were MSS/pMMR by MSI-PCR/IHC, and 257 (13.6%) had high TMB. With those arbitrary cut-off points, 10 (0.5%) MSS cases using NGS were discrepant with MSI-PCR or MMR IHC, and all were borderline MSI cases. The mean number of frameshift mutations was significantly higher in the MSI-H group (28.3) than in the borderline MSI (7.7) or MSS (1.3) groups (p 0.001). In conclusion, to facilitate therapeutic decision-making for NGS, cut-off points for MSI can be defined based on MSI-PCR/dMMR confirmation.

HOXD10 downregulation resulting from epigenetic changesas well as its role as a tumor suppressor have been reported in several cancers including hepatocellular carcinomas (HCCs). However, the prognostic role of HOXD10 expression in HCC tissue samples has not been evaluated.HOXD10 expression was investigated in 278 curatively resected HCC samples using immunohistochemistry and its effectiveness in predicting patient outcome was analyzed.Low expression of HOXD10 was observed in 82.7% of HCC samples, and this was associated with increased age, large tumor size and advanced stage.HOXD10 was an independent predictive factor for early tumor recurrence at less than 2 years. Patients with low HOXD10 expression showed shorter recurrence-free survival (RFS) (p=0.024) and disease-specific survival (DSS) (p=0.016) than those with high expression. Multivariate analysis confirmed that low HOXD10 expression was an independent predictor of shorter RFS (hazard ratio 1.873, p=0.006) and DSS (hazard ratio2.504, p=0.012) than high HOXD10 expression.The present study provides clinical evidence supporting the use of HOXD10 as a prognostic biomarker in curatively resected HCCs, and suggests that HOXD10 could also be a potential therapeutic target in HCC.

The tumor–stroma ratio (TSR) determined by pathologists is subject to intra- and inter-observer variability. We aimed to develop a computational quantification method of TSR using deep learning-based virtual cytokeratin staining algorithms. Patients with 373 advanced (stage III [n = 171] and IV [n = 202]) gastric cancers were analyzed for TSR. Moderate agreement was observed, with a kappa value of 0.623, between deep learning metrics (dTSR) and visual measurement by pathologists (vTSR) and the area under the curve of receiver operating characteristic of 0.907. Moreover, dTSR was significantly associated with the overall survival of the patients (P = 0.0024). In conclusion, we developed a virtual cytokeratin staining and deep learning-based TSR measurement, which may aid in the diagnosis of TSR in gastric cancer.

Programmed death-ligand 1 (PD-L1) immunohistochemistry (IHC), microsatellite instability (MSI), and tumor mutation burden (TMB) have been proposed as a predictive biomarker to predict response to immune checkpoint blockade (ICB). We aimed to find the relationship of PD-L1 IHC to TMB and MSI using a comprehensive cancer panel assay (CCPA) with &gt, 500 genes in advanced cancer patients. CCPA results from 588 archived tissue samples were analyzed for TMB and MSI. In seven samples, whole exome sequencing confirmed TMB with Pearson’s correlation coefficient of 0.972 and all MSI-high cases were validated by pentaplex PCR. Association of TMB and MSI with their corresponding PD-L1 IHC was analyzed. The median TMB value of 588 cases was 8.25 mutations (mut)/Mb (range 0–426.8) with different distributions among the tumor types, with high proportions of high-TMB (&gt, 10mut/Mb) in tumors from melanoma, colorectal, gastric, and biliary tract. The TMB values significantly correlated with PD-L1 expression, and this correlation was prominent in gastric and biliary tract cancers. Moreover, the MSI score, the proportion of unstable MSI sites to total assessed MSI sites, showed a significant correlation with the TMB values and PD-L1 scores. This study demonstrates that PD-L1 expression is significantly associated with TMB and MSI score and this correlation depends on the location of the primary tumor.