Hepatocellular carcinoma: Intratumoral EpCAM-positive cancer stem cell heterogeneity identifies high-risk tumor subtype

Abstract Background The translational interest in the intratumoral heterogeneity of hepatocellular carcinoma (HCC) has been increasing. The dismal prognosis of this pathology is linked to the features of the HCC harbouring cancer stem cells (CSC), represented by EpCAM-expression. However, the extent of the impact of intratumoral distribution of CSC-features, both on the recurrence after curative resection and on clinical outcome, remains unknown. To address this, we investigated the spatial heterogeneity of CSC-features with the aim of identifying the unique HCC patient subgroups amenable to adjuvant treatment. Methods We designed a tissue microarray (TMA) from patients who had received liver resection between 2011 and 2017. Tumor specimens were sampled at multiple locations (n = 3–8). EpCAM-positivity was assessed for intensity and proportion by applying a score dividing three groups: (i) negative (E−/−); (ii) heterogeneous (E−/+); and (iii) homogeneous (E+/+). The groups were further analysed with regard to time-to-recurrence (TTR) and recurrence-free-survival (RFS). Results We included 314 tumor spots from 69 patients (76.8% male, median age 66, liver cirrhosis/fibrosis 75.8%). The risk factors were alcohol abuse (26.2%), NASH (13.1%), HBV (15.5%), HCV (17.9%) and others (27.4%), representative of a typical Western cohort. E+/+ patients experienced significantly shorter TTR and RFS compared to E+/− and E−/− patients (TTR 5 vs. 19 months, p = 0.022; RFS 5 vs. 14 vs. 21 months, p = 0.016). Only homogeneous EpCAM-positivity correlated with higher AFP levels (> 400 ng/ml, p = 0.031). Conclusions Spatial heterogeneity of EpCAM-expression was markedly present in the cohort. Of note, only homogeneous EpCAM-expression correlated significantly with early recurrence, whereas heterogeneous EpCAM-expression was associated with clinical endpoints comparable to EpCAM-negativity. We identified a unique HCC subtype associated with a high risk of tumor recurrence.

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Hepatocellular carcinoma: Intratumoral EpCAM-positive cancer stem cell heterogeneity identifies high-risk tumor subtype

10.21203/rs.3.rs-54971/v3 ◽

2020 ◽

Author(s):

Jenny Krause ◽

Johann von Felden ◽

Christian Casar ◽

Thorben W. Fründt ◽

Johanna Galaski ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

High Risk ◽

Spatial Heterogeneity ◽

Early Recurrence ◽

Epcam Expression ◽

Clinical Endpoints ◽

Time To Recurrence ◽

Dismal Prognosis ◽

The Impact ◽

Intratumoral Distribution

Abstract Background: The translational interest in the intratumoral heterogeneity of hepatocellular carcinoma (HCC) has been increasing. The dismal prognosis of this pathology is linked to the features of the HCC harbouring cancer stem cells (CSC), represented by EpCAM-expression. However, the extent of the impact of intratumoral distribution of CSC-features, both on the recurrence after curative resection and on clinical outcome, remains unknown. To address this, we investigated the spatial heterogeneity of CSC-features with the aim of identifying the unique HCC patient subgroups amenable to adjuvant treatment.Methods: We designed a tissue microarray (TMA) from patients who had received liver resection between 2011 and 2017. Tumor specimens were sampled at multiple locations (n=3-8). EpCAM-positivity was assessed for intensity and proportion by applying a score dividing three groups: (i) negative (E-/-); (ii) heterogeneous (E-/+); and (iii) homogeneous (E+/+). The groups were further analysed with regard to time-to-recurrence (TTR) and recurrence-free-survival (RFS).Results: We included 314 tumor spots from 69 patients (76.8% male, median age 66 years, liver cirrhosis/fibrosis 75.8). The risk factors were alcohol abuse (26.2%), NASH (13.1%), HBV (15.5%), HCV (17.9%) and others (27.4%), representative of a typical Western cohort. E+/+ patients experienced significantly shorter TTR and RFS compared to E+/- (and E-/-) patients (TTR 5 vs. 19 months, p=0.022; RFS 5 vs. 14 vs. 21 months, p=0.016). Only homogeneous EpCAM-positivity correlated with higher AFP levels (>400 ng/ml, p=0.031).Conclusions: Spatial heterogeneity of EpCAM-expression was markedly present in the cohort. Of note, only homogeneous EpCAM-expression correlated significantly with early recurrence, whereas heterogeneous EpCAM-expression was associated with clinical endpoints comparable to EpCAM-negativity. We identified a unique HCC subtype associated with a high risk of tumor recurrence.

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Hepatocellular carcinoma: Intratumoral EpCAM-positive cancer stem cell heterogeneity identifies high-risk tumor subtype

10.21203/rs.3.rs-54971/v2 ◽

2020 ◽

Author(s):

Jenny Krause ◽

Johann von Felden ◽

Christian Casar ◽

Thorben W. Fründt ◽

Johanna Galaski ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Stem Cell ◽

High Risk ◽

Cancer Stem Cell ◽

Spatial Heterogeneity ◽

Early Recurrence ◽

Epcam Expression ◽

Clinical Endpoints ◽

Dismal Prognosis ◽

The Impact

Abstract Background: The translational interest in the intratumoral heterogeneity of hepatocellular carcinoma (HCC) has been increasing. The dismal prognosis of this pathology is linked to the features of the HCC harbouring cancer stem cell (CSC), represented by EpCAM-expression. However, the extent of the impact of intratumoral distribution of CSC-features both on the recurrence after curative resection, and on clinical outcome remains unknown. To address this, we investigated the spatial heterogeneity of CSC-features with the aim of identifying the unique HCC patient subgroups amenable to adjuvant treatment.Methods: We designed a tissue microarray (TMA) from patients who had received liver resection between 2011 and 2017. Tumor specimens were sampled at multiple locations (n=3-8). EpCAM-positivity was assessed for intensity and proportion by applying a score dividing three groups: (i) negative (E-/-); (ii) heterogeneous (E-/+); and (iii) homogeneous (E+/+). The groups were further analysed with regard to time-to-recurrence (TTR) and recurrence-free-survival (RFS).Results: We included 314 tumor spots from 69 patients (76.8% male, median age 66 years, liver cirrhosis/fibrosis 75.8). The risk factors were alcohol abuse (26.2%), NASH (13.1%), HBV (15.5%), HCV (17.9%) and others (27.4%), representative of a typical Western cohort. E+/+ patients experienced significantly shorter TTR and RFS compared to E+/- (and E-/-) patients (TTR 5 vs. 19 months, p=0.022; RFS 5 vs. 14 vs. 21 months, p=0.016). Only homogeneous EpCAM-positivity correlated with higher AFP levels (>400 ng/ml, p=0.031).Conclusions: Spatial heterogeneity of EpCAM-expression was markedly present in the cohort. Of note, only homogeneous EpCAM-expression correlated significantly with early recurrence, whereas heterogeneous EpCAM-expression was associated with clinical endpoints comparable to EpCAM-negativity. We identified a unique HCC subtype associated with a high risk of tumor recurrence.

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Clinical Impact of Intratumoral EpCAM-Positive Cancer Stem Cell Heterogeneity in Patients with Hepatocellular Carcinoma

10.21203/rs.3.rs-54971/v1 ◽

2020 ◽

Author(s):

Jenny Krause ◽

Johann von Felden ◽

Christian Casar ◽

Thorben W. Fründt ◽

Johanna Galaski ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Stem Cell ◽

Cancer Stem Cell ◽

Spatial Heterogeneity ◽

Adjuvant Treatment ◽

Early Recurrence ◽

Epcam Expression ◽

Clinical Endpoints ◽

Time To Recurrence ◽

Dismal Prognosis

Abstract Backgrounds & Aims: Intratumoural heterogeneity of hepatocellular carcinoma (HCC) is of increasing translational interest. Dismal prognosis is frequently linked to HCC harbouring cancer stem cell (CSC)-features, represented by EpCAM-expression. However, to what extent intratumoural distribution of CSC-features impacts on recurrence after curative resection remains unknown. Hence, we aimed to investigate the spatial heterogeneity of CSC-features and its impact on clinical outcome, identifying high-risk patients amenable to adjuvant treatment.Methods: We designed a tissue microarray (TMA) from patients, who received liver resection between 2011 and 2017. Tumour specimens were sampled at multiple locations (n=3-8). EpCAM-positivity was assessed for intensity and proportion by applying a score dividing three groups: negative (E-/-), heterogeneous-positive (E-/+), homogeneous-positive (E+/+). The groups were further analysed with respect to time-to-recurrence (TTR) and recurrence-free-survival (RFS).Results: We included 341 tumour spots from 75 patients (77% male, median age 66 years, liver cirrhosis/fibrosis 74.6%). Risk factors were alcohol abuse in 23.9%, NASH 16.3%, HBV 14.1%, HCV 17.4% and others 28.3%, representing a typical Western cohort. E+/+ patients experienced a significantly shorter TTR and RFS compared to E+/- (and E-/-) patients (TTR 5 vs. 19 months, p=0.017; RFS 5 vs. 14 vs. 18 months, p=0.016). Only homogeneous EpCAM-positivity correlated with higher AFP levels (>400 ng/ml, p=0.024).Conclusions: Spatial heterogeneity of EpCAM-expression was markedly present. Only homogeneously positive EpCAM-expression correlated significantly with early recurrence, whereas heterogeneous EpCAM-expression was associated with clinical endpoints comparable to EpCAM-negativity. Similar to colorectal cancer, high or low risk features for recurrence could be decisive for adjuvant treatment.

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Contrast-enhanced ultrasound–based ultrasomics score: a potential biomarker for predicting early recurrence of hepatocellular carcinoma after resection or ablation

British Journal of Radiology ◽

10.1259/bjr.20210748 ◽

2021 ◽

Author(s):

Hui Huang ◽

Si-min Ruan ◽

Meng-fei Xian ◽

Ming-de Li ◽

Mei-qing Cheng ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

High Risk ◽

Risk Group ◽

Early Recurrence ◽

High Risk Group ◽

Low Risk ◽

Primary Hepatocellular Carcinoma ◽

Contrast Enhanced Ultrasound ◽

Combined Model ◽

Contrast Enhanced

Objectives: This study aimed to construct a prediction model based on contrast-enhanced ultrasound (CEUS) ultrasomics features and investigate its efficacy in predicting early recurrence (ER) of primary hepatocellular carcinoma (HCC) after resection or ablation. Methods: This study retrospectively included 215 patients with primary HCC, who were divided into a developmental cohort (n = 139) and a test cohort (n = 76). Four representative images—grayscale ultrasound, arterial phase, portal venous phase and delayed phase —were extracted from each CEUS video. Ultrasomics features were extracted from tumoral and peritumoral area inside the region of interest. Logistic-regression was used to establish models, including a tumoral model, a peritumoral model and a combined model with additional clinical risk factors. The performance of the three models in predicting recurrence within 2 years was verified. Results: The combined model performed best in predicting recurrence within 2 years, with an area under the curve (AUC) of 0.845, while the tumoral model had an AUC of 0.810 and the peritumoral model one of 0.808. For prediction of recurrence-free survival, the 2 year cumulative recurrence rate was significant higher in the high-risk group (76.5%) than in the low-risk group (9.5%; p < 0.0001). Conclusion: These CEUS ultrasomics models, especially the combined model, had good efficacy in predicting early recurrence of HCC. The combined model has potential for individual survival assessment for HCC patients undergoing resection or ablation. Advances in knowledge: CEUS ultrasomics had high sensitivity, specificity and PPV in diagnosing early recurrence of HCC, and high efficacy in predicting early recurrence of HCC (AUC > 0.8). The combined model performed better than the tumoral ultrasomics model and peritumoral ultrasomics model in predicting recurrence within 2 years. Recurrence was more likely to occur in the high-risk group than in the low-risk group, with 2-year cumulative recurrence rates respectively 76.5% and 9.5% (p < 0.0001).

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Patients with polyclonal hepatocellular carcinoma are at a high risk of early recurrence and have a poor recurrence-free survival period

10.21203/rs.3.rs-782031/v1 ◽

2021 ◽

Author(s):

Masaki Kaibori ◽

Kazuko Sakai ◽

Hideyuki Matsushima ◽

Hisashi Kosaka ◽

Kosuke Matsui ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

High Risk ◽

Copy Number ◽

Snp Array ◽

Early Recurrence ◽

Copy Number Variations ◽

Survival Period ◽

Recurrence Free Survival ◽

Free Survival ◽

A Genome

Abstract Background/purpose of the study Tumor heterogeneity based on copy number variations is associated with the evolution of cancer and its clinical grade. Clonal composition (CC) represents the number of clones based on the distribution of B-allele frequency (BAF) obtained from a genome-wide single nucleotide polymorphism (SNP) array. A higher CC number represents a high degree of heterogeneity. We hypothesized and evaluated that the CC number in hepatocellular carcinoma (HCC) tissues might be associated with the clinical outcomes of patients. Methods Somatic mutation, whole transcriptome, and copy number variations of 36 frozen tissue samples of operably resected HCC tissues by targeted deepsequencing, RNAseq and SNP array. Results The samples were classified the heterogeneous tumors as poly-CC (n = 26) and the homogeneous tumors as mono-CC (n = 8). The patients with poly-CC had a higher rate of early recurrence and a significantly shorter recurrence-free survival period than the mono-CC patients (7.0 vs. not reached, p = 0.0084). No differences in pathogenic non-synonymous mutations, such as TP53, were observed between the two groups when targeted deep sequencing was applied. A transcriptome analysis showed that cell cycle-related pathways were enriched in the poly-CC tumors, compared with the mono-CC tumors. poly-CC HCC is highly proliferative and has a high risk of early recurrence. Conclusion CC is a candidate biomarker for predicting the risk of early postoperative recurrence and warrants further investigation.

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Identification of Novel Tumor Microenvironment-Related Long Non-coding RNAs to Predict the Prognosis for Hepatocellular Carcinoma

10.21203/rs.3.rs-877901/v1 ◽

2021 ◽

Author(s):

Shenglan Huang ◽

Jian Zhang ◽

Dan Li ◽

Xiaolan Lai ◽

Lingling Zhuang ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

High Risk ◽

Tumor Microenvironment ◽

Cox Regression ◽

Risk Group ◽

High Risk Group ◽

Low Risk ◽

Clinicopathological Parameters ◽

Kaplan Meier ◽

The Impact

Abstract Introduction: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors with poor prognosis. Tumor microenvironment (TME) plays a vital role in the tumor progression of HCC. Thus, we aimed to analyze the association of TME with HCC prognosis, and construct an TME-related lncRNAs signature for predicting the prognosis of HCC patients.Methods: We firstly assessed the stromal/immune /Estimate scores within the HCC microenvironment using the ESTIMATE algorithm based on TCGA database, and its associations with survival and clinicopathological parameters were also analyzed. Then, different expression lncRNAs were filtered out according to immune/stromal scores. Cox regression was performed to built an TME-related lncRNAs risk signature. Kaplan–Meier analysis was carried out to explored the prognostic values of the risk signature. Furthermore, we explored the biological functions and immune microenvironment feathers in high- and low risk groups. Lastly, we probed the association of the risk signature with the treatment responses to immune checkpoint inhibitors (ICIs) in HCC by comparing the immunophenoscore (IPS).Results: Stromal/immune /Estimate scores of HCC patients were obtained based on the ESTIMATE algorithm. The Kaplan-Meier curve analysis showed the high stromal/immune/ Estimate scores were significantly associated with better prognosis of the HCC patients. Then, six TME-related lncRNAs were screened for constructing the prognosis model. Kaplan-Meier survival curves suggested that HCC patients in high-risk group had worse prognosis than those with low-risk. ROC curve and Cox regression analyses demonstrated the signature could predict HCC survival exactly and independently. Function enrichment analysis revealed that some tumor- and immune-related pathways associated with HCC tumorigenesis and progression might be activated in high-risk group. We also discovered that some immune cells, which were beneficial to enhance immune responses towards cancer, were remarkably upregulated in low-risk group. Besides, there was closely correlation of immune checkmate inhibitors (ICIs) with the risk signature and the signature can be used to predict treatment response of ICIs.Conclusions: We analyzed the impact of the tumor microenvironment scores on the prognosis of patients with HCC. A novel TME-related prognostic risk signature was established, which may improve prognostic predictive accuracy and guide individualized immunotherapy for HCC patients.

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Post-operative AFP identifies a subset of patients at high risk of early recurrence in hepatitis b-associated hepatocellular carcinoma

HPB ◽

10.1016/j.hpb.2017.02.149 ◽

2017 ◽

Vol 19 ◽

pp. S102-S103

Author(s):

S. Bhagwandin ◽

Q. Wang ◽

W. Luan ◽

L. Warren ◽

M. Fiel ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

High Risk ◽

Hepatitis B ◽

Early Recurrence

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Clinicopathological Risk Factors for Recurrence within One Year after Initial Hepatectomy for Hepatocellular Carcinoma

The American Surgeon ◽

10.1177/000313481107700516 ◽

2011 ◽

Vol 77 (5) ◽

pp. 572-578 ◽

Cited By ~ 11

Author(s):

Michihiro Hayashi ◽

Tetsunosuke Shimizu ◽

Fumitoshi Hirokawa ◽

Yoshihiro Inoue ◽

Koji Komeda ◽

...

Keyword(s):

Risk Factors ◽

Hepatocellular Carcinoma ◽

High Risk ◽

Early Recurrence ◽

Intrahepatic Metastasis ◽

High Rate ◽

Preoperative Imaging ◽

Curative Intent ◽

Initial Hepatectomy ◽

Cancer Spread

Hepatocellular carcinoma (HCC) shows a high rate of recurrence after hepatectomy; predictive factors for early recurrence would help determine optimal therapeutic and management strategies. Among 163 patients with HCC undergoing hepatectomy with curative intent, 46 patients developed recurrence within 1 year. Clinicopathological data were retrospectively analyzed to identify predictive parameters for early recurrence. Survival rates in cases of recurrence within 1 year were worse than those of no recurrence within 1 year or recurrence after 1 year. Protein induced by vitamin K absence/antagonist II (PIVKA-II) greater than 150, positive fucosylated alpha-fetoprotein (L3-AFP), and deviancy from Milan criteria (MC) on preoperative imaging were associated with high risk of early recurrence and total number of these three risk factors predicted the survival. With multivariate analysis, 1) preoperatively, positive factors of two or more among three items of PIVKA-II, L3-AFP, and deviancy from MC; 2) and postoperatively, pathological cancer spread (microscopic vascular invasion and/or intrahepatic metastasis) both represented risks for early recurrence. A combination of three preoperative factors, PIVKA-II, L3-AFP, and MC status, in conjunction with the postoperative factor of cancer spread status represents a significant indicator for recurrence within 1 year. Improving the prognosis of patients with HCC would depend on how to adequately treat those at high risk of early recurrence.

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Patients with polyclonal hepatocellular carcinoma are at a high risk of early recurrence and have a poor recurrence-free survival period

Hepatology International ◽

10.1007/s12072-021-10278-4 ◽

2022 ◽

Author(s):

Masaki Kaibori ◽

Kazuko Sakai ◽

Hideyuki Matsushima ◽

Hisashi Kosaka ◽

Kosuke Matsui ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

High Risk ◽

Deep Sequencing ◽

Copy Number ◽

Snp Array ◽

Early Recurrence ◽

Copy Number Variations ◽

Recurrence Free Survival ◽

Free Survival ◽

Targeted Deep Sequencing

Abstract Background/purpose of the study Tumor heterogeneity based on copy number variations is associated with the evolution of cancer and its clinical grade. Clonal composition (CC) represents the number of clones based on the distribution of B-allele frequency (BAF) obtained from a genome-wide single nucleotide polymorphism (SNP) array. A higher CC number represents a high degree of heterogeneity. We hypothesized and evaluated that the CC number in hepatocellular carcinoma (HCC) tissues might be associated with the clinical outcomes of patients. Methods Somatic mutation, whole transcriptome, and CC number based on copy number variations of 36 frozen tissue samples of operably resected HCC tissues were analyzed by targeted deep sequencing, transcriptome analysis, and SNP array. Results The samples were classified into the heterogeneous tumors as poly-CC (n = 26) and the homogeneous tumors as mono-CC (n = 8). The patients with poly-CC had a higher rate of early recurrence and a significantly shorter recurrence-free survival period than the mono-CC patients (7.0 months vs. not reached, p = 0.0084). No differences in pathogenic non-synonymous mutations, such as TP53, were observed between the two groups when targeted deep sequencing was applied. A transcriptome analysis showed that cell cycle-related pathways were enriched in the poly-CC tumors, compared to the mono-CC tumors. Poly-CC HCC is highly proliferative and has a high risk of early recurrence. Conclusion CC is a possible candidate biomarker for predicting the risk of early postoperative recurrence and warrants further investigation.

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Portal vein infiltration in patients with hepatocellular carcinoma: The relevance of correct classification.

Journal of Clinical Oncology ◽

10.1200/jco.2017.35.15_suppl.e15651 ◽

2017 ◽

Vol 35 (15_suppl) ◽

pp. e15651-e15651

Author(s):

Arndt Weinmann ◽

Verena Steinle ◽

Sandra Koch ◽

Daniel Pinto dos Santos ◽

Jens Uwe Marquardt ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Liver Disease ◽

Portal Vein ◽

Liver Imaging ◽

Clinical Registry ◽

Main Trunk ◽

Cross Sectional ◽

Dismal Prognosis ◽

Tertiary Center ◽

The Impact

e15651 Background: Portal vein invasion (PVI) is has a significant impact on the prognosis of patients with hepatocellular carcinoma (HCC). Patients with PVI are classified as stage C in the BCLC score and systemic therapy is recommended. Patients with minor PVI are frequently misclassified due to radiological challenges in determining malignant PVI or non-adherence to guidelines. The concept of resection or TACE in limited PVI is sometimes followed with the assumption of a negligible influence on survival. Aim of this study is the reevaluation of PVI and the analysis of the impact of a misclassification. Methods: 763 patients with HCC of a total of 1413 were extracted from the clinical registry of our tertiary center as an ongoing effort to reevaluate the extent of PVI in all patients treated between 1/1/2000 and 12/31/2015. PVI was diagnosed by re-evaluating all available CT or MRI scans by an experienced liver imaging radiologist. PVI was documented using the Liver Cancer Study Group of Japan classification ranging from Vp0-Vp4: Vp0 = no PVI; Vp1 = segmental; Vp2 = right anterior or posterior PV; Vp3 = right or left PV; Vp4 = main trunk. The influence on survival was calculated for each BCLC stage. Results: 259 patients (pat) were classified with PVI. Median age at diagnosis was 65.3 years, 213 patients (82.2%) were male. Etiology of liver disease was alcohol (43.6%), viral hepatitis (29.8%), NASH (5.8%), and others (10.4%). No liver disease was present in 18 pat (6.9%). No liver cirrhosis (LCI) was present in 32 pat (12.4%). LCI was classified as Child Pugh stage A/B/C in 65 (25.1%)/109 (42.1%) and 52 (20.1%) of patients. BCLC classification prior to reevaluation in pat with new PVI was A/B/C/D in 9/13/164/71 of cases. Comparing the overall survival (OS) of pat initially classified as BCLC A with or without PVI was 21.3 months vs. 106.4 months (p = 0.001), in BCLC B the OS was 11.0 months vs. 37.7 months (p = 0.001). Conclusions: Even minor PVI leads to dismal prognosis. Meticulous evaluation of cross sectional imaging is crucial for the clinical management of patients with HCC. Once PVI has been diagnosed, such patients have to be classified as advanced stage. The guidelines should be followed closely, irrespective of the extent of PVI.

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