Utility of Serum YKL-40 as a Tumor-Specific Marker of Hepatobiliary Malignancies

( Ju Dong Yang ),( Eugene Kim ),( Rachel A. Pedersen ),( W. Ray Kim ),( Surakit Pungpapong ),( Lewis R. Roberts ) 대한소화기기능성질환·운동학회 2010 Gut and Liver Vol.4 No.4

Background/Aims: Serum YKL-40 has been linked to several human cancers. We investigated the potential role of serum YKL-40 as a marker of hepatobiliary malignancies. Methods: Archived serum samples of patients undergoing liver transplantation evaluation at the Mayo Clinic Rochester were used to measure YKL-40 levels. Patients were divided into three groups: hepatocellular carcinoma (HCC), cholangiocarcinoma (CCA), and end-stage liver disease (ESLD) without malignancies. The Model for ESLD (MELD) score was used to quantify the severity of liver disease. Results: The median serum YKL-40 level was highest in the ESLD group at 296 ng/mL, compared to 259 ng/mL in the HCC group and 80 ng/mL in the CCA group (p<0.01). There was a significant correlation between the MELD score and serum YKL-40 level (r=0.50, p<0.01). In a multivariate analysis, there was no significant difference in serum YKL-40 level between ESLD and HCC. CCA was associated with lower YKL-40 levels, a finding that was attributable to a lower prevalence of cirrhosis. Conclusions: The serum YKL-40 level has little utility as a cross-sectional screening tool for hepatobiliary malignancies, namely HCC and CCA. The role of YKL-40 as a surveillance marker in the follow-up of individual patients remains to be determined. (Gut Liver 2010;4:537-542)

Global patterns of hepatocellular carcinoma management from diagnosis to death: the BRIDGE Study

Park, Joong-Won,Chen, Minshan,Colombo, Massimo,Roberts, Lewis R,Schwartz, Myron,Chen, Pei-Jer,Kudo, Masatoshi,Johnson, Philip,Wagner, Samuel,Orsini, Lucinda S,Sherman, Morris Wiley-Blackwell Publishing 2015 Liver International Vol.35 No.9

<P><B>Background & Aims</B></P><P>Hepatocellular carcinoma (HCC) is the second most common cause of cancer deaths worldwide. The global HCC BRIDGE study was a multiregional, large-scale, longitudinal cohort study undertaken to improve understanding of real-life management of patients with HCC, from diagnosis to death.</P><P><B>Methods</B></P><P>Data were collected retrospectively from January 2005 to September 2012 by chart reviews of eligible patients newly diagnosed with HCC at participating institutions.</P><P><B>Results</B></P><P>Forty-two sites in 14 countries contributed final data for 18 031 patients. Asia accounted for 67% of patients, Europe for 20% and North America for 13%. As expected, the most common risk factor was hepatitis C virus in North America, Europe and Japan, and hepatitis B virus in China, South Korea and Taiwan. The most common Barcelona Clinic Liver Cancer stage at diagnosis was C in North America, Europe, China and South Korea, and A in Taiwan and Japan. Across all stages, first HCC treatment was most frequently transarterial chemoembolization in North America, Europe, China and South Korea, percutaneous ethanol injection or radiofrequency ablation in Japan and resection in Taiwan. Survival from first HCC treatment varied significantly by region, with median overall survival not reached for Taiwan and 60, 33, 31, 24 and 23 months for Japan, North America, South Korea, Europe and China respectively (<I>P</I> < 0.0001).</P><P><B>Conclusions</B></P><P>Initial results from the BRIDGE study confirm previously reported regional trends in patient demographic characteristics and HCC risk factors, document the heterogeneity of treatment approaches across regions/countries and underscore the need for earlier HCC diagnosis worldwide.</P>

Silencing of HSulf-2 expression in MCF10DCIS.com cells attenuate ductal carcinoma in situ progression to invasive ductal carcinoma <i>in vivo</i>

Khurana, Ashwani,McKean, Hiedi,Kim, Hyunseok,Kim, Sung-Hoon,Mcguire, Jacie,Roberts, Lewis R,Goetz, Matthew P,Shridhar, Viji BioMed Central 2012 Breast cancer research Vol.14 No.2

<P><B>Introduction</B></P><P>Ductal carcinoma <I>in situ </I>(DCIS) of the breast is a heterogeneous group of proliferative cellular lesions that have the potential to become invasive. Very little is known about the molecular alterations involved in the progression from DCIS to invasive ductal carcinoma (IDC). Heparan endosulfatase (HSulf-2) edits sulfate moieties on heparan sulfate proteoglycans (HSPGs) and has been implicated in modulating heparin binding growth factor signaling, angiogenesis and tumorigenesis. However, the role of HSulf-2 in breast cancer progression is poorly understood. MCF10DCIS.com cells (referred as MCF10DCIS) express HSulf-2 and form comedo type DCIS and progress to IDC when transplanted in immune-deficient mice and, therefore, is an ideal model to study breast cancer progression. We evaluated the role of HSulf-2 in progression from DCIS to IDC using mouse fat pad mammary xenografts.</P><P><B>Methods</B></P><P>Non-target control (NTC) and HSulf-2 knockdown in MCF10DCIS breast cancer cells were achieved by NTC shRNA and two different lentiviral shRNA against HSulf-2 respectively. Xenografts were established by injecting NTC and HSulf-2 deficient MCF10DCIS cells in mouse mammary fat pads. Xenografts were subjected to H&E staining for morphological analysis, TUNEL and Propidium iodide staining (to determine the extent of apoptosis), Western blot analysis and zymography.</P><P><B>Results</B></P><P>Using a mouse mammary fat pad derived xenograft model, we observed that compared to control treated xenografts, down-regulation of HSulf-2 was associated with significant delays in growth at Week 7 (<I>P-</I>value < 0.05). Histological examination of the tumors demonstrated substantial differences in comedo necrosis, with marked luminal apoptosis and up-regulation of apoptotic markers Bim, cleaved PARP and cleaved caspase 3 in HSulf-2 depleted xenografts. Furthermore, HSulf-2 depleted xenografts retained the basement membrane integrity with decreased activity and expression of matrix metalloproteinase 9 (MMP-9), an enzyme critical for degradation of extracellular matrix compared to nontargeted control.</P><P><B>Conclusion</B></P><P>Our data suggest that HSulf-2 expression may be critical for human breast cancer progression. Down-regulation of HSulf-2 leads to retention of comedo type DCIS and delays the progression of DCIS to IDC. Further studies are necessary to determine if therapeutic targeting of HSulf-2 expression might delay the progression of DCIS to IDC.</P>

Genes Associated with Recurrence of Hepatocellular Carcinoma: Integrated Analysis by Gene Expression and Methylation Profiling

양주동,설소영,임선희,김용훈,Zhifu Sun,이주석,Snorri S. Thorgeirsson,추인선,Lewis R. Roberts,강구정 대한의학회 2011 Journal of Korean medical science Vol.26 No.11

Gene expression is suppressed by DNA methylation. The goal of this study was to identify genes whose CpG site methylation and mRNA expression are associated with recurrence after surgical resection for hepatocellular carcinoma (HCC). Sixty-two HCCs were examined by both whole genome DNA methylation and transcriptome analysis. The Cox model was used to select genes associated with recurrence. A validation was performed in an independent cohort of 66 HCC patients. Among fifty-nine common genes, increased CpG site methylation and decreased mRNA expression were associated with recurrence for 12genes (Group A), whereas decreased CpG site methylation and increased mRNA expression were associated with recurrence for 25 genes (Group B). The remaining 22 genes were defined as Group C. Complement factor H (CFH) and myosin VIIA and Rab interacting protein (MYRIP) in Group A; proline/serine-rich coiled-coil 1 (PSRC1), meiotic recombination 11 homolog A (MRE11A), and myosin IE (MYO1E) in Group B; and autophagy-related protein LC3 A (MAP1LC3A), and NADH dehydrogenase 1 alpha subcomplex assembly factor 1 (NDUFAF1) in Group C were validated. In conclusion,potential tumor suppressor (CFH, MYRIP) and oncogenes (PSRC1, MRE11A, MYO1E) in HCC are reported. The regulation of individual genes by methylation in hepatocarcinogenesis needs to be validated.

Prognostic Subclasses of Intrahepatic Cholangiocarcinoma by Integrative Molecular - Clinical Analysis and Potential Targeted Approach

( Keun Soo Ahn ),( Koo Jeong Kang ),( Yu Na Kang ),( Yong Hoon Kim ),( Tae-seok Kim ),( Daniel O’ Brien ),( Lewis R Roberts ) 대한간학회 2018 춘·추계 학술대회 (KASL) Vol.2018 No.1

Aims: Although molecular characterization of intrahepatic cholangiocarcinoma(CCA) has been studied recently, integrative analysis between molecular and clinical characterization has not been established yet. We analyzed RNA sequencing data with annotated clinical data for clarifying genomic features of intrahepatic CCA, molecular specific clinical features and evaluating therapeutic potential based on molecular subtypes. Methods: We performed next generation RNA sequencing of 30 surgically resected intrahepatic CCA from Korean patients. RNA expression, variants and fusions were analyzed with clinical, pathologic features. RNA sequences from 32 intrahepatic CCA resected from USA were used for validation. Results: Patients were classified into 2 subclasses based on unsupervised clustering, which showed a significant difference 5- year survival. The validation cohort of USA data also revealed two subclasses with significant differences in survival. Two subclasses had different clinical and pathologic features for higher CEA and CA19-9 levels, underlying cholangitis and bile duct type pathology in the poor prognostic subclass and more frequent hepatitis and cholangiolar type of pathology in better prognostic subclass. On pathway analysis, liver related signatures were enriched in better prognosis subclass. In poor prognosis subclass, inflammation related pathways were enriched and KRAS mutation was more frequent. Cholangiocarcinoma cell lines which have similar gene expression pattern with better prognosis subclass were sensitive to gemcitabine. Conclusions: Two molecular subtypes of intrahepatic CCA with distinct clinical, biological and prognostic differences were identified. With clinical and pathological characteristics, molecular subtypes can be predicted and different signaling pathways of subtypes may lead to more rational targeted approaches to treatment.

The Functional Impact of HBV Integration into the Telomerase Reverse Transcriptase Promoter on Hepatocarcinogenesis

( Jeong Won Jang ),( Abdul M. Oseini ),( Ying Li ),( Catherine D. Moser ),( Karl J. Clark ),( Lewis R. Roberts ) 대한간학회 2016 춘·추계 학술대회 (KASL) Vol.2016 No.1

Aims: HBV integration into the cellular genome is frequently found in HCC and associated with genomic instability, leading to hepatocarcinogenesis. Telomerase-reverse transcriptase (TERT) promoter mutation was recently suggested to be gatekeeper in HCC.Interestingly, recent high-throughput DNA sequencing studies demonstrate that HBV integration is not random, with the highest frequency in telomere-related genes. The study aimed to investigate the molecular effect of HBV integration into the TERT promoter on hepatocarcinogenesis. Methods: Sixteen hepatoma/normal cell lines were sequenced for hTERT promoter mutation, followed by real-time RT-PCR for hTERT expression. Integration plasmid, pKT2C-HBx-TERTpmt-Luc containing HBx integrated into the various sites on hTERT promoter, was constructed and transfected into the cells. To generate genomic rearrangements associated with integration, HBx C-terminal truncation (aa27) and then hTERT hot spot mutation (-124C>T) were introduced in the HBx-hTERT fusion fragment by site-directed mutagenesis. The molecular functions of the HBx-hTERT integrants were examained by luciferase reporter and cell proliferation/apoptosis assays. Results: hTERT promoter mutation was detected in 7 (43.8%) of 16 cell lines. HCC cells with both HBV integration and hTERT promoter mutation showed a higher hTERT mRNA expression than those with either one alone or none. Although full-length HBx integration (Fl-HBx-TERT) exerts no or rather repressive effect on hTERT transcription, insertion of C-terminally truncated HBx (Tr-HBx-TERT) restored cis-activation of hTERT. The strongest effect was observed with further generation of hTERT promoter mutation (Tr-HBx-mt-TERT). Consistently, the HBx-hTERT integrants induced cell proliferation and inhibited apoptosis (caspases 3/7-dependent), with a general trend toward increasing effect from Fl-HBx-TERT to Tr-HBx-TERT and to Tr-HBx-mt-TERT. Conclusions: HBV integration can regulate the transcriptional activity of hTERT in cis and increase cell proliferation through anti-apoptotic actions, with a stronger oncogenic effect when HBx-truncation and hTERT promoter mutation are associated with integration. These findings provide insight into the implications of genomic rearrangements occurring at the viral-host junctions in integration-mediated hepatocarcinogenesis.

Free Paper Session : HCC ; A MELD-based risk score model for prediction of survival in patients with hepatocellular carcinoma (초)

( Ju Dong Yang ),( W. Ray Kim ),( Kyung Woo Park ),( Bo Hyun Kim ),( Schuyler O. Sanderson ),( Joseph J. Larson ),( Rachel A. Pedersen ),( Terry M. Therneau ),( Gregory J. Gores ),( Lewis R. Roberts ) 대한간학회 2011 Clinical and Molecular Hepatology(대한간학회지) Vol.17 No.3(S)

Persistence of intrahepatic hepatitis B virus DNA integration in patients developing hepatocellular carcinoma after hepatitis B surface antigen seroclearance

( Jeong Won Jang ),( Jin Seoub Kim ),( Hye Seon Kim ),( Kwon Yong Tak ),( Heechul Nam ),( Pil Soo Sung ),( Si Hyun Bae ),( Jong Young Choi ),( Seung Kew Yoon ),( Lewis R. Roberts ) 대한간학회 2021 Clinical and Molecular Hepatology(대한간학회지) Vol.27 No.1

Background/Aims: The role of hepatitis B virus (HBV) integration into the host genome in hepatocarcinogenesis following hepatitis B surface antigen (HBsAg) seroclearance remains unknown. Our study aimed to investigate and characterize HBV integration events in chronic hepatitis B (CHB) patients who developed hepatocellular carcinoma (HCC) after HBsAg seroclearance. Methods: Using probe-based HBV capturing followed by next-generation sequencing technology, HBV integration was examined in 10 samples (seven tumors and three non-tumor tissues) from seven chronic carriers who developed HCC after HBsAg loss. Genomic locations and patterns of HBV integration were investigated. Results: HBV integration was observed in six patients (85.7%) and eight (80.0%) of 10 tested samples. HBV integration breakpoints were detected in all of the non-tumor (3/3, 100%) and five of the seven (71.4%) tumor samples, with an average number of breakpoints of 4.00 and 2.43, respectively. Despite the lower total number of tumoral integration breakpoints, HBV integration sites in the tumors were more enriched within the genic area. In contrast, non-tumor tissues more often showed intergenic integration. Regarding functions of the affected genes, tumoral genes with HBV integration were mostly associated with carcinogenesis. At enrollment, patients who did not remain under regular HCC surveillance after HBsAg seroclearance had a large HCC, while those on regular surveillance had a small HCC. Conclusions: The biological functions of HBV integration are almost comparable between HBsAg-positive and HBsAg-serocleared HCCs, with continuing pro-oncogenic effects of HBV integration. Thus, ongoing HCC surveillance and clinical management should continue even after HBsAg seroclearance in patients with CHB. (Clin Mol Hepatol 2021;27:207-218)

Genes Associated with Recurrence of Hepatocellular Carcinoma: Integrated Analysis by Gene Expression and Methylation Profiling

Yang, Ju Dong,Seol, So-Young,Leem, Sun-Hee,Kim, Yong Hoon,Sun, Zhifu,Lee, Ju-Seog,Thorgeirsson, Snorri S.,Chu, In-Sun,Roberts, Lewis R.,Kang, Koo Jeong The Korean Academy of Medical Sciences 2011 JOURNAL OF KOREAN MEDICAL SCIENCE Vol.26 No.11

<P>Gene expression is suppressed by DNA methylation. The goal of this study was to identify genes whose CpG site methylation and mRNA expression are associated with recurrence after surgical resection for hepatocellular carcinoma (HCC). Sixty-two HCCs were examined by both whole genome DNA methylation and transcriptome analysis. The Cox model was used to select genes associated with recurrence. A validation was performed in an independent cohort of 66 HCC patients. Among fifty-nine common genes, increased CpG site methylation and decreased mRNA expression were associated with recurrence for 12 genes (Group A), whereas decreased CpG site methylation and increased mRNA expression were associated with recurrence for 25 genes (Group B). The remaining 22 genes were defined as Group C. Complement factor H (<I>CFH</I>) and myosin VIIA and Rab interacting protein (<I>MYRIP</I>) in Group A; proline/serine-rich coiled-coil 1 (<I>PSRC1</I>), meiotic recombination 11 homolog A (<I>MRE11A</I>), and myosin IE (<I>MYO1E</I>) in Group B; and autophagy-related protein LC3 A (<I>MAP1LC3A</I>), and NADH dehydrogenase 1 alpha subcomplex assembly factor 1 (<I>NDUFAF1</I>) in Group C were validated. In conclusion, potential tumor suppressor (<I>CFH</I>, <I>MYRIP</I>) and oncogenes (<I>PSRC1</I>, <I>MRE11A</I>, <I>MYO1E</I>) in HCC are reported. The regulation of individual genes by methylation in hepatocarcinogenesis needs to be validated.</P>

Integrative Genomic And Clinical Characteristics According to The Serum Tumor Markers in Hepatocellular Carcinoma

Keun Soo Ahn,Koo Jeong Kang,Yong Hoon Kim,Tae-Seok Kim,Sang-Jae Park,Seung Hoon Kim,Joong-Won Park,Lewis R Roberts 대한외과학회 2020 대한외과학회 학술대회 초록집 Vol.2020 No.11