Yoshito Ito

Genome-wide association of IL28B with response to pegylated interferon-α and ribavirin therapy for chronic hepatitis C

The recommended treatment for patients with chronic hepatitis C, pegylated interferon-α (PEG-IFN-α) plus ribavirin (RBV), does not provide sustained virologic response (SVR) in all patients. We report a genome-wide association study (GWAS) to null virological response (NVR) in the treatment of patients with hepatitis C virus (HCV) genotype 1 within a Japanese population. We found two SNPs near the gene IL28B on chromosome 19 to be strongly associated with NVR (rs12980275, P = 1.93 × 10−13, and rs8099917, 3.11 × 10−15). We replicated these associations in an independent cohort (combined P values, 2.84 × 10−27 (OR = 17.7; 95% CI = 10.0–31.3) and 2.68 × 10−32 (OR = 27.1; 95% CI = 14.6–50.3), respectively). Compared to NVR, these SNPs were also associated with SVR (rs12980275, P = 3.99 × 10−24, and rs8099917, P = 1.11 × 10−27). In further fine mapping of the region, seven SNPs (rs8105790, rs11881222, rs8103142, rs28416813, rs4803219, rs8099917 and rs7248668) located in the IL28B region showed the most significant associations (P = 5.52 × 10−28–2.68 × 10−32; OR = 22.3–27.1). Real-time quantitative PCR assays in peripheral blood mononuclear cells showed lower IL28B expression levels in individuals carrying the minor alleles (P = 0.015).

Difference in serum complement component C4a levels between hepatitis C virus carriers with persistently normal alanine aminotransferase levels or chronic hepatitis C

Certain hepatitis C virus (HCV) carriers exhibit persistently normal alanine aminotransferase (ALT) levels (PNALT) (≤30 IU/l) accompanied by normal platelet counts (≥15×104/μl); these individuals show milder disease activity and slower progression to cirrhosis. This study aimed to elucidate the characteristics of HCV carriers with PNALT using serum proteomics. The first group of subjects, who underwent clinical evaluation in the hospital, consisted of 19 HCV carriers with PNALT (PNALT-1) and 20 chronic hepatitis C (CHC-1) patients. The second group of subjects was part of a cohort study on the natural history of liver disease, and included 37 PNALT (PNALT-2) and 30 CHC (CHC-2) patients. Affinity bead-purified serum protein was subjected to matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis. Serum proteomics showed that 6 protein peaks with mass-to-charge ratios ranging from 1,000 to 3,000 differed significantly between the PNALT-1 and CHC-1 groups. Among these peaks, a 1738-m/z peak protein was identified as a fragment of complement component 4 (C4) and correlated significantly with serum C4a concentrations as determined by enzyme immunoassay. Serum C4a levels were also significantly higher in the PNALT-2 group compared to the CHC-2 group and healthy volunteers. Furthermore, in the PNALT-2 group, serum C4a levels negatively correlated with transaminase levels, but not with other biochemical tests, HCV core antigen levels, peripheral blood cell counts or serum hepatic fibrosis markers. This study indicates that host factors such as C4a not only differ between HCV carriers with PNALT and CHC, but that proteomic approaches could also contribute to the elucidation of factors in PNALT as more differences are discovered.

Novel single nucleotide polymorphisms of the cytokeratin 19 pseudogene are associated with primary biliary cirrhosis

Primary biliary cirrhosis (PBC) is characterized by chronic inflammation and destruction of intra-hepatic bile ducts. However, the pathogenesis of PBC has not been fully delineated. We examined whether patients with PBC harbor genomic mutations of the cytokeratin 19 (CK19) gene since that gene is specifically expressed in biliary epithelial cells. Thirty-six patients with PBC, 26 patients with other liver diseases, and 36 healthy volunteers were enrolled in this study, but there were no significant differences in the genomic sequence of the CK19 gene between those groups. On the other hand, novel single nucleotide polymorphisms (SNPs) of the CK19 pseudogene, C341T, T524G and A754G, were frequently detected in PBC patients. These results suggest that those novel SNPs of the CK19 pseudogene may be associated with PBC and may prove useful for predicting susceptibility to PBC.

Abstract 2539: MiR-96-5p functions as an oncogenic miRNA by inhibiting apoptosis in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the most aggressive cancers with high mortality worldwide. MicroRNAs (miRNAs) are small non-coding RNAs that have been used as cancer-related biomarkers and expected to be therapeutic agents. We performed genome-wide miRNA expression profiling of paired HCC tumors and non-tumorous liver tissues from patients with primary HCCs using the miRNA microarray (Agilent). We found that miR-96-5p was most significantly up-regulated in HCC tumors compared to non-tumor tissues. Although miR-96-5p is suggested to be an oncogenic miRNA, the function of miR-96-5p remains largely unknown. We identified the caspase-9 gene (CASP9) as a novel target of miR-96-5p, in addition to the forkhead box O1 gene (FOXO1) which is the known target of it. Caspase-9 protein is thought to play a central role in apoptosis and to be a tumor suppressor. Overexpression of miR-96-5p decreased caspase-9 protein expression and resulted in resistance to apoptosis induced by doxorubicin and UV in HCC cells. Our results suggested that miR-96-5p functions as an oncogenic miRNA by inhibiting apoptosis through decreasing caspase-9 expression in HCC. Citation Format: Naoto Iwai, Kohichiroh Yasui, Akira Tomie, Kei Teasaki, Tomoko Kitaichi, Osamu Dohi, Yasuyuki Gen, Yoshito Ito. MiR-96-5p functions as an oncogenic miRNA by inhibiting apoptosis in hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2539. doi:10.1158/1538-7445.AM2017-2539

Abstract 1326: Elevated levels of plasma VEGF associated with the attenuation of whole blood IFN-γ production and QOL impairment in patients with advanced gastric cancer

Introduction: Gastric cancer is one of the leading types of cancer and the second leading cause of cancer death in the world. Recently, the foci of treatment shift towards not only survival benefit but also better quality of life (QOL) in patients with advanced gastric cancer. Therefore, the development of an auxiliary tool to assess QOL is likely to be required. On the other hand, immune checkpoint blockade by anti-CTLA-4 antibody and anti-PD-1 antibody led to clinical breakthrough for the treatment of patients with advanced cancer. Early phase clinical trial of pembrolizumab in patients with advanced gastric cancer has already been conducted. Pretreatment circulating VEGF have been recently shown to associate with clinical outcome in patients treated with ipilimumab. Certain circulating cytokine levels have been shown to reflect tumor progression and have a prognostic value. Although, it remains obscure whether circulating cytokine levels affect QOL and immune function. In this study, we examined the correlation of circulating cytokine with QOL as well as parameters of immune function such as whole blood cytokine production and the number of peripheral Tregs in patients with advanced gastric cancer. Methods: Subjects comprised 31 patients with unresectable or recurrent gastric cancer. We evaluated plasma cytokine levels and whole blood cytokine production after phytohemagglutinin (PHA) stimulation using the bioplex array system. We also assessed the number of peripheral Tregs by flow cytometry. Health-related QOL was assessed using the Quality of Life Questionnaire (EORTC QLQ-C30). Results: Significant negative correlation was found between plasma VEGF levels and global health status scores (p = 0.0103) as well as physical functioning scale scores (p = 0.0006) and cognitive functioning scale scores (p = 0.0191). Some symptom scale scores (e.g. fatigue, appetite loss) were correlated with plasma VEGF levels. There was a negative correlation between plasma VEGF levels and whole blood IFN-γ production (p = 0.0002). Significant negative relationship existed between the number of peripheral Tregs and plasma IL-6 levels (p Conclusion: These data indicate that the evaluation of plasma VEGF level is likely to be useful to assess QOL such as global health status, physical functioning and cognitive functioning and elevated levels of plasma VEGF might associated with the attenuation of immune function in patients with advanced gastric cancer. Consequently, clinical utility of anti-VEGF therapy combined with immunotherapy is required to investigate in larger clinical studies in the future. Citation Format: Naoyuki Sakamoto, Takeshi Ishikawa, Tetsuya Okayama, Tomoyo Yasuda, Toshifumi Doi, Hideyuki Konishi, Satoshi Kokura, Mari Tanigawa, Kazuko Uno, Yuji Naito, Yoshito Ito, Toshikazu Yoshikawa. Elevated levels of plasma VEGF associated with the attenuation of whole blood IFN-γ production and QOL impairment in patients with advanced gastric cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1326. doi:10.1158/1538-7445.AM2015-1326

Abstract 3525: SOX2 promotes tumor growth via activation of the PI3K/Akt/mTORC1 signaling pathway in esophageal squamous cell carcinoma

Our previous study revealed that SOX2, a master regulator during embryogenesis, is an amplification target of 3q26.3 in esophageal squamous cell carcinoma (ESCC) and that SOX2 promotes ESCC cell proliferation. To identify the mechanisms by which SOX2 promotes proliferation of ESCC cells, we assayed multiple signaling pathways activated by SOX2 using a phosphoprotein array. We determined that SOX2 activated the AKT/mammalian target of rapamycin complex 1 (mTORC1) signaling pathway. Immunoblotting confirmed that SOX2 elevated levels of p-AKT and levels of p-p70S6K and p-4E-BP1 which are direct targets of mTORC1. Effects of SOX2 knockdown, including reduced levels of p-AKT and decreased ESCC cell viability, were reversed with constitutive activation of AKT with PTEN knockdown. SOX2 also promoted in vivo tumor growth of ESCC with AKT/mTORC1 activation in mouse xenografts. LY294002, a PI3K inhibitor, suppressed the ability of SOX2 to enhance tumor growth of ESCC by reducing cell proliferation. Furthermore, tissue microarray analysis showed a positive correlation between expression levels of SOX2 and p-AKT in 61 primary ESCC tumors. Our results suggest that SOX2 promotes ESCC tumor growth via activation of the PI3K/AKT / mTORC1 signaling pathway, which enhances cell proliferation. Citation Format: Yasuyuki Gen, Kohichiroh Yasui, Tomoko Kitaichi, Akira Tomie, Yoshito Ito. SOX2 promotes tumor growth via activation of the PI3K/Akt/mTORC1 signaling pathway in esophageal squamous cell carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3525. doi:10.1158/1538-7445.AM2014-3525

Development of a single expandable PEIT needle and its clinical application

The current mainstream percutaneous local treatment for HCC is RFA with supplementary PEIT. PEIT is sometimes employed for HCC in high-risk locations, such as HCC close to the heart. However, even with PEIT it is markedly difficult to treat HCC located at the back ofblood vessels deep in the liver. We developed a single expandable PEIT needle to treat difficult-to-puncture HCC. The tip of an inner needle can be expandable vertically to the long axis. We used this technique to successfully treat a case of HCC in S1 at the back of portal vein at the liver hilum. The needle is equipped with features absent in other devices and is expected to become a new option for percutaneous local therapy of HCC.

Endoscopically Treated Angiodysplasia of the Ileum

Abstract: Angiodysplasia has been recognized as one of the main causes of gastrointestinal bleeding, but few such lesions have been found, or treated by an endoscopic procedure, in the ileum. We endoscopically identified angiodysplasia in the ileum of an 85‐year‐old female who had tarry stools. Because she had severe aortic stenosis, we treated the lesion endoscopically by combining polidocanol injection and electrocoagulation with hot biopsy forceps. This method was considered to be safe and efficient, especially for lesions in the thin walled intestine.