Papers - KANAI Yae

Method for determining prognosis of endometrial cancer

Y Kanai, E Arai, D Aoki, N Susumu, W Yamagami, T Makabe

US Patent App. 17/298,863  2022

Quantification of DNA methylation for carcinogenic risk estimation in patients with non-alcoholic steatohepatitis

J Kuramoto, E Arai, M Fujimoto, Y Tian, Y Yamada, T Yotani, S Makiuchi, ...

Cancer Research 82 (12_Supplement), 5703-5703 (Clinical Epigenetics)  14 ( 1 ) 168 - 168 2022

ISSN  18687075

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Background: In recent years, non-alcoholic steatohepatitis (NASH) has become the main cause of hepatocellular carcinoma (HCC). As a means of improving the treatment of NASH-related HCCs based on early detection, this study investigated the feasibility of carcinogenic risk estimation in patients with NASH. Results: Normal liver tissue (NLT), non-cancerous liver tissue showing histological findings compatible with non-alcoholic fatty liver from patients without HCC (NAFL-O), non-cancerous liver tissue showing NASH from patients without HCC (NASH-O), non-cancerous liver tissue showing non-alcoholic fatty liver from patients with HCC (NAFL-W), non-cancerous liver tissue showing NASH from patients with HCC (NASH-W) and NASH-related HCC were analyzed. An initial cohort of 171 tissue samples and a validation cohort of 55 tissue samples were used. Genome-wide DNA methylation screening using the Infinium HumanMethylation450 BeadChip and DNA methylation quantification using high-performance liquid chromatography (HPLC) with a newly developed anion-exchange column were performed. Based on the Infinium assay, 4050 CpG sites showed alterations of DNA methylation in NASH-W samples relative to NLT samples. Such alterations at the precancerous NASH stage were inherited by or strengthened in HCC samples. Receiver operating characteristic curve analysis identified 415 CpG sites discriminating NASH-W from NLT samples with area under the curve values of more than 0.95. Among them, we focused on 21 CpG sites showing more than 85% specificity, even for discrimination of NASH-W from NASH-O samples. The DNA methylation status of these 21 CpG sites was able to predict the coincidence of HCC independently from histopathological findings such as ballooning and fibrosis stage. The methylation status of 5 candidate marker CpG sites was assessed using a HPLC-based system, and for 3 of them sufficient sensitivity and specificity were successfully validated in the validation cohort. By combining these 3 CpG sites including the ZC3H3 gene, NAFL-W and NASH-W samples from which HCCs had already arisen were confirmed to show carcinogenic risk with 95% sensitivity in the validation cohort. Conclusions: After a further prospective validation study using a larger cohort, carcinogenic risk estimation in liver biopsy specimens of patients with NASH may become clinically applicable using this HPLC-based system for quantification of DNA methylation.

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Cancer-specific glycosylation regulated by epigenomic alteration in renal cell carcinoma

Y Kitazume, E Arai, A Matsuda, K Ohara, AM Maeshima, T Yoshida, ...

Cancer Research 82 (12_Supplement), 5702-5702 82 ( 12 )  2022

ISSN  0008-5472

Potential DNA methylation biomarkers for the detection of clear cell renal cell carcinoma identified by a whole blood-based epigenome-wide association study

H Ohmomo, S Komaki, Y Sutoh, T Hachiya, K Ono, E Arai, H Fujimoto, ...

Epigenetics Communications 2 (1), 1-11  2022

Accepted

The oligodendroglial histological features are not independently predictive of patient prognosis in lower-grade gliomas

ES Pareira, M Shibuya, K Ohara, Y Nakagawa, T Kanazawa, ...

Brain tumor pathology, 1-9 (Brain Tumor Pathology)  39 ( 2 ) 79 - 87 2022

Accepted,  ISSN  14337398

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The relevance of oligodendroglial histological features to patient prognoses is controversial. 93 LrGGs resected for about 2 decades were re-assessed based on WHO2007 with special interest to pure oligodendroglial diagnosis (oligodendroglioma or anaplastic oligodendroglioma) and presence of CFO features. Those histological features, patients OS, and tumor chromosomal/genetic characteristics were correlated each other in each of the 3 IDH-1p/19q-based molecular groups. There was significant association between 1p19q status with the oligodendroglial histological diagnosis as well as presence of CFO in the entire cohort. The oligodendroglial diagnosis was associated with longer OS in IDHmut/codel group; however, this association was not significant in the multivariate analyses. In IDHmut/noncodel and IDH-wildtype groups, the oligodendroglial diagnosis was not associated with patient OS. Presence of CFO was not associated with patient OS in any molecular groups. Gain of 8q was associated with the oligodendroglial diagnosis in IDHmut/noncodel group. Neither the oligodendroglial diagnosis nor CFO was predictive for the methylation status of the MGMT gene in any molecular groups. The oligodendroglial histological features are not independently predictive of either patient prognosis or chemotherapeutic response in LrGGs, leaving the possibility of marginal favorable association only in IDHmut/codel tumors.

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DNA methylation status of the SPHK1 and LTB genes underlies the clinicopathological diversity of non-alcoholic steatohepatitis-related hepatocellular carcinomas

N Tsuda, Y Tian, M Fujimoto, J Kuramoto, S Makiuchi, H Ojima, M Gotoh, ...

Journal of Cancer Research and Clinical Oncology, 1-17 (Journal of Cancer Research and Clinical Oncology)  149 ( 8 ) 5109 - 5125 2022

ISSN  01715216

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Purpose: This study was performed to identify the DNA methylation profiles underlying the clinicopathological diversity of non-alcoholic steatohepatitis (NASH)-related hepatocellular carcinomas (HCCs). Methods: Genome-wide DNA methylation analysis of 88 liver tissue samples was performed using the Infinium assay. Results: Principal component analysis revealed that distinct DNA methylation profiles differing from such profiles in normal control liver tissue had already been established in non-cancerous liver tissue showing NASH, which is considered to be a precancerous condition. Hierarchical clustering separated 26 NASH-related HCCs into Cluster I (n = 8) and Cluster II (n = 18). Such epigenetic clustering was significantly correlated with histopathological diversity, i.e. poorer tumor differentiation, tumor steatosis and development of a scirrhous HCC component. Significant differences in DNA methylation levels between the two clusters were accumulated in molecular pathways participating in cell adhesion and cytoskeletal remodeling, as well as cell proliferation and apoptosis. Among tumor-related genes characterizing Clusters I and II, differences in the levels of DNA methylation and mRNA expression for the SPHK1, INHBA, LTB and PDE3B genes were correlated with poorer tumor differentiation. 5-Aza-2′-deoxycytidine treatment of HCC cells revealed epigenetic regulation of the SPHK1 and LTB genes. Knockdown experiments showed that SPHK1 promotes cell proliferation, represses apoptosis and enhances migration, whereas LTB enhances migration of HCC cells. DNA hypomethylation resulting in increased expression of SPHK1 and LTB in poorly differentiated HCCs may underlie the aggressive phenotype of such HCCs. Conclusion: These data indicate that DNA methylation profiles may determine the clinicopathological heterogeneity of NASH-related HCCs via alterations of tumor-related gene expression.

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The Japanese Society of Pathology Practical Guidelines on the handling of pathological tissue samples for cancer genomic medicine.

Y Hatanaka, T Kuwata, E Morii, Y Kanai, H Ichikawa, T Kubo, ...

Pathology International (Pathology International)  71 ( 11 ) 725 - 740 2021.10

Joint Work, Accepted,  ISSN  1320-5463

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Clinical cancer genomic testing based on next-generation sequencing can help select genotype-matched therapy and provide diagnostic and prognostic information. Pathological tissue from malignant tumors obtained during routine practice are frequently used for genomic testing. This article is aimed to standardize the proper handling of pathological specimens in practice for genomic medicine based on the findings established in “Guidelines on the handling of pathological tissue samples for genomic medicine (in Japanese)” published by The Japanese Society of Pathology (JSP) in 2018. The two-part practical guidelines are based on empirical data analyses; Part 1 describes the standard preanalytic operating procedures for tissue collection, processing, and storage of formalin-fixed paraffin-embedded (FFPE) samples, while Part 2 describes the assessment and selection of FFPE samples appropriate for genomic testing, typically conducted by a pathologist. The guidelines recommend that FFPE sample blocks be used within 3 years from preparation, and the tumor content should be ≥30% (minimum 20%). The empirical data were obtained from clinical studies performed by the JSP in collaboration with leading Japanese cancer genome research projects. The Japanese Ministry of Health, Labour, and Welfare (MHLW) recommended to comply with the JSP practical guidelines in implementing cancer genomic testing under the national health insurance system in over 200 MHLW-designated core and cooperative cancer genome medicine hospitals in Japan.

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Molecular classification of gastric cancer predicts survival in patients undergoing radical gastrectomy based on project HOPE

K Furukawa, K Hatakeyama, M Terashima, T Nagashima, K Urakami, ...

Gastric Cancer, 1-11 (Gastric Cancer)  25 ( 1 ) 138 - 148 2021.09

Joint Work, Accepted,  ISSN  1436-3291

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Background: Gastric cancer (GC) has been classified based on molecular profiling like The Cancer Genome Atlas (TCGA) and Asian Cancer Research Group (ACRG), and attempts have been made to establish therapeutic strategies based on these classifications. However, it is difficult to predict the survival according to these classifications especially in radically resected patients. We aimed to establish a new molecular classification of GC which predicts the survival in patients undergoing radical gastrectomy. Methods: The present study included 499 Japanese patients with advanced GC undergoing radical (R0/R1) gastrectomy. Whole-exome sequencing, panel sequencing, and gene expression profiling were conducted (High-tech Omics-based Patient Evaluation [Project HOPE]). We classified patients according to TCGA and ACRG subtypes, and evaluated the clinicopathologic features and survival. Then, we attempted to classify patients according to their molecular profiles associated with biological features and survival (HOPE classification). Results: TCGA and ACRG classifications failed to predict the survival. In HOPE classification, hypermutated (HMT) tumors were selected first as a distinctive feature, and T-cell-inflamed expression signature-high (TCI) tumors were then extracted. Finally, the remaining tumors were divided by the epithelial-mesenchymal transition (EMT) expression signature. HOPE classification significantly predicted the disease-specific and overall survival (p < 0.001 and 0.020, respectively). HMT + TCI showed the best survival, while EMT-high showed the worst survival. The HOPE classification was successfully validated in the TCGA cohort. Conclusions: We established a new molecular classification of gastric cancer that predicts the survival in patients undergoing radical surgery.

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【病理医が知っておくべき法令や指針】Q&A 剖検

若狹 朋子, 木ノ元 直樹, 種井 善一, 金井 弥栄, 柴原 純二

病理と臨床 ((株)文光堂)  39 ( 9 ) 904 - 908 2021.09

ISSN  0287-3745

Brain Pathology of 18 Supercentenarians Aged 110 And Over

Takao Masaki, Mihara Ban, Arai Yasumichi, Murakami Tomoyuki, Nose Soichiro, Hamaya Kazuo, Nishida Katsuya, Futamura Naonobu, Nakayama Yuji, Mizutani Masashi, Sano Terunori, Kanai Yae, Mimura Masaru, Okano Hideyuki, Hirose Nobuyoshi

JOURNAL OF NEUROPATHOLOGY AND EXPERIMENTAL NEUROLOGY (OXFORD UNIV PRESS INC)  80 ( 6 ) 594 - 594 2021.06

Joint Work,  ISSN  0022-3069

Demarcated redness associated with increased vascular density/size: a useful marker of flat-type dysplasia in patients with ulcerative colitis.

Ikebata A, Shimoda M, Okabayashi K, Uraoka T, Maehata T, Sugimoto S, Mutaguchi M, Naganuma M, Kameyama K, Yahagi N, Kanai T, Kitagawa Y, Kanai Y, Iwao Y

Endoscopy international open 9 ( 4 ) E552 - E561 2021.04

Research paper (scientific journal), Joint Work, Accepted,  ISSN  2364-3722

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Background and study aims  Recent advances in endoscopic equipment and diagnostic techniques have improved the detection of dysplasia in the inflamed mucosa of patients with ulcerative colitis (UC). However, it remains difficult to endoscopically identify flat-type dysplasia which has been formerly recognized as invisible dysplasia. Patients and methods  In this retrospective, single-center study, we endoscopically identified 10 cases of flat-type-predominant dysplasia by targeted biopsy among 38 intramucosal dysplasia lesions from patients with UC who underwent surgical or endoscopic resection from 2007 to 2017. Their endoscopic and histological features were examined, including color changes, intramucosal vascular density/size, and vascular endothelial growth factor (VEGF) expression. Results  All flat-type-predominant dysplasias were endoscopically recognized as demarcated red-colored areas and histologically diagnosed as low- (LGDs) or high-grade dysplasias (HGDs). Immunohistochemical examination using resected specimens revealed that flat-type dysplasia was characterized by significantly increased CD34-positive vascular density (LGDs, 1.7-fold, P < 0.01; HGDs, 2.2-fold, P  < 0.01) and size (LGDs, 1.03-fold, P  < 0.01; HGDs, 1.11-fold, P  < 0.01) in the mucosa, compared to adjacent non-neoplastic areas. Increased numbers of vessels were observed at the base of the mucosa in LGDs, whereas HGDs contained increased/enlarged vessels throughout the mucosa. Moreover, VEGF expression was elevated in all dysplastic epithelia. Conclusions  Demarcated red-colored areas, histologically characterized by an increased vascular density/size in the mucosa, are an endoscopic sign of formerly invisible flat-type dysplasia in patients with UC and should be considered for targeted biopsy. Prospective studies focusing on the mucosal color change for their early detection would be desirable in the future.

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Alterations of protein glycosylation and DNA methylation in urothelial carcinoma

Fujimoto Mao, Arai Eri, Matsuda Atsushi, Maeshima Akiko, Fujimoto Hiroyuki, Kuno Atsushi, Yoshida Teruhiko, Kanai Yae

CANCER SCIENCE 112   670 - 670 2021.02

Joint Work,  ISSN  1347-9032

Infiltration of TAMs is involved in tumor PD-L1 expression in early lung adenocarcinoma

Shima Toshiyuki, Shimoda Masayuki, Shigenobu Takao, Otsuka Takashi, Nishimura Tomoyasu, Emoto Katsura, Hayashi Yuichirou, Iwasaki Tatusrou, Abe Takayuki, Asamura Hisao, Kanai Yae

CANCER SCIENCE 112   397 - 397 2021.02

Joint Work, Accepted,  ISSN  1347-9032

Epigenomic alterations during developmental stages of non-inflammatory hepatocellular carcinoma of unknown etiology

Makiuchi Satomi, Arai Eri, Kuramoto Junko, Tian Ying, Tsuda Noboru, Ojima Hidenori, Takahashi Yoriko, Hiraoka Nobuyoshi, Yoshida Teruhiko, Kanai Yae

CANCER SCIENCE 112   674 - 674 2021.02

Joint Work,  ISSN  1347-9032

Epigenomic alterations and histological heterogeneity of non-alcoholic steatohepatitis-related hepatocellular carcinomas

Tsuda Noboru, Arai Eri, Kuramoto Junko, Tian Ying, Makiuchi Satomi, Ojima Hidenori, Takahashi Yoriko, Hiraoka Nobuyoshi, Yoshida Teruhiko, Kanai Yae

CANCER SCIENCE 112   671 - 671 2021.02

Joint Work,  ISSN  1347-9032

腸に限局した原発性AL型アミロイドーシスの4例

中島 真, 下田 将之, 岩男 泰, 川井田 みほ, 亀山 香織, 内木 宏延, 金井 弥栄

診断病理 ((一社)日本病理学会)  38 ( 1 ) 77 - 81 2021.01

Joint Work, Accepted,  ISSN  1345-6431

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消化管に限局した原発性アミロイドーシスは稀な疾患である。今回4例の腸限局性アミロイドーシスを経験した。いずれの症例も内視鏡的に大腸を中心とした多発性黄白色調粘膜下腫瘍様隆起として観察され、組織学的にはAL(λ)型アミロイド沈着を認めた。経過観察可能だった3例はいずれも生命予後良好で、進行は緩徐であった。また、非隆起部からアミロイド沈着初期像を検出するとともに、アミロイド沈着部にはCD34陽性細胞の軽度集簇が観察され、これらの所見はアミロイド沈着機序理解の新たな糸口となる可能性が示唆された。(著者抄録)

Multi-omic profiling of peritoneal metastases in gastric cancer identifies molecular subtypes and therapeutic vulnerabilities

Y Tanaka, F Chiwaki, S Kojima, M Kawazu, M Komatsu, T Ueno, S Inoue, ...

Nature Cancer, 1-16 (Nature Cancer)  2 ( 9 ) 962 - 977 2021

Joint Work

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Peritoneal metastasis, a hallmark of incurable advanced gastric cancer (GC), presently has no curative therapy and its molecular features have not been examined extensively. Here we present a comprehensive multi-omic analysis of malignant ascitic fluid samples and their corresponding tumor cell lines from 98 patients, including whole-genome sequencing, RNA sequencing, DNA methylation and enhancer landscape. We identify a higher frequency of receptor tyrosine kinase and mitogen-activated protein kinase pathway alterations compared to primary GC; moreover, approximately half of the gene alterations are potentially treatable with targeted therapy. Our analyses also stratify ascites-disseminated GC into two distinct molecular subtypes: one displaying active super enhancers (SEs) at the ELF3, KLF5 and EHF loci, and a second subtype bearing transforming growth factor-β (TGF-β) pathway activation through SMAD3 SE activation and high expression of transcriptional enhancer factor TEF-1 (TEAD1). In the TGF-β subtype, inhibition of the TEAD pathway circumvents therapy resistance, suggesting a potential molecular-guided therapeutic strategy for this subtype of intractable GC.

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Clinicopathological impacts of DNA methylation alterations on pancreatic ductal adenocarcinoma: prediction of early recurrence based on genome-wide DNA methylation profiling

Endo Y, Fujimoto M, Ito N, Takahashi Y, Kitago M, Gotoh M, Hiraoka N, Yoshida T, Kitagawa Y, Kanai Y, Arai E

Journal of Cancer Research and Clinical Oncology (Journal of Cancer Research and Clinical Oncology)  147 ( 5 ) 1341 - 1354 2021

Joint Work, Accepted,  ISSN  01715216

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© 2021, The Author(s). Purpose: The present study was conducted to clarify the clinicopathological impacts of DNA methylation alterations on pancreatic ductal adenocarcinoma (PDAC). Methods: Genome-wide DNA methylation screening was performed using the Infinium HumanMethylation450 BeadChip, and DNA methylation quantification was verified using pyrosequencing. We analyzed fresh-frozen tissues from an initial cohort (17 samples of normal control pancreatic tissue [C] from 17 patients without PDAC, and 34 samples of non-cancerous pancreatic tissue [N] and 82 samples of cancerous tissue [T] both obtained from 82 PDAC patients) and formalin-fixed paraffin-embedded T samples from 34 patients in a validation cohort. Results: The DNA methylation profiles of N samples tended to differ from those of C samples, and 91,907 probes showed significant differences in DNA methylation levels between C and T samples. Epigenetic clustering of T samples was significantly correlated with a larger tumor diameter and early recurrence (ER), defined as relapse within 6 months after surgery. Three marker CpG sites, applicable to formalin-fixed paraffin-embedded surgically resected materials regardless of their tumor cell content, were identified for prediction of ER. The sensitivity and specificity for detection of patients belonging to the ER group using a panel combining these three marker CpG sites, including a CpG site in the CDK14 gene, were 81.8% and 71.7% and 88.9% and 70.4% in the initial and validation cohorts, respectively. Conclusion: These findings indicate that DNA methylation alterations may have a clinicopathological impact on PDAC. Application of our criteria will ultimately allow prediction of ER after surgery to improve the outcome of PDAC patients.

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Evaluation of clinical formalin‐fixed paraffin‐embedded tissue quality for targeted‐bisulfite sequencing

H Ohmomo, S Komaki, K Ono, Y Sutoh, T Hachiya, E Arai, H Fujimoto, ...

Pathology international 71 ( 2 ) 135 - 140 2021

Accepted

DNA hypermethylation of the ZNF132 gene participates in the clinicopathological aggressiveness of ‘pan-negative’-type lung adenocarcinomas

K Hamada, Y Tian, M Fujimoto, Y Takahashi, T Kohno, K Tsuta, ...

Carcinogenesis 42 (2), 169-179  2021

Accepted