Academic literature on the topic 'Sawada, Yasuhiko'

Abstract Objective: To establish endometrial immortalized cell lines using the Sendai virus and analyze their characteristics. Methods: The study included patients with benign ovarian tumors and endometrioid carcinoma who had regular menstrual cycles. Endometrial cells were collected and separated into epithelial and non-epithelial cells using flowcytometry (FCM) (CD326 (EpCAM) antibody). Furthermore, the isolated cells were infected with Sendai virus carrying three immortalization genes (Bmi-1, hTERT, and SV40T) to create immortalized cell lines. The cells were characterized as epithelial cells by immunocytochemistry (ICC), and receptor expression was assessed by reverse transcription PCR (RT-PCR) and ICC. Non-epithelial cells from the patients with endometrial carcinoma were evaluated using ICC. Results: The immortalized cell lines were capable of more than 20 passages. Immortalized endometrial epithelial cells tested positive for anti-Keratin antibodies and negative for anti-Vimentin antibodies by ICC. Additionally, FCM detected these cells as positive for anti-EpCAM antibody, confirming that they were epithelial cells. RT-PCR confirmed mRNA expression of estrogen α-receptor and progesterone receptor after 20 passages and ICC confirmed protein expression of them after 10 passages, establishing an endometrial epithelial immortalized cell line. Non-epithelial cells isolated from patients with endometrial carcinoma tested positive for anti-α-smooth muscle actin and anti-Vimentin antibodies by ICC, indicating their characterization as cancer-associated fibroblasts. Conclusion: We successfully established an immortalized cell line of endometrial epithelial cells using Sendai virus and generated cancer-associated fibroblasts. Citation Format: Kohei Hikino, Hiroaki Komatsu, Genki Hichiwa, Kanako Kazuki, Yasuhiro Kazuki, Hiroyuki Kugoh, Masayo Okawa, Yuki Ida, Masayo Hosokawa, Mayumi Sawada, Akiko Kudoh, Shinya Sato, Fuminori Taniguchi. Establishment of immortalized endometrial cell lines using Sendai virus [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2791.

Abstract [Introduction] Proliferative lesions in the breast have been implicated in the development of breast cancer. Previous studies showed that some proliferative lesions and adjacent breast cancers shared common genetic alterations, suggesting that these originated from the same ancestral cell. However, the clonal structure of normal epithelia and their clonal history during evolution to cancer are poorly understood. In this study, we analyzed genetic profiles of normal epithelia and proliferative lesions in the cancer-borne breast to illustrate the clonal evolution of cancer from a normal epithelial cell. [Methods] Single cell-derived organoids (n=47) were established from breast milk of 4 healthy women aged 22–36 and normal breast tissue of 15 breast cancer patients aged 29–83 to evaluate somatic mutation rate in normal epithelial cells. Multiple normal lobules and proliferative lesions together with cancer lesions were collected using laser-capture micro-dissection (LCM) from fresh frozen (n=3) or formalin-fixed paraffin-embedded (n=5) surgical specimens in 9 premenopausal breast cancer patients. Somatic mutations and copy number alterations were evaluated using whole-genome sequencing. [Results] The mutation profile of single cell-derived organoids suggests that somatic mutations accumulate in normal mammary epithelial cells at a constant rate of 19.4/genome/year before menopause, and the mutation rate decreases to 6.9/genome/year after menopause. Parity was negatively associated with mutation number (-49.3 per life birth). In total, we analyzed 143 LCM samples, including those from 72 normal lobules, 43 proliferative lesions, and 19 non-invasive and 9 invasive cancer samples. Five cases showed a large expansion of proliferative lesions sharing a substantial number of somatic mutations with cancer. These lesions expanded over a distance of 35-90 mm, sharing tens to hundreds of mutations including those in breast cancer-related driver genes, such as PIK3CA, AKT1, GATA3, CBFB and PTEN, while harboring private mutations or copy number alterations of their own. Of interest, the cancers in 4 out of these 5 cases was luminal-A type invasive ducal carcinoma or ER-positive HER2-negative ductal carcinoma in situ, and characterized in common by the presence of der(1;16), concurrent whole-arm 1q gain and 16q loss, in both cancer and proliferative lesions. Phylogenetic analysis adapted with the mutation rate in normal cells predicted that der(1;16) had been acquired between puberty and early 20’s, and the common ancestors of non-cancerous and cancerous lesions emerged by early 30’s, >10 years earlier than at the time of cancer diagnosis. By contrast, analysis of non-cancerous lobules unrelated to cancer showed that der(1;16)-negative non-cancer clones that had emerged after puberty stayed within a single lobule or spatially confined to adjacent lobules and rarely expanded to a large area as observed for those carrying der(1;16), even if the clones had acquired mutations in driver genes such as PIK3CA and PIK3R1, which highlighted the role of der(1;16) in wide clonal expansion. [Conclusions] Our results suggest that in some breast cancer cases, particularly in those with der(1;16), a highly recurrent translocation accounting for the major subset of Luminal A breast cancer, the clones with the funder driver alterations expanded macroscopically long before the onset of cancer, in which further clonal evolutions recursively occur multi-focally, giving rise to multiple proliferative lesions and ultimately, invasive cancers. Our findings provide new insight into the early development of breast cancer. Citation Format: Tomomi Nishimura, Nobuyuki Kakiuchi, Kenichi Yoshida, Takaki Sakurai, Tatsuki R. Kataoka, Eiji Kondoh, Yoshitsugu Chigusa, Masahiko Kawai, Morio Sawada, Takuya Inoue, Yasuhide Takeuchi, Hirona Maeda, Satoko Baba, Yusuke Shiozawa, Ryunosuke Saiki, Masahiro M. Nakagawa, Yasuhito Nannya, Yotaro Ochi, Tomonori Hirano, Yukiko Inagaki-Kawata, Kosuke Aoki, Masahiro Hirata, Eiji Suzuki, Masahiro Takada, Masahiro Kawashima, Kosuke Kawaguchi, Kenichi Chiba, Yuichi Shiraishi, Junko Takita, Satoru Miyano, Masaki Mandai, Kengo Takeuchi, Hironori Haga, Masakazu Toi, Seishi Ogawa. Clonal evolution of mammary epithelial cells into breast cancers [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P5-13-04.

Abstract Background: In pathology, digitizing tissue slides has prompted a remarkable development in image analysis using deep learning. This technological advancement is anticipated to aid in pathological diagnosis and to enhance patient management. Deep learning-based image cytometry (DL-IC) enables accurate cell identification and counting and the acquisition of vast amounts of location information from tissue slides. DL-IC can capture information about the diverse and complex tumor immune microenvironment(TIME) and its constituent cells and help identify biomarkers to predict patient treatment efficacy and prognosis. This study will introduce a spatial interaction map and co-localization index (CLI) for the analysis of TIME using DL-IC. Materials and Methods: Cu-Cyto, a deep learning-based image analysis technology, was used in this study; bit-pattern kernel filtering technology, which can accurately count cells while avoiding the determination of multiple cell counts, was used by Cu-Cyto (Abe T, et al. Anticancer Res. 43:3755, 2023). First, the accuracy of cell counting using Cu-Cyto was evaluated. Second, tumor tissue slides with immunohistochemical (IHC) and hematoxylin-eosin (H&E) staining were prepared from surgical specimens of patients with rectal cancer who had undergone neoadjuvant chemoradiotherapy (NACRT), and the relationship between the co-localization index (CLI) of cancer cells and CD8+T cells and prognosis was investigated. CLI was defined to predict cell- cell interactions on the basis of the relative distances between different cell types (Nagasaka T. PCT/JP 2021/021455). Results: The performances of three versions of Cu-Cyto were evaluated according to their learning stages. In the early stage of learning, the F1 score for immunostained CD8+ T cells (0.343) was higher than that for non-immunostained cells (adenocarcinoma cells [0.040] and lymphocytes [0.002]). In the latest stage of learning, the F1 scores for adenocarcinoma cells, lymphocytes, and CD8+ T cells were 0.589, 0.889, and 0.911, respectively. Next, we examined the correlation of CLI between cancer cells and CD8+ T cells with prognosis: patients with a higher CLI significantly prolonged five-year disease-free survival (P=0.038), while there was no substantial difference in five-year overall survival (P=0.57). Conclusion]: Cu-Cyto performed well in cell determination. In particular, IHC was able to increase the learning efficiencies in the early stages of learning. The CLI calculated using Cu-Cyto ts an objective, reproducible, and innovative quantitative approach for assessing cell-cell interactions, which has been shown to be associated with recurrence-free survival in patients with rectal cancer after NACRT. Its performance is expected to improve even further with continuous learning, and the DL-IC can contribute to the implementation of precision oncology. Citation Format: Tomoki Abe, Kimihiro Yamashita, Toru Nagasaka, Tomosuke Mukoyama, Souichirou Miyake, Yasuhiro Ueda, Masayuki Ando, Yuki Okazoe, Takao Tsuneki, Yukari Adachi, Ryunosuke Konaka, Ryuichiro Sawada, Hironobu Goto, Hiroshi Hasegawa, Shingo Kanaji, Takeru Matsuda, Takumi Fukumoto, Yoshihiro Kakeji. Deep learning-based image cytometry and co-localization index in tumor immune microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3650.

Abstract Background and Aim: Immune checkpoint inhibitors (ICIs) have been widely used for the initial treatment of non-small cell lung cancer (NSCLC). Although both ICI monotherapy and the combination of ICI plus chemotherapy (chemo-ICI) are promising therapeutic options for patients with a programmed cell death ligand-1 tumor proportion score of 50% or higher (high PD-L1), the patient's clinical background and biomarkers differentiating the two options are still not well-defined. In our prior research, we found that a history of proton pump inhibitor (PPI) use independently have negative impact with significantly shorter overall survival (OS) and progression-free survival (PFS) in NSCLC patients harboring high PD-L1 with pembrolizumab monotherapy, but not with chemo-ICI. However, this OS data remain immature. Therefore, in our current study, we extended the follow-up period and conduct additional analysis including OS data to confirm this trend. Method: Advanced NSCLC patients with high PD-L1 who had received pembrolizumab or chemo-ICI as the first-line treatment between March 2017 and December 2020 at 13 hospitals in Japan were analyzed. Survival outcomes were estimated using the Kaplan- Meier method and were compared using the log-rank test. Results: A total of 425 patients with NSCLC were included in the study, with 271 patients (median [range] age, 72 [43-90] years; 215 [79%] men) receiving pembrolizumab monotherapy as their first-line treatment and 154 patients (median [range] age, 69 [36-86] years; 121 [79%] men) receiving chemo-ICI as their first-line treatment. The median follow-up duration was 22.8 months. Among patients with a history of PPI use, both median PFS (19.3 months vs. 6.8 months; HR, 0.53; 95% CI, 0.35-0.78; P = .003) and the median OS(not reached vs. 20.4 months; HR, 0.53; 95% CI, 0.34-0.85; P = .04) were significantly longer in the chemo-ICI group compared to the pembrolizumab monotherapy group. For patients without a history of PPI use, both median PFS (11.4 months vs. 10.8 months; HR, 0.99; 95% CI, 0.76-1.3; P = .36) and the median overall survival (42.2 months vs. 29.9 months, HR, 0.84; 95% CI, 0.62-1.16; P = .33) showed no significant differences between the groups. Discussion and Conclusion: This retrospective study showed that a concomitant treatment with PPI is poor prognostic factor in patients harboring high PD-L1 and treated with ICI monotherapy compared with chemo-ICI in Japanese cohort and this tendency is durable in long-term follow-up. These results indicate that when deciding on an ICI treatment, with or without chemotherapy, it is essential to take into account a concomitant treatment with PPI. Citation Format: Ryo Sawada, Hayato Kawachi, Tadaaki Yamada, Motohiro Tamiya, Yoshiki Negi, Yasuhiro Goto, Akira Nakao, Shinsuke Shiotsu, Takayuki Takeda, Asuka Okada, Taishi Harada, Koji Date, Yusuke Chihara, Isao Hasegawa, Takashi Kijima, Koichi Takayama. Impact of proton pump inhibitor on ICI with or without chemotherapy for NSCLC with high PD-L1 TPS [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3730.

Abstract Objective: We aimed to establish reversibly immortalized cell lines from human uterine and ovary cells using the Sendai virus (SeV) vector. The immortalized cells derive from normal and benign ovarian epithelial cells and endometrial epithelial cells. Furthermore, we sought to elucidate the mechanisms of carcinogenesis using the immortalized cell lines. Methods: Cells were collected at the time of surgery after obtaining patient consent. The cells used in this study were as follows: ovarian epithelial cells (normal epithelium, Ov n; normal epithelium with germline BRCA1 or BRCA2 mutation, Ov BRCA1 2; ovarian endometrioma, Ov endo; mucinous cystadenoma; Ov m), normal fallopian tube (FT) cells, and endometrial epithelium (normal epithelium, Em n). These cells were infected with temperature-sensitive SeV vectors carrying three immortalization genes, Bmi-1, hTERT, and SV40T.The presence of infection was confirmed through Green Fluorescent Protein (GFP) and Orange Fluorescent Protein (OFP). Immunoreactivity to the anti-human EpCAM antibody (a marker derived from epithelial carcinoma) in each SeV-infected cell was confirmed through flow cytometry. QH and multicolor FISH staining were performed for karyotyping of metaphase chromosomes in each cell line to determine chromosome number and structural abnormalities. Human transcriptome sequencing analysis was performed with NovaSeq 6000 (Illumina) using total RNA from each cell line. Some genes that showed significant expression in each cell line were subjected to real-time PCR (RT-PCR). Results: We established the immortalized cell lines from human uterine and ovarian tissues. SeV-infected cells exhibited GFP and OFP fluorescence, while non-infected cells did not. SeV infection allowed all primary cell lines to grow for 25 or more passages, while non-infected SeV cells lacked the proliferative capacity and showed senescence-like morphology. SeV-infected cells senesced in a temperature-dependent manner. Ov n SeV- infected cells ion causedshowed a small increase in chromosome structural abnormalities. But, Ov BRCA1 and 2 SeV-infected cells showed larger than thatand . Llong-term passaged cells did not show immune response to anti-human EpCAM antibodies in normal cells. Eleven Three genes were predominantly expressed in Ov BRCA1 and Ov BRCA2 cells and were not expressed in Ov n cells. SomeTwo out of ththe three genesem were found to be predominantly expressed in Ov endo cells compared to the expression in the Ovn cell line. Furthermore, RT-PCR results also indicated substantially higher expression of two genes in Ov BRCA1/2 cells and in Ov endo cells compared to the expression in Ov n cells. Conclusion: We succeeded in the reversible immortalization of endometrial and ovarian epithelial cells by using SeV infection. We identified several candidate genes that may be involved in the oncogenic mechanism of ovarian cancer associated with endometriosis or germline BRCA1 and BRCA2. Citation Format: Masayo Okawa, Hiroaki Komatsu, Kohei Hikino, Yuki Iida, Masayo Hosokawa, Mayumi Sawada, Akiko Kudoh, Jun Chikumi, Shinya Sato, Genki Hichiwa, Yasuhiro Kazuki, Kanako Kazuki, Fuminori Taniguchi, Mitsuo Oshimura, Tasuku Harada. Establishment and characterization of reversibly immortalized endometrial and ovarian epithelial cell lines using Sendai virus [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1214.

The ninth Best Paper Award 2018 ceremony was held at Hilltop Hotel, Tokyo, September 26, 2018, attended by the winners and IJAT Editorial Committee members. At the same time, the third Best Review Award 2018 has been decided by IJAT Editorial Committee. The Best Paper was severely selected from among 93 papers published in Vol.11, 2017, and Best Review was selected from 15 reviews published from 2016 to 2018. The Best Paper Award winner was given a certificate with a nearly US$1,000 honorarium, and the Best Review Award winner was given a certificate with commemorative shield. We congratulate the winners and sincerely wish for their future success. The Best Paper Award 2018 Integrated Chatter Monitoring Based on Sensorless Cutting Force/Torque Estimation in Parallel Turning by Yuki Yamada, Takashi Kadota, Shinya Sakata, Junji Tachibana, Kenichi Nakanishi, Manabu Sawada, and Yasuhiro Kakinuma Int. J. of Automation Technology, Vol.11 No.2, pp. 215-225, March 2017 The Best Review Award 2018 “Industrie 4.0” and Smart Manufacturing – A Review of Research Issues and Application Examples by Klaus-Dieter Thoben, Stefan Wiesner, and Thorsten Wuest Int. J. of Automation Technology, Vol.11 No.1, pp. 4-16, January 2017