nature portfolio Acta Medica Okayama 2399-3642 6 1 2023 NFYA promotes malignant behavior of triple-negative breast cancer in mice through the regulation of lipid metabolism 596 EN Nobuhiro Okada Graduate School of Interdisciplinary Science & Engineering in Health Systems, Okayama University Chihiro Ueki Graduate School of Interdisciplinary Science & Engineering in Health Systems, Okayama University Masahiro Shimazaki Laboratory for Malignancy Control Research, Medical Innovation Center, Kyoto University Goki Tsujimoto Graduate School of Interdisciplinary Science & Engineering in Health Systems, Okayama University Susumu Kohno Division of Oncology and Molecular Biology, Cancer Research Institute, Kanazawa University Hayato Muranaka Division of Oncology and Molecular Biology, Cancer Research Institute, Kanazawa University Kiyotsugu Yoshikawa Faculty of Pharmaceutical Sciences, Doshisha Women�fs College of Liberal Arts Chiaki Takahashi Division of Oncology and Molecular Biology, Cancer Research Institute, Kanazawa University Two splicing variants exist in NFYA that exhibit high expression in many human tumour types. The balance in their expression correlates with prognosis in breast cancer, but functional differences remain unclear. Here, we demonstrate that NFYAv1, a long-form variant, upregulates the transcription of essential lipogenic enzymes ACACA and FASN to enhance the malignant behavior of triple-negative breast cancer (TNBC). Loss of the NFYAv1-lipogenesis axis strongly suppresses malignant behavior in vitro and in vivo, indicating that the NFYAv1-lipogenesis axis is essential for TNBC malignant behavior and that the axis might be a potential therapeutic target for TNBC. Furthermore, mice deficient in lipogenic enzymes, such as Acly, Acaca, and Fasn, exhibit embryonic lethality; however, Nfyav1-deficient mice exhibited no apparent developmental abnormalities. Our results indicate that the NFYAv1-lipogenesis axis has tumour-promoting effects and that NFYAv1 may be a safe therapeutic target for TNBC. No potential conflict of interest relevant to this article was reported.

Frontiers Media SA Acta Medica Okayama 2296-634X 10 2022 NFYA promotes the anti-tumor effects of gluconeogenesis in hepatocellular carcinoma through the regulation of PCK1 expression 983599 EN Goki Tsujimoto Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Rin Ito Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Kei Yoshikawa Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Chihiro Ueki Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Nobuhiro Okada Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Reprogramming of glucose metabolism occurs in many human tumor types, and one of these, gluconeogenesis, is known to exhibit anti-tumor effects in hepatocellular carcinoma (HCC). The transcription factor NFYA regulates gluconeogenesis in the normal liver tissue, but the function of the NFYA-gluconeogenesis axis in cancer and the functional differences of NFYA splicing variants in the regulation of gluconeogenesis is still unclear. Here, we demonstrate that NFYAv2, the short-form variant, upregulates the transcription of a gluconeogenic enzyme PCK1. We further reveal that its regulation induces high ROS levels and energy crisis in HCC and promotes cell death. These indicate that the NFYAv2-gluconeogenesis axis has enhanced anti-tumor effects in HCC, suggesting that the axis may be a potential therapeutic target for HCC. Furthermore, Nfyav1-deficient mice, spontaneously overexpressing Nfyav2, had no increasing gluconeogenesis in the liver. Taken together, our results reveal NFYAv2-gluconeogenesis axis has anti-tumor effects and the potential for NFYAv2 to be a safer therapeutic target for HCC. No potential conflict of interest relevant to this article was reported.

NATURE PUBLISHING GROUP Acta Medica Okayama 2045-2322 10 1 2020 Signaling Inhibitors Accelerate the Conversion of mouse iPS Cells into Cancer Stem Cells in the Tumor Microenvironment 9955 EN Juan Du Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Yanning Xu Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Saki Sasada Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Aung Ko Ko Oo Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Ghmkin Hassan Laboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Hafizah Mahmud Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Apriliana Cahya Khayrani Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Md Jahangir Alam Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Kazuki Kumon Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Ryo Uesaki Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Said M. Afify Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Hager M. Mansour Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Neha Nair Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Maram H. Zahra Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Akimasa Seno Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Nobuhiro Okada Laboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Ling Chen Department of Pathology, Tianjin Central Hospital of Gynecology Obstetrics Ting Yan Department of Pathology, Shanxi Key Laboratory of Carcinogenesis and Translational Research on Esophageal Cancer, Shanxi Medical University Masaharu Seno Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Cancer stem cells (CSCs) are a class of cancer cells characterized by self-renewal, differentiation and tumorigenic potential. We previously established a model of CSCs by culturing mouse induced pluripotent stem cells (miPSCs) for four weeks in the presence of a conditioned medium (CM) of cancer cell lines, which functioned as the tumor microenvironment. Based on this methodology of developing CSCs from miPSCs, we assessed the risk of 110 non-mutagenic chemical compounds, most of which are known as inhibitors of cytoplasmic signaling pathways, as potential carcinogens. We treated miPSCs with each compound for one week in the presence of a CM of Lewis lung carcinoma (LLC) cells. However, one-week period was too short for the CM to convert miPSCs into CSCs. Consequently, PDO325901 (MEK inhibitor), CHIR99021 (GSK-3 beta inhibitor) and Dasatinib (Abl, Src and c-Kit inhibitor) were found to confer miPSCs with the CSC phenotype in one week. The tumor cells that survived exhibited stemness markers, spheroid formation and tumorigenesis in Balb/c nude mice. Hence, we concluded that the three signal inhibitors accelerated the conversion of miPSCs into CSCs. Similarly to our previous study, we found that the PI3K-Akt signaling pathway was upregulated in the CSCs. Herein, we focused on the expression of relative genes after the treatment with these three inhibitors. Our results demonstrated an increased expression of pik3ca, pik3cb, pik3r5 and pik3r1 genes indicating class IA PI3K as the responsible signaling pathway. Hence, AKT phosphorylation was found to be up-regulated in the obtained CSCs. Inhibition of Erk1/2, tyrosine kinase, and/or GSK-3 beta was implied to be involved in the enhancement of the PI3K-AKT signaling pathway in the undifferentiated cells, resulting in the sustained stemness, and subsequent conversion of miPSCs into CSCs in the tumor microenvironment. No potential conflict of interest relevant to this article was reported.

Amber Publication Acta Medica Okayama 2347-2367 8 1 2020 Upregulated CCL20 and CCR6 in Cancer Stem Cells Converted from Mouse iPS Cells 200 207 EN Juan Du Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Akimasa Seno Laboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama Universityalth Systems Saki Sasada Graduate School of Natural Science and Technology, Okayama University Yanning Xu Department of Pathology, Tianjin Central Hospital of Gynecology Obstetrics Aung Ko Ko Oo Graduate School of Natural Science and Technology, Okayama University Ghmkin Hassan Graduate School of Natural Science and Technology, Okayama University Shunsuke Ueno Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Said M. Afify Graduate School of Natural Science and Technology, Okayama University Maram H Zahra Graduate School of Natural Science and Technology, Okayama University Nobuhiro Okada Graduate School of Natural Science and Technology, Okayama University Ling Chen Department of Pathology, Tianjin Central Hospital of Gynecology Obstetrics Xiaoying Fu School of Integrative Medicine, Tianjin University of Traditional Chinese Medicine Heizo Tokutaka SOM Japan Ting Yan The Hong Kong University of Science and Technology Medical Center, Shenzhen Peking University Masaharu Seno Graduate School of Natural Science and Technology, Okayama University Background: Cancer stem cells (CSCs) as a class of malignant cancer cells play an important role in tumor progression. Previous studies by our group have demonstrated the establishment of the model of CSCs converting mouse iPS cells (miPSCs) into CSCs by treating the miPSCs with a conditioned medium (CM) of Lewis Lung Carcinoma (LLC) cells with or without the nonmutagenic chemical compounds. CSCs converted from miPSCs developed highly malignant adenocarcinoma when subcutaneously transplanted into the nude mice.</br> Methods: The miPSCs were treated with each compound for 1 week in the presence of a CM of LLC cells. We evaluated the gene expression in the resultant CSCs comparing that in miPSCs by microarray analysis. And the expression of chemokine (C-C motif) ligand 20 (CCL20) and C-C chemokine receptor type 6 (CCR6) in converted cells were evaluated by rt-qPCR. The CCR6 expression in converted cells and primary cells were determined by flow cytometry.</br> Results: As the result, the expression of CCL20 was found upregulated in the presence of CM supplemented with PD0325901. Then we assessed the expression of CCR6, which was considered to be stimulated by CCL20. Then the expression of CCR6 was also found up-regulated. Interestingly, IL17A expression was also observed only in the CSCs from the primary tumor implying the effect of tumor microenvironment. Moreover, significantly high level of CCR6 was showed in flow cytometric analysis.</br> Conclusion: These results suggest that a model of CSCs with CCL20-CCR6 autocrine loop was obtained as the result of the conversion of iPSCs. This CSC should be a good model to study targeting CCR6 as a G protein-coupled receptor (GPCR). No potential conflict of interest relevant to this article was reported.

MDPI Acta Medica Okayama 1422-0067 20 5 2019 Targeting Ovarian Cancer Cells Overexpressing CD44 with Immunoliposomes Encapsulating Glycosylated Paclitaxel 1042 EN Apriliana Cahya Khayrani Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Hafizah Mahmud Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Maram H. Zahra Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Aung Ko Ko Oo Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Miharu Oze Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Juan Du Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Md Jahangir Alam Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Said M. Afify Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Hagar A. Abu Quora Laboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Tsukasa Shigehiro Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Anna Sanchez Calle Division of Molecular and Cellular Medicine, National Cancer Center Research Institute Nobuhiro Okada Laboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Akimasa Seno Laboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University Koki Fujita Ensuiko Sugar Refining Co., Ltd. Hiroki Hamada Faculty of Science, Okayama University of Science Yuhki Seno Graduate School of Pharmaceutical Science, Tokushima University Tadakatsu Mandai Faculty of Life Science, Kurashiki University of Science and the Arts Masaharu Seno Department of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama University Paclitaxel (PTX) is one of the front-line drugs approved for the treatment of ovarian cancer. However, the application of PTX is limited due to the significant hydrophobicity and poor pharmacokinetics. We previously reported target-directed liposomes carrying tumor-selective conjugated antibody and encapsulated glycosylated PTX (gPTX-L) which successfully overcome the PTX limitation. The tubulin stabilizing activity of gPTX was equivalent to that of PTX while the cytotoxic activity of gPTX was reduced. In human ovarian cancer cell lines, SK-OV-3 and OVK18, the concentration at which cell growth was inhibited by 50% (IC50) for gPTX range from 15⁻20 nM, which was sensitive enough to address gPTX-L with tumor-selective antibody coupling for ovarian cancer therapy. The cell membrane receptor CD44 is associated with cancer progression and has been recognized as a cancer stem cell marker including ovarian cancer, becoming a suitable candidate to be targeted by gPTX-L therapy. In this study, gPTX-loading liposomes conjugated with anti-CD44 antibody (gPTX-IL) were assessed for the efficacy of targeting CD44-positive ovarian cancer cells. We successfully encapsulated gPTX into liposomes with the loading efficiency (LE) more than 80% in both of gPTX-L and gPTX-IL with a diameter of approximately 100 nm with efficacy of enhanced cytotoxicity in vitro and of convenient treatment in vivo. As the result, gPTX-IL efficiently suppressed tumor growth in vivo. Therefore gPTX-IL could be a promising formulation for effective ovarian cancer therapies. No potential conflict of interest relevant to this article was reported.