BPB Reports

2024 - Vol. 7 No. 4

2024 - Vol. 7

Regular Article
The Evaluation of Lipid Analysis for PXB-Cells LA as a Human Non-Alcoholic Fatty Liver Disease Model Vol.7, No.4, p.147-156
Masaki Takahashi , Sayaka Tomatsu , Mutsumi Inamatsu , Nami Yoshikawa , Keishi Hata , Masakazu Kakuni
Received: May 22, 2024
Accepted: August 05, 2024
Released: August 28, 2024
Abstract Full Text PDF[2M]

Fatty liver can progress into serious conditions, and the number of patients with fatty liver disease has risen globally in recent years. Various lipid metabolism disorders can cause fatty liver, and in vitro models, such as hepatoma cell lines, have been utilized in research related to lipid metabolism disorders, including the development of treatment strategies. We previously demonstrated that fresh hepatocytes (PXB-cells®) from chimeric mice with humanized livers display lipid metabolism similar to that of normal human hepatocytes. Additionally, we developed PXB-cells Lipid Analysis (PXB-cells LA) as a model of non-alcoholic fatty liver disease (NAFLD). PXB-cells LA exhibited increased levels of intracellular lipid droplets and lipids, especially triglycerides, compared to PXB-cells. Additionally, albumin secretion, drug metabolism, bile excretion transporters, mitochondria-derived oxidative phosphorylation, and intracellular adenosine triphosphate levels were attenuated in PXB-cells LA, while inflammatory marker levels were elevated. Collectively, these findings indicate hepatic dysfunction. Additionally, PXB-cells LA showed a fractional profile with a peak in very low-density lipoproteins, similar to PXB-cells. PXB-cells LA also secreted lipoproteins with a higher triglyceride content, associated with NAFLD. Taken together, these results suggest that PXB-cells LA is a useful cellular model of human NAFLD.

Regular Article
Potential Anti-Cancer Therapeutic Agents: Effects of VHH Antibodies on Mesothelin Binding Affinity in Cancer Vol.7, No.4, p.141-146
Yoshimi Shimizu , Katsuhiro Isoda , Kaho Shinohara , Miki Nakayama , Ikuko Taira , Yuichiro Taira
Received: April 28, 2024
Accepted: August 08, 2024
Released: August 22, 2024
Abstract Full Text PDF[2M]

Mesothelin (MSLN) is a glycosyl phosphatidyl inositol-anchored glycoprotein involved in the carcinogenesis and metastasis of some cancers such as ovarian cancer, pancreatic cancer, and mesothelioma. The interaction between cancer antigen 125 (CA125) and MSLN enhances tumor metastasis. MSLNs are highly expressed in cancer and are therefore promising targets for cancer therapy. Variable domains of heavy chain of heavy chain (VHH; also known as nanobodies) monoclonal antibodies (mAbs) from alpacas exhibit unique properties such as high affinity, low immunogenicity, and thermal stability. Herein, we aimed to investigate the effects of anti-MSLN VHH mAbs on MSLN binding affinity. Two anti-MSLN VHH mAb clones (MT1A1 and MT3C2), targeting the extracellular domain of human MSLN, were used. MT1A1 and MT3C2 are specific to human and mouse MSLN. MT1A1 and MT3C2 bound to MSLN are expressed on ovarian, pancreatic, and mesothelioma cancer cells. These antibodies recognize the region (296–390) at the N-terminal of MSLN on the cell surface and relying on their conformation-dependent recognition. MT1A1 and MT3C2 significantly inhibited the MSLN-CA125 interaction. Overall, our results suggest that MT1A1 and MT3C2 are promising anticancer agents.

Regular Article
Adverse Effects Induced by Osimertinib Based on the Dose per Body Constitutional Parameters: A Retrospective Observational Study Vol.7, No.4, p.132-140
Masaya Nagamizu , Yuji Hotta , Issei Morozumi , Daigaku Nakamura , Masayuki Hori , Yuto Otsuka , Ryuhei Takemoto , Yasuhiro Horita , Eri Wakita , Nobuyuki Morishita , Masahiro Kondo , Yoko Furukawa‐Hibi , Kazunori Kimura
Received: June 04, 2024
Accepted: August 10, 2024
Released: August 22, 2024
Abstract Full Text PDF[460K]

Background: Osimertinib is a third-generation epidermal growth factor receptor tyrosine kinase inhibitor treatment for non-small-cell lung cancer. Area under the blood concentration-time curve (AUC) of osimertinib can be related to the development of adverse events (AEs). Furthermore, the dose-per-body constitutional parameters (BCPs), such as body weight, body surface area (BSA), or lean body mass (LBM), are closely related to the AUC. However, BCPs other than weight have not been considered in clinical trials. Therefore, in this retrospective study, we investigated the association between doses per BCP (Dose/BCPs) and AEs of osimertinib. Method: Forty-two patients who received osimertinib between January 2010 and December 2020 were investigated. Differences in Dose/BCPs were compared between patients with and without AEs. Results: Among the patients, 54.8%, 38.1%, and 28.6% developed thrombocytopenia, neutropenia, and leukopenia, respectively. The Dose/BSA and Dose/LBM were significantly higher in patients who developed leukopenia than in those who did not (p < 0.05). The cutoff values of Dose/BSA and Dose/LBM associated with leukopenia were 50.0 mg/m2 and 1.86 mg/kg, respectively. Conclusion: This study suggests that Dose/BCPs are associated with the development of leukopenia. Now, fixed dose regardless of BCPs is approved on almost of oral molecular targeting agents. However, the patients with low BCPs can be also received these medications in clinical practices. Therefore, although the sample size is small, the results of this study suggests the potential risk on the fixed dose. Moreover, Dose/BSA or Dose/LBM may be a useful parameter for assessing the risk of leukopenia with osimertinib.

Regular Article
Tapinarof, a Novel Topical Therapeutic Aryl Hydrocarbon Receptor Agonist, Suppresses Atopic Dermatitis-like Skin Inflammation in Mice Vol.7, No.4, p.123-131
Toshiki Urashima , Yoshiaki Katsuda , Hiromi Yoshiuchi , Shin Ebihara , Yuichi Shinozaki , Tomoko Kato , Taishi Shimazaki , Yuzo Yasui , Yasuko Mera , Noriko Konishi
Received: March 22, 2024
Accepted: July 19, 2024
Released: August 20, 2024
Abstract Full Text PDF[2M]

Tapinarof is a non-steroidal, small molecule aryl hydrocarbon receptor (AhR) agonist that is being developed for the treatment of atopic dermatitis (AD). AD is a chronic inflammatory skin disease mediated by type 2 helper T (Th2) cells, characterized by impaired epidermal differentiation and skin barrier function. Tapinarof has been reported to regulate target gene expression through activation of AhR, improve skin barrier function, and exhibit an antioxidant effect. In this study, we investigated the pharmacological properties of tapinarof and its efficacy in mice with AD-like skin inflammation induced by 2, 4-dinitrofluorobenzene (DNFB). Tapinarof induced the mRNA expression of CYP1A1, an indicator of AhR activation, induced the mRNA expression of NAD(P)H: quinone oxidoreductase (NQO1), an antioxidant enzyme in human peripheral blood mononuclear cells (PBMCs) stimulated with T-cell activators. It also suppressed the production of interleukin (IL)-4, a Th2 cytokine. In mice with AD-like dermatitis, topical administration of tapinarof promoted the expression of Cyp1a1 and Nqo1 in the skin. It suppressed IL-4 production, the ear swelling, and histopathological changes. Tapinarof also suppressed an increase in transepidermal water loss (TEWL), an indicator of skin barrier function. These results indicate that tapinarof suppresses AD-like skin dermatitis and suggest that a variety of pharmacological actions, including an antioxidant effect, inhibition of Th2 cytokine, and improvement in barrier function, are involved.

Regular Article
Pharmacological Properties of Tapinarof in Mice as a Novel Topical Agent for Plaque Psoriasis Vol.7, No.4, p.116-122
Toshiki Urashima , Yoshiaki Katsuda , Hiromi Yoshiuchi , Shin Ebihara , Yuichi Shinozaki , Tomoko Kato , Taishi Shimazaki , Yuzo Yasui , Yasuko Mera , Noriko Konishi
Received: May 04, 2024
Accepted: July 17, 2024
Released: August 06, 2024
Abstract Full Text PDF[1M]

Tapinarof is a non-steroidal, small molecule aryl hydrocarbon receptor (AhR) agonist that has demonstrated clinical efficacy and safety in patients with plaque psoriasis. In this study, we investigated the effects of tapinarof on interleukin (IL)-23-induced psoriasis-like dermatitis, which is a direct reflection of IL-23/type 17 helper T (Th17) axis activation considered pivotal in the pathogenesis of psoriasis, or normal skin in mice to elucidate its pharmacological properties. In mice with dermatitis, topical administration of tapinarof induced AhR activation, promoted the expression of an antioxidant molecule, and suppressed Th17 cytokine production in the skin, resulting in the reduction of skin swelling and histopathological changes. In normal mice, tapinarof did not induce the skin thinning observed with dexamethasone. These findings suggest that tapinarof represents a new topical treatment option for psoriasis carrying a lower risk of skin atrophy.

Regular Article
Establishment of a Reporter System to Monitor FcRγ-Dependent Activation of NFAT Vol.7, No.4, p.110-115
Arisa Morikawa , Kaho Togawa , Reina Mukainaka , Takuma Kitano , Saotomo Itoh , Shinsuke Taki , Shigeaki Hida
Received: March 27, 2023
Accepted: September 22, 2023
Released: August 02, 2024
Abstract Full Text PDF[832K]

Basophils are recognized as effectors of type 2 immune responses, producing IL-4 in response to various stimuli such as IL-3 and papain in addition to IgE. In this study, we have established a novel cell-based reporter system that can monitor the activation of the transcription factor NFAT using retroviral vectors. Using this system, we examined whether papain, which is known to induce IL-4 in an FcRγ-dependent manner, could be used to identify its receptor. We created a chimeric receptor in which the extracellular and transmembrane portions are CD8 and the cytoplasmic domain is FcRγ. This chimeric receptor was able to induce GFP in RBL by cross-linking with anti-CD8 antibody. Furthermore, we found that this chimeric receptor can function as a papain receptor, as GFP was upregulated by papain stimulation. On the other hand, statins were able to suppress the expression of GFP by IgE crosslinking. This reporter system can be used to transduce candidate receptors and examine their papain receptor activity using GFP expression as an indicator, and is therefore a useful system that can be used for expression-cloning of unknown papain receptors and for the study of various inhibitors.

Report
Microbicidal Effect of Sodium Chlorite in Combination with Caffeine at a Neutral pH Vol.7, No.4, p.106-109
Kiyoshi Konishi , Takekatsu Shibata
Received: April 22, 2024
Accepted: July 16, 2024
Released: August 01, 2024
Abstract Full Text PDF[686K]

Sodium chlorite (NaClO2) is applied as a disinfectant for the sanitization of food and environmental surfaces. HClO2 (pKa = 1.86) is believed to be the main species responsible for the antimicrobial effects, whereas NaClO2 solution ([HClO2] / [ClO2-] = 10-5.64 at pH 7.5) shows only weak antimicrobial activity at neutral pH conditions. However, NaClO2 solution at a neutral pH has the advantages of having very low reactivity to organic materials and high stability for long-term storage at around room temperature. In our previous screening of food additives, phytochemicals, and related compounds, we found that caffeine could strongly promote the antimicrobial effects of NaClO2 solution at a neutral pH. Caffeine is a purine alkaloid found in nearly 100 plant species that has very weak antibacterial properties against many bacteria. In the present study, we evaluated the antimicrobial activity of NaClO2 in combination with caffeine against Acinetobacter baumannii, Pseudomonas aeruginosa, Staphylococcus aureus, and Candida albicans strains. We found that caffeine strongly potentiated the antimicrobial action of NaClO2 in all four strains, indicating that this combination has potential as a disinfectant.

Report
Cycloalliin, an Organosulfur Compound in Garlic, Inhibits EMT and Invasion of the A549 Non-Small Cell Lung Cancer Cell Line Vol.7, No.4, p.101-105
Ayumi Sasaki , Yoshifumi Yamaya , Noriyuki Notoya , Makoto Nishizuka
Received: June 21, 2024
Accepted: July 19, 2024
Released: August 01, 2024
Abstract Full Text PDF[1M]

Non-small cell lung cancer (NSCLC) accounts for 80%–90% of all lung cancers. The metastasis of NSCLC has a considerable impact on prognosis and systemic status. Cycloalliin, an organosulfur compound found in garlic, is known for its health benefits on cardiovascular disease and obesity. However, the role of cycloalliin in cancer cell invasion and metastasis remains unknown. In this study, we investigated the effect of cycloalliin on the epithelial-to-mesenchymal transition (EMT) and invasiveness of the NSCLC cell line A549. Cycloalliin inhibited transforming growth factor (TGF)-β-induced EMT and invasive potential of A549 cells. Furthermore, we found that cycloalliin suppressed Smad3 phosphorylation and expression of Snail, a transcription factor that promotes EMT, during the early stages of TGF-β-mediated EMT. This study provides valuable insights into the inhibitory potential of cycloalliin on EMT, suggesting that this compound may have a therapeutic role against EMT in NSCLC cells.

Report
TIC10/ONC201 Enhances Phosphate Uptake in the Human Neuroblastoma Cell Line SH-SY5Y Vol.7, No.4, p.96-100
Junya Murata , Kazuki Ohuchi , Yuto Murayama , Ayane Mishima , Hisaka Kurita , Isao Hozumi , Masatoshi Inden
Received: May 21, 2024
Accepted: July 09, 2024
Released: August 01, 2024
Abstract Full Text PDF[1M]

The type-III sodium-dependent phosphate transporters, SLC20A1 and SLC20A2, are distributed throughout the body, including the central nervous system. Various neurodegenerative diseases, including primary basal ganglia calcification (PBGC), involve the disruption of phosphate homeostasis. Patients with PBGC harbor a mutated SLC20A2. Previously, we demonstrated that the phosphate transport activity of SLC20A2 was involved in PBGC pathology. Thus, we hypothesized the activation of phosphate transport as one of the therapeutic targets for PBGC. It was previously reported that SLC20A1 and SLC20A2 were increased at vascular smooth muscle cell of ATF4-overexpression mice. This study investigated the effect of TIC10/ONC201, a potential activator of ATF4, on phosphate transport in SH-SY5Y, a neuronal cell model. Treatment with 3 µM TIC10, which did not cause cell death, increased phosphate uptake along with the ATF4 and SLC20A1 but not SLC20A2. Treatment with 3 µM TIC10 also enhanced phosphate uptake in SLC20A2-knockdowned cells but not SLC20A1-knockdowned cells. In conclusion, TIC10 enhanced phosphate uptake in SH-SY5Y cells via SLC20A1 but not SLC20A2.