207 results on '"Sakurai F"'
Search Results
2. Upregulation of RECK gene expression by small double-stranded RNA targeting the promoter region
- Author
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Sakurai, F, Nanjo, Y, Okamoto, S, Tachibana, M, and Mizuguchi, H
- Published
- 2014
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3. Efficient antitumor effects of carrier cells loaded with a fiber-substituted conditionally replicating adenovirus on CAR-negative tumor cells
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Iguchi, K, Sakurai, F, Tomita, K, Katayama, K, Yamaguchi, T, Kawabata, K, Tagawa, M, Kawabata, M, Shirakawa, T, and Mizuguchi, H
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- 2012
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4. Apoptosis-Resistant and Highly Functional Islet Cell Sheets for Transplantation.: Abstract# C1561
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Hanayama, H., Ohashi, K., Utoh, R., Ise, K., Shimizu, T., Yamato, M., Mizuguchi, H., Sakurai, F., Okano, T., and Gotoh, M.
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- 2014
5. Development of fiber-substituted adenovirus vectors containing foreign peptides in the adenovirus serotype 35 fiber knob
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Matsui, H, Sakurai, F, Kurachi, S, Tashiro, K, Sugio, K, Kawabata, K, Yamanishi, K, and Mizuguchi, H
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- 2009
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6. Adenovirus serotype 35 vector-mediated transduction following direct administration into organs of nonhuman primates
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Sakurai, F, Nakamura, S-i, Akitomo, K, Shibata, H, Terao, K, Kawabata, K, Hayakawa, T, and Mizuguchi, H
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- 2009
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7. Interaction of penton base Arg-Gly-Asp motifs with integrins is crucial for adenovirus serotype 35 vector transduction in human hematopoietic cells
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Murakami, S, Sakurai, F, Kawabata, K, Okada, N, Fujita, T, Yamamoto, A, Hayakawa, T, and Mizuguchi, H
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- 2007
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8. Positive and negative regulation of adenovirus infection by CAR-like soluble protein, CLSP
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Kawabata, K, Tashiro, K, Sakurai, F, Osada, N, Kusuda, J, Hayakawa, T, Yamanishi, K, and Mizuguchi, H
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- 2007
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9. Fiber-modified adenovirus vectors containing the TAT peptide derived from HIV-1 in the fiber knob have efficient gene transfer activity
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Kurachi, S, Tashiro, K, Sakurai, F, Sakurai, H, Kawabata, K, Yayama, K, Okamoto, H, Nakagawa, S, and Mizuguchi, H
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- 2007
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10. Downregulation of human CD46 by adenovirus serotype 35 vectors
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Sakurai, F, Akitomo, K, Kawabata, K, Hayakawa, T, and Mizuguchi, H
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- 2007
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11. Characterization of capsid-modified adenovirus vectors containing heterologous peptides in the fiber knob, protein IX, or hexon
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Kurachi, S, Koizumi, N, Sakurai, F, Kawabata, K, Sakurai, H, Nakagawa, S, Hayakawa, T, and Mizuguchi, H
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- 2007
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12. Adenovirus serotype 35 vector-mediated transduction into human CD46-transgenic mice
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Sakurai, F, Kawabata, K, Koizumi, N, Inoue, N, Okabe, M, Yamaguchi, T, Hayakawa, T, and Mizuguchi, H
- Published
- 2006
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13. Optimization of adenovirus serotype 35 vectors for efficient transduction in human hematopoietic progenitors: comparison of promoter activities
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Sakurai, F, Kawabata, K, Yamaguchi, T, Hayakawa, T, and Mizuguchi, H
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- 2005
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14. Efficient gene transfer into human CD34+ cells by an adenovirus type 35 vector
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Sakurai, F, Mizuguchi, H, and Hayakawa, T
- Published
- 2003
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15. The role of tissue macrophages in the induction of proinflammatory cytokine production following intravenous injection of lipoplexes
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Sakurai, F, Terada, T, Yasuda, K, Yamashita, F, Takakura, Y, and Hashida, M
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- 2002
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16. Interaction between DNA–cationic liposome complexes and erythrocytes is an important factor in systemic gene transfer via the intravenous route in mice: the role of the neutral helper lipid
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Sakurai, F, Nishioka, T, Saito, H, Baba, T, Okuda, A, Matsumoto, O, Taga, T, Yamashita, F, Takakura, Y, and Hashida, M
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- 2001
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17. A MicroRNA Derived from Adenovirus Virus-Associated RNAII Promotes Virus Infection via Posttranscriptional Gene Silencing
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Wakabayashi, K., primary, Machitani, M., additional, Tachibana, M., additional, Sakurai, F., additional, and Mizuguchi, H., additional
- Published
- 2019
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18. P1.03-006 Clinicopathological Features and Poor Outcome for ALK Inhibitors of Squamous Cell Lung Cancer with ALK-Rearrangement
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Motomura, H., primary, Watanabe, J., additional, Togo, S., additional, Sumiyoshi, I., additional, Namba, Y., additional, Suina, K., additional, Mizuno, T., additional, Kadoya, K., additional, Iwai, M., additional, Nagaoka, T., additional, Sasaki, S., additional, Hayashi, T., additional, Uekusa, T., additional, Abe, K., additional, Urata, Y., additional, Sakurai, F., additional, Mizuguchi, H., additional, Kato, S., additional, and Takahashi, K., additional
- Published
- 2017
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19. NF-κB promotes leaky expression of adenovirus genes in a replication-incompetent adenovirus vector
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Machitani, M., primary, Sakurai, F., additional, Wakabayashi, K., additional, Nakatani, K., additional, Shimizu, K., additional, Tachibana, M., additional, and Mizuguchi, H., additional
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- 2016
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20. Proposal of A New Structure Thermal Vacuum Sensor with Diode-Thermistors Combined with a Micro-Air-Bridge Heater
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Kimura, M., Sakurai, F., Ohta, H., Terada, T., and Publishing Association, EDA
- Subjects
thermistor ,[INFO.INFO-AR] Computer Science [cs]/Hardware Architecture [cs.AR] ,microheater ,silicon ,temperature sensor ,pn diode ,Vacuum sensor - Abstract
A new structure thermal vacuum sensor with two pn junction diodes, Th-A and Th-B, as a high sensitive temperature sensor working like a thermistor and a micro heater formed on a micro air‒bridge is proposed. The micro air‒bridge is separated into two regions of A and B. The Th-A measures the temperature of the micro-heater formed on the region A and the Th-B does that of the region B which is connected to the micro-heater with thermal resistance. The diode thermistor, Th-C, formed on the SOI substrate is provided to measure the ambient temperature Tc. Vacuum pressure is measured based on heat dissipation change from the heated micro air‒bridge to ambient gas. It is demonstrated that this sensor will expand the measuring vacuum range into about two orders or more wider range (2x10-3-1x105 Pa) than the traditional thermal vacuum sensor, such as Pirani vacuum sensor, and has very fast response and low power consumption.
- Published
- 2005
21. Efficient antitumor effects of carrier cells loaded with a fiber-substituted conditionally replicating adenovirus on CAR-negative tumor cells
- Author
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Iguchi, K, primary, Sakurai, F, additional, Tomita, K, additional, Katayama, K, additional, Yamaguchi, T, additional, Kawabata, K, additional, Tagawa, M, additional, Kawabata, M, additional, Shirakawa, T, additional, and Mizuguchi, H, additional
- Published
- 2011
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22. Effects of Sophy β-glucan on growth performance, carcass traits, meat composition, and immunological responses of Peking ducks
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Tang, X.Y., primary, Gao, J.S., additional, Yuan, F., additional, Zhang, W.X., additional, Shao, Y.J., additional, Sakurai, F., additional, and Li, Z.D., additional
- Published
- 2011
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23. Adenovirus serotype 35 vector-mediated transduction following direct administration into organs of nonhuman primates
- Author
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Sakurai, F, primary, Nakamura, S-i, additional, Akitomo, K, additional, Shibata, H, additional, Terao, K, additional, Kawabata, K, additional, Hayakawa, T, additional, and Mizuguchi, H, additional
- Published
- 2008
- Full Text
- View/download PDF
24. Characterization of capsid-modified adenovirus vectors containing heterologous peptides in the fiber knob, protein IX, or hexon
- Author
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Kurachi, S, primary, Koizumi, N, additional, Sakurai, F, additional, Kawabata, K, additional, Sakurai, H, additional, Nakagawa, S, additional, Hayakawa, T, additional, and Mizuguchi, H, additional
- Published
- 2006
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- View/download PDF
25. Efficient gene transfer into human CD34+ cells by an adenovirus type 35 vector.
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Sakurai, F, Mizuguchi, H, and Hayakawa, T
- Subjects
- *
GENETIC transformation , *HEMATOPOIETIC stem cells , *ADENOVIRUSES , *COXSACKIEVIRUSES , *GENE therapy - Abstract
Efficient gene transfer into human hematopoietic stem cells (HSCs) is the most important requirement for gene therapy of hematopoietic disorders and for study of the hematopoietic system. An adenovirus (Ad) vector based on the Ad serotype 5 (Ad5) is known to transduce HSCs, including CD34[SUP+] cells, with very low efficiency because of low-level expression of its primary receptor, coxsackievirus and adenovirus receptor (CAR). In the present study, we developed a recombinant Ad vector composed of the whole Ad serotype 35 (Ad35), which recognizes an unidentified receptor different from CAR for its infection. A transduction study showed that the Ad35-based vectors exhibit a higher transduction efficiency in human CD34[SUP+] cells than the conventional Ad5 vectors and the Ad5F35 vectors, which are fiber-substituted Ad5 vectors containing Ad35 fiber proteins. The mean of fluorescence intensity in the CD34[SUP+] cells transduced with the Ad35 vectors was 12-76 and 1.4-3 times higher than that in the cells transduced with the Ad5 and Ad5F35 vectors, respectively. The percentages of green fluorescent protein (GFP)-positive CD34[SUP+] cells by transduction with Ad35, Ad5, and Ad5F35 vectors expressing GFP at 300 PFU/cell were 53%, 5%, and 52%, respectively, suggesting that Ad35 vectors mediate a more efficient gene transfer into human CD34[SUP+] cells than Ad5 and Ad5F35 vectors, although the percentage of transduced cells was similar between Ad35 and Ad5F35 vectors. The Ad vector based on Ad35 could be very useful in gene therapy for blood disorders and gene transfer experiments using HSCs. [ABSTRACT FROM AUTHOR]
- Published
- 2003
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26. Recent improvements in the acid plant at Naoshima smelter.
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Sakurai F., Third International Symposium held at the TMS Annual Meeting Seattle, Washington 17-Feb-0220-Feb-02, Yagishita S., Sakurai F., Third International Symposium held at the TMS Annual Meeting Seattle, Washington 17-Feb-0220-Feb-02, and Yagishita S.
- Abstract
The sulphuric acid plants at the Mitsubishi process plant in Naoshima, Japan, have been in operation for more than 30 years. The plants were modernised between 1999 and 2001 by renewing some of the main equipment, including the drying tower and gas heat exchangers, and by introducing new technology. Electricity consumption was substantially reduced by decreasing the volume of gas treated, the use of a low-temperature activity catalyst in the first converter pass and optimization of the main gas blower runner. The oxygen plant was replaced and enlarged. Anode production increased from 240 000 to 280 000 tonnes per year with an associated increase in sulphuric acid production from 550 000 to 660 000 tonnes per year., The sulphuric acid plants at the Mitsubishi process plant in Naoshima, Japan, have been in operation for more than 30 years. The plants were modernised between 1999 and 2001 by renewing some of the main equipment, including the drying tower and gas heat exchangers, and by introducing new technology. Electricity consumption was substantially reduced by decreasing the volume of gas treated, the use of a low-temperature activity catalyst in the first converter pass and optimization of the main gas blower runner. The oxygen plant was replaced and enlarged. Anode production increased from 240 000 to 280 000 tonnes per year with an associated increase in sulphuric acid production from 550 000 to 660 000 tonnes per year.
27. A microRNA derived from adenovirus virus-associated RNAII promotes virus infection via post-transcriptional gene silencing.
- Author
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Wakabayashi, K., Machitani, M., Tachibana, M., Sakurai, F., and Mizuguchi, H.
- Subjects
- *
MICRORNA , *ADENOVIRUS diseases , *GENE silencing , *VIRAL disease treatment , *RNA polymerases , *ANTIVIRAL agents , *VIRAL replication , *VIRUSES - Abstract
The adenovirus (Ad) serotype 5 genome encodes two non-coding small RNAs (virus-associated RNAs: VA-RNAI and II), which are approximately 160nt-long RNAs transcribed by RNA polymerase III. It is well-known that VA-RNAI supports Ad infection via the inhibition of double-stranded RNA-dependent protein kinase (PKR), which recognizes double-stranded RNA and acts as an antiviral system. Recent studies revealed that VA-RNAs are processed into VA-RNA-derived microRNAs (miRNAs) (mivaRNAI, II); however, we and another group recently demonstrated that mivaRNAI does not promote Ad replication. On the other hand, the roles of VA-RNAII and VA-RNAII-derived miRNA (mivaRNAII) in Ad replication have remained to be clarified. In this study, we demonstrate mivaRNAII-mediated promotion of Ad replication. Transfection with chemically synthesized 3'-mivaRNAII-138, one of the most abundant mivaRNAII, significantly enhanced Ad replication, while the other species of mivaRNAII did not. We identified 8 putative target genes of 3'-mivaRNAII-138 by microarray analysis and in silico analysis. Among the 8 candidates, knockdown of the cullin4A (CUL4A) gene, which encodes a component of the ubiquitin ligase complex, most significantly enhanced Ad replication. CUL4A expression was significantly suppressed by 3'-mivaRNAII-138 via post-transcriptional gene silencing, indicating that CUL4A is a target gene of 3'-mivaRNAII-138 and mivaRNAII functions as a viral miRNA promoting Ad infection. It has been reported that CUL4A is involved in degradation of c-Jun, which acts as a transcription factor in the Jun-N-terminal kinase (JNK) signaling cascade. Treatment with JNK inhibitors dramatically suppressed Ad replication, suggesting that mivaRNAII-mediated down-regulation of CUL4A enhanced JNK signaling, and thereby promoted Ad infection. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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28. Novel conditionally replicating adenovirus-mediated efficient detection of circulating tumor cells in lung cancer patients.
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Ikemoto S, Sakurai F, Tokuoka S, Yamashita T, Takayama K, Hoshi K, Okabe T, Sumiyoshi I, Togo S, Takahashi K, Tachibana M, and Mizuguchi H
- Subjects
- Humans, Adenoviridae physiology, Green Fluorescent Proteins genetics, Green Fluorescent Proteins metabolism, Tumor Cells, Cultured, Cell Line, Tumor, Neoplastic Cells, Circulating pathology, Lung Neoplasms genetics, Lung Neoplasms pathology
- Abstract
Circulating tumor cells (CTCs) are present in the blood of cancer patients from the early stage of cancer development, and their presence has been correlated with patient prognosis and treatment responses. Accordingly, CTCs have been attracting attention as a novel biomarker for early detection of cancer and monitoring of treatment responses. However, since patients typically have only a few CTCs per milliliter of blood, development of an accurate and highly sensitive CTC detection method is crucial. We previously developed a CTC detection method using a novel conditionally replicating adenovirus (Ad) that expresses green fluorescence protein (GFP) in a tumor cell-specific manner by expressing the E1 gene using a tumor-specific human telomerase reverse transcriptase (hTERT) promoter (rAdF35-142T-GFP). CTCs were efficiently detected using rAdF35-142T-GFP, but GFP expression levels in the CTCs and production efficiencies of rAdF35-142T-GFP were relatively low. In this study, in order to overcome these problems, we developed four types of novel GFP-expressing conditionally replicating Ads and examined their ability to visualize CTCs in the blood samples of lung cancer patients. Among the four types of novel recombinant Ads, the novel conditionally replicating Ad containing the 2A peptide and the GFP gene downstream of the E1A gene and the adenovirus death protein (ADP) gene in the E3 region (rAdF35-E1-2A-GFP-ADP) mediated the highest number of GFP-positive cells in the human cultured tumor cell lines. Titers of rAdF35-E1-2A-GFP-ADP were significantly higher (about 4-fold) than those of rAdF35-142T-GFP. rAdF35-E1-2A-GFP-ADP and rAdF35-142T-GFP efficiently detected CTCs in the blood of lung cancer patients at similar levels. GFP+/CD45- cells (CTCs) were found in 10 of 17 patients (58.8%) for both types of recombinant Ads., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Ikemoto et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
- Published
- 2023
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29. miR-27b targets MAIP1 to mediate lipid accumulation in cultured human and mouse hepatic cells.
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Sakai E, Imaizumi T, Suzuki R, Taracena-Gándara M, Fujimoto T, Sakurai F, and Mizuguchi H
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- Animals, Humans, Mice, Hepatocytes metabolism, Lipid Metabolism genetics, Lipids, Phosphoprotein Phosphatases metabolism, MicroRNAs genetics, MicroRNAs metabolism, Non-alcoholic Fatty Liver Disease genetics, Non-alcoholic Fatty Liver Disease metabolism
- Abstract
Non-alcoholic liver disease (NAFLD) is a condition caused by excessive fat accumulation in the liver and developed via multiple pathways. miR-27b has been suggested to play crucial roles in the development of NAFLD, assuming via targeting genes involved in lipid catabolism and anabolism. However, other pathways regulated by miR-27b are largely unknown. Here we show that lipid accumulation was induced in miR-27b-transfected human and mouse hepatic cells and that knockdowns of three miR-27b-target genes, β-1,4-galactosyltransferase 3 (B4GALT3), matrix AAA peptidase interacting protein 1 (MAIP1) and PH domain and leucine rich repeat protein phosphatase 2 (PHLPP2), induced lipid accumulation. We also show that B4GALT3 and MAIP1 were direct targets of miR-27b and overexpression of MAIP1 ameliorated miR-27b-induced lipid accumulation. In addition, we show that hepatic Maip1 expression declined in mice fed a high-fat diet, suggesting the involvement of decreased Maip1 expression in the condition of fatty liver. Overall, we identified MAIP1/miR-27b axis as a mediator of hepatic lipid accumulation, a potential therapeutic target for NAFLD., (© 2023. The Author(s).)
- Published
- 2023
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30. Modified method for differentiation of myeloid-derived suppressor cells in vitro enhances immunosuppressive ability via glutathione metabolism.
- Author
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Zhou H, Xie Z, Morikawa N, Sakurai F, Mizuguchi H, Okuzaki D, Okada N, and Tachibana M
- Abstract
Myeloid-derived suppressor cells (MDSCs), which accumulate in tumor bearers, are known to suppress anti-tumor immunity and thus promote tumor progression. MDSCs are considered a major cause of resistance against immune checkpoint inhibitors in patients with cancer. Therefore, MDSCs are potential targets in cancer immunotherapy. In this study, we modified an in vitro method of MDSC differentiation. Upon stimulating bone marrow (BM) cells with granulocyte-macrophage colony-stimulating factor in vitro , we obtained both lymphocyte antigen 6G positive (Ly-6G
+ ) and negative (Ly-6G- ) MDSCs (collectively, hereafter referred to as conventional MDSCs), which were non-immunosuppressive and immunosuppressive, respectively. We then found that MDSCs differentiated from Ly-6G- BM (hereafter called 6G- BM-MDSC) suppressed T-cell proliferation more strongly than conventional MDSCs, whereas the cells differentiated from Ly-6G+ BM (hereafter called 6G+ BM-MDSC) were non-immunosuppressive. In line with this, conventional MDSCs or 6G- BM-MDSC, but not 6G+ BM-MDSC, promoted tumor progression in tumor-bearing mice. Moreover, we identified that activated glutathione metabolism was responsible for the enhanced immunosuppressive ability of 6G- BM-MDSC. Finally, we showed that Ly-6G+ cells in 6G- BM-MDSC, which exhibited weak immunosuppression, expressed higher levels of Cybb mRNA, an immunosuppressive gene of MDSCs, than 6G+ BM-MDSC. Together, these data suggest that the depletion of Ly-6G+ cells from the BM cells leads to differentiation of immunosuppressive Ly-6G+ MDSCs. In summary, we propose a better method for MDSC differentiation in vitro . Moreover, our findings contribute to the understanding of MDSC subpopulations and provide a basis for further research on MDSCs., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Authors.)- Published
- 2022
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31. Effects of pre-existing anti-adenovirus antibodies on transgene expression levels and therapeutic efficacies of arming oncolytic adenovirus.
- Author
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Ono R, Nishimae F, Wakida T, Sakurai F, and Mizuguchi H
- Subjects
- Humans, Mice, Animals, Adenoviridae genetics, Genetic Vectors genetics, Seroepidemiologic Studies, Transgenes, Antibodies, Viral, Luciferases genetics, Antibodies, Neutralizing genetics, Adenoviridae Infections genetics, Adenoviruses, Human genetics, Neoplasms therapy, Neoplasms genetics
- Abstract
Oncolytic adenoviruses (OAds), most of which are based on species C human adenovirus serotype 5 (Ad5) (OAd5), have recently received much attention as potential anticancer agents. High seroprevalence of anti-Ad5 neutralizing antibodies is a major hurdle for Ad5-based gene therapy. However, the impacts of anti-Ad5 neutralizing antibodies on OAd5-mediated transgene expression in the tumor and antitumor effects remain to be fully elucidated. In this study, we examined the impact of anti-Ad5 neutralizing antibodies on the OAd5-mediated antitumor effects and OAd5-mediated transgene expression. The luciferase expression of OAd-tAIB-Luc, which contains the cytomegalovirus promoter-driven luciferase gene, was inhibited in human cultured cells in the presence of human serum. Although the inhibitory effects of human serum possessing the low anti-Ad5 neutralizing antibody titers were overcome by long-term infection, the in vitro tumor cell lysis activities of OAd-tAIB-Luc were entirely attenuated by human serum containing the high titers of anti-Ad5 neutralizing antibodies. OAd-tAIB-Luc-mediated luciferase expression in the subcutaneous tumors 3 days after administration and tumor growth suppression levels following intratumoral administration were significantly lower in mice possessing the high titers of anti-Ad5 neutralizing antibodies, compared to those in control mice. These results suggested that pre-existing anti-Ad5 antibodies attenuated both transgene expression and potential antitumor effects of OAd5 following intratumoral administration., (© 2022. The Author(s).)
- Published
- 2022
- Full Text
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32. Liver-specific overexpression of lipoprotein lipase improves glucose metabolism in high-fat diet-fed mice.
- Author
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Shimizu K, Nishimuta S, Fukumura Y, Michinaga S, Egusa Y, Hase T, Terada T, Sakurai F, Mizuguchi H, Tomita K, and Nishinaka T
- Subjects
- Animals, Diet, High-Fat, Glucose metabolism, Lipoprotein Lipase genetics, Lipoprotein Lipase metabolism, Liver metabolism, Mice, Mice, Inbred C57BL, Triglycerides metabolism, Diabetes Mellitus, Type 2 metabolism, Insulin Resistance physiology
- Abstract
The liver is the main organ that regulates lipid and glucose metabolism. Ectopic lipid accumulation in the liver impairs insulin sensitivity and glucose metabolism. Lipoprotein lipase (LPL), mainly expressed in the adipose tissue and muscle, is a key enzyme that regulates lipid metabolism via the hydrolysis of triglyceride in chylomicrons and very-low-density lipoproteins. Here, we aimed to investigate whether the suppression level of hepatic lipid accumulation via overexpression of LPL in mouse liver leads to improved metabolism. To overexpress LPL in the liver, we generated an LPL-expressing adenovirus (Ad) vector using an improved Ad vector that exhibited considerably lower hepatotoxicity (Ad-LPL). C57BL/6 mice were treated with Ad vectors and simultaneously fed a high-fat diet (HFD). Lipid droplet formation in the liver decreased in Ad-LPL-treated mice relative to that in control Ad vector-treated mice. Glucose tolerance and insulin resistance were remarkably improved in Ad-LPL-treated mice compared to those in control Ad vector-treated mice. The expression levels of fatty acid oxidation-related genes, such as peroxisome proliferator-activated receptor α, carnitine palmitoyltransferase 1, and acyl-CoA oxidase 1, were 1.7-2.0-fold higher in Ad-LPL-treated mouse livers than that in control Ad-vector-treated mouse livers. Furthermore, hepatic LPL overexpression partly maintained mitochondrial content in HFD-fed mice. These results indicate that LPL overexpression in the livers of HFD-fed mice attenuates the accumulation of lipid droplets in the liver and improves glucose metabolism. These findings may enable the development of new drugs to treat metabolic syndromes such as type 2 diabetes mellitus and non-alcoholic fatty liver disease., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2022
- Full Text
- View/download PDF
33. Real-world efficacy of sequential nivolumab for metastatic renal cancer after first-line molecular targeting therapy.
- Author
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Obinata D, Funakoshi D, Sakurai F, Yoshizawa T, Mochida J, Yamaguchi K, and Takahashi S
- Subjects
- Humans, Molecular Targeted Therapy, Nivolumab, Retrospective Studies, Antineoplastic Agents, Immunological therapeutic use, Carcinoma, Renal Cell pathology, Kidney Neoplasms pathology
- Abstract
This study aimed to clarify the real-world efficacy of sequential nivolumab for treating metastatic renal cancer after first-line molecular targeting therapy. Patients were divided into two groups (2014-2016 and 2017-2020) according to the year when they started primary treatment with molecular targeted drugs (MTDs). We compared the overall survival of patients and investigated a contributing factor for survival. The mean duration of overall survival was significantly longer in the 2017-2020 group (44.0 months) than in the 2014-2016 group (8.5 months). Univariate analysis showed that nivolumab treatment was a significant prognostic factor (P = .0021). Patients treated with nivolumab as second-line therapy had a significantly higher 5-year survival rate compared to that of other patients (70% vs 32%). In addition, the time from commencement of MTDs to switch to nivolumab was significantly shorter in the 2017-2020 group compared to the 2014-2016 group (8.94 vs 34.12 months, P = .03). In our study, cases with first-line MTDs had markedly prolonged outcomes after the 2017 guideline update, and sequential nivolumab with prompt switching to nivolumab was an important factor., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2022 the Author(s). Published by Wolters Kluwer Health, Inc.)
- Published
- 2022
- Full Text
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34. Tumor-targeted fluorescence labeling systems for cancer diagnosis and treatment.
- Author
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Tazawa H, Shigeyasu K, Noma K, Kagawa S, Sakurai F, Mizuguchi H, Kobayashi H, Imamura T, and Fujiwara T
- Subjects
- Cell Line, Tumor, Epithelial-Mesenchymal Transition, Fluorescence, Humans, Tissue Distribution, Neoplasms diagnosis, Neoplasms therapy, Telomerase metabolism
- Abstract
Conventional imaging techniques are available for clinical identification of tumor sites. However, detecting metastatic tumor cells that are spreading from primary tumor sites using conventional imaging techniques remains difficult. In contrast, fluorescence-based labeling systems are useful tools for detecting tumor cells at the single-cell level in cancer research. The ability to detect fluorescent-labeled tumor cells enables investigations of the biodistribution of tumor cells for the diagnosis and treatment of cancer. For example, the presence of fluorescent tumor cells in the peripheral blood of cancer patients is a predictive biomarker for early diagnosis of distant metastasis. The elimination of fluorescent tumor cells without damaging normal tissues is ideal for minimally invasive treatment of cancer. To capture fluorescent tumor cells within normal tissues, however, tumor-specific activated target molecules are needed. This review focuses on recent advances in tumor-targeted fluorescence labeling systems, in which indirect reporter labeling using tumor-specific promoters is applied to fluorescence labeling of tumor cells for the diagnosis and treatment of cancer. Telomerase promoter-dependent fluorescence labeling using replication-competent viral vectors produces fluorescent proteins that can be used to detect and eliminate telomerase-positive tumor cells. Tissue-specific promoter-dependent fluorescence labeling enables identification of specific tumor cells. Vimentin promoter-dependent fluorescence labeling is a useful tool for identifying tumor cells that undergo epithelial-mesenchymal transition (EMT). The evaluation of tumor cells undergoing EMT is important for accurately assessing metastatic potential. Thus, tumor-targeted fluorescence labeling systems represent novel platforms that enable the capture of tumor cells for the diagnosis and treatment of cancer., (© 2022 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.)
- Published
- 2022
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- View/download PDF
35. miR-27b antagonizes BMP signaling in early differentiation of human induced pluripotent stem cells.
- Author
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Lim J, Sakai E, Sakurai F, and Mizuguchi H
- Subjects
- CRISPR-Cas Systems, Endoderm metabolism, Humans, Mesoderm metabolism, MicroRNAs genetics, RNA Interference, Bone Morphogenetic Proteins genetics, Bone Morphogenetic Proteins metabolism, Cell Differentiation genetics, Gene Expression Regulation, Developmental, Induced Pluripotent Stem Cells cytology, Induced Pluripotent Stem Cells metabolism, Signal Transduction
- Abstract
Human induced pluripotent stem (hiPS) cells are feasible materials for studying the biological mechanisms underlying human embryogenesis. In early embryogenesis, definitive endoderm and mesoderm are differentiated from their common precursor, mesendoderm. Bone morphogenetic protein (BMP) signaling is responsible for regulating mesendoderm and mesoderm formation. Micro RNAs (miRNAs), short non-coding RNAs, broadly regulate biological processes via post-transcriptional repression. The expression of miR-27b, which is enriched in somatic cells, has been reported to increase through definitive endoderm and hepatic differentiation, but little is known about how miR-27b acts during early differentiation. Here, we used miR-27b-inducible hiPS cells to investigate the roles of miR-27b in the undifferentiated and early-differentiated stages. In undifferentiated hiPS cells, miR-27b suppressed the expression of pluripotency markers [alkaline phosphatase (AP) and nanog homeobox (NANOG)] and cell proliferation. Once differentiation began, miR-27b expression repressed phosphorylated SMAD1/5, the mediators of the BMP signaling, throughout definitive endoderm differentiation. Consistent with the above findings, miR-27b overexpression downregulated BMP-induced mesendodermal marker genes [Brachyury, mix paired-like homeobox 1 (MIXL1) and eomesodermin (EOMES)], suggesting that miR-27b had an inhibitory effect on early differentiation. Collectively, our findings revealed a novel antagonistic role of miR-27b in the BMP signaling pathway in the early differentiation of hiPS cells., (© 2021. The Author(s).)
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- 2021
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36. A TGFβ Signaling Inhibitor, SB431542, Inhibits Reovirus-mediated Lysis of Human Hepatocellular Carcinoma Cells in a TGFβ-independent Manner.
- Author
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Ishigami I, Shuwari N, Kaminade T, Mizuguchi H, and Sakurai F
- Subjects
- Carcinoma, Hepatocellular genetics, Carcinoma, Hepatocellular pathology, Carcinoma, Hepatocellular virology, Cell Survival drug effects, Epoxy Compounds, Humans, Liver Neoplasms genetics, Liver Neoplasms pathology, Liver Neoplasms virology, Orthoreovirus, Mammalian drug effects, Orthoreovirus, Mammalian genetics, Pyrazoles pharmacology, Quinolines pharmacology, RNA, Double-Stranded genetics, Signal Transduction drug effects, Transforming Growth Factor beta1 antagonists & inhibitors, Tyrosine analogs & derivatives, Tyrosine genetics, Benzamides pharmacology, Carcinoma, Hepatocellular drug therapy, Dioxoles pharmacology, Liver Neoplasms drug therapy, Transforming Growth Factor beta1 genetics
- Abstract
Background/aim: Oncolytic reovirus, which is a non-enveloped virus possessing a 10-segmented double-stranded RNA genome, has been anticipated as a novel class of antitumor agent. Hepatocellular carcinoma (HCC) is considered to be a target suitable for reovirus-mediated virotherapy. Transforming growth factor (TGF)-β plays an important role in the pathogenesis of HCC. TGF-β-signaling inhibitors have proceeded to clinical trials as potential antitumor agents for HCC. On the other hand, TGF-β is involved in induction of expression of cathepsins B and L, which are important for reovirus infection. It remains to be examined whether TGF-β signaling inhibitors affect reovirus-mediated lysis of HCC cells. The aim of this study was to evaluate the effects of TGF-β-signaling inhibitors on tumor cell lysis efficiency of reovirus in human HCC cells., Materials and Methods: Reovirus was added to four types of human HCC cell lines pretreated with one of three TGF-β type I receptor inhibitors: SB431542, A-83-01, or galunisertib (LY2157299). Cell viability, virus genome copy numbers, and virus protein expression were evaluated following reovirus infection., Results: SB431542 significantly inhibited reovirus-mediated killing of human HCC cell lines, while A-83-01 and galunisertib did not inhibit., Conclusion: These data indicate that SB431542 inhibited reovirus-mediated lysis of human HCC cells in a TGF-β signaling-independent manner., (Copyright © 2021 International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)
- Published
- 2021
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37. The infectivity of progeny adenovirus in the presence of neutralizing antibody.
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Hirai T, Sato A, Koizumi N, Kurioka Y, Suzuki Y, Kano J, Yamakawa M, Nomura T, Fujii M, Sakurai F, Mizuguchi H, Watanabe Y, and Utoguchi N
- Subjects
- Genes, Reporter, Genetic Vectors, HEK293 Cells, Humans, Adenovirus Infections, Human virology, Adenoviruses, Human pathogenicity, Antibodies, Neutralizing immunology, Virulence immunology
- Abstract
Human adenoviruses (Ads), common pathogens that cause upper respiratory and gastrointestinal infections, are blocked by neutralizing antibodies (nAbs). However, Ads are not fully eliminated even in hosts with nAbs. In this study, we assessed the infectivity of progeny Ad serotype 5 (Ad5) in the presence of nAb. The infectivity of Ad5 was evaluated according to the expression of the Ad genome and reporter gene. Infection by wild-type Ad5 and Ad5 vector continued to increase until 3 days after infection even in the presence of nAb. We established an assay for determining the infection levels of progeny Ad5 using a sorting system with magnetic beads and observed little difference in progeny Ad5 counts in the presence and absence of nAb 1 day after infection. Moreover, progeny Ad5 in the presence of nAb more effectively infected coxsackievirus and adenovirus receptor (CAR)-positive cells than CAR-negative cells. We investigated the function of fiber proteins, which are the binding partners of CAR, during secondary infection, observing that fibre proteins spread from infected cells to adjacent cells in a CAR-dependent manner. In conclusion, this study revealed that progeny Ad5 could infect cells even in the presence of nAb, differing from the common features of the Ad5 infection cycle. Our findings may be useful for developing new therapeutic agents against Ad infection.
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- 2021
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38. Optimization of an E1A Gene Expression Cassette in an Oncolytic Adenovirus for Efficient Tumor Cell Killing Activity.
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Sakurai F, Nishimae F, Takayama K, and Mizuguchi H
- Subjects
- Adenoviridae genetics, Apoptosis, Cell Line, Tumor, Gene Expression, HCT116 Cells, HEK293 Cells, HeLa Cells, Hep G2 Cells, Human Umbilical Vein Endothelial Cells, Humans, MCF-7 Cells, Oncolytic Virotherapy, Oncolytic Viruses genetics, Oncolytic Viruses physiology, Promoter Regions, Genetic, Virus Replication, Adenoviridae physiology, Adenovirus E1A Proteins genetics, Sequence Deletion, Survivin genetics, Telomerase genetics
- Abstract
Background/aim: Oncolytic adenoviruses (OAds) have attracted much attention as novel anticancer therapeutics. The proper design of an expression cassette containing the E1A gene, which is indispensable for self-replication of the Ad genome, is crucial for efficient tumor cell-specific infection of an OAd. Various types of oncolytic adenoviruses (OAds) possessing different types of the E1A gene expression cassettes have been developed, but their oncolytic activities and safety profiles have not been systematically evaluated. Herein we examined the oncolytic activities and safety profiles of five types of OAds possessing different types of the E1A gene expression cassette in order to optimize the E1A gene expression cassette for development of an efficient and safe OAd., Materials and Methods: We prepared five types of OAds containing different types of E1 gene expression cassettes, and examined the oncolytic activities and safety profiles of the OAds., Results: Among the OAds examined, OAd-Δ24, which had a 24-bp deletion in the E1A gene, mediated the most efficient oncolytic activities against the human tumor cell lines, although OAd-Δ24 showed slightly higher cytotoxicity to normal human cells than the other OAds., Conclusion: These results provide important clues for the development of safe and efficient OAds., (Copyright © 2021 International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)
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- 2021
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39. Efficient antitumor effects of a novel oncolytic adenovirus fully composed of species B adenovirus serotype 35.
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Ono R, Takayama K, Sakurai F, and Mizuguchi H
- Abstract
Oncolytic adenoviruses (OAds) are among the most promising oncolytic viruses. Almost all oncolytic adenoviruses are composed of human adenovirus serotype 5 (Ad5) (OAd5). However, expression of the primary infection receptor for Ad5, coxsackievirus-adenovirus receptor (CAR), often declines on malignant tumor cells, resulting in inefficient infection in CAR-negative tumor cells. In addition, at least 80% of adults have neutralizing antibodies against Ad5. In this study, we developed a novel OAd fully composed of OAd35. OAd35 recognizes CD46, which is ubiquitously expressed on almost all human cells and is often upregulated on malignant tumor cells, as an infection receptor. Moreover, 20% or fewer adults have neutralizing antibodies against Ad35. OAd35 mediated efficient cell lysis activities at levels similar to OAd5 in CAR-positive tumor cells, while OAd35 showed higher levels of cell lysis activities than OAd5 in CAR-negative tumor cells. Anti-Ad5 serum significantly inhibited in vitro tumor cell lysis activities of OAd5, whereas OAd35 exhibited comparable levels of in vitro tumor cell lysis activities in the presence of anti-Ad5 and naive serum. OAd35 significantly suppressed growth of the subcutaneous CAR-positive and CAR-negative tumors following intratumoral administration. These results indicated that OAd35 is a promising alternative oncolytic virus for OAd5., Competing Interests: F.S. and H.M. have the potential to receive patent royalities from Oncolys BioPharm, Inc. The other authors declare no conflicts of interest., (© 2021 The Author(s).)
- Published
- 2021
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40. Transduction Properties of an Adenovirus Vector Containing Sequences Complementary to a Liver-Specific microRNA, miR-122a, in the 3'-Untranslated Region of the E4 Gene in Human Hepatocytes from Chimeric Mice with Humanized Liver.
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Sakurai F, Tsukamoto T, Ono R, Nishimae F, Shiota A, Iizuka S, Shimizu K, Sakai E, Ishida Y, Tateno C, Chayama K, and Mizuguchi H
- Subjects
- 3' Untranslated Regions genetics, Animals, Genetic Therapy methods, Genetic Vectors genetics, HEK293 Cells, Hepatocytes, Humans, Mice, Promoter Regions, Genetic, Transplantation Chimera, Adenoviridae genetics, Adenovirus E4 Proteins genetics, MicroRNAs genetics, Transduction, Genetic methods
- Abstract
Replication-incompetent adenovirus (Ad) vectors are promising gene delivery vehicles, especially for hepatocytes, due to their superior hepatic tropism; however, in vivo application of an Ad vector often results in hepatotoxicity, mainly due to the leaky expression of Ad genes from the Ad vector genome. In order to reduce the Ad vector-induced hepatotoxicity, we previously developed an Ad vector containing the sequences perfectly complementary to a liver-specific microRNA (miRNA), miR-122a, in the 3'-untranslated region (UTR) of the E4 gene. This improved Ad vector showed a significant reduction in the leaky expression of Ad genes and hepatotoxicity in the mouse liver and primary mouse hepatocytes; however, the safety profiles and transduction properties of this improved Ad vector in human hepatocytes remained to be elucidated. In this study, we examined the transgene expression and safety profiles of Ad vectors with miR-122a-targeted sequences in the 3'-UTR of the E4 gene in human hepatocytes from chimeric mice with humanized liver. The transgene expression levels of Ad vectors with miR-122a-targeted sequences in the 3'-UTR of the E4 gene were significantly higher than those of the conventional Ad vectors. The leaky expression levels of Ad genes of Ad vectors with miR-122a-targeted sequences in the 3'-UTR of the E4 gene in the primary human hepatocytes were largely reduced, compared with the conventional Ad vectors, resulting in an improvement in Ad vector-induced cytotoxicity. These data indicated that this improved Ad vector was a superior gene delivery vehicle without severe cytotoxicity for not only mouse hepatocytes but also human hepatocytes.
- Published
- 2021
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41. Ablation of IL-17A leads to severe colitis in IL-10-deficient mice: implications of myeloid-derived suppressor cells and NO production.
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Tachibana M, Watanabe N, Koda Y, Oya Y, Kaminuma O, Katayama K, Fan Z, Sakurai F, Kawabata K, Hiroi T, and Mizuguchi H
- Subjects
- Animals, Body Weight, Inflammation immunology, Interleukin-10 deficiency, Interleukin-17 deficiency, Mice, Mice, Inbred C57BL, Mice, Knockout, Nitric Oxide analysis, Nitric Oxide immunology, Nitric Oxide Synthase Type II deficiency, Nitric Oxide Synthase Type II immunology, Colitis immunology, Interleukin-10 immunology, Interleukin-17 immunology, Myeloid-Derived Suppressor Cells immunology, Nitric Oxide biosynthesis
- Abstract
IL-10 is an immune regulatory cytokine and its genetic defect leads to gastrointestinal inflammation in humans and mice. Moreover, the IL-23/Th17 axis is known to be involved in these inflammatory disorders. IL-17A, a representative cytokine produced by Th17 cells, has an important role for the pathological process of inflammatory diseases. However, the precise function of IL-17A in inflammatory bowel disease (IBD) remains controversial. In this study, we evaluated the effect of IL-17A on colitis in IL-10-deficient (Il10-/-) mice. Mice lacking both IL-10 and IL-17A (Il10-/-Il17a-/-) suffered from fatal wasting and manifested more severe colitis compared with Il10-/-Il17a+/- mice. Moreover, we found that CD11b+Gr-1+ myeloid-derived suppressor cells (MDSCs) accumulated in the bone marrow, spleen and peripheral blood of Il10-/-Il17a-/- mice. These MDSCs highly expressed inducible nitric oxide synthase (iNOS) (Nos2) and suppressed the T-cell response in vitro in a NOS-dependent manner. In correlation with these effects, the concentration of nitric oxide was elevated in the serum of Il10-/-Il17a-/- mice. Surprisingly, the severe colitis observed in Il10-/-Il17a-/- mice was ameliorated in Il10-/-Il17a-/-Nos2-/- mice. Our findings suggest that IL-17A plays suppressive roles against spontaneous colitis in Il10-/- mice in an iNOS-dependent manner and inhibits MDSC differentiation and/or proliferation., (© The Japanese Society for Immunology. 2019. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2020
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42. Development of a Novel Oncolytic Adenovirus Expressing a Short-hairpin RNA Against Cullin 4A.
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Wakabayashi K, Sakurai F, Ono R, Fujiwara T, and Mizuguchi H
- Subjects
- Animals, Cell Line, Tumor, Cell Survival genetics, Gene Expression, Gene Knockdown Techniques, Humans, Mice, Oncolytic Virotherapy, RNA Interference, Transduction, Genetic, Adenoviridae genetics, Cullin Proteins genetics, Genetic Vectors genetics, Oncolytic Viruses genetics, RNA, Small Interfering genetics
- Abstract
Background: Arming of an oncolytic adenovirus (OAd) by inserting expression cassettes of therapeutic transgenes into the OAd genome is a promising approach to enhance the therapeutic effects of an OAd. Ideally, this approach would simultaneously promote the replication of an OAd in tumor cells and transgene product-mediated antitumor effects by expressing therapeutic transgenes. We previously demonstrated that knockdown of cullin 4A (CUL4A), which is an E3 ubiquitin ligase, significantly promoted adenovirus replication by increasing the c-JUN protein level. In addition, previous studies reported that CUL4A was highly expressed in various types of tumor, and was involved in tumor growth and metastasis., Materials and Methods: In this study, we developed a novel OAd expressing a short-hairpin RNA (shRNA) against CUL4A (OAd-shCUL4A)., Results: OAd-shCUL4 mediated higher levels of cytotoxic effects on various types of human tumor cell than a conventional OAd. Higher levels of OAd genome copy numbers were found in the tumor cells for OAd-shCUL4A, compared with a conventional OAd., Conclusion: OAd-shCUL4A showed efficient antitumor effects by both enhancing OAd replication and inhibiting tumor cell growth., (Copyright© 2020, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)
- Published
- 2020
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43. Efficient generation of adenovirus vectors carrying the Clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR associated proteins (Cas)12a system by suppressing Cas12a expression in packaging cells.
- Author
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Tsukamoto T, Sakai E, Nishimae F, Sakurai F, and Mizuguchi H
- Subjects
- Adenoviridae genetics, CRISPR-Cas Systems, Cell Proliferation, Gene Editing, Genetic Vectors physiology, HEK293 Cells cytology, HEK293 Cells virology, Humans, Viral Load, Adenoviridae physiology, CRISPR-Associated Proteins genetics, RNA, Guide, CRISPR-Cas Systems genetics
- Abstract
Clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR associated proteins (Cas) 9 system is a powerful tool for genome editing and still being aggressively improved. Cas12a, a recently discovered Cas9 ortholog, is expected to become complementary to Cas9 due to its unique characteristics. Previously we attempted to establish an adenovirus (Ad) vector-mediated delivery of CRISPR-Cas12a system since Ad vector is widely used for gene transfer in basic researches and medical applications. However, we found difficulties preparing of Ad vectors at an adequate titer. In this study, we have developed Ad vectors that conditionally express Cas12a either by a tetracycline-controlled promoter or a hepatocyte specific promoter to avoid putative inhibitory effects of Cas12a. These vectors successfully proliferated in packaging cells, HEK293 cells, and were recovered at high titers. We have also developed packaging cells that express shRNA for Cas12a to suppress expression of Cas12a. Using the cells, the Ad vector directing constitutive expression of Cas12a proliferated efficiently and was successfully recovered at a high titer. Overall, we improved recovery of Ad vectors carrying CRISPR-Cas12a system, thus provided them as a tool in genome editing researches., (Copyright © 2019 Elsevier B.V. All rights reserved.)
- Published
- 2019
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44. miR-27b-mediated suppression of aquaporin-11 expression in hepatocytes reduces HCV genomic RNA levels but not viral titers.
- Author
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Sakurai F, Hashimoto R, Inoue C, Wakabayashi K, Tsukamoto T, Imaizumi T, Andres TGM, Sakai E, Itsuki K, Sakamoto N, Wakita T, and Mizuguchi H
- Subjects
- Cell Line, Gene Expression Regulation, Gene Knockdown Techniques, Genome, Viral, Humans, RNA, Viral genetics, Transfection, Viral Load, Aquaporins genetics, Hepacivirus genetics, Hepatocytes virology, MicroRNAs genetics, RNA, Viral analysis
- Abstract
Background: MicroRNAs (miRNAs) have gained much attention as cellular factors regulating hepatitis C virus (HCV) infection. miR-27b has been shown to regulate HCV infection in the hepatocytes via various mechanisms that have not been fully elucidated. In this study, therefore, we examined the mechanisms of miR-27b-mediated regulation of HCV infection., Methods: In silico screening analysis, transfection with miR-27b mimic, and a cell-based reporter assay were performed to identify miR-27b target genes. Cell cultured-derived HCV (HCVcc) was added to Huh7.5.1 cells knocked down for aquaporin (AQP) 11 (AQP11) and overexpressing AQP11. HCV replication levels were evaluated by real-time RT-PCR analysis of HCVcc genome., Results: Infection of Huh7.5.1 cells with HCVcc resulted in significant elevation in miR-27b expression levels. In silico analysis revealed that AQP11, which is an AQP family member and is mainly localized in the endoplasmic reticulum (ER), was a candidate for a target gene of miR-27b. Transfection of a miR-27b mimic significantly reduced AQP11 expression, but a cell-based reporter assay demonstrated that miR-27b did not suppress the expression of a reporter gene containing the 3'-untranslated region of the AQP11 gene, suggesting that miR-27b indirectly suppressed AQP11 expression. AQP11 expression levels were significantly reduced by infection with HCVcc in Huh7.5.1 cells. Knockdown and over-expression of AQP11 significantly reduced and increased HCVcc genome levels in the cells following infection, respectively, however, AQP11 knockdown did not show significant effects on the HCVcc titers in the culture supernatants., Conclusions: These results indicated that HCV infection induced a miR-27b-mediated reduction in AQP11 expression, leading to a modest reduction in HCV genome levels in the cells, not HCV titers in the culture supernatants.
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- 2019
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45. FGF signal is not required for hepatoblast differentiation of human iPS cells.
- Author
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Toba Y, Kiso A, Nakamae S, Sakurai F, Takayama K, and Mizuguchi H
- Subjects
- Bone Morphogenetic Proteins metabolism, Cell Differentiation physiology, Endoderm cytology, Humans, Induced Pluripotent Stem Cells metabolism, Receptors, Fibroblast Growth Factor metabolism, Recombinant Proteins metabolism, Fibroblast Growth Factors metabolism, Hepatocytes metabolism, Induced Pluripotent Stem Cells physiology
- Abstract
Human induced pluripotent stem cell-derived hepatocyte-like cells are expected to be utilized in pharmaceutical research and regenerative medicine. In general, human induced pluripotent stem (iPS) cells are differentiated into hepatocyte-like cells through definitive endoderm cells and hepatoblast-like cells using various growth factors that are essential for liver development. Although recombinant bone morphogenetic proteins (BMPs) and fibroblast growth factors (FGFs) are widely used in the hepatoblast differentiation, hepatoblast differentiation process has not been fully modified. In this study, we examined the roles of BMPs and FGFs in the hepatoblast differentiation from human iPS cells. Surprisingly, the gene expression levels of hepatoblast markers were upregulated by the removal of FGFs. In addition, the percentages of hepatoblast markers-positive cells were increased by the removal of FGFs. Furthermore, the hepatocyte differentiation potency was also significantly increased by the removal of FGFs. To examine whether FGF signals are completely unnecessary for the hepatoblast differentiation, the expression levels of endogenous FGF ligands and receptors were examined. The definitive endoderm cells highly expressed the FGF ligand, FGF2, and the FGF receptor, FGFR1. To examine the role of endogenous FGF signals, an FGFR inhibitor was treated during the hepatoblast differentiation. The hepatoblast differentiation was promoted by using FGFR inhibitor, suggesting that endogenous FGF signals are also unnecessary for the hepatoblast differentiation. In conclusion, we found that FGF signals are not essential for hepatoblast differentiation. We believe that our finding will be useful for generating functional hepatocyte-like cells for medical applications.
- Published
- 2019
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46. Case of caval lobular capillary hemangioma mimicking tumor thrombus.
- Author
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Hashimoto S, Obinata D, Yamaguchi K, Sakurai F, Yoshida T, Yoshizawa T, Matsui T, Mochida J, Masuda S, and Takahashi S
- Abstract
Introduction: We presented a rare case of caval lobular capillary hemangioma., Case Presentation: A 66-year-old female visited our department complaint with shadow defect in vena cava of right renal hilum appeared on computed tomography for periodically checking 3 years after radical hysterectomy with bilateral ovariectomy. Abdominal computed tomography identified a shadow defect of 35 mm in diameter in the inferior vena cava continuing posteriorly to a 35 mm mass of retroperitoneum. During the total removal of this lesion, we identified the lesion was connected to right ovarian vein. The specimen consisted of microcapillaries which formed reticular structure. Immunostaining of specimens identified positive CD31, CD34, and Factor 8 in all cells. Ki67 antibody was positive at 2-3% of all cells. These findings suggested the tumor was intravenous lobular papillary hemangioma., Conclusion: This is the first report of intravenous lobular papillary hemangioma originated from right ovarian vein and extended to inferior vena cava., Competing Interests: The authors declare no conflict of interest., (© 2019 The Authors. IJU Case Reports published by John Wiley & Sons Australia, Ltd on behalf of the Japanese Urological Association.)
- Published
- 2019
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47. Systemically Administered Reovirus-Induced Downregulation of Hypoxia Inducible Factor-1α in Subcutaneous Tumors.
- Author
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Hotani T, Mizuguchi H, and Sakurai F
- Abstract
Reovirus, which possesses a 10-segmented double-stranded RNA genome, mediates superior antitumor effects via not only virus replication in a tumor cell-specific manner but also other mechanisms distinct from virus replication. Several groups, including ours, reported the reovirus-mediated downregulation of hypoxia inducible factor-1α (HIF-1α) following infection in cultured tumor cells; however, it remained to be clarified whether reovirus downregulates the expression of HIF-1α and its target genes in tumor-bearing hosts. We found that reovirus induced significant downregulation of protein levels of HIF-1α and its target genes in the subcutaneous tumors at 120 h post-systemic administration. Expression of reovirus capsid protein σ3 was found in the pimonidazole-positive hypoxic area in the tumor. Significant levels of tumor cell apoptosis were not found in the tumors of reovirus-treated mice at this time point, suggesting that reovirus-mediated tumor cell killing did not largely contribute to the downregulation of HIF-1α protein levels in the tumors. UV-inactivated reovirus did not induce downregulation of HIF-1α expression in the tumors, indicating that virus replication was indispensable for downregulation of HIF-1α expression in the subcutaneous tumors. This study provides important information for the development of reovirus-mediated virotherapy against various types of tumors.
- Published
- 2018
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48. Efficient Generation of Small Intestinal Epithelial-like Cells from Human iPSCs for Drug Absorption and Metabolism Studies.
- Author
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Negoro R, Takayama K, Kawai K, Harada K, Sakurai F, Hirata K, and Mizuguchi H
- Subjects
- Animals, Biomarkers metabolism, Caco-2 Cells, Carrier Proteins pharmacology, Cell Differentiation drug effects, Dexamethasone pharmacology, Epidermal Growth Factor pharmacology, Epithelial Cells drug effects, Gene Expression Regulation drug effects, Heterocyclic Compounds, 3-Ring pharmacology, Humans, Imidazoles pharmacology, Induced Pluripotent Stem Cells drug effects, Induced Pluripotent Stem Cells metabolism, Insulin-Like Growth Factor I pharmacology, Maleimides pharmacology, Mice, Pyridines pharmacology, Thrombospondins pharmacology, Wnt3A Protein pharmacology, Epithelial Cells metabolism, Induced Pluripotent Stem Cells cytology, Intestinal Absorption drug effects, Intestine, Small cytology, Pharmaceutical Preparations metabolism
- Abstract
The small intestine plays an important role in the absorption and metabolism of oral drugs. In the current evaluation system, it is difficult to predict the precise absorption and metabolism of oral drugs. In this study, we generated small intestinal epithelial-like cells from human induced pluripotent stem cells (hiPS-SIECs), which could be applied to drug absorption and metabolism studies. The small intestinal epithelial-like cells were efficiently generated from human induced pluripotent stem cell by treatment with WNT3A, R-spondin 3, Noggin, EGF, IGF-1, SB202190, and dexamethasone. The gene expression levels of small intestinal epithelial cell (SIEC) markers were similar between the hiPS-SIECs and human adult small intestine. Importantly, the gene expression levels of colonic epithelial cell markers in the hiPS-SIECs were much lower than those in human adult colon. The hiPS-SIECs generated by our protocol exerted various SIEC functions. In conclusion, the hiPS-SIECs can be utilized for evaluation of drug absorption and metabolism., (Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2018
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49. Antibodies against adenovirus fiber and penton base proteins inhibit adenovirus vector-mediated transduction in the liver following systemic administration.
- Author
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Tomita K, Sakurai F, Iizuka S, Hemmi M, Wakabayashi K, Machitani M, Tachibana M, Katayama K, Kamada H, and Mizuguchi H
- Subjects
- Adenoviridae genetics, Animals, Female, Gene Dosage genetics, Genetic Vectors genetics, Mice, Mice, Inbred C57BL, Plasmids genetics, Transduction, Genetic methods, Adenoviridae immunology, Antibodies, Viral immunology, Capsid Proteins immunology, Liver metabolism
- Abstract
Pre-existing anti-adenovirus (Ad) neutralizing antibodies (AdNAbs) are a major barrier in clinical gene therapy using Ad vectors and oncolytic Ads; however, it has not been fully elucidated which Ad capsid protein-specific antibodies are involved in AdNAb-mediated inhibition of Ad infection in vivo. In this study, mice possessing antibodies specific for each Ad capsid protein were prepared by intramuscular electroporation of each Ad capsid protein-expressing plasmid. Ad vector-mediated hepatic transduction was efficiently inhibited by more than 100-fold in mice immunized with a fiber protein-expressing plasmid or a penton base-expressing plasmid. An Ad vector pre-coated with FX before administration mediated more than 100-fold lower transduction efficiencies in the liver of warfarinized mice immunized with a fiber protein-expressing plasmid or a penton base-expressing plasmid, compared with those in the liver of warfarinized non-immunized mice. These data suggest that anti-fiber protein and anti-penton base antibodies bind to an Ad vector even though FX has already bound to the hexon, and inhibit Ad vector-mediated transduction. This study provides important clues for the development of a novel Ad vector that can circumvent inhibition with AdNAbs.
- Published
- 2018
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50. Clinical features of squamous cell lung cancer with anaplastic lymphoma kinase (ALK)-rearrangement: a retrospective analysis and review.
- Author
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Watanabe J, Togo S, Sumiyoshi I, Namba Y, Suina K, Mizuno T, Kadoya K, Motomura H, Iwai M, Nagaoka T, Sasaki S, Hayashi T, Uekusa T, Abe K, Urata Y, Sakurai F, Mizuguchi H, Kato S, and Takahashi K
- Abstract
Anti-anaplastic lymphoma kinase (ALK)-targeted therapy dramatically improves therapeutic responses in patients with ALK-rearranged lung adenocarcinoma (Ad-LC). A few cases of squamous cell lung carcinoma (Sq-LC) with ALK rearrangement have been reported; however, the clinicopathological features and clinical outcomes following treatment with ALK inhibitors are unknown. We addressed this in the present study by retrospectively comparing the clinical characteristics of five patients with ALK-rearranged Sq-LC with those of patients with ALK-rearranged Ad-LC and by evaluating representative cases of ALK inhibitor responders and non-responders. The prevalence of ALK rearrangement in Sq-LCs was 1.36%. Progression-free survival (PFS) after initial treatment with crizotinib was significantly shorter in Sq-LC than in Ad-LC with ALK rearrangement ( p = 0.033). Two ALK rearrangements assayed by fluorescence in situ hybridization (FISH)-positive/immunohistochemistry-negative cases did not respond to crizotinb, and PFS decreased following alectinib treatment of ALK-rearranged Sq-LC ( p = 0.045). A rebiopsy revealed that responders to ceritinib harbored the L1196M mutation, which causes resistance to other ALK inhibitors. However, non-responders were resistant to all ALK inhibitors, despite the presence of ALK rearrangement in FISH-positive circulating tumor cells and circulating free DNA and absence of the ALK inhibitor resistance mutation. These results indicate that ALK inhibitors remain a reasonable therapeutic option for ALK-rearranged Sq-LC patients who have worse outcomes than ALK-rearranged Ad-LC patients and that resistance mechanisms are heterogeneous. Additionally, oncologists should be aware of the possibility of ALK-rearranged Sq-LC based on clinicopathological features, and plan second-line therapeutic strategies based on rebiopsy results in order to improve patient outcome., Competing Interests: CONFLICTS OF INTEREST The authors declare no conflicts of interest.
- Published
- 2018
- Full Text
- View/download PDF
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