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Article: Enhanced reactivity of Li

Ueno, Hiroshi / Yamazaki, Yu / Okada, Hiroshi / Misaizu, Fuminori / Kokubo, Ken / Sakurai, Hidehiro

2024 Volume 20, Page(s) 653–660

Abstract: Lithium ion-endohedral fullerene ( ... ...

Abstract	Lithium ion-endohedral fullerene (Li
Language	English
Publishing date	2024-03-25
Publishing country	Germany
Document type	Journal Article
ZDB-ID	2192461-2
ISSN	1860-5397
ISSN	1860-5397
DOI	10.3762/bjoc.20.58
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Article ; Online: Involvement of Protein Kinase R in Double-Stranded RNA-Induced Proteasomal Degradation of Hypoxia Inducible Factor-1α.

Hotani, Takuma / Nakagawa, Kanako / Tsukamoto, Tomohito / Mizuguchi, Hiroyuki / Sakurai, Fuminori

Inflammation

2023 Volume 46, Issue 6, Page(s) 2332–2342

Abstract: Hypoxia inducible factor-1α (HIF-1α) is a crucial therapeutic target in various diseases, including cancer and fibrosis. We previously demonstrated that transfection with double-stranded RNA (dsRNA), including polyI:C and the dsRNA genome of mammalian ... ...

Abstract	Hypoxia inducible factor-1α (HIF-1α) is a crucial therapeutic target in various diseases, including cancer and fibrosis. We previously demonstrated that transfection with double-stranded RNA (dsRNA), including polyI:C and the dsRNA genome of mammalian orthoreovirus, resulted in significant reduction in HIF-1α protein levels in cultured cells; however, it remained to be elucidated how dsRNA induced down-regulation of HIF-1α protein levels. In this study, we examined the mechanism of dsRNA-mediated down-regulation of HIF-1α protein levels. We found that among the various cellular factors involved in dsRNA-mediated innate immunity, knockdown and knockout of protein kinase R (PKR) significantly restored HIF-1α protein levels in dsRNA-transfected cells, indicating that PKR was involved in dsRNA-mediated down-regulation of HIF-1α. Proteasome inhibitors significantly restored the HIF-1α protein levels in dsRNA-transfected cells. Ubiquitination levels of HIF-1α were increased by transfection with dsRNA. These findings indicated that degradation of HIF-1α in a ubiquitin-proteasome pathway was promoted in a PKR-dependent manner following dsRNA transfection. Expression of not only HIF-1α but also several proteins, including CDK4 and HER2, was down-regulated following dsRNA transfection. These data provide important clues for elucidation of the mechanism of dsRNA-mediated cellular toxicity, as well as for therapeutic application of dsRNA.
MeSH term(s)	Animals ; Humans ; Cell Hypoxia ; Down-Regulation ; eIF-2 Kinase/genetics ; eIF-2 Kinase/metabolism ; Hypoxia ; Hypoxia-Inducible Factor 1, alpha Subunit/genetics ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism ; RNA, Double-Stranded/metabolism ; Ubiquitination
Chemical Substances	eIF-2 Kinase (EC 2.7.11.1) ; Hypoxia-Inducible Factor 1, alpha Subunit ; RNA, Double-Stranded ; EIF2AK1 protein, human (EC 2.7.11.1)
Language	English
Publishing date	2023-08-24
Publishing country	United States
Document type	Journal Article
ZDB-ID	434408-x
ISSN	1573-2576 ; 0360-3997
ISSN (online)	1573-2576
ISSN	0360-3997
DOI	10.1007/s10753-023-01881-8
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Article ; Online: Adenovirus Vector With ADP Gene Induces Cytopathic Effects in HEK293 Cells Without Significant Elevation of Virus Titers.

Shiota, Aoi / Nishimae, Fumitaka / Sakurai, Fuminori / Mizuguchi, Hiroyuki

Anticancer research

2022 Volume 42, Issue 4, Page(s) 1719–1727

Abstract: Background/aim: Efficient production of adenovirus vectors is crucial for their clinical use. Adenovirus death protein (ADP), which is encoded in the E3 region of the adenovirus genome, is involved in host-cell lysis and the subsequent release of ... ...

Abstract	Background/aim: Efficient production of adenovirus vectors is crucial for their clinical use. Adenovirus death protein (ADP), which is encoded in the E3 region of the adenovirus genome, is involved in host-cell lysis and the subsequent release of progeny virus; however, the ADP gene is often removed from the adenovirus vector genome. Materials and methods: We have developed adenovirus vectors that possess the ADP gene and maintain a relatively large insertion capacity for foreign genes by deleting the partial E3 region. Adenovirus vector-mediated transgene expression levels and virus titers were examined. Results: The adenovirus vectors maintaining the ADP gene showed cytopathic effect earlier than conventional adenovirus vector without the ADP gene following treatment of HEK293 cells, although there were no significant differences in total virus titers. Conclusion: The adenovirus vectors possessing the ADP gene showed efficient spread of progeny virus infection following transduction in HEK293 cells.
MeSH term(s)	Adenosine Diphosphate ; Adenoviridae/genetics ; Genetic Vectors/genetics ; HEK293 Cells ; Humans ; Viral Load
Chemical Substances	Adenosine Diphosphate (61D2G4IYVH)
Language	English
Publishing date	2022-03-28
Publishing country	Greece
Document type	Journal Article
ZDB-ID	604549-2
ISSN	1791-7530 ; 0250-7005
ISSN (online)	1791-7530
ISSN	0250-7005
DOI	10.21873/anticanres.15648
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Article: Efficient antitumor effects of a novel oncolytic adenovirus fully composed of species B adenovirus serotype 35.

Ono, Ryosuke / Takayama, Kosuke / Sakurai, Fuminori / Mizuguchi, Hiroyuki

Molecular therapy oncolytics

2021 Volume 20, Page(s) 399–409

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- ... ...

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
Language	English
Publishing date	2021-01-26
Publishing country	United States
Document type	Journal Article
ISSN	2372-7705
ISSN	2372-7705
DOI	10.1016/j.omto.2021.01.015
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Article ; Online: Treatment of Human Pancreatic Cancers Following Local and Systemic Administration of Oncolytic Adenovirus Serotype 35.

Ono, Ryosuke / Takayama, Kosuke / Onishi, Rika / Tokuoka, Sora / Sakurai, Fuminori / Mizuguchi, Hiroyuki

Anticancer research

2023 Volume 43, Issue 2, Page(s) 537–546

Abstract: Background/aim: Oncolytic adenoviruses (Ads) (OAds) are gaining attention as an effective remedy for pancreatic cancer. Most OAds are based on human Ad serotype 5 (Ad5) (OAd5); however, two major drawbacks of OAd5 have been reported. Expression of ... ...

Abstract	Background/aim: Oncolytic adenoviruses (Ads) (OAds) are gaining attention as an effective remedy for pancreatic cancer. Most OAds are based on human Ad serotype 5 (Ad5) (OAd5); however, two major drawbacks of OAd5 have been reported. Expression of coxsackievirus-adenovirus receptor, a primary infection receptor of Ad5, is often decreased on malignant tumor cells, including pancreatic cancers. More than 60% of adults have neutralizing antibodies against Ad5. Previously, we developed an OAd composed of Ad serotype 35 (Ad35) (OAd35). Ad35 recognizes CD46, which is often up-regulated on pancreatic cancers. In addition, only 20% or fewer adults have anti-Ad35 neutralizing antibodies. Materials and methods: We examined the tumor cell lysis activities of OAd35 in the four human pancreatic cancer cell lines in the presence and absence of human serum. The tumor growth suppression effects of OAd35 after local and systemic administration were evaluated in nude mice bearing human pancreatic tumors. Results: OAd35 showed higher levels of tumor cell lysis activities than OAd5 in the human pancreatic cancer cell lines AsPC-1 and BxPC-3. Although the in vitro tumor cell lysis activities of OAd5 against MIA PaCa-2 and PANC-1 cells were strongly attenuated in the presence of human serum, OAd35 mediated comparable levels of tumor cell lysis in the presence and absence of human serum. Systemic administration of OAd5 did not mediate significant growth inhibition against the subcutaneous BxPC-3 tumor. On the other hand, OAd35 significantly suppressed tumor growth. Conclusion: OAd35 would be suitable as an alternative anticancer agent for pancreatic cancer.
MeSH term(s)	Mice ; Animals ; Humans ; Serogroup ; Mice, Nude ; Adenoviridae/genetics ; Pancreatic Neoplasms/therapy ; Antibodies, Neutralizing ; Cell Line, Tumor ; Oncolytic Virotherapy ; Oncolytic Viruses/genetics ; Genetic Vectors
Chemical Substances	Antibodies, Neutralizing
Language	English
Publishing date	2023-01-25
Publishing country	Greece
Document type	Journal Article
ZDB-ID	604549-2
ISSN	1791-7530 ; 0250-7005
ISSN (online)	1791-7530
ISSN	0250-7005
DOI	10.21873/anticanres.16190
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Article ; Online: Effects of pre-existing anti-adenovirus antibodies on transgene expression levels and therapeutic efficacies of arming oncolytic adenovirus.

Ono, Ryosuke / Nishimae, Fumitaka / Wakida, Takuro / Sakurai, Fuminori / Mizuguchi, Hiroyuki

Scientific reports

2022 Volume 12, Issue 1, Page(s) 21560

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 ... ...

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.
MeSH term(s)	Humans ; Mice ; Animals ; Adenoviridae/genetics ; Genetic Vectors/genetics ; Seroepidemiologic Studies ; Adenoviridae Infections/genetics ; Transgenes ; Adenoviruses, Human/genetics ; Antibodies, Viral ; Luciferases/genetics ; Neoplasms/therapy ; Neoplasms/genetics ; Antibodies, Neutralizing/genetics
Chemical Substances	Antibodies, Viral ; Luciferases (EC 1.13.12.-) ; Antibodies, Neutralizing
Language	English
Publishing date	2022-12-13
Publishing country	England
Document type	Journal Article
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-022-26030-3
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: miR-27b antagonizes BMP signaling in early differentiation of human induced pluripotent stem cells.

Lim, Jaeeun / Sakai, Eiko / Sakurai, Fuminori / Mizuguchi, Hiroyuki

Scientific reports

2021 Volume 11, Issue 1, Page(s) 19820

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. ...

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.
MeSH term(s)	Bone Morphogenetic Proteins/genetics ; Bone Morphogenetic Proteins/metabolism ; CRISPR-Cas Systems ; Cell Differentiation/genetics ; Endoderm/metabolism ; Gene Expression Regulation, Developmental ; Humans ; Induced Pluripotent Stem Cells/cytology ; Induced Pluripotent Stem Cells/metabolism ; Mesoderm/metabolism ; MicroRNAs/genetics ; RNA Interference ; Signal Transduction
Chemical Substances	Bone Morphogenetic Proteins ; MIRN27b microRNA, human ; MicroRNAs
Language	English
Publishing date	2021-10-06
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-021-99403-9
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Article ; Online: Optimization of an E1A Gene Expression Cassette in an Oncolytic Adenovirus for Efficient Tumor Cell Killing Activity.

Sakurai, Fuminori / Nishimae, Fumitaka / Takayama, Kosuke / Mizuguchi, Hiroyuki

Anticancer research

2021 Volume 41, Issue 2, Page(s) 773–782

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 ... ...

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.
MeSH term(s)	Adenoviridae/genetics ; Adenoviridae/physiology ; Adenovirus E1A Proteins/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 ; Sequence Deletion ; Survivin/genetics ; Telomerase/genetics ; Virus Replication
Chemical Substances	Adenovirus E1A Proteins ; BIRC5 protein, human ; Survivin ; TERT protein, human (EC 2.7.7.49) ; Telomerase (EC 2.7.7.49)
Language	English
Publishing date	2021-01-23
Publishing country	Greece
Document type	Journal Article
ZDB-ID	604549-2
ISSN	1791-7530 ; 0250-7005
ISSN (online)	1791-7530
ISSN	0250-7005
DOI	10.21873/anticanres.14829
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Article ; Online: Adenovirus vector-based vaccine for infectious diseases.

Sakurai, Fuminori / Tachibana, Masashi / Mizuguchi, Hiroyuki

Drug metabolism and pharmacokinetics

2021 Volume 42, Page(s) 100432

Abstract: Replication-incompetent adenovirus (Ad) vectors have been widely used as gene delivery vehicles in both gene therapy studies and basic studies for gene function analysis due to their highly advantageous properties, which include high transduction ... ...

Abstract	Replication-incompetent adenovirus (Ad) vectors have been widely used as gene delivery vehicles in both gene therapy studies and basic studies for gene function analysis due to their highly advantageous properties, which include high transduction efficiencies, relatively large capacities for transgenes, and high titer production. In addition, Ad vectors induce moderate levels of innate immunity and have relatively high thermostability, making them very attractive as potential vaccine vectors. Accordingly, it is anticipated that Ad vectors will be used in vaccines for the prevention of infectious diseases, including Ebola virus disease and acquired immune deficiency syndrome (AIDS). Much attention is currently focused on the potential use of an Ad vector vaccine for coronavirus disease 2019 (COVID-19). In this review, we describe the basic properties of an Ad vector, Ad vector-induced innate immunity and immune responses to Ad vector-produced transgene products. Development of novel Ad vectors which can overcome the drawbacks of conventional Ad vector vaccines and clinical application of Ad vector vaccines to several infectious diseases are also discussed.
MeSH term(s)	Adenoviridae/genetics ; Adenovirus Vaccines ; COVID-19 ; Communicable Diseases ; Genetic Vectors/genetics ; Humans ; SARS-CoV-2 ; Vaccines
Chemical Substances	Adenovirus Vaccines ; Vaccines
Language	English
Publishing date	2021-11-12
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	2095748-8
ISSN	1880-0920 ; 1347-4367 ; 0916-1139
ISSN (online)	1880-0920
ISSN	1347-4367 ; 0916-1139
DOI	10.1016/j.dmpk.2021.100432
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Article ; Online: miR-27b antagonizes BMP signaling in early differentiation of human induced pluripotent stem cells

Jaeeun Lim / Eiko Sakai / Fuminori Sakurai / Hiroyuki Mizuguchi

Scientific Reports, Vol 11, Iss 1, Pp 1-

2021 Volume 10

Abstract: 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, ... ...

Abstract	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.
Keywords	Medicine ; R ; Science ; Q
Subject code	500
Language	English
Publishing date	2021-10-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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