Language：English   Publishing type：Research paper (scientific journal)   Publisher：Biochemical and Biophysical Research Communications  

There is no effective therapy for peritoneal carcinomatosis derived from gastric cancer. An ideal conditionally replicating adenovirus (CRA) that selectively replicates in and kills cancer cells has not been developed for gastric cancer-derived peritoneal carcinomatosis. Using our platform technology of CRA regulated and treating tumors with multiple factors (m-CRA), we generated two types of survivin-responsive m-CRAs, Surv.m-CRA-CMVp and Surv.m-CRA-CEAp, consisting of E1A downstream of the survivin promoter, and the mutated E1B gene downstream of the human cytomegalovirus immediate early gene enhancer/promoter and carcinoembryonic antigen promoter, respectively. Survivin mRNA was expressed at high and undetectable levels in two gastric cancer cells and eleven normal cells, respectively. Carcinoembryonic antigen was expressed at high and very low levels in MKN-45 gastric cancer and normal PrEC cells, respectively, and was not detected in other cell types. While both Surv.m-CRA-CEAp and Surv.m-CRA-CMVp exhibited potent cytotoxic effects on MKN-45 cells in vitro, Surv.m-CRA-CEAp significantly reduced cytotoxicity to normal cells compared to Surv.m-CRA-CMVp. Control mice that received an intraperitoneal injection of MKN-45 cells gradually lost body weight and died of peritoneal carcinomatosis within 98 days. In contrast, all mice receiving Surv.m-CRA-CEAp or Surv.m-CRA-CMVp-infected MKN-45 cells increased their body weight and survived 120 days. In conclusion, the triple-regulated Surv.m-CRA-CEAp enhances cancer specificity (i.e., safety) without reducing the potent therapeutic effect for carcinoembryonic antigen-positive gastric cancer-derived peritoneal carcinomatosis. The modified E1B promoter strategy of CRA facilitates the development of novel CRAs for the effective and safe treatment of a variety of refractory cancers.

DOI： 10.1016/j.bbrc.2024.150894

Scopus

PubMed

Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：Translational Research  

In general, ensuring safety is the top priority of a new modality. Although oncolytic virus armed with an immune stimulatory transgene (OVI) showed some promise, the strategic concept of simultaneously achieving maximum effectiveness and minimizing side effects has not been fully explored. We generated a variety of survivin-responsive “conditionally replicating adenoviruses that can target and treat cancer cells with multiple factors (m-CRAs)” (Surv.m-CRAs) armed with the granulocyte-macrophage colony-stimulating factor (GM-CSF) transgene downstream of various promoters using our m-CRA platform technology. We carefully analyzed both therapeutic and adverse effects of them in the in vivo syngeneic Syrian hamster cancer models. Surprisingly, an intratumor injection of a conventional OVI, which expresses the GM-CSF gene under the constitutively and strongly active “cytomegalovirus enhancer and β-actin promoter”, provoked systemic and lethal GM-CSF circulation and shortened overall survival (OS). In contrast, a new conceptual type of OVI, which expressed GM-CSF under the cancer-predominant and mildly active E2F promoter or the moderately active “Rous sarcoma virus long terminal repeat”, not only abolished lethal adverse events but also prolonged OS and systemic anti-cancer immunity. Our study revealed a novel concept that optimal expression levels of an immune stimulatory transgene regulated by a suitable upstream promoter is crucial for achieving high safety and maximal therapeutic effects simultaneously in OVI therapy. These results pave the way for successful development of the next-generation OVI and alert researchers about possible problems with ongoing clinical trials.

DOI： 10.1016/j.trsl.2024.07.002

Scopus

PubMed

Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：Drug Delivery System  

Replication-defective adenovirus(Ad) vectors played a pivotal role in the advancement of in vivo cancer gene therapy research during the 1990s. The development of conditionally replicated Ad (CRA), i.e. the representative oncolytic virus (OV), commenced during the 2000s, and research and clinical applications of CRAs armed with immune genes have been active in recent years. We developed a platform technology that can efficiently generate diverse types of “CRAs that can specifically target tumors with multiple factors” (m-CRAs) and survivin-responsive m-CRAs (Surv.m-CRAs) as innovative anticancer agents. Surv.m-CRA-1, which does not carry a therapeutic gene, is currently undergoing a multicenter, Phase II investigator-initiated clinical trial for early approval for malignant bone tumors. Moreover, we have recently identified a novel concept, “the need for optimal expression levels of immune transgene by optimal promoters,” in OVs armed with immune genes, to achieve both maximal therapeutic effects and the highest level of safety. In this review, we show Ad virology, the aforementioned points and discuss the potential for the next generation of CRAs.

DOI： 10.2745/dds.39.248

Scopus

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1186/1479-5876-12-27

Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.bbrc.2009.08.075

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1089/zeb.2009.0596

Scopus

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1073/pnas.0806976105

Language：English   Publishing type：Research paper (scientific journal)   Publisher：Journal of Phycology  

We investigated the phylogenetic relationships and the taxonomy of 20 sarcinoid strains by their morphological features and using 18S rDNA sequence data. Nineteen strains of Chlorosarcinopsis (Gerneck) Herndon and allied genera were divided into four groups (A-D). Group A, comprising eight species, was concordant with the description of Chlorosarcinopsis and divided into four polyphyletic subgroups. Group B included two species of Chlorosarcinopsis and two strains of Desmotetra stigmatica (Deason) Deason et Floyd, all of which had parallel thylakoid membranes in pyrenoid matrices, and zoospores bearing apical stigma and subapical flagellar apparatus. The definition of Desmotetra Deason et Floyd was emended to include these features, and Chlorosarcinopsis delicata S. Watanabe was transferred to this genus. Group C comprised Chlorosarcina stigmatica Deason (ASIB.T105), for which we proposed Sarcinochlamys gen. nov., a genus distinct from Chlorosarcina Gerneck in having walled zoospores. Group D, comprising six species, corresponded to Neochlorosarcina and was divided into two subgroups. The presence of thin cell walls in Neochlorosarcina S. Watanabe was ascertained as a valid feature for circumscribing the sarcinoid genera. The physiological experiments on the species of Chlorosarcinopsis and Chlorosarcina by Groover and Bold (1968) were assessed based on the phylogenetic results. Groups (A, B, D) were roughly characterized by these features: algal mass color, utilization of selected nitrogen compounds and carbon sources under light/dark and aerobic/anaerobic conditions, and requirement for vitamin B12. Molecular analysis revealed that Chlorosphaeropsis alveolata Herndon had closer affinity with solitary Protosiphon Klebs and Spongiochloris Starr than with sarcinoid members. © 2005 Phycological Society of America.

DOI： 10.1111/j.1529-8817.2006.00196.x

Scopus

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1038/nature01646

Scopus

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1128/MCB.23.8.2699-2708.2003

Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：Biochemical and Biophysical Research Communications  

Cyclin E-Cdk2 is an evolutionary conserved cyclin-dependent kinase (CDK) complex that drives the G1 to S phase transition of the cell cycle. A novel cDNA encoding a HECT family protein also containing RCC1-like repeats was isolated by a yeast two-hybrid screening using both cyclin E and its inhibitor p21. The protein product of this cDNA, Ceb1, interacts with various cyclin subunits of CDKs in mammalian cells. Expression of Ceb1 is specifically detected in testis and ovary and is highly elevated when the functions of the tumor suppressor proteins, p53 and RB, are compromised by mutations or viral oncoproteins. The present results suggest that Ceb1 may play a critical role when its expression and the CDK activity are upregulated by inactivation of p53 and RB.

DOI： 10.1006/bbrc.1999.1777

Scopus

Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1006/bbrc.1999.1516

Scopus

Language：English   Publishing type：Research paper (scientific journal)   Publisher：FEBS Letters  

The N-terminus of MDM2 proto-oncoprotein interacts with p53 and down modulates p53 activity by inhibiting transcriptional activity and promoting p53 degradation. MDMX is structurally related to MDM2 and also binds to p53. However, the function of MDMX has not been clarified yet. We found that MDM2 hetero-oligomerized with MDMX through their C-terminal RING finger domains. Yeast two-hybrid analysis revealed that the hetero-oligomerization between MDMX and MDM2 was more stable than the homo-oligomerization of each protein. MDM2 has been shown to be degraded by the ubiquitin-proteasome pathway, while MDMX was a stable protein. Interaction of MDMX with MDM2 through the C-terminal RING finger domains resulted in inhibiting degradation of MDM2. These data indicate that MDMX functions as a regulator of MDM2. Copyright (C) 1999 Federation of European Biochemical Societies.

DOI： 10.1016/S0014-5793(99)00254-9

Scopus

Language：English   Publishing type：Research paper (scientific journal)   Publisher：Leukemia  

We cloned a novel adaptor protein, APS (adaptor molecule containing Pleckstrin homology (PH) and Src Homology-2 (SH2) domains), which was tyrosine phosphorylated in response to c-kit or B cell receptor stimulation. Here, we report that APS was tyrosine phosphorylated by Janus kinase-2 (JAK2) at its C-terminal tyrosine residue and interacted with c-Cbl. Forced expression of APS in an erythropoietin (EPO)-dependent hematopoietic cell line resulted in reduced activation of STAT5 but not cell proliferation in response to EPO. APS bound to the phosphorylated tyrosine residue, Y343 of the erythropoietin receptor cytoplasmic domain. Co-expression of APS and c-Cbl, but not expression of either alone inhibited EPO-dependent STAT5 activation in 293 cells. This required the C-terminal phosphorylation site, as well as PH and SH2 domains of APS. Therefore, one of the major functions of APS is in recruitment of c-Cbl into the receptor/JAK complex, thereby inhibiting JAK signaling activity.

DOI： 10.1038/sj.leu.2401397

Scopus

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1006/bbrc.1997.7484

Scopus

Language：English   Publishing type：Research paper (scientific journal)   Publisher：Nature  

The proliferation and differentiation of cells of many lineages are regulated by secreted proteins known as cytokines. Cytokines exert their biological effect through binding to cell-surface receptors that are associated with one or more members of the JAK family of cytoplasmic tyrosine kinases. Cytokine-induced receptor dimerization leads to the activation of JAKs, rapid tyrosine-phosphorylation of the cytoplasmic domains, and subsequent recruitment of various signaling proteins, including members of the STAT family of transcription factors, to the receptor complex. Using the yeast two-hybrid system, we have now isolated a new SH2-domain-containing protein, JAB, which is a JAK-binding protein that interacts with the Jak2 tyrosine-kinase JH1 domain. JAB is structurally related to CIS, a cytokine- inducible SH2 protein. Interaction of JAB with Jak1, Jak2 or Jak3 markedly reduces their tyrosine-kinase activity and suppresses the tyrosine- phosphorylation and activation of STATs. JAB and CIS appear to function as negative regulators in the JAK signalling pathway.

DOI： 10.1038/43213

Scopus

Language：English   Publishing type：Research paper (scientific journal)   Publisher：Blood  

We searched for immediate early cytokine responsive genes and isolated a novel gene, CIS (Cytokine Inducible SH2 containing protein) that is induced in hematopoietic cells by a subset of cytokines including interleukin-2 (IL- 2), IL-3, and erythropoietin (EPO). The mutant IL-2 receptor that fails to activate STAT5 could not induce CIS, suggesting that STATS is involved in the cytokine-inducible expression of CIS. We cloned the 5'-flanking region of the CIS gene and found that about 200 bases upstream of the transcription- initiation site contain four potential STATS binding sites (MGF boxes). Luciferase reporter assays showed that these MGF boxes were essential for EPO-dependent promoter activity. Expression of STAT5 and the EPO receptor in HEK293 cells conferred EPO-dependent activation of the CIS promoter. These data indicate that CIS is a target of the JAK-STAT5 pathway of cytokine receptors. CIS contains an SH2 domain and binds to tyrosine-phosphorylated EPO and IL-3 receptors. In HEK293 cells expressing STAT5 and the EPO receptor, EPO-dependent tyrosine phosphorylation of STAT5, as well as EPO- dependent CIS-promoter activation, was suppressed when CIS was coexpressed. Moreover, the induction of oncostatin M, another STAT5 target, as well as the tyrosine-phosphorylation of STAT5, were partially suppressed by CIS expression in Ba/F3 cells. Thus, CIS is a feedback modulator of STAT5; its expression is induced by STAT5 and it negatively modulates STAT5 activation.

DOI： 10.1182/blood.v89.9.3148

Scopus

Language：English   Publishing type：Research paper (scientific journal)   Publisher：John Wiley & Sons  

DOI： 10.1111/j.1440-1835.1996.tb00037.x

Event date： 2024.3

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：沖縄  

Event date： 2024.3

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：沖縄  

Event date： 2022.10

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：福岡  

Event date： 2022.10

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：福岡  

Event date： 2022.10

Language：English   Presentation type：Oral presentation (general)  

Event date： 2022.3

Language：Japanese   Presentation type：Symposium, workshop panel (nominated)  

Venue：東京・オンライン  

Event date： 2021.10

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：大分・オンライン  

Event date： 2021.9

Language：Japanese   Presentation type：Oral presentation (general)  

Event date： 2020.10

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：鹿児島・オンライン  

Event date： 2020.10

Language：Japanese   Presentation type：Symposium, workshop panel (nominated)  

Venue：鹿児島・オンライン  

Event date： 2019.11

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：東京  

Event date： 2019.11

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：福岡  

Event date： 2019.3

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：兵庫  

Event date： 2018.12

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：東京  

Event date： 2018.6

Language：English   Presentation type：Poster presentation  

Venue：Melbourne, Australia  

Event date： 2017.10

Language：Japanese   Presentation type：Symposium, workshop panel (nominated)  

Venue：種子島、鹿児島県  

Event date： 2017.7

Language：Japanese   Presentation type：Symposium, workshop panel (nominated)  

Venue：岡山  

Event date： 2017.3

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：長崎県長崎市  

Event date： 2017.3

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：長崎県長崎市  

Event date： 2017.3

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：長崎県長崎市  

Event date： 2017.3

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：宮城県仙台市  

Event date： 2017.3

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：宮城県仙台市  

Event date： 2016.12

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：東京都  

Event date： 2016.11 - 2016.12

Language：Japanese   Presentation type：Poster presentation  

Venue：神奈川県横浜市  

Event date： 2016.11 - 2016.12

Language：Japanese   Presentation type：Poster presentation  

Venue：神奈川県横浜市  

Event date： 2016.11 - 2016.12

Language：English   Presentation type：Poster presentation  

Venue：神奈川県横浜市  

Event date： 2016.8

Language：English   Presentation type：Poster presentation  

Venue：Hong Kong  

Event date： 2016.7

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：東京  

Event date： 2016.7

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：東京  

Event date： 2016.6

Language：English   Presentation type：Poster presentation  

Venue：San Francisco  

Event date： 2016.6

Language：English   Presentation type：Poster presentation  

Venue：San Francisco  

Event date： 2016.5

Language：English   Presentation type：Poster presentation  

Venue：Washington, DC  

Event date： 2016.3

Language：Japanese   Presentation type：Symposium, workshop panel (public)  

Venue：福島県郡山市  

Event date： 2016.3

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：大阪  

Event date： 2016.3

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：大阪  

Event date： 2015.10

Language：English   Presentation type：Poster presentation  

Event date： 2015.10

Language：English   Presentation type：Poster presentation  

Event date： 2015.7

Language：English   Presentation type：Poster presentation  

Venue：大阪  

Event date： 2015.7

Language：Japanese   Presentation type：Oral presentation (general)  

Venue：大阪  

Human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs), are promising sources of material for use in cell transplantation therapy. However, the risk of formation of tumors, including teratomas and cancers originating from contaminating undifferentiated and transformed cells, represents the most critical obstacle to the safe clinical application of hPSC-based regenerative medicine. Multiple approaches have been taken to improve safety by reducing the risk of carcinogenesis. However, most previous studies focused on improving generation of hiPSCs by eliminating potential oncogenic factors, such as the oncogene c-myc, or by integrating the reprogramming transgenes into chromosomes. Although these sorts of strategies, classified here as the first safety approach, reduce the reprogramming-associated oncogenic potential of hiPSCs, they cannot completely eliminate tumorigenic potentials due to the intrinsic characteristics of hPSCs. We previously developed a novel method (adenoviral conditional targeting) that securely isolated target cells from other cell types and undifferentiated hPSCs (Mol Ther. 14(5): 673-683. 2006 ; Cover & News in the issue of the journal). This method, which can increase the efficacy and safety of hPSC-based regenerative medicine by decreasing tumorigenicity, is classified here as the second safety approach. Strategies that can directly target and kill, rather than merely inhibit, tumorigenic cells are classified here as the third safety approach (See Ide’s presentation). Conditionally replicating adenoviruses (CRAs), also called oncolytic adenoviruses, can selectively replicate in and kill cancer cells; consequently, CRAs represent attractive anticancer drugs. Previously, we developed a method for generating CRAs that can target cancers with multiple cancer-specific factors (m-CRAs); this approach further increased cancer specificity without reducing the anticancer effects. We also demonstrated that among candidate m-CRAs, survivin-responsive m-CRA (Surv.m-CRA) is one of the most promising anticancer agents, in two respects: superior cancer specificity (i.e., safety) and therapeutic efficacy relative to clinically tested telomerase reverse transcriptase (TERT)-responsive m-CRAs (Tert.m-CRAs), and strong anticancer effects against currently incurable cancer stem cells. Based on our careful and extensive preclinical studies, we have a plan to start the investigator initiated first-in-human clinical trial of Surv.m-CRA for malignant orthopedic tumors after J-IND approval, hopefully, this year (as presented in a preclinical symposium session). Here, we demonstrate that m-CRAs may also be useful as novel antitumorigenic agents in hPSC-based therapy. We show that the survivin promoter was more active in undifferentiated hPSCs than the TERT promoter, whereas both promoters were minimally active in differentiated normal cells. Accordingly, Surv.m-CRA killed undifferentiated hPSCs more efficiently than Tert.m-CRA; both m-CRAs exhibited efficient viral replication and cytotoxicity in undifferentiated hPSCs, but not in co-cultured differentiated normal cells. Pre-infection of hPSCs with Surv.m-CRA or Tert.m-CRA abolished in vivo teratoma formation in a dose- dependent manner following hPSC implantation into mice. Thus, we demonstrate that m-CRAs, in particular Surv.m-CRAs, are potentially useful as both potent anticancer drugs and as novel antitumorigenic agents in hPSC-based regenerative medicine; in the latter context, the viruses act by specifically killing contaminating undifferentiated hPSCs.

Event date： 2015.3

Language：Japanese  

Venue：神奈川  

国内学会

Event date： 2015.3

Language：Japanese  

Venue：神奈川  

国内学会

Event date： 2015.1

Language：Japanese  

Venue：鹿児島  

研究会

Event date： 2014.3

Language：Japanese  

Venue：京都  

国内学会

Event date： 2014.1

Language：Japanese  

Venue：鹿児島  

研究会

Event date： 2014.1

Language：Japanese  

Venue：鹿児島  

研究会

Event date： 2013.1

Language：Japanese  

Venue：鹿児島  

研究会

Event date： 2012.6

Language：Japanese  

Venue：熊本  

国内学会

Event date： 2012.6

Language：Japanese  

Venue：神奈川  

国内学会

Event date： 2012.6

Language：Japanese  

Venue：神奈川  

国内学会

Event date： 2012.6

Language：Japanese  

Venue：神奈川  

国内学会

Event date： 2012.1

Language：Japanese  

Venue：鹿児島  

研究会

Event date： 2010.12

Language：Japanese  

Venue：長崎  

国内学会

Event date： 2010.7

Language：Japanese  

Venue：栃木  

国内学会

Event date： 2010.3

Language：Japanese  

Venue：広島  

国内学会

Event date： 2010.3

Language：English  

Venue：福岡  

国際学会

Language：Japanese   Presentation type：Poster presentation