Kyushu University Academic Staff Educational and Research Activities Database
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Arihiro Kano Last modified date:2018.06.26



Graduate School
Undergraduate School


E-Mail
Homepage
http://www.cm.kyushu-u.ac.jp/dv01/dmstj.html
Academic Degree
Ph. D.
Field of Specialization
Cell Biology, Molecular Biology, Polymer Chemistry
Outline Activities
1. Interferon-gamma (IFN-gamma), an inflammatory cytokine, is a potent apoptosis inducer for hepatocytes. We have shown that the IFN-gamma-induced apoptosis is mediated by anti-tumor factors, such as p53 and IRF-1. We are now developing new type drugs on the basis of the mechanism of IFN-gamma-induced apoptosis, other than the basic study of the apoptosis. These results would contribute to understand the machinery of inflammatory diseases of liver and establish the method of remedy for them. We study sinusoidal endothelial cells of liver (LSEC). We have revealed that the activation of immune cells are controlled by LSEC. This finding has let us work on developing of the methods for treatment to allergies, against pollen, egg, and so on, by the technique of delivery of allergen to LSEC, and through the study of the function of LSEC. And yet we study the mechanisms of LSEC to control immune responses.
2. Delivery carriers for therapeutic agents, including gene and nucleic acid medicine, have the great ability to provide revolutionary disease treatments. A large number of studies have been performed for this aim using liposome, micelle, gel, polymer, and virus-mediated techniques. Liposome and micelle are self-assembled and size-controllable nano particles. These properties are preferable to encapsulate polar or lipophilic drugs, whereas liposome and micelle are often not stable enough in blood circulation. Unlike the lipid carrier, hydrophilic polymeric material itself normally determines the size of the complex with its lords. Therefore molecular weight and steric effects of polymeric material should be considered. Poly-L-lysine (PLL) is a biocompatible polymer consisted with cationic peptides. Such cationic polymers interact with negatively charged DNA or RNA and form a complex, so-called polyplex. Therefore cationic polymers have been widely studied for gene therapy as a non-viral carrier. Although positive charges are needed for polyplex formation and the interaction to cell membranes to be internalized into the cell, these are also bound to plasma proteins and quickly cleared from the circulation. In this study, we employed PLL as a carrier backbone and grafted hydrophilic linear polymer as a side chain to prevent unwanted interactions and provide specific targeting activities. For these objects, we chose hyaluronan (HA), a linear poly saccharide that is known to be metabolized in liver, and poly(ethylene glycol) (PEG) to examine the effect of hydrophilic side chains grafted on PLL as a carrier for drugs and genes. Additionally, the ε-amine group of PLL is thought to be a reactive moiety for interaction to other therapeutic agents, not only for ionic interaction to DNA or RNA. Based on this consideration, we also examined the effect of ε-amine group on biodistribution of PLL-g-PEG after the intravenous injection in mice.
Research
Research Interests
  • Cancer selective cytotoxicity of mitochondrial inhibitor and its application
    keyword : Mitochondrial inhibitor, Glycolysis, ATP, Warburg Effect, Cancer
    2014.04~2021.06.
  • Search for immunoregulatory substances by splenocyte culture system
    keyword : Immune Suppression, Splenocytes, LPS, Liver Sinusoidal Endothelial cells, Cancer cells
    2008.04~2021.06.
  • Complex formation of the tumor-directed graft polymer and anticancer agents and its application to the cancer therapy
    keyword : water soluble macromolecule, DDS, photo dynamic therapy
    2008.04~2013.03Delivery carriers for therapeutic agents, including gene and nucleic acid medicine, have the great ability to provide revolutionary disease treatments. A large number of studies have been performed for this aim using liposome, micelle, gel, polymer, and virus-mediated techniques. Liposome and micelle are self-assembled and size-controllable nano particles. These properties are preferable to encapsulate polar or lipophilic drugs, whereas liposome and micelle are often not stable enough in blood circulation. Unlike the lipid carrier, hydrophilic polymeric material itself normally determines the size of the complex with its lords. Therefore molecular weight and steric effects of polymeric material should be considered. Poly-L-lysine (PLL) is a biocompatible polymer consisted with cationic peptides. Such cationic polymers interact with negatively charged DNA or RNA and form a complex, so-called polyplex. Therefore cationic polymers have been widely studied for gene therapy as a non-viral carrier. Although positive charges are needed for polyplex formation and the interaction to cell membranes to be internalized into the cell, these are also bound to plasma proteins and quickly cleared from the circulation. In this study, we employed PLL as a carrier backbone and grafted hydrophilic linear polymer as a side chain to prevent unwanted interactions and provide specific targeting activities. For these objects, we chose hyaluronan (HA), a linear poly saccharide that is known to be metabolized in liver, and poly(ethylene glycol) (PEG) to examine the effect of hydrophilic side chains grafted on PLL as a carrier for drugs and genes. Additionally, the ε-amine group of PLL is thought to be a reactive moiety for interaction to other therapeutic agents, not only for ionic interaction to DNA or RNA. Based on this consideration, we also examined the effect of ε-amine group on biodistribution of PLL-g-PEG after the intravenous injection in mice..
  • Analysis of the dynamics of water soluble macromolecules in vivo and application to tumor therapy
    keyword : water soluble macromolecule, DDS, EPR
    2007.04~2011.03Delivery carriers for therapeutic agents, including gene and nucleic acid medicine, have the great ability to provide revolutionary disease treatments. A large number of studies have been performed for this aim using liposome, micelle, gel, polymer, and virus-mediated techniques. Liposome and micelle are self-assembled and size-controllable nano particles. These properties are preferable to encapsulate polar or lipophilic drugs, whereas liposome and micelle are often not stable enough in blood circulation. Unlike the lipid carrier, hydrophilic polymeric material itself normally determines the size of the complex with its lords. Therefore molecular weight and steric effects of polymeric material should be considered. Poly-L-lysine (PLL) is a biocompatible polymer consisted with cationic peptides. Such cationic polymers interact with negatively charged DNA or RNA and form a complex, so-called polyplex. Therefore cationic polymers have been widely studied for gene therapy as a non-viral carrier. Although positive charges are needed for polyplex formation and the interaction to cell membranes to be internalized into the cell, these are also bound to plasma proteins and quickly cleared from the circulation. In this study, we employed PLL as a carrier backbone and grafted hydrophilic linear polymer as a side chain to prevent unwanted interactions and provide specific targeting activities. For these objects, we chose hyaluronan (HA), a linear poly saccharide that is known to be metabolized in liver, and poly(ethylene glycol) (PEG) to examine the effect of hydrophilic side chains grafted on PLL as a carrier for drugs and genes. Additionally, the ε-amine group of PLL is thought to be a reactive moiety for interaction to other therapeutic agents, not only for ionic interaction to DNA or RNA. Based on this consideration, we also examined the effect of ε-amine group on biodistribution of PLL-g-PEG after the intravenous injection in mice..
  • Delivery of siRNA to tumors by passive targetting
    keyword : siRNA, PLL-PEG, EPR
    2005.04~2009.10Recently, it has been believed that the liver plays an important role in the immune system. For example, we demonstrated that the liver sinusoidal endothelial cells (LSEC) suppress the activation of T cells. Then we keep studying the machinery of this function of LSEC, and try to control the immune system through LSEC..
  • The role of liver in immune response and its control
    keyword : liver, liver sinusoidal endothelial cell, immunology, hyaluronan
    2003.04Recently, it has been believed that the liver plays an important role in the immune system. For example, we demonstrated that the liver sinusoidal endothelial cells (LSEC) suppress the activation of T cells. Then we keep studying the machinery of this function of LSEC, and try to control the immune system through LSEC..
  • Analysis of apoptosis induced by IFN-gamma
    keyword : Hepatocyte, Interferon-gamma, Apoptosis, SV40 large T antigen,
    2002.04Primary cultured hepatocytes are highly sensitive to IFN-gamma, and undergo apoptosis by IFN-gamma. We previously reported that this apoptotic signal is dominantly through IRF-1. However, further investigation is limited, because molecular techniques are not well applied to the primary hepatocytes due to low growth ability and gene transfection efficiency. To overcome this problem, we established cell lines from primary hepatocytes by SV40-T encoding retrovirus-mediated immortalization and characterized IFN-gammaミinduced apoptosis in these cell lines. Hepatocytes were isolated from C57BL/6 mice by two-step collagenase digestion method and cultured in DMEM with FCS, EGF and insulin. The isolated hepatocytes were treated with the retrovirus encoding SV40-T and neomycin phosphotransferase, and selected with neomycin. To obtain IFN-gamma sensitive and insensitive clones, further cloning procedure was applied to the immortalized hepatocytes. The cloned cell line, designated as HepaB6TC5, was IFN-gamma sensitive, and underwent apoptotic cell death within 72 h from IFN-gamma treatment, as a similar manner to the primary hepatocytes. The IFN-gamma-induced apoptosis in HepaB6TC5 was characterized by DNA fragmentation, and increased caspase-3 like activity. We are investigating further mechanisms of apoptosis in HepaB6TC5..
  • Conditional knockout mice of STAT3
    keyword : STAT-3, Knockout mouse, Endothelial cell, Endotoxin, Immune suppression
    1998.09~2002.02In a pathogenic environment, animals have to balance their responses to the pathogens between an effective immune defense and a controlled immune tolerance. Here we report the anti-inflammatory role of endothelial cells in systemic innate immunity controlled by Stat3. Since Stat3 knockout resulted in early embryonic lethality, we generated a conditional Stat3 knockout mouse using cre-loxP system. The Cre recombinase was under the control of a Tie2 promoter highly specific to endothelial cells. The endothelium specific Stat3 knockout mice were born and appeared normal developmentally. However, the mice were hypersusceptible to lipopolysaccharide (LPS) challenge. 60 % of the knockout mice died in 48 hrs after the LPS treatment whereas none of the wild type liter mates died. Massive leukocytes infiltration and tissue damage were observed in the liver in the knockout mice. Furthermore enhanced TNF-α and IFN-γ concentration in the serum was observed in knockout mice. To address the cellular mechanism underlying the anti-inflammatory functions of endothelial cells in innate immunity, we isolated liver sinusoidal endothelial cells (LSEC) and performed an in vitro study. We showed that the wild type endothelial cells strongly suppressed INF-γ production of splenocytes, which is at least partially mediated by a soluble factor(s), while this suppressive function was completely lost in Stat3 knockout LSEC despite their tube forming function was intact. These results suggest that the endothelium plays an essential role in anti-inflammatory damages at systemic level with the suppression of the INF-γ production, regulated by Stat3..
Academic Activities
Papers
1. Arihiro Kano, Tumor cell secretion of soluble factor(s) for specific immunosuppression, SCIENTIFIC REPORTS, 10.1038/srep08913, 5, 2015.03.
2. Kano, A., Taniwaki, Y., Nakamura, I., Shimada, N., Moriyama, K., Maruyama, A., Tumor delivery of Photofrin® by PLL-g-PEG for photodynamic therapy, J Control Release, 167, 32, 315-321, 2013.03.
3. Tsutomu Ishihara, Arihiro Kano, Kentaro Obara, Minako Saito, Xuesi Chen, Tae Gwan Park,Toshihiro Akaike, Atsushi Maruyama, Nuclear localization and antisense effect of PNA internalized by ASGP-R-mediated endocytosis with protein/DNA conjugates, J. Control. Release, 155, 1, 34-39, 2011.10.
4. Kano A., Wolfgang M.J., Gao Q., Jacoby J., Chai G.X., Hansen W., Iwamoto Y, Pober J.S., Flavell R.A., Fu XY., Endothelial cells require STAT3 for protection against endotoxin-induced inflammation., J Exp Med., 198, 1517-1525, 2003.01.
5. Welte T., Zhang S.S., Wang T., Zhang Z., Hesslein D.G., Yin Z., Kano A., Iwamoto Y., Li E., Craft J.E., Bothwell A.L., Fikrig E., Koni P.A., Flavell R.A., Fu XY., STAT3 deletion during hematopoiesis causes Crohn's disease-like pathogenesis and lethality, Proc Natl Acad Sci U S A., 100, 1879-84, 2003.01.
6. Kano A., Haruyama T., Akaike T., Watanabe Y., IRF-1 is an essential mediator in IFN-gamma-induced cell cycle arrest and apoptosis of primary cultured hepatocytes. Biochem Biophys Res Commun., Biochem Biophys Res Commun., 121, 677-683., 1999.01.
Presentations
1. Studies of tumor models using syngeneic transplantation have advanced our understanding of
tumor immunity, including both immune surveillance and evasion. Murine mammary carcinoma 4T1
cells secrete immunosuppressive soluble factors as demonstrated in splenocyte culture. Cultured
primary splenocytes secrete IFN-γ, which was strikingly elevated when the cells were isolated from
4T1 tumor-bearing mice. The secretion of IFN-γ peaked at a week after 4T1 inoculation and then
declined. This reduction may be due to the relatively decreased lymphocytes and increased
granulocytes in a spleen accompanied by splenomegaly with the 4T1 engraftment. Additionally, IFN-
γ production was further suppressed with the addition of the conditioned media from 4T1 cells to
the splenocyte culture. This suppressive effect was more evident in the splenocytes isolated from
mice that had 4T1 tumors for a longer period of time and was not observed in the conditioned
medium either from CT26 cells or with splenocytes isolated from CT26 tumor-bearing mice. These
results suggest that the IFN-γ suppression is 4T1 tumor-specific. The soluble factor(s) in the 4T1-
conditoned media was a protein between 10 to 100 kDa. The cytokine tip assay demonstrated
several known cytokines that negatively regulate immune responses and may be candidates for this
immunosuppression activity..
Membership in Academic Society
  • Japanese Society of Interferon & Cytokine Research
  • Japanese Society for Immunology
  • The Molecular Biology Society of Japan
  • The Japanese Biochemical Society
Educational
Educational Activities
Plan, teaching, and marking of the practice for under graduate students.
Teaching and guiding for the thesis for master degree and graduation.
Other Educational Activities
  • 2010.06, The 9th China-Japan-Korea Foresight Joint Symposium on Gene Delivery and the International Workshop on Biomaterials 2010 was held in Changchun. This symposium was financially supported by Japan Society for the Promotion of Science. I worked on it as an executive committee in the Japanese side. Fourteen Japanese resercher and one graduate student were sent to this sympoium. .
  • 2010.07, Young Exchange Seminar for A3 Foresight Program that was held in Korea Advanced Institute of Science and Technology, Daejeon, Korea, from July 4 to 7, 2010, by the financial support of Japan Society for the Promotion of Science, JSPS. For this 4-day meeting, 10 graduate students, including from Kumamoto university were sent. I essentially contributed to this project. .
Social
Professional and Outreach Activities
Based on an agreement among JSPS, Korea Science and Engineering Foundation(KOSEF) and National Natural Science Foundation of China(NSFC), this program supports joint research conducted by researchers of Japan, China and Korea. The three countries(A3) work as consortium in advancing leading-edge research with an aim to establishing a top- level research hub in Asia.(Started from FY2005.).