https://kyushu-u.elsevierpure.com/en/persons/arihiro-kano
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https://arihirokano.wordpress.com/english/
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Ph. D.

No

Cell Biology, Molecular Biology, Cancer Biology

03years06months

1. Interferon-gamma (IFN-gamma), an inflammatory cytokine and T-cell activator, was found to induce apoptosis in parenchymal cells, which play a major role in the liver, and the cancer suppressor factors p53 and IRF-1 have been implicated in the mechanism of this action. I have also focused on STATs (Signal Transducers and Activators of Transcription), which are responsible for the intracellular signalling of inflammatory cytokines, and are analysing their functions and conducting applied research targeting them. The results of these studies may lead to a better understanding of the pathogenesis of fulminant or chronic hepatitis, which is thought to be caused by an abnormal immune system, and to its treatment. In addition, he has conducted research focusing on sinusoidal endothelial cells in the liver and found that these cells suppress the excessive activation of immune cells.
2. It is now widely accepted that while the immune system monitors cancer and prevents its emergence and proliferation (Immune Surveillance), cancer evades the surveillance system in various ways. Indeed, it has been reported that cancer suppresses the immune system in various ways, including reduced expression of cancer antigens and human leukaemic antigen (HLA), loss of cell death-inducing signals and co-stimulatory molecules, induction of regulatory T cells and undifferentiated bone marrow-derived cells, and production of immunosuppressive cytokines. In addition, immune checkpoint inhibitors that suppress regulatory T cells have been commercialised for the treatment of malignant melanoma, and their remarkable efficacy is still being used in a growing number of indications. I found that many cancer cells have an activity to inhibit IFN-γ production through the co-culture of mouse spleen cells with various cancer cells and the analysis of the culture supernatant. In a study using splenocytes from mice transplanted with 4T1 breast cancer cells, I found that the inhibition of IFN-γ production by the culture supernatant of cancer cells increased with the passage of more days after transplantation, i.e. with the growth of the tumour. Therefore, a search for immunosuppressive substances secreted by 4T1 cells was conducted. As a result of the experiments, 18 candidate proteins have been found so far. Functional analysis of the main immuno-related functions of these will be carried out in the future.
3 Adenine Nucleotide Translocase (ANT) is a protein present in the inner mitochondrial membrane that mediates the exchange reaction between cytosolic ADP and mitochondria-synthesised ATP and is also reported to be involved in the execution of apoptosis. ANTs comprise a family of four subtypes in humans, but the relationship of these subtypes to apoptosis in particular remains unclear. We are therefore analysing the mechanisms of ANT subtypes and apoptosis execution using siRNA and genome editing techniques.

Research Interests

• Photo dynamic therapy by Cancer receptor-ligand DDS
  keyword : Photo Dynamic Therapy, DDS, cancer, receptor, ligand
  2021.06.
• Study of mitochondria ADP/ATP translocase
  keyword : ADP, ATP, cancer, apoptosis
  2021.11.
• Targeting on immunosuppresive cells by photo dynamic probes for cancer therapy
  keyword : Immune Suppression, Spleen, Cancer cells、Photo Dynamic Therapy
  2020.04～2021.06.
• Search for immunoregulatory substances by splenocyte culture system
  keyword : Immune Suppression, Splenocytes, LPS, Liver Sinusoidal Endothelial cells, Cancer cells
  2008.04.
• Cancer selective cytotoxicity of mitochondrial inhibitor and its application
  keyword : Mitochondrial inhibitor, Glycolysis, ATP, Warburg Effect, Cancer
  2014.04～2019.06.
• 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
  2003.04～2009.03Recently, 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
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 Activities
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. .