Nakano LAb. for immunoregulation & HEALTHY AGING
PI: Toshiaki Nakano
Professor
Graduate Institute of Clinical Medical Sciences,
Chang Gung University College of Medicine, Taoyuan, Taiwan
Liver Transplantation Center,
Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan
appointment
Professor
Aug. 2022 to present
Graduate Institute of Clinical Medical Sciences,
Chang Gung University College of Medicine
Associate Professor
Aug. 2013 to Jul. 2022
Graduate Institute of Clinical Medical Sciences,
Chang Gung University College of Medicine
Associate Research Fellow (Joint Appointment)
Jan. 2016 to present
Graduate Institute of Clinical Medical Sciences,
Chang Gung University College of Medicine
Assistant Professor
Aug. 2009 to Jul. 2013
Department of Surgery,
Kaohsiung Chang Gung Memorial Hospital
Assistant Research Fellow
Jan. 2008 to Jul. 2009
Department of Medical Research,
Kaohsiung Chang Gung Memorial Hospital
Post-doctoral Fellow
Apr. 2004 to Dec. 2007
Department of Surgery,
Kaohsiung Chang Gung Memorial Hospital
Education
Cluster III (Chemistry, Biotechnology and Process Engineering), Fuculty of Enginnering, Hiroshima University, Japan.
1995 - 1999
Bachelor of Engineering
Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Japan.
1999-2001
Master of Science
Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Japan.
2001-2004
Ph.D. of Science
Research topics
Extracellular Nuclear Antigens as Alermins
Nuclear histone H1, which previously was thought to function only as a nuclear factor that is involved in the formation of higher-order chromatin structures and plays a vital role in transcriptional regulation, was recently highlighted as an important player for regulation of innate and adaptive immunity. First discover of histone H1 as an immune regulator in transplant immunology was published in 2004 (Nakano T, et al. Transplantation 2004). In addition, another nuclear antigen, high-mobility group box 1 (HMGB1), was also discovered as a nuclear weapon in the immune arsenal (Lotze MT, et al. Nat Rev Immunol 2005). Specific antibodies against nuclear histone H1 and HMGB1 are highly expressed not only in experimental models of liver allograft tolerance but also in the patient who accept the donor liver without immunosuppressive therapy (termed as operational tolerance) (Nakano T, et al. Transplantation 2007). Our further study strongly suggest that sensitivity to nuclear antigens such as histone H1 may be one of the key factors determining the acceptance or rejection of donor liver grafts (Nakano T, et al. Transplant Immunol 2008).
Histone H1 blockage by anti-histone H1 antibody downregulated the intracellular activation of mitogen-activated protein kinases (MAPKs), IkB and NF-kB of dendritic cells (DCs), and inhibited DC activity in the proliferation of CD4+ T cells. In contrast, the addition of histone H1 upregulated major histocompatibility complex class II (MHC II), the CD80 and CD86 surface markers of DCs. These results suggest that the translocation of histone H1 from nuclei to cytoplasm and the release of their own histone H1 are necessary for the maturation of DCs and the activation for T-lymphocytes (Hsu LW, Goto S, Nakano T, et al. Clin Exp Immunol 2008). We speculate that the induction of autoantibody against nuclear antigens may be an “ultimate weapon” to prevent a breakdown of the immune system. In deed, nuclear histone H1 and HMGB1 were detectable in the serum after liver transplantation (Nakano T*, et al. Clin Dev Immunol 2013) and the active induction of de novo autoimmune hepatitis after liver transplantation could prolong liver allograft survival (Nakano T*, et al. Clin Dev Immunol 2013).
In addition to the impact of histone H1 and corresponding autoantibody in transplant rejection and tolerance, we have recently demonstrated the correlation between circulating histone H1 and OVA-specific IgE levels in a mouse model of OVA-induced allergic rhinitis (Nakano T*, et al. PLoS One 2016). The elevation of histone H1 may cause non-specific allergic response and it must be one of molecular targets for diagnosis and therapeutics.
Extracellular GAPDH for Immunomodulation
GAPDH was accidentally identified as one of the target antigens for anti-histone H1 antibody on T cell surface (see above project). GAPDH, which is constitutively expressed in the cytosol, is a glycolytic pathway-related enzyme and is best known as a housekeeping molecule. GAPDH reversibly catalyzes the phosphorylation of glyceraldehyde-3-phosphate into 1,3-bisphosphoglycerate, which is essential for energy metabolism. To explore the impact of GAPDH on inflammatory response, we have established LPS-induced sepsis-related severe acute lung injury model. Briefly, 10 mg/kg of GAPDH was pre-injected before LPS administration and evaluated the survival rate. Surprisingly, we have demonstrated the prevention of LPS-induced septic death by GAPDH pre-treatment (Takaoka Y, Goto S, Nakano T*, et al. Sci Rep 2014).
To further explore mechanical insights of GAPDH on prevention of LPS-induced septic death, we have explored the impact of GAPDH on macrophages, one of important players at the onset of sepsis and acute lung injury. Macrophages have two opposite phenotypes, which include pro-inflammatory (M1) and anti-inflammatory (M2) statuses. We speculated that GAPDH treatment modulate M1/M2 macrophage polarization. In deed, GAPDH treatment significantly reduced TNF-α production, while it enhanced IL-10 production in LPS-stimulated macrophages. Heat-inactivated GAPDH failed to suppress TNF-α production, suggesting the involvement of enzyme activity. In the course of glycolysis, GAPDH activity produces reduced nicotinamide adenine dinucleotide (NADH). GAPDH treatment enhanced the intracellular level of NADH in LPS-stimulated macrophages similar to IL-4-induced M2 macrophages. Correspondingly, NAD+/NADH ratio was significantly reduced by GAPDH treatment as compared with LPS/IFN-γ-induced M1 macrophages. These results suggest the involvement of NADH formation on M2 macrophage polarization. LPS stimulation may induce proinflammatory cytokines, while it may induce the secretion of GAPDH in macrophages. Extracellular GAPDH modulates intracellular NAD+/NADH balance, resulting in the promotion of anti-inflammatory M2 macrophages for termination of inflammatory response (Nakano T*, et al. Biofactors 2018).
microRNAs for Prediction/Diagnosis of Post-transplant Complications
Liver transplantation is widely accepted as an effective therapeutic modality for end-stage liver diseases. However, post-transplant complications are common in the early and long-term period and contribute to significant morbidity and mortality. Therefore, how we precisely predict/diagnose post-transplant complications such as liver graft dysfunction, acute rejection (AR), infection and hepatitis or hepatocellular carcinoma (HCC) recurrence is a quite important issue for the transplantation society.
In our previous study, we compared microRNA expression profiles between naïve and AR livers at day 7 after OLT with short- (<14 days, donor DA liver into LEW recipient) and long-term (>60 days, donor DA liver into PVG recipient) survival fates (Nakano T*, et al. OMICS 2017). The microarray analysis revealed that the levels of miR-301a in the lethal AR livers were significantly higher than in naïve and tolerogenic AR livers. The reduced expression of miR-301a in inflamed livers such as Concanavalin A (Con A)-induced autoimmune hepatitis and non-alcoholic steatohepatitis (NASH) suggested a difference between AR and inflammation in terms of miR-301a-mediated molecular events.
Recently, tumor-derived exosomes are highlighted for diagnostic and therapeutic targets. We speculate that some HCC patients possess unique components in circulating exosomes responsible for HCC development and recurrence. However, the mode of action of circulating exosomes on HCC development and recurrence is not fully understood. In our recent study, we have confirmed the impact of circulating exosomes on HCC development in a rat model of HCC (Nakano T*, et al. Am J Transplant 2019). Microarray revealed unique microRNA profile in HCC serum exosomes and miR-92b was selected as one of highly expressing microRNAs. In addition, in situ hybridization revealed highly intense staining for miR-92b in HCC tissues. Clinically, circulating exosomal miR-92b may be potential biomarkers for early prediction of post-transplant HCC recurrence.
Phototherapy and Vitamin D3 in Non-alcoholic Fatty Liver Disease
Obesity and hepatic steatosis are becoming increasingly common medical problems in the developed world population, and it has been reported that some of potential donor livers present some degree of steatosis. Hepatic steatosis includes alcoholic and non-alcoholic fatty liver disease (NAFLD). Additionally, NAFLD includes both simple fatty liver and non-alcoholic steatohepatitis (NASH), defined by the presence of lobular necroinflammatory activity with or without the presence of perisinusoidal fibrosis on liver biopsy. To date, the only reliable method of differentiating simple steatosis from NASH is by liver biopsy, which is costly and carries some risk to patients.
Vitamin D is recognized as the “sunshine vitamin.” Most humans depend on sun exposure to satisfy their requirements for vitamin D, but it can be obtained from fortified food, oily fish and vitamin D supplements. Solar ultraviolet B photons are absorbed by 7-dehydro-cholesterol in the skin, leading to its transformation to pre-vitamin D3, which is rapidly converted to vitamin D3. Once formed, vitamin D3 is metabolized in the liver to 25-hydroxyvitamin D3 (25(OH)D3; calcidiol) and then in the kidney to its biologically active form, 1a, 25-dihydroxyvitamin D3 (1,25(OH)2D3; calcitriol). Targher et al. recently reported that NAFLD patients have a marked decrease in serum 25(OH)D3 concentration, which is closely associated with histopathological features of NAFLD (Targher G, et al. Nutr Metab Cardiovasc Dis 2007). However, much remains to be learned about the effects of phototherapy and vitamin D status on the progress of NAFLD. To explore the therapeutic effects of phototherapy on the progress of NASH, artificial sunlight (color temperature 5500 K, color rendition indexes >90; Chang Gung Biotechnology, Taipei, Taiwan) was used for 12 hr/day with animals including choline-deficient and iron-supplemented L-amino acid-defined (CDAA) diet-induced and an obesity-related NASH models.
In our present study, we have demonstrated that serum apolipoprotein E and low molecular weight-adiponectin levels were gradually reduced and reached the lowest level at fatty liver/NASH stage both in CDAA diet-induced NASH model and in genetically obese model. Phototherapy ameliorated hepatocyte apoptosis, inflammation, fibrosis and insulin/leptin resistance caused by CDAA diet with alteration of the levels of lipid transfer/metabolic proteins and elevation of the circulating active form of vitamin D3. Vitamin D3 supplementation ameliorated NASH progression in CDAA diet-induced NASH model. However, phototherapy failed to ameliorate the obesity and steatosis, suggesting that phototherapy may possess anti-inflammatory/fibrotic activity rather than anti-obesity/steatotic activity.
These results suggest that serum lipid transfer/metabolic proteins and vitamin D3 status may be effective biomarkers for non-invasive diagnosis of NASH progression, and that phototherapy may be a good complementary therapy for NASH because of its regulation of lipid transfer/metabolic proteins and vitamin D3. This study (Nakano T, et al. J Hepatol 2011) is the first confirmation of therapeutic potency of sunlight therapy and Vitamin D3 supplementation in the animal model of fatty liver disease, which clearly builds the basis for subsequent human therapeutic trials in NAFLD. This finding is highlighted as an Editorial (Geier A. J Hepatol 2011).
Phototherapy and Fecal Microbiota Transplantation in Food Allergies
In our previous study, we have demonstrated the reduced expression of vitamin D3in non-alcoholic steatohepatitis (NASH), and phototherapy with the artificial sunlight (full spectrum, color temperature 5500 K, color rendition index (CRI)>90Ra; Chang Gung Biotechnology, Taipei, Taiwan) and vitamin D3 supplementation ameliorated NASH progression in rats (Nakano T, et al. J Hepatol 2011). In our recent study, we applied phototherapy in food allergies (FAs) because of the increasing prevalence of FAs in westernized countries (Chen PJ, et al. NPJ Biofilms Microbiomes 2021). There is currently no cure for FAs, and the traditional treatment is the avoidance of allergenic foods. Otherwise patients must carry a self-injectable form of epinephrine for emergency treatment of anaphylaxis. Here, we proposed applying phototherapy as an alternative approach for the prevention and treatment of FA-like allergic diarrhea.
Although the mode of action of phototherapy was not fully clarified in this study, one of the potential mechanisms underlining the beneficial impact of phototherapy could be the improvement in FA-associated vitamin D3insufficiency. Another potential mechanism of phototherapy in the prevention of FA could be the modulation of gut microbiota. We performed fecal microbiota transplantation (FMT) to demonstrate functional roles of microbiota composition in FA. Briefly, FA-associated dysbiosis could directly transfer FA symptoms in naïve BALB/c mice. On the other hand, the replacement of FA-associated bacteria with healthy microbiota may be a promising strategy for FA therapeutics. We identified the genus Lachnospiraceae_NK4A136_group (phylum Firmicutes) as an FA-associated microbacteria. In addition, one of the beneficial bacterial species, Parabacteroides goldsteinii (phylum Bacteroidetes), was enriched in phototherapy-treated mice. Although further studies are necessary, these unique microbes may play some roles in FA development and prevention.
Ongoing projects
The Ministry of Science and Technology (MOST)
Aug. 2020-Mar. 2022
Development of innovative diagnostic procedures for post-liver transplant patients: at-home health screening by detection of nuclear antigen release and following molecular diagnosis for precision medicine (MOST109-2320-B-182-014)
Chang Gung Medical Foundation
Sep. 2022-
Vitamin D3 and alarming as molecular targets for prevention and amelioration of food allergy
(CMRPD8L1121)
Call For Papers
-Mar. 31, 2023
International Journal of Molecular Sciences
Special Issue: The Road to Tolerance: Molecular Biomarkers for Prediction and Diagnosis of Post-transplant Complications in Liver Transplantation
lab members and collaborators
Ms. Hui-Peng Tseng (Senior Research Assistant, CGU)
Dr. Hui-Ying Liu (Dept. of Urology, Kaohsiung CGMH, Ph.D. students in CGU)
Prof. Chao-Long Chen (Superintendent Emeritus, Liver Transplantation Center, Kaohsiung CGMH)
Prof. Yu-Fan Cheng (Dept. of Diagnostic Radiology, Liver Transplantation Center, Kaohsiung CGMH)
Dr. Chien-Chih Chen (Dept. of Psychiatry, Kaohsiung CGMH)
Dr. King-Wah Chiu (Division of Hepato-Gastroenterology, Kaohsiung CGMH)
Group Photo
Basic Research Group
Liver Transplantation Center
Kaohsiung Chang Gung Memorial Hospital
international collaborators
Histone H1 and Immune Regulation, Antioxidant and Allergic Regulation, Phototherapy
Hiroshima University, Japan
Prof. Seiji Kawamoto
Josai International University, Japan
Prof. Shigeru Goto
Prof. Takeshi Goto
Prof. Naoya Ohmori
Oita University, Japan
Prof. Seigo Kitano (President)
Prof. Masafumi Inomata
