A.              Personal Statement

I have been studying FceRI-mediated mast cell biology and signal transduction for the last 25 years. During this period, I have acquired all necessary expertise for the advanced study of IgE, FceRI signaling, mast cell biology, and allergy disease models. My laboratory has been supported by NIH and institutional/foundations fundings since 1990. Our papers have been published by Nature Immunology, Nature Reviews Immunology, Immunity, Cancer Cell, Journal of Experimental Medicine, Journal of Clinical Investigation, Journal of Allergy and Clinical Immunology, PNAS, Cell Reports, Journal of Immunology, Journal of Biological Chemistry, Trends in Immunology, Current Opinion in Immunology, etc. as research breakthroughs and led to invited talks at a Major Symposium at an AAI meeting, AAAA&I, FASEB and Aegean conferences as well as platform presentations at other national and international meetings. I served as an Associate Editor and a Section Editor for the Journal of Immunology and as a board member of several research journals, and as a reviewer for many major research journals. In addition, I recently edited a Frontiers Special Topic for Frontiers in Immunology (the official journal of the International Union of Immunological Societies) and a Special Series issue for the Open Allergy Journal. I also organized the FASEB Conference “IgE and Allergy, 50 Years & Onward” earlier this year (July 24-29, 2016) to celebrate the 50^th anniversary of the discovery of IgE by gathering more than 110 allergy researchers. I am an elected member of Collegium Internationale Allergologicum. As a Co-PI of a T32 Training grant together with David Broide, UC San Diego, I hosted the monthly journal club meeting at the La Jolla Institute for Allergy and Immunology. In addition to the NIH Study Sections and other NIH review panels listed below, I have been serving as a reviewer of grants for Agency for Science, Technology and Research’s Biomedical Research Council (Singapore), Arthritis National Research Foundation, Asthma UK, British Lung Foundation, German-Israeli Foundation for Scientific Research and Development, JAPAN Health Sciences Foundation, The Israel Science Foundation, The Wellcome Trust, Deutsche Forschungsgemeinschaft, de l’Agence Nationale de la Recherche (ANR) and Swedish Research Council. I also serve as a Consultant for The Japan Prize, The Science and Technology Foundation of Japan and as a reviewer of research programs at the Campus for Research Excellence and Technological Enterprise (CREATE) for the National Research Foundation of Singapore.

                        In 2012, my group found that histamine-releasing factor (HRF) binds directly to about 25% of the tested monoclonal IgE and IgG molecules via their Fab region. We also demonstrated that HRF inhibitors, which blocked HRF-Ig interactions, suppress the mast cell- and IgE-dependent inflammation in multiple models of asthma and food allergy. As HRF’s role in skin inflammation was revealed in passive cutaneous anaphylaxis, it is highly likely that HRF plays a similar critical role in causing atopic dermatitis, at least in a subset of the patients. Separate from our HRF study, we established a highly efficient model of atopic dermatitis using the dermatitis-prone mouse strain NC/Nga. As this model dermatitis depends on high-affinity IgE receptors, FceRI, and is sensitive to corticosteroids (e.g., dexamethasone), and HRF activates mast cells and basophils via aggregation of IgE-bound FceRI, it should serve as an ideal model in order to test efficacy of the HRF inhibitory peptides. To highlight its utility, our murine model has been used to reveal the cellular and molecular mechanisms by which atopic dermatitis patients are rendered susceptible to severe viral infections as exemplified by eczema vaccinatum and eczema herpeticum. During the course of our HRF study, we have developed highly sensitive methods to quantify HRF-reactive IgE and HRF-reactive IgG in human and mouse blood, which will allow us to extend our mouse study to human applications smoothly in the future.

B. Positions and Honors

Professional Experience:

1981-1984       Assist. Prof., Dept. of Physiological Chem. and Nutrition, Faculty of Medicine, Univ. of Tokyo

1984-1987       Visiting Fellow, Lab. of Cell. and Mol. Biol., National Cancer Institute (NCI), Bethesda, MD

1987-1987                   Visiting Associate, Lab. of Cell. and Mol. Biol., NCI, Bethesda, MD

1987-1988                   Visiting Associate, Lab. of Cell. Develop. and Oncology, NIDR/NIH, Bethesda, MD

1988-1990                   Assist. Professor, Dept. of Med. Chem., Faculty of Medicine, Kyoto University

1990-1994                   Assist. Member, La Jolla Institute for Allergy and Immunology, La Jolla, CA

1994-2000                   Associate Member, La Jolla Institute for Allergy and Immunology, La Jolla, CA

2003-2009       Adjunct Professor, Department of Medicine, University of California, San Diego, School of Medicine, La Jolla, CA

2007-pres.       Visiting Professor, Chiba University, School of Medicine, Chiba, Japan

2000-pres.                   Professor, La Jolla Institute for Allergy and Immunology, La Jolla, CA (Title of ‘Member’ was                                                                    changed to ‘Professor’ in 2011)

Professional Activities:

1991-pres.                   Member: American Association of Immunologists

1992-pres.                   Reviewer: Science, Nat. Immunol., Cancer Cell, Immunity, J. Exp. Med., J Clin. Invest., PNAS

2002-2006                   Associate Editor, Journal of Immunology

2004-pres.                   Elected Member, Collegium Internationale Allergologicum

2008-pres.                   Editorial Board, The Open Autoimmunity Journal

2009-2013                   Section Editor, Journal of Immunology

2011-pres.                   Editorial Board, World Journal of Immunology

2011-pres.                   Editorial Board, World Journal of Translational Medicine

2011-pres.                   Guest Editor for The Open Allergy Journal

2011-pres.                   Guest Editor for Frontiers in Immunology

2015-pres.                   Medical-Scientific Council, Asthma and Allergy Foundation of America

Recent Grant/Program Reviews (since 2006):

2006                            Lung Cellular, Molecular, and Immunobiology Study Section, NIH

2006-pres.                   Advisory Review Committee, Arthritis National Research Foundation

2006                            British Lung Foundation

2007-2010                   The Welcome Trust

2007                            Immunology IRG Special Study Section ZRG1 HAI-K (08) F, NIH

2008                            Hypersensitivity, Autoimmune, and Immune-med. Diseases Study Section, NIH

2008                            Agency for Science, Technology and Research’s (A*STAR) Biomedical Research Council                                                (BMRC) in Singapore

2008                            Cellular and Molecular Immunology-A (CMIA) Study Section, NIH

2010                            Lung Cellular, Molecular, and Immunobiology Study Section, NIH

2011                            Cellular and Molecular Immunology Study Section ZRG1 IMM-G (02), NIH

2011                            Microbiology, Immunology and Infection Panel, French National Research Agency

2012                            National Medical Research Council, Ministry of Health, Singapore

2013-2014                   Deutsche Forschungsgemeinschaft

2014                            Allergy, Immunology, and Transplantation Committee, NIAID/NIH

2014                            Special Emphasis Panel ZRG1 MOSS-T12, NIH

2015                Institute of Biomedical Sciences, Academia Sinica, Taiwan

2015                Campus for Research Excellence and Technological Enterprise (CREATE), National Research Foundation, Singapore

2015                de l’Agence Nationale de la Recherche (ANR), France

2015                Special Emphasis Panel ZRG1 MOSS-C02, NIH

2016                RFA-AR-16-001 Special Emphasis Panel/NIAMS-NIH

2016                Swedish Research Council

2016                            Special Emphasis Panel/SRG 2017/01 ZAI1 JTS-I(J3) 1, NIAID/NIH (2016)

C. Contribution to Science

I have been interested in immunology and cancer from the beginning of my scientific career.  After molecular cloning of immunoglobulin m gene during the course of the graduate school at The University of Tokyo (the mentor Tasuku Honjo), I discovered a novel Src family protein-tyrosine kinase (PTK), Fyn, during my postdoctoral stint (the mentor Stuart A. Aaronson) at the National Institutes of Health. These findings led me to study the normal function of PTKs and their networks in immunological settings. 

1. We have been using the high-affinity IgE receptor (FceRI) signaling in mast cells as a model system. Following our co-discovery of two Tec family PTKs, Btk and Itk, in mast cells, we revealed crucial roles of these kinases in FceRI-mediated activation and apoptosis. By introducing retroviral transduction into the field of mast cell research, we established a standard methodology to investigate the FceRI signaling system.

1.     Yamada, N., Kawakami, Y., Kimura, H., Fukamachi, H., Baier, G., Altman, A., Kato, T., Inagaki, Y., and Kawakami, T. Structure and expression of novel protein-tyrosine kinases, Emb and Emt, in hematopoietic cells. Biochem. Biophys. Res. Comm. 192:231-240, 1993.

2.     Kawakami, Y., Yao, L., Miura, T., Tsukada, S., Witte, O. N., and Kawakami, T. Tyrosine phosphorylation and activation of Bruton tyrosine kinase (Btk) upon FceRI cross-linking. Mol. Cell. Biol. 14:5108-5113, 1994.

3.     Kawakami, Y., Miura, T., Bissonnette, R., Hata, D., Khan, W. N., Kitamura, T., Maeda-Yamamoto, M., Hartman, S. E., Yao, L., Alt, F. W., and Kawakami, T. Bruton’s tyrosine kinase regulates apoptosis and JNK/SAPK kinase activity. Proc. Natl. Acad. Sci. USA 94:3938-3942, 1997.

4.     Hata, D., Kawakami, Y., Inagaki, N., Lantz, C. S., Kitamura, T., Khan, W. N., Tashiro, M., Maeda-Yamamoto, M., Miura, T., Han, W., Hartman, S. E., Yao, L., Nagai, H., Goldfeld, A. E., Alt, F. W., Galli, S. J., Witte, O. N., and Kawakami, T. Involvement of Bruton’s tyrosine kinase in FceRI-dependent mast cell degranulation and cytokine production. J. Exp. Med. 187:1235-1247, 1998.

2. During the course of the above-mentioned study, we found interactions between Btk and protein kinase C (PKC)-b and terreic acid as a novel Btk inhibitor that blocked Btk-PKC-b interactions. Our study also showed that PKC-b and Akt are essential for degranulation and cytokine production via activation of several transcription factors. These studies established how cytokine production is regulated by FceRI-stimulated mast cells.

1.     Yao, L., Kawakami, Y., and Kawakami, T. The pleckstrin homology domain of Btk tyrosine kinase interacts with protein kinase C. Proc. Natl. Acad. Sci. USA 91:9175-9179, 1994

2.     Kitaura, J., Asai, K., Maeda-Yamamoto, M., Kawakami, Y., Kikkawa, U., and Kawakami, T. Akt-dependent cytokine production in mast cells. J. Exp. Med. 192:729-739, 2000.

3.     Kawakami, Y., Kitaura, J., Hartman, S. E., Lowell, C. A., Siraganian, R. P., and Kawakami, T. Regulation of protein kinase CbI by two protein-tyrosine kinases, Btk and Syk. Proc. Natl. Acad. Sci. USA. 97:7423-7428, 2000.

4.     Kawakami, Y., Kitaura, J., Yao, L., McHenry, R. W., Kawakami, Y., Newton, A. C., Kang, S., Kato, R. M., Leitges, M., Rawlings, D. J., and Kawakami, T. A Ras activation pathway dependent on Syk phosphorylation of protein kinase C. Proc. Natl. Acad. Sci. USA. 100:9470-9475, 2003.

3. As our knowledge of the FceRI-mediated signal transduction has grown for the last two decades, the need to study in vivo roles of this pathway in disease settings encouraged us to develop novel in vivo models of allergic diseases. We devised a novel procedure to induce atopic dermatitis (AD)-like skin lesions in mice using a house dust mite extract and staphylococcal enterotoxin B. This model allowed us to show that mast cells and T cells, but not B cells or eosinophils, are required for the full expression of dermatitis. Its application to develop a model to study the susceptibility of AD patients to vaccinia virus revealed the weakened ability of NK cells to control this virus in AD-bearing mice, which recapitulates eczema vaccinatum, a complication sometimes seen in severe cases of human AD. This model was also useful to develop or confirm the efficacy of certain compounds and monoclonal anti-viral antibodies to fight against eczema vaccinatum.

1.     Kawakami, Y., Yumoto, K., and Kawakami, T. ­­An improved mouse model of atopic dermatitis and suppression of skin lesions by an inhibitor of Tec family kinases. Allergol. International 56: 403-409, 2007.

2.     Kawakami, Y., Tomimori, Y., Yumoto, K., Hasegawa, S., Ando, T., Tagaya, Y., Crotty, S., and Kawakami, T. Inhibition of NK cell activity by IL-17 allows vaccinia virus to induce severe skin lesions in a mouse model of eczema vaccinatum. J. Exp. Med. 206:1219-1225, 2009.

3.     Tomimori, Y., Kawakami, Y., McCausland, M.M., Ando, T., Koriazova, L., Kato, S., Kawakami, T., and Crotty, S. Protective murine and human monoclonal antibodies against eczema vaccinatum. Antivir. Ther. 16:67-75, 2011.

4.     Kawakami, Y., Ando, T., Lee, J.-R., Kim, G, Kawakami, Y., Nakasaki, T., Nakasaki, M., Matsumoto, K., Choi, Y.S., and Kawakami, T. Defective NK cell activity in a mouse model of eczema herpeticum. J. Allergy Clin. Immunol. I http://dx.doi.org/10.1016/j.jaci.2016.06.034

4. Given our previous studies on crosstalks between PTKs-dependent and G protein-coupled receptor-dependent signals in FceRI-stimulated mast cells, we have been characterizing phospholipase C (PLC)-b isoforms. In addition to its role in an early FceRI signal transduction, we found that deficiency in PLC-b3 results in myeloproliferative neoplasm (MPN) and AD. We demonstrated that a novel multi-molecular complex, SPS complex, that contains SHP-1 (SH2 domain-containing protein phosphatase 1), PLC-b3 and Stat5, regulates Stat5 activity. Increased Stat5 activity due to the loss of PLC-b3 was crucial for the leukemogenesis of MPN as well as the development of AD. This mechanism seems important in the pathogenesis of Burkitt’s lymphoma, chronic lymphocytic leukemia, and human AD.

1.     Xiao, W., Hong, H., Kawakami, Y., Kato, Y., Wu, D., Yasudo, H., Kimura, A., Kubagawa, H., Bertoli, L. F., Davis, R. S., Chau, L. A., Madrenas. J., Hsia, C. C., Xenocostas. A., Kipps, T. J., Hennighausen, L., Iwama, A., Nakauchi, H., and Kawakami, T. Tumor suppression by phospholipase C-b3 via SHP-1-mediated dephosphorylation of Stat5. Cancer Cell 16: 161-171, 2009.

2.     Xiao, W., Ando, T., Wang, H., Kawakami, Y., and Kawakami, T. Lyn- and PLC-b3-dependent regulation of SHP-1 phosphorylation controls Stat5 activity and myelomonocytic leukemia-like disease. Blood 116:6003-6013, 2010.

3.     Xiao, W., Kashiwakura, J., Hong, H., Yasudo, H., Ando, T., Maeda-Yamamoto, M., Wu, D., Kawakami, Y., and Kawakami, T. Phospholipase C-b3 regulates FceRI-mediated mast cell activation by recruiting the protein phosphatase SHP-1. Immunity 34:893-904, 2011.

4.     Ando, T., Xiao, W., Gao, P., Namiranian, S., Matsumoto, K., Tomimori, Y., Hong, H., Yamashita, H., Kimura, M., Kashiwakura, J., Hata, T.R., Izuhara, K., Gurish, M.F., Roers, A., Rafaels, N.M., Barnes, K.C., Jamora, C., Kawakami, Y., and Kawakami, T. Critical role for mast-cell Stat5 activity in skin inflammation. Cell Reports 6:366-376, 2014.

5. Since our finding of ‘monomeric IgE effects’ on mast cell survival in 2001, we have shown that IgEs exhibit a tremendous heterogeneity in their ability to induce mast cell activation, with highly cytokinergic (HC) IgEs at one end of the spectrum and poorly cytokinergic (PC) IgEs at the other: HC IgEs can induce strong survival promotion, degranulation, cytokine production, and migration very efficiently, whereas PC IgEs do so inefficiently. Intrigued by a similar dichotomy for the requirement of a certain type of IgE to prime basophils and mast cells in response to histamine releasing factor (HRF), a cytokine-like factor secreted into body fluids during allergic reactions, we recently found that a subset (~25%) of IgE and IgG molecules directly interact with HRF. HRF-reactive IgE together with HRF could activate mast cells in vitro. By mapping the binding sites on both HRF and IgE/IgG molecules, we developed competitive inhibitors of HRF-IgE (or IgG) interactions, which inhibited IgE+HRF-induced mast cell activation. Using these inhibitors, we could show that HRF promotes allergic inflammation in in vivo mouse models of passive cutaneous anaphylaxis and asthma. Thus, our study discovered the long sought receptors for HRF and enabled us to investigate the role of HRF in allergic diseases and potential utility of HRF inhibitors as therapeutics. Based on these studies, our current proposal will establish HRF inhibitors as efficacious means to prevent and treat food allergy.

1.     Asai, K., Kitaura, J., Kawakami, Y., Yamagata, N, Tsai, M., Carbone, D. P., Liu, F. –T., Galli, S. J., and Kawakami, T. Regulation of mast cell survival by IgE. Immunity 14:791-800, 2001.

2.     Kitaura, J., Song, J., Tsai, M., Asai, K., Maeda-Yamamoto, M., Mocsai, A., Kawakami, Y., Liu, F. -T., Lowell, C. A., Barisas, B. G., Galli, S. J., and Kawakami, T. Highly or poorly cytokinergic IgE molecules mediate a spectrum of effects on mast cell survival and activation. Proc. Natl. Acad. Sci. USA 100:12911-12916, 2003.

3.     Kitaura, J., Kinoshita, T., Matsumoto, M., Chung, S., Kawakami, Y., Leitges, M., Wu, D., Lowell, C. L., and Kawakami, T. IgE- and IgE+Ag-mediated mast cell migration in an autocrine/paracrine fashion. Blood 105: 3222-3229, 2005.

4.     Kashiwakura, J.*, Ando, T.*, Matsumoto, K., Kimura, M., Zajonc, D.M., Ozeki, T., Siraganian, R.P., Broide, D., Kawakami, Y., and Kawakami, T. Histamine-releasing factor has a proinflammatory role in mouse models of asthma and allergy. J. Clin. Invest. 122:218-228, 2012. (*equal contributions)

Complete List of Published Work in My Bibliography:

http://www.ncbi.nlm.nih.gov/sites/myncbi/toshiaki.kawakami.1/bibliography/41163406/public/?sort=date&direction=ascending

D. Research Support

Ongoing Research Support

R01 AR064418-01A1             PI: T. Kawakami                                                    04/01/14-03/31/19

Mast cell Stat5-regulatory pathway in atopic dermatitis   

Major goals: 1) To investigate the role of the mast cell Stat5-regulatory pathway in skin inflammation. 2) To investigate the regulation of the Th2 cytokines (mast cells)-periostin (fibroblasts)-TSLP (keratinocytes) vicious cycle by SPS components. 3) To investigate the role of PLC-b3 and other SPS components in human mast cell biology.

R01 HL124283-01                  PI: T. Kawakami                                                    08/01/14-05/31/18

Interaction of histamine-releasing factor with immunoglobulins in asthma  

Major goals: 1) To identify the molecular signature of HRF-reactive IgE and IgG molecules. 2) To analyze structural details of HRF-Ig interactions at the atomic level. 3) To evaluate the role of HRF in asthma models with abundant HRF-Ig interactions. 4) To study HRF-related molecules.

R21 AI 115534-01                               PI: T. Kawakami                                                          12/01/2014-11/30/2016

Anaphylaxis and IgE heterogeneity

Major goals: 1) To analyze the paired IgE heavy and light chain sequences in IgE⁺ B cells to find

distinct structural features in IgE molecules between severely and mildly anaphylactic mice. 2) To investigate their abilities to exhibit highly cytokinergic vs. poorly cytokinergic property, to bind HRF, and to induce food allergy.

R41AI124734-01                                 PI: T. Kawakami & T. Maruyama            08/01/2016-07/31/2017

Blockade of histamine-releasing factor as a new prophylactic and therapeutic strategy for food allergy

Major goals: 1) To optimize HRF-inhibitory peptides as an anti-food allergy drug. 2) To investigate the mechanisms by which HRF inhibitors suppress food allergy.

Institutional Funds                                                                                         

01/01/17-12/31/17                  La Jolla Institute for Allergy and Immunology                                                           

The amount, which is determined annually on a per need basis, represents Institutional support form the Scientific Director for a one-year term to the Principal Investigator in his capacity as a Professor of the Division of Cell Biology. These funds support research activities and other costs of operating the Division that are not covered by outside grants sources.