論文 - 籠谷　勇紀

Efficient production of human neutrophils from iPSCs that prevent murine lethal infection with immune cell recruitment

Miyauchi M., Ito Y., Nakahara F., Hino T., Nakamura F., Iwasaki Y., Kawagoshi T., Koya J., Yoshimi A., Arai S., Kagoya Y., Kurokawa M.

Blood （Blood）  138 （ 24 ） 2555 - 2569 2021年09月

査読有り,  ISSN  00064971

　概要を見る

Neutrophils play an essential role in innate immune responses to bacterial and fungal infections, and loss of neutrophil function can increase the risk of acquiring lethal infections in clinical settings. Here, we show that engineered neutrophil-primed progenitors derived from human induced pluripotent stem cells can produce functional neutrophil-like cells at a clinically applicable scale that can act rapidly in vivo against lethal bacterial infections. Using 5 different mouse models, we systematically demonstrated that these neutrophil-like cells migrate to sites of inflammation and infection and increase survival against bacterial infection. In addition, we found that these human neutrophil-like cells can recruit murine immune cells. This system potentially provides a straight-forward solution for patients with neutrophil deficiency: an off-the-shelf neutrophil transfusion. This platform should facilitate the administration of human neutrophils for a broad spectrum of physiological and pathological conditions.

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Affinity-matured HLA class II dimers for robust staining of antigen-specific CD4⁺ T cells

Sugata K., Matsunaga Y., Yamashita Y., Nakatsugawa M., Guo T., Halabelian L., Ohashi Y., Saso K., Rahman M.A., Anczurowski M., Wang C.H., Murata K., Saijo H., Kagoya Y., Ly D., Burt B.D., Butler M.O., Mak T.W., Hirano N.

Nature Biotechnology （Nature Biotechnology）  39 （ 8 ） 958 - 967 2021年08月

研究論文（学術雑誌）, 査読有り,  ISSN  1087-0156

　概要を見る

Peptide–major histocompatibility complex (pMHC) multimers enable the detection of antigen-specific T cells in studies ranging from vaccine efficacy to cancer immunotherapy. However, this technology is unreliable when applied to pMHC class II for the detection of CD4+ T cells. Here, using a combination of molecular biological and immunological techniques, we cloned sequences encoding human leukocyte antigen (HLA)-DP, HLA-DQ and HLA-DR molecules with enhanced CD4 binding affinity (with a Kd of 8.9 ± 1.1 µM between CD4 and affinity-matured HLA-DP4) and produced affinity-matured class II dimers that stain antigen-specific T cells better than conventional multimers in both in vitro and ex vivo analyses. Using a comprehensive library of dimers for HLA-DP4, which is the most frequent HLA allele in many ancestry groups, we mapped 103 HLA-DP4-restricted epitopes derived from diverse tumor-associated antigens and cloned the cognate T-cell antigen receptor (TCR) genes from in vitro-stimulated CD4+ T cells. The availability of affinity-matured class II dimers across HLA-DP, HLA-DQ and HLA-DR alleles will aid in the investigation of human CD4+ T-cell responses.

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Genetic Ablation of HLA Class I, Class II, and the T-cell Receptor Enables Allogeneic T Cells to Be Used for Adoptive T-cell Therapy

Yuki Kagoya, Tingxi Guo, Brian Yeung, Kayoko Saso, Mark Anczurowski, Chung-Hsi Wang, Kenji Murata, Kenji Sugata, Hiroshi Saijo, Yukiko Matsunaga, Yota Ohashi, Marcus O. Butler, Naoto Hirano

Cancer Immunology Research （American Association for Cancer Research (AACR)）  8 （ 7 ） 926 - 936 2020年07月

筆頭著者, 査読有り,  ISSN  2326-6066

　概要を見る

Adoptive immunotherapy can induce sustained therapeutic effects in some cancers. Antitumor T-cell grafts are often individually prepared in vitro from autologous T cells, which requires an intensive workload and increased costs. The quality of the generated T cells can also be variable, which affects the therapy's antitumor efficacy and toxicity. Standardized production of antitumor T-cell grafts from third-party donors will enable widespread use of this modality if allogeneic T-cell responses are effectively controlled. Here, we generated HLA class I, HLA class II, and T-cell receptor (TCR) triple-knockout (tKO) T cells by simultaneous knockout of the B2M, CIITA, and TRAC genes through Cas9/sgRNA ribonucleoprotein electroporation. Although HLA-deficient T cells were targeted by natural killer cells, they persisted better than HLA-sufficient T cells in the presence of allogeneic peripheral blood mononuclear cells (PBMC) in immunodeficient mice. When transduced with a CD19 chimeric antigen receptor (CAR) and stimulated by tumor cells, tKO CAR-T cells persisted better when cultured with allogeneic PBMCs compared with TRAC and B2M double-knockout T cells. The CD19 tKO CAR-T cells did not induce graft-versus-host disease but retained antitumor responses. These results demonstrated the benefit of HLA class I, HLA class II, and TCR deletion in enabling allogeneic-sourced T cells to be used for off-the-shelf adoptive immunotherapy.

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Landscape mapping of shared antigenic epitopes and their cognate TCRs of tumor-infiltrating T lymphocytes in melanoma

Kenji Murata, Munehide Nakatsugawa, Muhammed A Rahman, Linh T Nguyen, Douglas G Millar, David T Mulder, Kenji Sugata, Hiroshi Saijo, Yukiko Matsunaga, Yuki Kagoya, Tingxi Guo, Mark Anczurowski, Chung-Hsi Wang, Brian D Burt, Dalam Ly, Kayoko Saso, Alexandra Easson, David P Goldstein, Michael Reedijk, Danny Ghazarian, Trevor J Pugh, Marcus O Butler, Tak W Mak, Pamela S Ohashi, Naoto Hirano

eLife （eLife Sciences Publications, Ltd）  9 2020年04月

査読有り

　概要を見る

HLA-restricted T cell responses can induce antitumor effects in cancer patients. Previous human T cell research has largely focused on the few HLA alleles prevalent in a subset of ethnic groups. Here, using a panel of newly developed peptide-exchangeable peptide/HLA multimers and artificial antigen-presenting cells for 25 different class I alleles and greater than 800 peptides, we systematically and comprehensively mapped shared antigenic epitopes recognized by tumor-infiltrating T lymphocytes (TILs) from eight melanoma patients for all their class I alleles. We were able to determine the specificity, on average, of 12.2% of the TILs recognizing a mean of 3.1 shared antigen-derived epitopes across HLA-A, B, and C. Furthermore, we isolated a number of cognate T cell receptor genes with tumor reactivity. Our novel strategy allows for a more complete examination of the immune response and development of novel cancer immunotherapy not limited by HLA allele prevalence or tumor mutation burden.

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Chaperones of the class I peptide-loading complex facilitate the constitutive presentation of endogenous antigens on HLA-DP84GGPM87

Mark Anczurowski, Kenji Sugata, Yukiko Matsunaga, Yuki Yamashita, Chung-Hsi Wang, Tingxi Guo, Kenji Murata, Hiroshi Saijo, Yuki Kagoya, Kayoko Saso, Marcus O. Butler, Naoto Hirano

Journal of Autoimmunity 102   114 - 125 2019年08月

筆頭著者, 査読有り,  ISSN  0896-8411

　概要を見る

Recent work has delineated key differences in the antigen processing and presentation mechanisms underlying HLA-DP alleles encoding glycine at position 84 of the DPβ chain (DP84GGPM87). These DPs are unable to associate with the class II-associated Ii peptide (CLIP) region of the invariant chain (Ii) chaperone early in the endocytic pathway, leading to continuous presentation of endogenous antigens. However, little is known about the chaperone support involved in the loading of these endogenous antigens onto DP molecules. Here, we demonstrate the proteasome and TAP dependency of this pathway and reveal the ability of HLA class I to compete with DP84GGPM87 for the presentation of endogenous antigens, suggesting that shared subcellular machinery may exist between the two classes of HLA. We identify physical interactions of prototypical class I-associated chaperones with numerous DP alleles, including TAP2, tapasin, ERp57, calnexin, and calreticulin, using a conventional immunoprecipitation and immunoblot approach and confirm the existence of these interactions in vivo through the use of the BioID2 proximal biotinylation system in human cells. Based on immunological assays, we then demonstrate the ability of each of these chaperones to facilitate the presentation of endogenously derived, but not exogenously derived, antigens on DP molecules. Considering previous genetic and clinical studies linking DP84GGPM87 to disease frequency and severity in autoimmune disease, viral infections, and cancer, we suggest that the above chaperones may form the molecular basis of these observable clinical differences through facilitating the presentation of endogenously derived antigens to CD4+ T cells.

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Arginine methylation of FOXP3 is crucial for the suppressive function of regulatory T cells

Yuki Kagoya, Hiroshi Saijo, Yukiko Matsunaga, Tingxi Guo, Kayoko Saso, Mark Anczurowski, Chung-Hsi Wang, Kenji Sugata, Kenji Murata, Marcus O. Butler, Cheryl H. Arrowsmith, Naoto Hirano

Journal of Autoimmunity （Elsevier BV）  97   10 - 21 2019年02月

筆頭著者, 最終著者, 責任著者, 査読有り,  ISSN  0896-8411

　概要を見る

Forkhead box transcription factor 3 (FOXP3) plays a pivotal role in the suppressive function of regulatory T cells. In addition to mRNA levels, FOXP3 activity can also be controlled by posttranslational mechanisms, which have not been studied in a comprehensive manner. Through extensive screening using selective inhibitors, we demonstrate that the inhibition of type I protein arginine methytransferases (PRMTs) attenuates the suppressive functions of regulatory T cells. FOXP3 undergoes methylation on arginine residues at positions 48 and 51 by interacting with protein arginine methyltransferase 1 (PRMT1). The inhibition of arginine methylation confers gene expression profiles representing type I helper T cells to FOXP3+ T cells, which results in attenuated suppressive activity. A methylation-defective mutant of FOXP3 displays less potent activity to suppress xenogeneic graft-versus-host disease in vivo. These results elucidate an important role of arginine methylation to enhance FOXP3 functions and are potentially applicable to modulate regulatory T cell functions.

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DOT1L inhibition attenuates graft-versus-host disease by allogeneic T cells in adoptive immunotherapy models

Yuki Kagoya, Munehide Nakatsugawa, Kayoko Saso, Tingxi Guo, Mark Anczurowski, Chung-Hsi Wang, Marcus O. Butler, Cheryl H. Arrowsmith, Naoto Hirano

Nature Communications （Springer Science and Business Media LLC）  9 （ 1 ）  2018年12月

筆頭著者, 査読有り

　概要を見る

Adoptive T-cell therapy is a promising therapeutic approach for cancer patients. The use of allogeneic T-cell grafts will improve its applicability and versatility provided that inherent allogeneic responses are controlled. T-cell activation is finely regulated by multiple signaling molecules that are transcriptionally controlled by epigenetic mechanisms. Here we report that inhibiting DOT1L, a histone H3-lysine 79 methyltransferase, alleviates allogeneic T-cell responses. DOT1L inhibition reduces miR-181a expression, which in turn increases the ERK phosphatase DUSP6 expression and selectively ameliorates low-avidity T-cell responses through globally suppressing T-cell activation-induced gene expression alterations. The inhibition of DOT1L or DUSP6 overexpression in T cells attenuates the development of graft-versus-host disease, while retaining potent antitumor activity in xenogeneic and allogeneic adoptive immunotherapy models. These results suggest that DOT1L inhibition may enable the safe and effective use of allogeneic antitumor T cells by suppressing unwanted immunological reactions in adoptive immunotherapy.

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Mechanisms underlying the lack of endogenous processing and CLIP-mediated binding of the invariant chain by HLA-DP84Gly

Mark Anczurowski, Yuki Yamashita, Munehide Nakatsugawa, Toshiki Ochi, Yuki Kagoya, Tingxi Guo, Chung-Hsi Wang, Muhammed A. Rahman, Kayoko Saso, Marcus O. Butler, Naoto Hirano

Scientific Reports （Springer Science and Business Media LLC）  8 （ 1 ）  2018年12月

査読有り

　概要を見る

While the principles of classical antigen presentation via MHC class II are well-established, the mechanisms for the many routes of cross-presentation by which endogenous antigens become associated with class II molecules are not fully understood. We have recently demonstrated that the single amino acid polymorphism HLA-DPβ84Gly (DP84Gly) is critical to abrogate class II invariant chain associated peptide (CLIP) region-mediated binding of invariant chain (Ii) to DP, allowing endoplasmic reticulum (ER)-resident endogenous antigens to constitutively associate with DP84Gly such as DP4. In this study, we demonstrate that both the CLIP and N-terminal non-CLIP Ii regions cooperatively generate an Ii conformation that cannot associate with DP84Gly via the CLIP region. We also demonstrate the ability of DP4 to efficiently process and present antigens encoded in place of CLIP in a chimeric Ii, regardless of wild type Ii and HLA-DM expression. These data highlight the complex interplay between DP polymorphisms and the multiple Ii regions that cooperatively regulate this association, ultimately controlling the presentation of endogenous antigens on DP molecules. These results may also offer a mechanistic explanation for recent studies identifying the differential effects between DP84Gly and DP84Asp as clinically relevant in human disease.

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A novel chimeric antigen receptor containing a JAK–STAT signaling domain mediates superior antitumor effects

Yuki Kagoya, Shinya Tanaka, Tingxi Guo, Mark Anczurowski, Chung-Hsi Wang, Kayoko Saso, Marcus O Butler, Mark D Minden, Naoto Hirano

Nature Medicine 24 （ 3 ） 352 - 359 2018年03月

筆頭著者, 査読有り,  ISSN  1078-8956

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A Subset of Human Autoreactive CD1c-Restricted T Cells Preferentially Expresses TRBV4-1+TCRs

Tingxi Guo, Ming Yin Koo, Yuki Kagoya, Mark Anczurowski, Chung-Hsi Wang, Kayoko Saso, Marcus O. Butler, Naoto Hirano

The Journal of Immunology （The American Association of Immunologists）  200 （ 2 ） 500 - 511 2018年01月

査読有り,  ISSN  0022-1767

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HLA-DP84Gly constitutively presents endogenous peptides generated by the class I antigen processing pathway

Yuki Yamashita, Mark Anczurowski, Munehide Nakatsugawa, Makito Tanaka, Yuki Kagoya, Ankit Sinha, Kenji Chamoto, Toshiki Ochi, Tingxi Guo, Kayoko Saso, Marcus O. Butler, Mark D. Minden, Thomas Kislinger, Naoto Hirano

Nature Communications （Springer Science and Business Media LLC）  8 （ 1 ）  2017年08月

最終著者, 責任著者, 査読有り

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Key Residues at Third CDR3β Position Impact Structure and Antigen Recognition of Human Invariant NK TCRs

Kenji Chamoto, Tingxi Guo, Stephen W. Scally, Yuki Kagoya, Mark Ancruzowski, Chung-Hsi Wang, Muhammed A. Rahman, Kayoko Saso, Marcus O. Butler, Priscilla P. L. Chiu, Jean-Philippe Julien, Naoto Hirano

The Journal of Immunology （The American Association of Immunologists）  198 （ 3 ） 1056 - 1065 2017年02月

査読有り,  ISSN  0022-1767

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Transient stimulation expands superior antitumor T cells for adoptive therapy

Yuki Kagoya, Munehide Nakatsugawa, Toshiki Ochi, Yuchen Cen, Tingxi Guo, Mark Anczurowski, Kayoko Saso, Marcus O. Butler, Naoto Hirano

JCI Insight （American Society for Clinical Investigation）  2 （ 2 ）  2017年01月

筆頭著者, 査読有り

　概要を見る

Adoptive cell therapy is a potentially curative therapeutic approach for patients with cancer. In this treatment modality, antitumor T cells are exponentially expanded in vitro prior to infusion. Importantly, the results of recent clinical trials suggest that the quality of expanded T cells critically affects their therapeutic efficacy. Although anti-CD3 mAb-based stimulation is widely used to expand T cells in vitro, a protocol to generate T cell grafts for optimal adoptive therapy has yet to be established. In this study, we investigated the differences between T cell stimulation mediated by anti-CD3/CD28 mAb-coated beads and cell-based artificial antigen-presenting cells (aAPCs) expressing CD3/CD28 counter-receptors. We found that transient stimulation with cell-based aAPCs, but not prolonged stimulation with beads, resulted in the superior expansion of CD8+ T cells. Transiently stimulated CD8+ T cells maintained a stem cell-like memory phenotype and were capable of secreting multiple cytokines significantly more efficiently than chronically stimulated T cells. Importantly, the chimeric antigen receptor-engineered antitumor CD8+ T cells expanded via transient stimulation demonstrated superior persistence and antitumor responses in adoptive immunotherapy mouse models. These results suggest that restrained stimulation is critical for generating T cell grafts for optimal adoptive immunotherapy for cancer.

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Generating <em>De Novo</em> Antigen-specific Human T Cell Receptors by Retroviral Transduction of Centric Hemichain

Tingxi Guo, Toshiki Ochi, Munehide Nakatsugawa, Yuki Kagoya, Mark Anczurowski, Chung-Hsi Wang, Muhammed A. Rahman, Kayoko Saso, Marcus O. Butler, Naoto Hirano

Journal of Visualized Experiments （MyJove Corporation）   （ 116 ）  2016年10月

査読有り

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BET bromodomain inhibition enhances T cell persistence and function in adoptive immunotherapy models

Yuki Kagoya, Munehide Nakatsugawa, Yuki Yamashita, Toshiki Ochi, Tingxi Guo, Mark Anczurowski, Kayoko Saso, Marcus O. Butler, Cheryl H. Arrowsmith, Naoto Hirano

Journal of Clinical Investigation （American Society for Clinical Investigation）  126 （ 9 ） 3479 - 3494 2016年08月

筆頭著者, 査読有り,  ISSN  0021-9738

　概要を見る

Adoptive immunotherapy is a potentially curative therapeutic approach for patients with advanced cancer. However, the in vitro expansion of antitumor T cells prior to infusion inevitably incurs differentiation towards effector T cells and impairs persistence following adoptive transfer. Epigenetic profiles regulate gene expression of key transcription factors over the course of immune cell differentiation, proliferation, and function. Using comprehensive screening of chemical probes with defined epigenetic targets, we found that JQ1, an inhibitor of bromodomain and extra-terminal motif (BET) proteins, maintained CD8+ T cells with functional properties of stem cell-like and central memory T cells. Mechanistically, the BET protein BRD4 directly regulated expression of the transcription factor BATF in CD8+ T cells, which was associated with differentiation of T cells into an effector memory phenotype. JQ1-treated T cells showed enhanced persistence and antitumor effects in murine T cell receptor and chimeric antigen receptor gene therapy models. Furthermore, we found that histone acetyltransferase p300 supported the recruitment of BRD4 to the BATF promoter region, and p300 inhibition similarly augmented antitumor effects of the adoptively transferred T cells. These results demonstrate that targeting the BRD4-p300 signaling cascade supports the generation of superior antitumor T cell grafts for adoptive immunotherapy.

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CD4+ and CD8+ TCRβ repertoires possess different potentials to generate extraordinarily high-avidity T cells

Munehide Nakatsugawa, Muhammed A. Rahman, Yuki Yamashita, Toshiki Ochi, Piotr Wnuk, Shinya Tanaka, Kenji Chamoto, Yuki Kagoya, Kayoko Saso, Tingxi Guo, Mark Anczurowski, Marcus O. Butler, Naoto Hirano

Scientific Reports （Springer Science and Business Media LLC）  6 （ 1 ）  2016年07月

査読有り

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BAALC potentiates oncogenic ERK pathway through interactions with MEKK1 and KLF4

K Morita, Y Masamoto, K Kataoka, J Koya, Y Kagoya, H Yashiroda, T Sato, S Murata, M Kurokawa

Leukemia （Springer Science and Business Media LLC）  29 （ 11 ） 2248 - 2256 2015年11月

査読有り,  ISSN  0887-6924

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Targeted gene correction of RUNX1 in induced pluripotent stem cells derived from familial platelet disorder with propensity to myeloid malignancy restores normal megakaryopoiesis

Hiromitsu Iizuka, Yuki Kagoya, Keisuke Kataoka, Akihide Yoshimi, Masashi Miyauchi, Kazuki Taoka, Keiki Kumano, Takashi Yamamoto, Akitsu Hotta, Shunya Arai, Mineo Kurokawa

Experimental Hematology （Elsevier BV）  43 （ 10 ） 849 - 857 2015年10月

査読有り,  ISSN  0301-472X

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Central nervous system relapse in patients with diffuse large B-cell lymphoma: analysis of incidence and prognostic factors

Masahiro Uni, Yuki Kagoya, Yasuhito Nannya, Fumihiko Nakamura, Mineo Kurokawa

Leukemia & Lymphoma （Informa UK Limited）  56 （ 6 ） 1869 - 1871 2015年06月

査読有り,  ISSN  1042-8194

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骨髄性白血病幹細胞におけるNF-κB活性

籠谷 勇紀

臨床血液 56 （ 4 ） 384 - 391 2015年04月

筆頭著者, 最終著者, 責任著者, 査読有り