2022/10/01 更新

写真a

マルヤマ タケシ
丸山 剛
所属
附属機関・学校 高等研究所
職名
准教授(任期付)

兼担

  • 理工学術院   先進理工学部

経歴

  • 2021年
    -
    継続中

    早稲田大学・高等研究所 准教授

  • 2018年
    -
    2021年

    早稲田大学・高等研究所 講師

  • 2016年
    -
    2020年

    戦略的創造研究推進事業 さきがけ 研究員(兼任)

  • 2015年
    -
    2018年

    北海道大学 遺伝子病制御研究所 分子腫瘍分野 助教

  • 2017年
     
     

    早稲田大学先進理工学部生命医科学科 非常勤講師

  • 2015年
     
     

    北海道大学 遺伝子病制御研究所 分子腫瘍分野 特任助教 エーザイ研究所(神戸)

  • 2013年
    -
    2015年

    日本学術振興会・海外特別研究員

  • 2013年
    -
    2015年

    MITホワイトヘッド研究所

  • 2011年
    -
    2013年

    東京大学大学院薬学系研究科・特任研究員

  • 2009年
    -
    2011年

    日本学術振興会・学振特別研究員

▼全件表示

 

研究分野

  • 腫瘍生物学

論文

  • ASKA technology-based pull-down method reveals a suppressive effect of ASK1 on the inflammatory NOD-RIPK2 pathway in brown adipocytes.

    Saki Takayanagi, Kengo Watanabe, Takeshi Maruyama, Motoyuki Ogawa, Kazuhiro Morishita, Mayumi Soga, Tomohisa Hatta, Tohru Natsume, Tomoya Hirano, Hiroyuki Kagechika, Kazuki Hattori, Isao Naguro, Hidenori Ichijo

    Scientific reports   11 ( 1 ) 22009 - 22009  2021年11月  [国際誌]

     概要を見る

    Recent studies have shown that adipose tissue is an immunological organ. While inflammation in energy-storing white adipose tissues has been the focus of intense research, the regulatory mechanisms of inflammation in heat-producing brown adipose tissues remain largely unknown. We previously identified apoptosis signal-regulating kinase 1 (ASK1) as a critical regulator of brown adipocyte maturation; the PKA-ASK1-p38 axis facilitates uncoupling protein 1 (UCP1) induction cell-autonomously. Here, we show that ASK1 suppresses an innate immune pathway and contributes to maintenance of brown adipocytes. We report a novel chemical pull-down method for endogenous kinases using analog sensitive kinase allele (ASKA) technology and identify an ASK1 interactor in brown adipocytes, receptor-interacting serine/threonine-protein kinase 2 (RIPK2). ASK1 disrupts the RIPK2 signaling complex and inhibits the NOD-RIPK2 pathway to downregulate the production of inflammatory cytokines. As a potential biological significance, an in vitro model for intercellular regulation suggests that ASK1 facilitates the expression of UCP1 through the suppression of inflammatory cytokine production. In parallel to our previous report on the PKA-ASK1-p38 axis, our work raises the possibility of an auxiliary role of ASK1 in brown adipocyte maintenance through neutralizing the thermogenesis-suppressive effect of the NOD-RIPK2 pathway.

    DOI PubMed

  • Epithelial cells remove precancerous cells by cell competition via MHC class I-LILRB3 interaction.

    Shiyu Ayukawa, Nagisa Kamoshita, Jun Nakayama, Ryohei Teramoto, Novalia Pishesha, Kenji Ohba, Nanami Sato, Kei Kozawa, Hikari Abe, Kentaro Semba, Nobuhito Goda, Yasuyuki Fujita, Takeshi Maruyama

    Nature immunology   22 ( 11 ) 1391 - 1402  2021年11月  [国際誌]

     概要を見る

    Epithelial cells have an ability termed 'cell competition', which is an immune surveillance-like function that extrudes precancerous cells from the epithelial layer, leading to apoptosis and clearance. However, it remains unclear how epithelial cells recognize and extrude transformed cells. Here, we discovered that a PirB family protein, leukocyte immunoglobulin-like receptor B3 (LILRB3), which is expressed on non-transformed epithelial cells, recognizes major histocompatibility complex class I (MHC class I) that is highly expressed on transformed cells. MHC class I interaction with LILRB3 expressed on normal epithelial cells triggers an SHP2-ROCK2 pathway that generates a mechanical force to extrude transformed cells. Removal of transformed cells occurs independently of natural killer (NK) cell or CD8+ cytotoxic T cell-mediated activity. This is a new mechanism in that the immunological ligand-receptor system generates a mechanical force in non-immune epithelial cells to extrude precancerous cells in the same epithelial layer.

    DOI PubMed

  • Cell competition in vertebrates - a key machinery for tissue homeostasis.

    Takeshi Maruyama, Yasuyuki Fujita

    Current opinion in genetics & development   72   15 - 21  2021年10月  [国際誌]

     概要を見る

    Cell competition is a process by which cells with different properties compete with each other for survival and space, and consequently suboptimal/abnormal cells are often eliminated from, in particular, epithelial tissues. In the last few years, cell competition studies have been developing at an explosive speed, and the molecular mechanisms of cell competition have been considerably revealed. For instance, upon cell competition, loser cells are eliminated from tissues via a variety of loser phenotypes, including apoptosis, cell differentiation, and cell death-independent extrusion. In addition, upstream regulatory mechanisms for the induction of these phenotypes have been elucidated. Furthermore, it has become evident that cell competition is involved in various physiological and pathological processes and thus is a crucial and indispensable homeostatic machinery that is required for embryonic development and prevention of diseases and ageing. Moreover, cell competition now has a profound impact on other research fields such as regenerative medicine. In this review, we will summarize the development of these recent studies, especially focusing on cell competition in vertebrates.

    DOI PubMed

  • The CD44/COL17A1 pathway promotes the formation of multilayered, transformed epithelia.

    Kei Kozawa, Miho Sekai, Kenji Ohba, Shoko Ito, Hiroaki Sako, Takeshi Maruyama, Mai Kakeno, Takanobu Shirai, Keisuke Kuromiya, Tomoko Kamasaki, Koki Kohashi, Shinya Tanaka, Susumu Ishikawa, Nanami Sato, Shota Asano, Hironori Suzuki, Nobuyuki Tanimura, Yohei Mukai, Noriko Gotoh, Mishie Tanino, Shinya Tanaka, Ken Natsuga, Tomoyoshi Soga, Tomonori Nakamura, Yukihiro Yabuta, Mitinori Saitou, Takahiro Ito, Kenkyo Matsuura, Makoto Tsunoda, Toyone Kikumori, Tadashi Iida, Yasuyuki Mizutani, Yuki Miyai, Kozo Kaibuchi, Atsushi Enomoto, Yasuyuki Fujita

    Current biology : CB   31 ( 14 ) 3086 - 3097  2021年05月  [国際誌]

     概要を見る

    At the early stage of cancer development, oncogenic mutations often cause multilayered epithelial structures. However, the underlying molecular mechanism still remains enigmatic. By performing a series of screenings targeting plasma membrane proteins, we have found that collagen XVII (COL17A1) and CD44 accumulate in RasV12-, Src-, or ErbB2-transformed epithelial cells. In addition, the expression of COL17A1 and CD44 is also regulated by cell density and upon apical cell extrusion. We further demonstrate that the expression of COL17A1 and CD44 is profoundly upregulated at the upper layers of multilayered, transformed epithelia in vitro and in vivo. The accumulated COL17A1 and CD44 suppress mitochondrial membrane potential and reactive oxygen species (ROS) production. The diminished intracellular ROS level then promotes resistance against ferroptosis-mediated cell death upon cell extrusion, thereby positively regulating the formation of multilayered structures. To further understand the functional role of COL17A1, we performed comprehensive metabolome analysis and compared intracellular metabolites between RasV12 and COL17A1-knockout RasV12 cells. The data imply that COL17A1 regulates the metabolic pathway from the GABA shunt to mitochondrial complex I through succinate, thereby suppressing the ROS production. Moreover, we demonstrate that CD44 regulates membrane accumulation of COL17A1 in multilayered structures. These results suggest that CD44 and COL17A1 are crucial regulators for the clonal expansion of transformed cells within multilayered epithelia, thus being potential targets for early diagnosis and preventive treatment for precancerous lesions.

    DOI PubMed

  • ZAK Inhibitor PLX4720 Promotes Extrusion of Transformed Cells via Cell Competition.

    Takeshi Maruyama, Ayana Sasaki, Sayuri Iijima, Shiyu Ayukawa, Nobuhito Goda, Keisuke Tazuru, Norikazu Hashimoto, Takashi Hayashi, Kei Kozawa, Nanami Sato, Susumu Ishikawa, Tomoko Morita, Yasuyuki Fujita

    iScience   23 ( 7 ) 101327 - 101327  2020年07月  [査読有り]  [国際誌]

     概要を見る

    Previous studies have revealed that, at the initial step of carcinogenesis, transformed cells are often eliminated from epithelia via cell competition with the surrounding normal cells. In this study, we performed cell competition-based high-throughput screening for chemical compounds using cultured epithelial cells and confocal microscopy. PLX4720 was identified as a hit compound that promoted apical extrusion of RasV12-transformed cells surrounded by normal epithelial cells. Knockdown/knockout of ZAK, a target of PLX4720, substantially enhanced the apical elimination of RasV12 cells in vitro and in vivo. ZAK negatively modulated the accumulation or activation of multiple cell competition regulators. Moreover, PLX4720 treatment promoted apical elimination of RasV12-transformed cells in vivo and suppressed the formation of potentially precancerous tumors. This is the first report demonstrating that a cell competition-promoting chemical drug facilitates apical elimination of transformed cells in vivo, providing a new dimension in cancer preventive medicine.

    DOI PubMed

  • The COX-2/PGE2 pathway suppresses apical elimination of RasV12-transformed cells from epithelia.

    Nanami Sato, Yuta Yako, Takeshi Maruyama, Susumu Ishikawa, Keisuke Kuromiya, Suzumi M Tokuoka, Yoshihiro Kita, Yasuyuki Fujita

    Communications biology   3 ( 1 ) 132 - 132  2020年03月  [査読有り]  [国際誌]

     概要を見る

    At the initial stage of carcinogenesis, when RasV12-transformed cells are surrounded by normal epithelial cells, RasV12 cells are apically extruded from epithelia through cell competition with the surrounding normal cells. In this study, we demonstrate that expression of cyclooxygenase (COX)-2 is upregulated in normal cells surrounding RasV12-transformed cells. Addition of COX inhibitor or COX-2-knockout promotes apical extrusion of RasV12 cells. Furthermore, production of Prostaglandin (PG) E2, a downstream prostanoid of COX-2, is elevated in normal cells surrounding RasV12 cells, and addition of PGE2 suppresses apical extrusion of RasV12 cells. In a cell competition mouse model, expression of COX-2 is elevated in pancreatic epithelia harbouring RasV12-exressing cells, and the COX inhibitor ibuprofen promotes apical extrusion of RasV12 cells. Moreover, caerulein-induced chronic inflammation substantially suppresses apical elimination of RasV12 cells. These results indicate that intrinsically or extrinsically mediated inflammation can promote tumour initiation by diminishing cell competition between normal and transformed cells.

    DOI PubMed

  • Graphene/Au Hybrid Antenna Coil Exfoliated with Multi-Stacked Graphene Flakes for Ultra-Thin Biomedical Devices

    Yuma Tetsu, Yusuke Kido, Meiting Hao, Shinji Takeoka, Takeshi Maruyama, Toshinori Fujie

    ADVANCED ELECTRONIC MATERIALS   6 ( 2 )  2020年02月  [査読有り]

     概要を見る

    Flexible electronics with organic substrates have been developed for bio-conformable devices and soft robotics. Although biodegradable polymers are preferred substrates for biomedical applications, they have poor heat durability, which precludes printing of conductive lines that require annealing at high temperatures (>250 degrees C). The fabrication of an ultra-flexible, inkjet-printed antenna coil with a resistivity of 4.30 x 10(-5) omega-cm is reported. It involves annealing of a graphene/Au antenna coil printed on a glass substrate and transferring onto a 182-nm-thick poly(D, L-lactic acid) nanosheet by exfoliation of multi-stacked graphene flakes. Then, a light-emitting device, powered wirelessly, even in the rounded, twisted, or attached states, is fabricated by mounting a blue LED chip on the nanosheet antenna coil. The self-deploying device is also stored in a water-soluble capsule, injected into a silicone bag, released from the dissolved capsule, and operated wirelessly. This work facilitates the hybridization of conductive lines and biodegradable polymers on ultra-flexible biomaterials for the biomedical application of flexible electronics.

    DOI

  • Accumulation of the myosin-II-spectrin complex plays a positive role in apical extrusion of Src-transformed epithelial cells.

    Mikio Takagi, Masaya Ikegawa, Takashi Shimada, Susumu Ishikawa, Mihoko Kajita, Takeshi Maruyama, Tomoko Kamasaki, Yasuyuki Fujita

    Genes to cells : devoted to molecular & cellular mechanisms   23 ( 11 ) 974 - 981  2018年11月  [査読有り]  [国際誌]

     概要を見る

    At the initial stage of carcinogenesis, transformation occurs in single cells within the epithelium. Recent studies have revealed that the newly emerging transformed cells are often apically eliminated from epithelial tissues. However, the underlying molecular mechanisms of this cancer preventive phenomenon still remain elusive. In this study, we first demonstrate that myosin-II accumulates in Src-transformed cells when they are surrounded by normal epithelial cells. Knock-down of the heavy chains of myosin-II substantially diminishes apical extrusion of Src cells, suggesting that accumulated myosin-II positively regulates the apical elimination of transformed cells. Furthermore, we have identified β-spectrin as a myosin-II-binding protein under the coculture of normal and Src-transformed epithelial cells. β-spectrin is also accumulated in Src cells that are surrounded by normal cells, and the β-spectrin accumulation is regulated by myosin-II. Moreover, knock-down of β-spectrin significantly suppresses apical extrusion of Src cells. Collectively, these results indicate that accumulation of the myosin-II-spectrin complex plays a positive role in apical extrusion of Src-transformed epithelial cells. Further elucidation of the molecular mechanisms of apical extrusion would lead to the establishment of a novel type of cancer preventive medicine.

    DOI PubMed

  • The paxillin-plectin-EPLIN complex promotes apical elimination of RasV12-transformed cells by modulating HDAC6-regulated tubulin acetylation.

    Nobuhiro Kasai, Ailijiang Kadeer, Mihoko Kajita, Sayaka Saitoh, Susumu Ishikawa, Takeshi Maruyama, Yasuyuki Fujita

    Scientific reports   8 ( 1 ) 2097 - 2097  2018年02月  [査読有り]  [国際誌]

     概要を見る

    Recent studies have revealed that newly emerging RasV12-transformed cells are often apically extruded from the epithelial layer. During this cancer preventive process, cytoskeletal proteins plectin and Epithelial Protein Lost In Neoplasm (EPLIN) are accumulated in RasV12 cells that are surrounded by normal cells, which positively regulate the apical elimination of transformed cells. However, the downstream regulators of the plectin-EPLIN complex remain to be identified. In this study, we have found that paxillin binds to EPLIN specifically in the mix culture of normal and RasV12-transformed cells. In addition, paxillin is accumulated in RasV12 cells surrounded by normal cells. Paxillin, plectin and EPLIN mutually influence their non-cell-autonomous accumulation, and paxillin plays a crucial role in apical extrusion of RasV12 cells. We also demonstrate that in RasV12 cells surrounded by normal cells, acetylated tubulin is accumulated. Furthermore, acetylation of tubulin is promoted by paxillin that suppresses the activity of histone deacetylase (HDAC) 6. Collectively, these results indicate that in concert with plectin and EPLIN, paxillin positively regulates apical extrusion of RasV12-transformed cells by promoting microtubule acetylation. This study shed light on the unexplored events occurring at the initial stage of carcinogenesis and would potentially lead to a novel type of cancer preventive medicine.

    DOI PubMed

  • Cell competition in mammals - novel homeostatic machinery for embryonic development and cancer prevention.

    Takeshi Maruyama, Yasuyuki Fujita

    Current opinion in cell biology   48   106 - 112  2017年10月  [査読有り]  [国際誌]

     概要を見る

    In the multi-cellular community, cells with different properties often compete with each other for survival and space. This process is named cell competition and was originally discovered in Drosophila. Recent studies have revealed that comparable phenomena also occur in mammals under various physiological and pathological conditions. Within the epithelium, normal cells often recognize the presence of the neighboring transformed cells and actively eliminate them from the epithelium; a process termed EDAC (Epithelial Defense Against Cancer). Furthermore, physical force can play a crucial role in the intercellular recognition and elimination of loser cells during cell competition. Further studies are expected to reveal a variety of roles of cell competition in embryonic development and human diseases.

    DOI PubMed

  • Cell competition with normal epithelial cells promotes apical extrusion of transformed cells through metabolic changes.

    Kon S, Ishibashi K, Katoh H, Kitamoto S, Shirai T, Tanaka S, Kajita M, Ishikawa S, Yamauchi H, Yako Y, Kamasaki T, Matsumoto T, Watanabe H, Egami R, Sasaki A, Nishikawa A, Kameda I, Maruyama T, Narumi R, Morita T, Sasaki Y, Enoki R, Honma S, Imamura H, Oshima M, Soga T, Miyazaki JI, Duchen MR, Nam JM, Onodera Y, Yoshioka S, Kikuta J, Ishii M, Imajo M, Nishida E, Fujioka Y, Ohba Y, Sato T, Fujita Y

    Nature cell biology   19 ( 5 ) 530 - 541  2017年05月  [査読有り]  [国際誌]

    DOI PubMed

  • Rab5-regulated endocytosis plays a crucial role in apical extrusion of transformed cells.

    Sayaka Saitoh, Takeshi Maruyama, Yuta Yako, Mihoko Kajita, Yoichiro Fujioka, Yusuke Ohba, Nobuhiro Kasai, Natsu Sugama, Shunsuke Kon, Susumu Ishikawa, Takashi Hayashi, Tomohiro Yamazaki, Masazumi Tada, Yasuyuki Fujita

    Proceedings of the National Academy of Sciences of the United States of America   114 ( 12 ) E2327-E2336 - E2336  2017年03月  [査読有り]  [国際誌]

     概要を見る

    Newly emerging transformed cells are often eliminated from epithelial tissues. Recent studies have revealed that this cancer-preventive process involves the interaction with the surrounding normal epithelial cells; however, the molecular mechanisms underlying this phenomenon remain largely unknown. In this study, using mammalian cell culture and zebrafish embryo systems, we have elucidated the functional involvement of endocytosis in the elimination of RasV12-transformed cells. First, we show that Rab5, a crucial regulator of endocytosis, is accumulated in RasV12-transformed cells that are surrounded by normal epithelial cells, which is accompanied by up-regulation of clathrin-dependent endocytosis. Addition of chlorpromazine or coexpression of a dominant-negative mutant of Rab5 suppresses apical extrusion of RasV12 cells from the epithelium. We also show in zebrafish embryos that Rab5 plays an important role in the elimination of transformed cells from the enveloping layer epithelium. In addition, Rab5-mediated endocytosis of E-cadherin is enhanced at the boundary between normal and RasV12 cells. Rab5 functions upstream of epithelial protein lost in neoplasm (EPLIN), which plays a positive role in apical extrusion of RasV12 cells by regulating protein kinase A. Furthermore, we have revealed that epithelial defense against cancer (EDAC) from normal epithelial cells substantially impacts on Rab5 accumulation in the neighboring transformed cells. This report demonstrates that Rab5-mediated endocytosis is a crucial regulator for the competitive interaction between normal and transformed epithelial cells in mammals.

    DOI PubMed

  • Engineered erythrocytes covalently linked to antigenic peptides can protect against autoimmune disease.

    Novalia Pishesha, Angelina M Bilate, Marsha C Wibowo, Nai-Jia Huang, Zeyang Li, Rhogerry Deshycka, Djenet Bousbaine, Hojun Li, Heide C Patterson, Stephanie K Dougan, Takeshi Maruyama, Harvey F Lodish, Hidde L Ploegh

    Proceedings of the National Academy of Sciences of the United States of America   114 ( 12 ) 3157 - 3162  2017年03月  [査読有り]  [国際誌]

     概要を見る

    Current therapies for autoimmune diseases rely on traditional immunosuppressive medications that expose patients to an increased risk of opportunistic infections and other complications. Immunoregulatory interventions that act prophylactically or therapeutically to induce antigen-specific tolerance might overcome these obstacles. Here we use the transpeptidase sortase to covalently attach disease-associated autoantigens to genetically engineered and to unmodified red blood cells as a means of inducing antigen-specific tolerance. This approach blunts the contribution to immunity of major subsets of immune effector cells (B cells, CD4+ and CD8+ T cells) in an antigen-specific manner. Transfusion of red blood cells expressing self-antigen epitopes can alleviate and even prevent signs of disease in experimental autoimmune encephalomyelitis, as well as maintain normoglycemia in a mouse model of type 1 diabetes.

    DOI PubMed

  • Plectin is a novel regulator for apical extrusion of RasV12-transformed cells.

    Ailijiang Kadeer, Takeshi Maruyama, Mihoko Kajita, Tomoko Morita, Ayana Sasaki, Atsuko Ohoka, Susumu Ishikawa, Masaya Ikegawa, Takashi Shimada, Yasuyuki Fujita

    Scientific reports   7   44328 - 44328  2017年03月  [査読有り]  [国際誌]

     概要を見る

    Several lines of evidence have revealed that newly emerging transformed cells are often eliminated from the epithelium, though the underlying molecular mechanisms of this cancer preventive phenomenon still remain elusive. In this study, using mammalian cell culture systems we have identified plectin, a versatile cytoskeletal linker protein, as a novel regulator for apical extrusion of RasV12-transformed cells. Plectin is accumulated in RasV12 cells when they are surrounded by normal epithelial cells. Similarly, cytoskeletal proteins tubulin, keratin, and Epithelial Protein Lost In Neoplasm (EPLIN) are also accumulated in the transformed cells surrounded by normal cells. Knockdown or functional disruption of one of these molecules diminishes the accumulation of the others, indicating that the accumulation process of the individual protein mutually depends on each other. Furthermore, plectin-knockdown attenuates caveolin-1 (Cav-1) enrichment and PKA activity in RasV12 cells and profoundly suppresses the apical extrusion. These results indicate that the plectin-microtubules-EPLIN complex positively regulates apical elimination of RasV12-transformed cells from the epithelium in a coordinated fashion. Further development of this study would open a new avenue for cancer preventive medicine.

    DOI PubMed

  • Increasing the efficiency of precise genome editing with CRISPR-Cas9 by inhibition of nonhomologous end joining (vol 33, pg 538, 2015)

    Takeshi Maruyama, Stephanie K. Dougan, Matthias C. Truttmann, Angelina M. Bilate, Jessica R. Ingram, Hidde L. Ploegh

    NATURE BIOTECHNOLOGY   34 ( 2 ) 210 - 210  2016年02月  [査読有り]

    DOI

  • Graphene Oxide Nanosheets Modified with Single-Domain Antibodies for Rapid and Efficient Capture of Cells.

    Guan-Yu Chen, Zeyang Li, Christopher S Theile, Neelkanth M Bardhan, Priyank V Kumar, Joao N Duarte, Takeshi Maruyama, Ali Rashidfarrokh, Angela M Belcher, Hidde L Ploegh

    Chemistry (Weinheim an der Bergstrasse, Germany)   21 ( 48 ) 17178 - 83  2015年11月  [査読有り]  [国際誌]

     概要を見る

    Peripheral blood can provide valuable information on an individual's immune status. Cell-based assays typically target leukocytes and their products. Characterization of leukocytes from whole blood requires their separation from the far more numerous red blood cells.1 Current methods to classify leukocytes, such as recovery on antibody-coated beads or fluorescence-activated cell sorting require long sample preparation times and relatively large sample volumes.2 A simple method that enables the characterization of cells from a small peripheral whole blood sample could overcome limitations of current analytical techniques. We describe the development of a simple graphene oxide surface coated with single-domain antibody fragments. This format allows quick and efficient capture of distinct WBC subpopulations from small samples (∼30 μL) of whole blood in a geometry that does not require any specialized equipment such as cell sorters or microfluidic devices.

    DOI PubMed

  • Pre-emptive Quality Control Protects the ER from Protein Overload via the Proximity of ERAD Components and SRP.

    Hisae Kadowaki, Atsushi Nagai, Takeshi Maruyama, Yasunari Takami, Pasjan Satrimafitrah, Hironori Kato, Arata Honda, Tomohisa Hatta, Tohru Natsume, Takashi Sato, Hirofumi Kai, Hidenori Ichijo, Hideki Nishitoh

    Cell reports   13 ( 5 ) 944 - 56  2015年11月  [査読有り]  [国際誌]

     概要を見る

    Cells possess ER quality control systems to adapt to ER stress and maintain their function. ER-stress-induced pre-emptive quality control (ER pQC) selectively degrades ER proteins via translocational attenuation during ER stress. However, the molecular mechanism underlying this process remains unclear. Here, we find that most newly synthesized endogenous transthyretin proteins are rerouted to the cytosol without cleavage of the signal peptide, resulting in proteasomal degradation in hepatocytes during ER stress. Derlin family proteins (Derlins), which are ER-associated degradation components, reroute specific ER proteins, but not ER chaperones, from the translocon to the proteasome through interactions with the signal recognition particle (SRP). Moreover, the cytosolic chaperone Bag6 and the AAA-ATPase p97 contribute to the degradation of ER pQC substrates. These findings demonstrate that Derlins-mediated substrate-specific rerouting and Bag6- and p97-mediated effective degradation contribute to the maintenance of ER homeostasis without the need for translocation.

    DOI PubMed

  • Disruption of Sphingolipid Biosynthesis Blocks Phagocytosis of Candida albicans.

    Fikadu G Tafesse, Ali Rashidfarrokhi, Florian I Schmidt, Elizaveta Freinkman, Stephanie Dougan, Michael Dougan, Alexandre Esteban, Takeshi Maruyama, Karin Strijbis, Hidde L Ploegh

    PLoS pathogens   11 ( 10 ) e1005188  2015年10月  [査読有り]  [国際誌]

     概要を見る

    The ability of phagocytes to clear pathogens is an essential attribute of the innate immune response. The role of signaling lipid molecules such as phosphoinositides is well established, but the role of membrane sphingolipids in phagocytosis is largely unknown. Using a genetic approach and small molecule inhibitors, we show that phagocytosis of Candida albicans requires an intact sphingolipid biosynthetic pathway. Blockade of serine-palmitoyltransferase (SPT) and ceramide synthase-enzymes involved in sphingolipid biosynthesis- by myriocin and fumonisin B1, respectively, impaired phagocytosis by phagocytes. We used CRISPR/Cas9-mediated genome editing to generate Sptlc2-deficient DC2.4 dendritic cells, which lack serine palmitoyl transferase activity. Sptlc2-/- DC2.4 cells exhibited a stark defect in phagocytosis, were unable to bind fungal particles and failed to form a normal phagocytic cup to engulf C. albicans. Supplementing the growth media with GM1, the major ganglioside present at the cell surface, restored phagocytic activity of Sptlc2-/- DC2.4 cells. While overall membrane trafficking and endocytic pathways remained functional, Sptlc2-/- DC2.4 cells express reduced levels of the pattern recognition receptors Dectin-1 and TLR2 at the cell surface. Consistent with the in vitro data, compromised sphingolipid biosynthesis in mice sensitizes the animal to C. albicans infection. Sphingolipid biosynthesis is therefore critical for phagocytosis and in vivo clearance of C. albicans.

    DOI PubMed

  • Increasing the efficiency of precise genome editing with CRISPR-Cas9 by inhibition of nonhomologous end joining.

    Takeshi Maruyama, Stephanie K Dougan, Matthias C Truttmann, Angelina M Bilate, Jessica R Ingram, Hidde L Ploegh

    Nature biotechnology   33 ( 5 ) 538 - 42  2015年05月  [査読有り]  [国際誌]

     概要を見る

    Methods to introduce targeted double-strand breaks (DSBs) into DNA enable precise genome editing by increasing the rate at which externally supplied DNA fragments are incorporated into the genome through homologous recombination. The efficiency of these methods is limited by nonhomologous end joining (NHEJ), an alternative DNA repair pathway that competes with homology-directed repair (HDR). To promote HDR at the expense of NHEJ, we targeted DNA ligase IV, a key enzyme in the NHEJ pathway, using the inhibitor Scr7. Scr7 treatment increased the efficiency of HDR-mediated genome editing, using Cas9 in mammalian cell lines and in mice for all four genes examined, up to 19-fold. This approach should be applicable to other customizable endonucleases, such as zinc finger nucleases and transcription activator-like effector nucleases, and to nonmammalian cells with sufficiently conserved mechanisms of NHEJ and HDR.

    DOI PubMed

  • The E2 ubiquitin-conjugating enzyme UBE2J1 is required for spermiogenesis in mice.

    Paul-Albert Koenig, Peter K Nicholls, Florian I Schmidt, Masatoshi Hagiwara, Takeshi Maruyama, Galit H Frydman, Nicki Watson, David C Page, Hidde L Ploegh

    The Journal of biological chemistry   289 ( 50 ) 34490 - 502  2014年12月  [査読有り]  [国際誌]

     概要を見る

    ER-resident proteins destined for degradation are dislocated into the cytosol by components of the ER quality control machinery for proteasomal degradation. Dislocation substrates are ubiquitylated in the cytosol by E2 ubiquitin-conjugating/E3 ligase complexes. UBE2J1 is one of the well-characterized E2 enzymes that participate in this process. However, the physiological function of Ube2j1 is poorly defined. We find that Ube2j1(-/-) mice have reduced viability and fail to thrive early after birth. Male Ube2j1(-/-) mice are sterile due to a defect in late spermatogenesis. Ultrastructural analysis shows that removal of the cytoplasm is incomplete in Ube2j1(-/-) elongating spermatids, compromising the release of mature elongate spermatids into the lumen of the seminiferous tubule. Our findings identify an essential function for the ubiquitin-proteasome-system in spermiogenesis and define a novel, non-redundant physiological function for the dislocation step of ER quality control.

    DOI PubMed

  • GPR107, a G-protein-coupled receptor essential for intoxication by Pseudomonas aeruginosa exotoxin A, localizes to the Golgi and is cleaved by furin.

    Fikadu G Tafesse, Carla P Guimaraes, Takeshi Maruyama, Jan E Carette, Stephen Lory, Thijn R Brummelkamp, Hidde L Ploegh

    The Journal of biological chemistry   289 ( 35 ) 24005 - 18  2014年08月  [査読有り]  [国際誌]

     概要を見る

    A number of toxins, including exotoxin A (PE) of Pseudomonas aeruginosa, kill cells by inhibiting protein synthesis. PE kills by ADP-ribosylation of the translation elongation factor 2, but many of the host factors required for entry, membrane translocation, and intracellular transport remain to be elucidated. A genome-wide genetic screen in human KBM7 cells was performed to uncover host factors used by PE, several of which were confirmed by CRISPR/Cas9-gene editing in a different cell type. Several proteins not previously implicated in the PE intoxication pathway were identified, including GPR107, an orphan G-protein-coupled receptor. GPR107 localizes to the trans-Golgi network and is essential for retrograde transport. It is cleaved by the endoprotease furin, and a disulfide bond connects the two cleaved fragments. Compromising this association affects the function of GPR107. The N-terminal region of GPR107 is critical for its biological function. GPR107 might be one of the long-sought receptors that associates with G-proteins to regulate intracellular vesicular transport.

    DOI PubMed

  • Engineered red blood cells as carriers for systemic delivery of a wide array of functional probes.

    Jiahai Shi, Lenka Kundrat, Novalia Pishesha, Angelina Bilate, Chris Theile, Takeshi Maruyama, Stephanie K Dougan, Hidde L Ploegh, Harvey F Lodish

    Proceedings of the National Academy of Sciences of the United States of America   111 ( 28 ) 10131 - 6  2014年07月  [査読有り]  [国際誌]

     概要を見る

    We developed modified RBCs to serve as carriers for systemic delivery of a wide array of payloads. These RBCs contain modified proteins on their plasma membrane, which can be labeled in a sortase-catalyzed reaction under native conditions without inflicting damage to the target membrane or cell. Sortase accommodates a wide range of natural and synthetic payloads that allow modification of RBCs with substituents that cannot be encoded genetically. As proof of principle, we demonstrate site-specific conjugation of biotin to in vitro-differentiated mouse erythroblasts as well as to mature mouse RBCs. Thus modified, RBCs remain in the bloodstream for up to 28 d. A single domain antibody attached enzymatically to RBCs enables them to bind specifically to target cells that express the antibody target. We extend these experiments to human RBCs and demonstrate efficient sortase-mediated labeling of in vitro-differentiated human reticulocytes.

    DOI PubMed

  • Roquin-2 promotes ubiquitin-mediated degradation of ASK1 to regulate stress responses.

    Takeshi Maruyama, Toshihiro Araki, Yosuke Kawarazaki, Isao Naguro, Susanne Heynen, Pedro Aza-Blanc, Ze'ev Ronai, Atsushi Matsuzawa, Hidenori Ichijo

    Science signaling   7 ( 309 ) ra8  2014年01月  [査読有り]  [国際誌]

     概要を見る

    Apoptosis signal-regulating kinase 1 (ASK1, also known as MAP3K5) mediates reactive oxygen species (ROS)-induced cell death. When activated by ROS, ASK1 ultimately becomes ubiquitinated and degraded by the proteasome, a process that is antagonized by the ubiquitin-specific protease USP9X. Using a functional siRNA (small interfering RNA) screen in HeLa cells, we identified Roquin-2 (also called RC3H2) as an E3 ubiquitin ligase required for ROS-induced ubiquitination and degradation of ASK1. In cells treated with H2O2, knockdown of Roquin-2 promoted sustained activation of ASK1 and the downstream stress-responsive kinases JNK (c-Jun amino-terminal kinase) and p38 MAPK (mitogen-activated protein kinase), and led to cell death. The nematode Caenorhabditis elegans produces ROS as a defense mechanism in response to bacterial infection. In C. elegans, mutation of the gene encoding the Roquin-2 ortholog RLE-1 promoted accumulation of the activated form of the ASK1 ortholog NSY-1 and conferred resistance to infection by the bacteria Pseudomonas aeruginosa. Thus, these data suggest that degradation of ASK1 mediated by Roquin-2 is an evolutionarily conserved mechanism required for the appropriate regulation of stress responses, including pathogen resistance and cell death.

    DOI PubMed

  • Type I interferon imposes a TSG101/ISG15 checkpoint at the Golgi for glycoprotein trafficking during influenza virus infection.

    Sumana Sanyal, Joseph Ashour, Takeshi Maruyama, Arwen F Altenburg, Juan Jose Cragnolini, Angelina Bilate, Ana M Avalos, Lenka Kundrat, Adolfo García-Sastre, Hidde L Ploegh

    Cell host & microbe   14 ( 5 ) 510 - 21  2013年11月  [査読有り]  [国際誌]

     概要を見る

    Several enveloped viruses exploit host pathways, such as the cellular endosomal sorting complex required for transport (ESCRT) machinery, for their assembly and release. The influenza A virus (IAV) matrix protein binds to the ESCRT-I complex, although the involvement of early ESCRT proteins such as Tsg101 in IAV trafficking remain to be established. We find that Tsg101 can facilitate IAV trafficking, but this is effectively restricted by the interferon (IFN)-stimulated protein ISG15. Cytosol from type I IFN-treated cells abolished IAV hemagglutinin (HA) transport to the cell surface in infected semi-intact cells. This inhibition required Tsg101 and could be relieved with deISGylases. Tsg101 is itself ISGylated in IFN-treated cells. Upon infection, intact Tsg101-deficient cells obtained by CRISPR-Cas9 genome editing were defective in the surface display of HA and for infectious virion release. These data support the IFN-induced generation of a Tsg101- and ISG15-dependent checkpoint in the secretory pathway that compromises influenza virus release.

    DOI PubMed

  • CHIP-dependent termination of MEKK2 regulates temporal ERK activation required for proper hyperosmotic response.

    Takeshi Maruyama, Hisae Kadowaki, Noriaki Okamoto, Atsushi Nagai, Isao Naguro, Atsushi Matsuzawa, Hiroshi Shibuya, Keiji Tanaka, Shigeo Murata, Kohsuke Takeda, Hideki Nishitoh, Hidenori Ichijo

    The EMBO journal   29 ( 15 ) 2501 - 14  2010年08月  [査読有り]  [国際誌]

     概要を見る

    The extracellular signal-regulated kinase (ERK) pathway is an important signalling pathway that regulates a large number of cellular processes, including proliferation, differentiation and gene expression. Hyperosmotic stress activates the ERK pathway, whereas little is known about the regulatory mechanisms and physiological functions of ERK activation in hyperosmotic response. Here, we show that MAPK/ERK kinase kinase 2 (MEKK2), a member of the MAPKKK family, mediated the specific and transient activation of ERK, which was required for the induction of aquaporin 1 (AQP1) and AQP5 gene expression in response to hyperosmotic stress. Moreover, we identified the E3 ubiquitin ligase carboxyl terminus of Hsc70-interacting protein (CHIP) as a binding partner of MEKK2. Depletion of CHIP by small-interference RNA or gene targeting attenuated the degradation of MEKK2 and prolonged the ERK activity. Interestingly, hyperosmolality-induced gene expression of AQP1 and AQP5 was suppressed by CHIP depletion and was reversed by inhibition of the prolonged phase of ERK activity. These findings show that transient activation of the ERK pathway, which depends not only on MEKK2 activation, but also on CHIP-dependent MEKK2 degradation, is crucial for proper gene expression in hyperosmotic stress response.

    DOI PubMed

  • USP14 inhibits ER-associated degradation via interaction with IRE1alpha.

    Atsushi Nagai, Hisae Kadowaki, Takeshi Maruyama, Kohsuke Takeda, Hideki Nishitoh, Hidenori Ichijo

    Biochemical and biophysical research communications   379 ( 4 ) 995 - 1000  2009年02月  [査読有り]  [国際誌]

     概要を見る

    Accumulation of unfolded proteins within the endoplasmic reticulum (ER) lumen induces ER stress. Eukaryotic cells possess the ER quality control systems, the unfolded protein response (UPR), to adapt to ER stress. IRE1alpha is one of the ER stress receptors and mediates the UPR. Here, we identified ubiquitin specific protease (USP) 14 as a binding partner of IRE1alpha. USP14 interacted with the cytoplasmic region of IRE1alpha, and the endogenous interaction between USP14 and IRE1alpha was inhibited by ER stress. Overexpression of USP14 inhibited the ER-associated degradation (ERAD) pathway, and USP14 depletion by small interfering RNA effectively activated ERAD. These findings suggest that USP14 is a novel player in the UPR by serving as a physiological inhibitor of ERAD under the non-stressed condition.

    DOI PubMed

  • ALS-linked mutant SOD1 induces ER stress- and ASK1-dependent motor neuron death by targeting Derlin-1.

    Hideki Nishitoh, Hisae Kadowaki, Atsushi Nagai, Takeshi Maruyama, Takanori Yokota, Hisashi Fukutomi, Takuya Noguchi, Atsushi Matsuzawa, Kohsuke Takeda, Hidenori Ichijo

    Genes & development   22 ( 11 ) 1451 - 64  2008年06月  [査読有り]  [国際誌]

     概要を見る

    Mutation in Cu/Zn-superoxide dismutase (SOD1) is a cause of familial amyotrophic lateral sclerosis (ALS). Mutant SOD1 protein (SOD1(mut)) induces motor neuron death, although the molecular mechanism of SOD1(mut)-induced cell death remains controversial. Here we show that SOD1(mut) specifically interacted with Derlin-1, a component of endoplasmic reticulum (ER)-associated degradation (ERAD) machinery and triggered ER stress through dysfunction of ERAD. SOD1(mut)-induced ER stress activated the apoptosis signal-regulating kinase 1 (ASK1)-dependent cell death pathway. Perturbation of binding between SOD1(mut) and Derlin-1 by Derlin-1-derived oligopeptide suppressed SOD1(mut)-induced ER stress, ASK1 activation, and motor neuron death. Moreover, deletion of ASK1 mitigated the motor neuron loss and extended the life span of SOD1(mut) transgenic mice. These findings demonstrate that ER stress-induced ASK1 activation, which is triggered by the specific interaction of Derlin-1 with SOD1(mut), is crucial for disease progression of familial ALS.

    DOI PubMed

▼全件表示

Misc

  • がん変異細胞排除のためのMHC-I/AltR相互作用を介した上皮細胞の偏向性移動

    寺本龍平, 中山淳, 鴨下渚, 丸山剛

    日本分子生物学会年会プログラム・要旨集(Web)   43rd  2020年

    J-GLOBAL

  • 上皮細胞層における哺乳類細胞競合現象の観察方法

    丸山 剛, 藤田 恭之

    実験医学   36 ( 4 ) 575 - 583  2018年

    担当区分:筆頭著者

  • 正常上皮細胞によるがん細胞の排除

    丸山 剛, 藤田 恭之

    生体の科学   67 ( 2 ) 95 - 100  2016年

  • CRISPR/Cas9でHDR効率を高める方法

    丸山 剛

    実験医学   34 ( 1 ) 93 - 101  2016年

  • 小胞体(ER)ストレス誘導性アポトーシスの分子メカニズム

    丸山 剛, 西頭 英起, 一條 秀憲

    血管医学   9 ( 4 ) 331 - 336  2008年

受賞

  • MSD(メルク・アンド・カンパニー)生命科学財団

    2021年  

  • 三菱財団

    2020年  

  • テルモ生命科学振興財団

    2019年  

  • SGHがん研究助成

    2019年  

  • 研究助成・奨励金

    2018年10月   内藤記念科学振興財団  

    受賞者: 丸山 剛

  • ビジョナリーリサーチ(スタート)研究助成

    2017年   武田科学記念財団  

    受賞者: 丸山 剛

  • 研究助成

    2017年   加藤記念バイオサイエンス振興財団  

    受賞者: 丸山 剛

  • 研究助成

    2015年   小野がん研究助成基金  

    受賞者: 丸山 剛

▼全件表示

共同研究・競争的資金等の研究課題

  • 超早期がん状態を投影する脳波シグナルの特定

    日本学術振興会  科学研究費助成事業 挑戦的研究(開拓)

    研究期間:

    2022年06月
    -
    2025年03月
     

    丸山 剛, 関口 寛人, 瀬戸川 将, 大川 宜昭

  • 基盤B 異常細胞のMHC-Iシグナルにより惹起される上皮細胞の免疫細胞様機能

    日本学術振興会(JSPS)  科学研究費助成事業 基盤研究(B)

    研究期間:

    2021年
    -
    2023年
     

  • AMED prime 時空間的異常センシングによるがん変異細胞除去修復

    日本医療研究開発機構研究費(AMED)  革新的先端研究開発事業・PRIME

    研究期間:

    2019年
    -
    2023年
     

  • 基盤B がん変異細胞の抗原提示と正常上皮細胞による認識排除機構

    JSPS  基盤研究(B)

    研究期間:

    2018年
    -
    2021年
     

    丸山 剛

  • 萌芽 生体内に注射針にて注入・展開・動作可能なナノ薄膜状アンテナの開発

    JSPS  挑戦的研究(萌芽)

    研究期間:

    2017年
    -
    2020年
     

  • さきがけ 光操作型ー生体内不均一変異細胞誘導と変異細胞の挙動解明

    JST  さきがけ

    研究期間:

    2016年
    -
    2020年
     

    丸山 剛

  • AMED 革新がん 細胞競合による変異細胞排除機構を応用した新規がん予防・治療薬の開発

    AMED  革新的がん医療実用化研究事業

    研究期間:

    2016年
    -
    2018年
     

    丸山 剛

  • がん細胞から正常細胞への細胞外プッシュミーアウトシグナル関連分子の同定

    JSPS  研究活動スタート支援

    研究期間:

    2015年
    -
    2016年
     

    丸山 剛

▼全件表示

特定課題研究

  • 正常細胞による硬さおよび抗原提示認識によるがん変異細胞排除機構

    2020年  

     概要を見る

    上皮組織細胞層にがん変異細胞が生じたとき、それを取り囲む周辺上皮細胞はこの変異細胞を駆逐する。しかしながら、どのような細胞間コミュニケーション機構が上皮細胞の抗腫瘍能を惹起しているかについては多くが不明である。変異細胞に隣接する上皮細胞はAltR(Suboptimalalteration recognizing protein)の発現を亢進させ、このAltRによってMHC-Iを認識することを見出した。さらに正常細胞内での詳細なシグナルも解析し、変異細胞を排除することを解明した。さらに、マウス肺における化学発がんモデルでは肺における上皮細胞由来のadenomaの発生を抑制する。これらのことから、これまで未解明であった上皮細胞の腫瘍能を促進する機構を解明した。

  • 異常細胞の抗原提示と正常上皮細胞による認識排除機構

    2019年  

     概要を見る

    我々は機能未知である受容体膜タンパク質(Suboptimal alteration recognizing protein:AltRと命名)に注目した。AltRはそのドメイン構成から免疫系に関わると予測されてきたが、同受容体を活性化するリガンドは不明であり、上皮細胞にはほとんど発現しない。しかし興味深いことに、AltRはがん変異細胞の硬さ依存的に上皮細胞でも誘導されることを見出した。硬さにより誘導されたAltRはRasV12の発現依存的に形質膜に移行促進された抗原提示関連膜タンパク質を認識する。このように、異常細胞を認識することで、正常細胞の排除能を惹起する。このことは、非免疫系である上皮細胞は抗原提示変化を認識できるという、免疫細胞に類似した機能が備わっていることを世界で初めて示した例である。

  • 異常細胞の抗原提示と正常上皮細胞による認識排除機構

    2018年  

     概要を見る

    イヌの細胞であるMDCK細胞には4種の抗原提示関連膜タンパク質Epithelial antigen associated protein (ECA)が存在する。この内、ECA1が変異―正常細胞間シグナルに必須であることがわかった。さらに、受容体タンパク質としてECA1受容体(ECAR)を同定している。重要なことに、ECA1もしくはECARの細胞外ドメインのリコンビナントタンパク質は、変異細胞の排除を促進および抑制した。このことから、変異細胞側のECA1が上流に位置し、正常細胞側のECARにシグナルを受け渡していることが示唆された。変異細胞側のリガンドタンパク質の同定および正常細胞側の受容体の同定、この大きな2つの点について計画通りの進行状況である。

 

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