Faculty Profiles - ENOMOTO, Atsushi

Dissection of the function of CD109 in lung adenocarcinoma

Tetsuro Taki, Shinji Mii, Yukihiro Shiraki, Atsushi Enomoto, Masahide Takahashi

CANCER SCIENCE Vol. 109 page： 1407 - 1407 2018.12

Identification of cancer restraining CAF

Mizutani, Y; Enomoto, A; Takahashi, M

CANCER SCIENCE Vol. 109 page： 1396 - 1396 2018.12

Maintenance of Undifferentiated Mesenchymal Stromal Cells-Like Fibroblast by Meflin is Crucial for Heart Repair and Homeostasis

Hara, A; Enomoto, A; Murohara, T

CIRCULATION Vol. 138 2018.11

ABERRANT ACTIVE-ENHANCERS ASSOCIATED WITH DOWNREGULATION OF HDAC1-RET FINGER PROTEIN COMPLEX OVERCOME CHEMORESISTANCE IN GLIOBLASTOMA

Hirano, M; Ranjit, M; Ohka, F; Aoki, K; Yamamichi, A; Kato, T; Matsuo, K; Enomoto, A; Takahashi, M; Wakabayashi, T; Natsume, A

NEURO-ONCOLOGY Vol. 20 page： 111 - 111 2018.11

Girdin/GIV Regulates Collective Cancer Cell Migration by Controlling Cell Adhesion and Cytoskeletal Organization. Reviewed International journal

Wang X, Enomoto A, Weng L, Mizutani Y, Abudureyimu S, Esaki N, Tsuyuki Y, Chen C, Mii S, Asai N, Haga H, Ishida S, Yokota K, Akiyama M, Takahashi M

Cancer science Vol. 109 ( 11 ) page： 3643 - 3656 2018.11

Language：English Publishing type：Research paper (scientific journal)

Pathological observations show that cancer cells frequently invade the surrounding stroma in collective groups rather than through single cell migration. Here, we studied the role of the actin-binding protein Girdin, a specific regulator of collective migration of neuroblasts in the brain, in collective cancer cell migration. We found that Girdin was essential for the collective migration of the skin cancer cell line A431 on collagen gels as well as their fibroblast-led collective invasion in an organotypic culture model. We provide evidence that Girdin binds to β-catenin that plays important roles in the Wnt signaling pathway and in E-cadherin-mediated cell-cell adhesion. Girdin-depleted cells displayed scattering and impaired E-cadherin-specific cell-cell adhesion. Importantly, Girdin depletion led to impaired cytoskeletal association of the β-catenin complex, which was accompanied by changes in the supracellular actin cytoskeletal organization of cancer cell cohorts on collagen gels. Although the underlying mechanism is unclear, this observation is consistent with the established role of the actin cytoskeletal system and cell-cell adhesion in the collective behavior of cells. Finally, we showed the correlation of the expression of Girdin with that of the components of the E-cadherin complex and the differentiation of human skin cancer. Collectively, our results suggest that Girdin is an important modulator of the collective behavior of cancer cells.

DOI： 10.1111/cas.13795

Trefoil factor 1 inhibits epithelial-mesenchymal transition of pancreatic intraepithelial neoplasm. Reviewed

Large cell neuroendocrine carcinoma of the breast showing triple negative, basal-like subtype Reviewed

Kawasaki T, Hasebe T, Ichihara S, Oiwa M, Sato Y, Morita T, Hayashi T, Kato A, Sugiyama K, Nozawa K, Enomoto A, Takahashi M, Horibe K, Naoe T, Osaki A, Saeki T

VIRCHOWS ARCHIV Vol. 473 page： S208 2018.9

Yamaguchi J, Yokoyama Y, Kokuryo T, Ebata T, Enomoto A, Nagino M

The Journal of clinical investigation Vol. 128 ( 8 ) page： 3619 - 3629 2018.8

The actin-binding protein Girdin/GIV regulates collective cancer cell migration by controlling cell adhesion and cytoskeletal organization

Wang, XZ; Enomoto, A; Weng, L; Haga, H; Ishida, S; Takahashi, M

CANCER RESEARCH Vol. 78 ( 13 ) 2018.7

CD109 deficiency induces osteopenia with an osteoporosis-like phenotype in vivo. Reviewed International journal

Shinji Mii, Akiyoshi Hoshino, Atsushi Enomoto, Yoshiki Murakumo, Masako Ito, Akira Yamaguchi, Masahide Takahashi

Genes to cells : devoted to molecular & cellular mechanisms Vol. 23 ( 7 ) page： 590 - 598 2018.7

Language：English Publishing type：Research paper (scientific journal)

Osteoporosis is a global public health problem that is increasing along with an aging population. A major determinant of osteoporosis is high bone turnover, which results from osteoclast activation. CD109 is a glycosylphosphatidylinositol-anchored glycoprotein, a deficiency that leads to a psoriasis-like skin inflammation in mice. Although the expression of CD109 has been reported in mouse pre-osteoclast cells, its function in osteoclasts in vivo remains largely unknown. To investigate the physiological role of CD109 in bone metabolism, we analyzed bones from wild-type and CD109-deficient adult mice. Micro-computed tomography analysis of the femur (thigh bone) showed that bone volume was lower in CD109-deficient mice than in wild-type mice. Bone histomorphometric analysis showed not only a reduction in bone volume but also an increase in bone turnover in CD109-deficient mice as compared with wild-type mice. Additionally, we measured serum levels of several markers of bone turnover and found a significant increase in the N-terminal telopeptide of type I collagen, a bone resorption marker, as well as alkaline phosphatase, a bone formation marker, in CD109-deficient mice. These results indicate that CD109 deficiency induces a high-turnover, osteoporosis-like phenotype, which suggests that CD109 plays a role in bone metabolism in vivo.

DOI： 10.1111/gtc.12593

INHIBITION OF MTORC1 THROUGH AMINO ACID TRANSPORTER CD98/LAT1 MAINTAINS THE STEMNESS OF GLIOMA STEM CELL

Han, YP; Enomoto, A; Takahashi, M; Ma, J

NEURO-ONCOLOGY Vol. 20 page： 180 - 180 2018.6

Essential Role of Linx/Islr2 in the Development of the Forebrain Anterior Commissure Reviewed

Shaniya Abudureyimu, Naoya Asai, Atsushi Enomoto, Liang Weng, Hiroki Kobayashi, Xiaoze Wang, Chen Chen, Shinji Mii, Masahide Takahashi

Scientific Reports Vol. 8 ( 1 ) page： 7292 2018.5

Language：English Publishing type：Research paper (scientific journal) Publisher：Nature Publishing Group

Linx is a member of the leucine-rich repeat and immunoglobulin family of membrane proteins which has critical roles in the development of the peripheral nervous system and forebrain connectivity. A previous study showed that Linx is expressed in projection neurons in the cortex and in cells that comprise the passage to the prethalamus that form the internal capsule, indicating the involvement of Linx in axon guidance and cell-cell communication. In this study, we found that Linx-deficient mice develop severe hydrocephalus and die perinatally by unknown mechanisms. Importantly, mice heterozygous for the linx gene exhibited defects in the development of the anterior commissure in addition to hydrocephalus, indicating haploinsufficiency of the linx gene in forebrain development. In N1E-115 neuroblastoma cells and primary cultured hippocampal neurons, Linx depletion led to impaired neurite extension and an increase in cell body size. Consistent with this, but of unknown significance, we found that Linx interacts with and upregulates the activity of Rho-kinase, a modulator of many cellular processes including cytoskeletal organization. These data suggest a role for Linx in the regulation of complex forebrain connectivity, and future identification of its extracellular ligand(s) will help clarify this function.

DOI： 10.1038/s41598-018-24064-0

Regulation of keratin 5/14 intermediate filaments by CDK1, Aurora-B, and Rho-kinase Reviewed

金属接着凍結法による脂肪を含む凍結標本作製の試み

牛田かおり, 内山孝蔵, 浅井直也, 榎本篤, 高橋雅英

病理と臨床 Vol. 36 ( 5 ) page： 493 - 497 2018.5

Hironori Inaba, Daishi Yamakawa, Yasuko Tomono, Atsushi Enomoto, Shinji Mii, Kousuke Kasahara, Hidemasa Goto, Masaki Inagaki

Biochemical and Biophysical Research Communications Vol. 498 ( 3 ) page： 544 - 550 2018.4

Language：English Publishing type：Research paper (scientific journal) Publisher：Elsevier B.V.

We previously reported that vimentin, GFAP, and desmin (type III intermediate filament [IF] proteins) are mitotically phosphorylated by CDK1, Aurora-B, and Rho-kinase. This phosphorylation is critical for efficient separation of these IFs and completion of cytokinesis. Keratin 5 (K5) and K14 form a heterodimer, which constitutes IF network in basal layer cells of stratified squamous epithelia. Here, we report that the solubility of K5/K14 increased in mitosis. The in vitro assays revealed that three mitotic kinases phosphorylate K5 more than K14. We then identified Thr23/Thr144, Ser30, and Thr159 on murine K5 as major phosphorylation sites for CDK1, Aurora-B, and Rho-kinase, respectively. Using site- and phosphorylation-state-specific antibodies, we demonstrated that K5-Thr23 was phosphorylated in entire cytoplasm from prometaphase to metaphase, whereas K5-Ser30 phosphorylation occurred specifically at the cleavage furrow from anaphase to telophase. Efficient K5/K14-IF separation was impaired by K5 mutations at the sites phosphorylated by these mitotic kinases. K5-Thr23 phosphorylation was widely detected in dividing K5-positive cells of murine individuals. These results suggested that mitotic reorganization of K5/K14-IF network is governed largely through K5 phosphorylation by CDK1, Aurora-B, and Rho-kinase.

DOI： 10.1016/j.bbrc.2018.03.016

Critical role of rabphilin-3A in the pathophysiology of experimental lymphocytic neurohypophysitis Reviewed

Yoshinori Yasuda, Shintaro Iwama, Atsushi Kiyota, Hisakazu Izumida, Kohtaro Nakashima, Naoko Iwata, Yoshihiro Ito, Yoshiaki Morishita, Motomitsu Goto, Hidetaka Suga, Ryoichi Banno, Atsushi Enomoto, Masahide Takahashi, Hiroshi Arima, Yoshihisa Sugimura

Journal of Pathology Vol. 244 ( 4 ) page： 469 - 478 2018.4

Language：English Publishing type：Research paper (scientific journal) Publisher：John Wiley and Sons Ltd

Autoimmune hypophysitis (AH) is thought to be an autoimmune disease characterized by lymphocytic infiltration of the pituitary gland. Among AH pathologies, lymphocytic infundibulo-neurohypophysitis (LINH) involves infiltration of the neurohypophysis and/or the hypothalamic infundibulum, causing central diabetes insipidus resulting from insufficiency of arginine vasopressin secretion. The pathophysiological and pathogenetic mechanisms underlying LINH are largely unknown. Clinically, differentiating LINH from other pituitary diseases accompanied by mass lesions, including tumours, has often been difficult, because of similar clinical manifestations. We recently reported that rabphilin-3A is an autoantigen and that anti-rabphilin-3A antibodies constitute a possible diagnostic marker for LINH. However, the involvement of rabphilin-3A in the pathogenesis of LINH remains to be elucidated. This study was undertaken to explore the role of rabphilin-3A in lymphocytic neurohypophysitis and to investigate the mechanism. We found that immunization of mice with rabphilin-3A led to neurohypophysitis. Lymphocytic infiltration was observed in the neurohypophysis and supraoptic nucleus 1 month after the first immunization. Mice immunized with rabphilin-3A showed an increase in the volume of urine that was hypotonic as compared with control mice. Administration of a cocktail of monoclonal anti-rabphilin-3A antibodies did not induce neurohypophysitis. However, abatacept, which is a chimeric protein that suppresses T-cell activation, decreased the number of T cells specific for rabphilin-3A in peripheral blood mononuclear cells (PBMCs). It ameliorated lymphocytic infiltration of CD3+ T cells in the neurohypophysis of mice that had been immunized with rabphilin-3A. Additionally, there was a linear association between the number of T cells specific for rabphilin-3A in PBMCs and the number of CD3+ T cells infiltrating the neurohypophysis. In conclusion, we suggest that rabphilin-3A is a pathogenic antigen, and that T cells specific for rabphilin-3A are involved in the pathogenesis of neurohypophysitis in mice. Copyright © 2018 Pathological Society of Great Britain and Ireland. Published by John Wiley &amp
Sons, Ltd.

DOI： 10.1002/path.5046

Development of a method to preliminarily embed tissue samples using low melting temperature fish gelatin before sectioning: A technical note Reviewed

Kaori Ushida, Naoya Asai, Kozo Uchiyama, Atsushi Enomoto, Masahide Takahashi

Pathology International Vol. 68 ( 4 ) page： 241 - 245 2018.4

Language：English Publishing type：Research paper (scientific journal) Publisher：Blackwell Publishing

Embedding of tissue samples that maintains a desired orientation is critical for preparing sections suitable for diagnosis and study objectives. Methods to prepare tissue sections include: (i) paraffin embedding or snap-freezing followed by microtome or cryostat sectioning
and (ii) agarose embedding followed by cutting on a vibrating microslicer. Although these methods are useful for routine laboratory work, preparation of small and fragile tissues such as mouse organs, small human biopsy samples, and cultured floating spheres is difficult and requires special skills. In particular, tissue specimen orientation can be lost during embedding in molds and subsequent sectioning. Here, we developed a method using low melting temperature (LM) gelatin either alone or mixed with agarose to preliminarily embed collected tissues that are either prefixed or unfixed, followed by conventional fixation, paraffin embedding, freezing, and sectioning. The advantage of the method is that the LM gelatin and its mixture with agarose can be handled at room temperature but quickly hardens at 4°C, which allows embedding, trimming, and arranging of small and fragile tissues in a desired orientation and are compatible with traditional stainings. Thus, this method can have various laboratory applications and can be modified according to the needs of each laboratory.

DOI： 10.1111/pin.12652

Negative regulation of amino acid signaling by MAPK-regulated 4F2hc/Girdin complex. Reviewed International journal

Liang Weng, Yi-Peng Han, Atsushi Enomoto, Yasuyuki Kitaura, Shushi Nagamori, Yoshikatsu Kanai, Naoya Asai, Jian An, Maki Takagishi, Masato Asai, Shinji Mii, Takashi Masuko, Yoshiharu Shimomura, Masahide Takahashi

PLoS biology Vol. 16 ( 3 ) page： e2005090 2018.3

Language：English Publishing type：Research paper (scientific journal)

Amino acid signaling mediated by the activation of mechanistic target of rapamycin complex 1 (mTORC1) is fundamental to cell growth and metabolism. However, how cells negatively regulate amino acid signaling remains largely unknown. Here, we show that interaction between 4F2 heavy chain (4F2hc), a subunit of multiple amino acid transporters, and the multifunctional hub protein girders of actin filaments (Girdin) down-regulates mTORC1 activity. 4F2hc interacts with Girdin in mitogen-activated protein kinase (MAPK)- and amino acid signaling-dependent manners to translocate to the lysosome. The resultant decrease in cell surface 4F2hc leads to lowered cytoplasmic glutamine (Gln) and leucine (Leu) content, which down-regulates amino acid signaling. Consistently, Girdin depletion augments amino acid-induced mTORC1 activation and inhibits amino acid deprivation-induced autophagy. These findings uncovered the mechanism underlying negative regulation of amino acid signaling, which may play a role in tightly regulated cell growth and metabolism.

DOI： 10.1371/journal.pbio.2005090