TY - JOUR
T1 - Identification of GPI-anchored protein LYPD1 as an essential factor for odontoblast differentiation in tooth development
AU - Fu, Yao
AU - Miyazaki, Kanako
AU - Chiba, Yuta
AU - Funada, Keita
AU - Yuta, Tomomi
AU - Tian, Tian
AU - Mizuta, Kanji
AU - Kawahara, Jumpei
AU - Zhang, Ling
AU - Martin, Daniel
AU - Iwamoto, Tsutomu
AU - Takahashi, Ichiro
AU - Fukumoto, Satoshi
AU - Yoshizaki, Keigo
N1 - Funding Information:
This study was supported by a Grant-in-Aid from the Japan Society for the Promotion of Science (JSPS) KAKENHI ( JP18H03012 , JP21K19610 , JP21H03150 to K. Y., JP17H01606 to S. F., and JP20K18786 to K. M.), JST FOREST Program ( JPMJFR2013 to K. Y.), and JST SPRING (Grant Number JPMJSP2136 ). K. Y. was supported by Takeda Science Foundation . Y. F. was supported by an Otsuka-Toshimi Scholarship .
Funding Information:
We appreciate the technical assistance from The Research Support Center, Research Center for Human Disease Modeling, Kyushu University Graduate School of Medical Sciences. This work utilized the computational resources of the NIH HPC Biowulf cluster ( http://hpc.nih.gov ). We would like to thank NIDCD/NIDCR Genomics and Computational Biology Core authors: E. T. Boger (National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda, Maryland, USA); B. Choudhury (National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda, Maryland, USA); D. Martin (National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA); C. Zheng (National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA); Z. Wei (National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA); R. J. Morell (National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda, Maryland, USA). We also wish to thank Editage ( www.editage.com ) for English language editing. This work was supported by funds from the NIDCD , National Institutes of Health Division of Intramural Research ( ZIC DC000086 to the G. C. B. C.), NIDCR , National Institutes of Health Transfer Core Facility ( ZIC DE000744-04 ), Veterinary Resources Core ( ZIC DE000740-05 ), and the combined technical research core facility ( ZIC DE000729-09 ). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Funding Information:
We appreciate the technical assistance from The Research Support Center, Research Center for Human Disease Modeling, Kyushu University Graduate School of Medical Sciences. This work utilized the computational resources of the NIH HPC Biowulf cluster (http://hpc.nih.gov). We would like to thank NIDCD/NIDCR Genomics and Computational Biology Core authors: E. T. Boger (National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda, Maryland, USA); B. Choudhury (National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda, Maryland, USA); D. Martin (National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA); C. Zheng (National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA); Z. Wei (National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA); R. J. Morell (National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda, Maryland, USA). We also wish to thank Editage (www.editage.com) for English language editing. This work was supported by funds from the NIDCD, National Institutes of Health Division of Intramural Research (ZIC DC000086 to the G. C. B. C.), NIDCR, National Institutes of Health Transfer Core Facility (ZIC DE000744-04), Veterinary Resources Core (ZIC DE000740-05), and the combined technical research core facility (ZIC DE000729-09). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Y. F. K. M. and K. Y. conceptualization; Y. F. K. M. and Y. C. methodology; K. M. Y. C. and K. Y. validation; K. F. T. Y. T. T. K. M. J. K. L. Z. and D. M. formal analysis; Y. F. and K. M. investigation; Y. F. K. M. and Y. C. data curation; Y. F. and K. M. writing–original draft; Y. C. K. Y. and I. T. writing–review & editing; T. I. I. T. S. F. and K. Y. supervision; S. F. and K. Y. project administration; S. F. and K. Y. funding acquisition. This study was supported by a Grant-in-Aid from the Japan Society for the Promotion of Science (JSPS) KAKENHI (JP18H03012, JP21K19610, JP21H03150 to K. Y. JP17H01606 to S. F. and JP20K18786 to K. M.), JST FOREST Program (JPMJFR2013 to K. Y.), and JST SPRING (Grant Number JPMJSP2136). K. Y. was supported by Takeda Science Foundation. Y. F. was supported by an Otsuka-Toshimi Scholarship.
Publisher Copyright:
© 2023 The Authors
PY - 2023/5
Y1 - 2023/5
N2 - Lipid rafts are membrane microdomains rich in cholesterol, sphingolipids, glycosylphosphatidylinositol-anchored proteins (GPI-APs), and receptors. These lipid raft components are localized at the plasma membrane and are essential for signal transmission and organogenesis. However, few reports have been published on the specific effects of lipid rafts on tooth development. Using microarray and single-cell RNA sequencing methods, we found that a GPI-AP, lymphocyte antigen-6/Plaur domain-containing 1 (Lypd1), was specifically expressed in preodontoblasts. Depletion of Lypd1 in tooth germ using an ex vivo organ culture system and in mouse dental pulp (mDP) cells resulted in the inhibition of odontoblast differentiation. Activation of bone morphogenetic protein (BMP) signaling by BMP2 treatment in mDP cells promoted odontoblast differentiation via phosphorylation of Smad1/5/8, while this BMP2-mediated odontoblast differentiation was inhibited by depletion of Lypd1. Furthermore, we created a deletion construct of the C terminus containing the omega site in LYPD1; this site is necessary for localizing GPI-APs to the plasma membrane and lipid rafts. We identified that this site is essential for odontoblast differentiation and morphological change of mDP cells. These findings demonstrated that LYPD1 is a novel marker of preodontoblasts in the developing tooth; in addition, they suggest that LYPD1 is important for tooth development and that it plays a pivotal role in odontoblast differentiation by regulating Smad1/5/8 phosphorylation through its effect as a GPI-AP in lipid rafts.
AB - Lipid rafts are membrane microdomains rich in cholesterol, sphingolipids, glycosylphosphatidylinositol-anchored proteins (GPI-APs), and receptors. These lipid raft components are localized at the plasma membrane and are essential for signal transmission and organogenesis. However, few reports have been published on the specific effects of lipid rafts on tooth development. Using microarray and single-cell RNA sequencing methods, we found that a GPI-AP, lymphocyte antigen-6/Plaur domain-containing 1 (Lypd1), was specifically expressed in preodontoblasts. Depletion of Lypd1 in tooth germ using an ex vivo organ culture system and in mouse dental pulp (mDP) cells resulted in the inhibition of odontoblast differentiation. Activation of bone morphogenetic protein (BMP) signaling by BMP2 treatment in mDP cells promoted odontoblast differentiation via phosphorylation of Smad1/5/8, while this BMP2-mediated odontoblast differentiation was inhibited by depletion of Lypd1. Furthermore, we created a deletion construct of the C terminus containing the omega site in LYPD1; this site is necessary for localizing GPI-APs to the plasma membrane and lipid rafts. We identified that this site is essential for odontoblast differentiation and morphological change of mDP cells. These findings demonstrated that LYPD1 is a novel marker of preodontoblasts in the developing tooth; in addition, they suggest that LYPD1 is important for tooth development and that it plays a pivotal role in odontoblast differentiation by regulating Smad1/5/8 phosphorylation through its effect as a GPI-AP in lipid rafts.
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U2 - 10.1016/j.jbc.2023.104638
DO - 10.1016/j.jbc.2023.104638
M3 - Article
C2 - 36963497
AN - SCOPUS:85152231566
SN - 0021-9258
VL - 299
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 5
M1 - 104638
ER -