title: Gelatin scaffold combined with bone morphogenetic protein-4 induces odontoblast-like cell differentiation involving integrin profile changes, autophagy-related gene 10, and Wnt5 sequentially in human induced pluripotent stem cells.
abstract: While human induced pluripotent stem (hiPS) cells have potential use in regenerative medicine, there are no reports on odontoblastic differentiation of hiPS cells. In the current study, to examine integrin profiles and explore the early signaling cascade of odontoblastic differentiation in hiPS cells, we investigated the regulation of autophagy-related gene (Atg) and wingless/int1 (Wnt) signaling in gelatin scaffold (GS) combined with bone morphogenetic protein (BMP)-4 (GS/BMP-4)-mediated odontoblastic differentiation. Following GS/BMP-4 treatment, there was a dramatic loss of α3 and α6 integrins, and reciprocal strong induction of α1 integrin expression in the differentiated cells. GS/BMP-4 increased the mRNA and protein levels of Atg10, Lrp5/Fzd9 (an Atg10 receptor), and Wnt5 together with the amount of autophagosomes and autophagic fluxes. Treatment with siRNAs against Atg10 and Wnt5a individually suppressed the GS/BMP-4-induced increase in odontoblastic differentiation. The odontoblastic phenotype was inhibited by chloroquine, but increased after treatment with rapamycin (an autophagy enhancer). Taken together with our previous findings, we have replicated our results from the rodent system in a novel human system. We have revealed a unique sequential cascade involving Atg10, Wnt5a, α1 integrin, and matrix metalloproteinase-3 in GS/BMP-4-induced differentiation of hiPS cells into odontoblast-like cells at a relatively early stage.
authors.affiliation: Department of Endodontics, School of Dentistry, Aichi Gakuin University, Nagoya, Aichi, 464-8651 Japan.
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Ozeki
authors.affiliation: Department of Endodontics, School of Dentistry, Aichi Gakuin University, Nagoya, Aichi, 464-8651 Japan.
authors.forename: Naoko
authors.initials: N
authors.lastname: Hase
authors.affiliation: Department of Endodontics, School of Dentistry, Aichi Gakuin University, Nagoya, Aichi, 464-8651 Japan.
authors.forename: Naoya
authors.initials: N
authors.lastname: Higuchi
authors.affiliation: Department of Endodontics, School of Dentistry, Aichi Gakuin University, Nagoya, Aichi, 464-8651 Japan.
authors.forename: Taiki
authors.initials: T
authors.lastname: Hiyama
authors.affiliation: Department of Endodontics, School of Dentistry, Aichi Gakuin University, Nagoya, Aichi, 464-8651 Japan.
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Yamaguchi
authors.affiliation: Department of Endodontics, School of Dentistry, Aichi Gakuin University, Nagoya, Aichi, 464-8651 Japan.
authors.forename: Rie
authors.initials: R
authors.lastname: Kawai
authors.affiliation: Department of Endodontics, School of Dentistry, Aichi Gakuin University, Nagoya, Aichi, 464-8651 Japan.
authors.forename: Toru
authors.initials: T
authors.lastname: Matsumoto
authors.affiliation: Department of Endodontics, School of Dentistry, Aichi Gakuin University, Nagoya, Aichi, 464-8651 Japan.
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Nakata
authors.affiliation: Department of Integrative Education of Pharmacy, School of Pharmacy, Aichi Gakuin University, Nagoya, Aichi, 464-8650 Japan. Electronic address: makio@dpc.agu.ac.jp.
authors.forename: Makio
authors.initials: M
authors.lastname: Mogi
mesh_terms: D000071183:Autophagy-Related Proteins / Q000235:genetics*; D055415:Bone Morphogenetic Protein 4 / Q000008:administration & dosage; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics*; D005780:Gelatin / Q000008:administration & dosage / Q000737:chemistry; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects*; D039001:Integrin alpha Chains / Q000235:genetics*; D019278:Matrix Metalloproteinase 3 / Q000235:genetics*; D009804:Odontoblasts / Q000187:drug effects; D034741:RNA, Small Interfering / Q000008:administration & dosage; D044968:Regenerative Medicine; D015398:Signal Transduction / Q000187:drug effects; D020123:Sirolimus / Q000008:administration & dosage; D054457:Tissue Scaffolds; D033921:Vesicular Transport Proteins / Q000235:genetics*; D000071818:Wnt-5a Protein / Q000235:genetics*
chemical_list: D000071183:Autophagy-Related Proteins; C525809:BMP4 protein, human; D055415:Bone Morphogenetic Protein 4; D039001:Integrin alpha Chains; D034741:RNA, Small Interfering; D033921:Vesicular Transport Proteins; C085389:WNT5A protein, human; D000071818:Wnt-5a Protein; D005780:Gelatin; C581901:ATG10 protein, human; D019278:Matrix Metalloproteinase 3; D020123:Sirolimus
keywords: Atg10; Cell differentiation; Human induced pluripotent stem cell; Matrix metalloproteinase-3; Odontoblast; Wnt5