Single cell RNA sequencing reveals human tooth type identity and guides in vitro hiPSC derived odontoblast differentiation (iOB)
Over 90% of the U.S. adult population suffers from tooth structure loss due to caries. Most of the mineralized tooth structure is composed of dentin, a material produced and mineralized by ectomesenchyme derived cells known as odontoblasts. Clinicians, scientists, and the general public share the de...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-07-01
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| Series: | Frontiers in Dental Medicine |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fdmed.2023.1209503/full |
| _version_ | 1797635375371386880 |
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| author | Sesha Hanson-Drury Sesha Hanson-Drury Sesha Hanson-Drury Anjali P. Patni Anjali P. Patni Anjali P. Patni Anjali P. Patni Deborah L. Lee Deborah L. Lee Deborah L. Lee Ammar Alghadeer Ammar Alghadeer Ammar Alghadeer Ammar Alghadeer Yan Ting Zhao Yan Ting Zhao Yan Ting Zhao Devon Duron Ehnes Devon Duron Ehnes Devon Duron Ehnes Vivian N. Vo Vivian N. Vo Vivian N. Vo Sydney Y. Kim Sydney Y. Kim Druthi Jithendra Druthi Jithendra Ashish Phal Ashish Phal Ashish Phal Natasha I. Edman Natasha I. Edman Natasha I. Edman Natasha I. Edman Thomas Schlichthaerle Thomas Schlichthaerle David Baker David Baker David Baker Jessica E. Young Jessica E. Young Julie Mathieu Julie Mathieu Hannele Ruohola-Baker Hannele Ruohola-Baker Hannele Ruohola-Baker Hannele Ruohola-Baker Hannele Ruohola-Baker |
| author_facet | Sesha Hanson-Drury Sesha Hanson-Drury Sesha Hanson-Drury Anjali P. Patni Anjali P. Patni Anjali P. Patni Anjali P. Patni Deborah L. Lee Deborah L. Lee Deborah L. Lee Ammar Alghadeer Ammar Alghadeer Ammar Alghadeer Ammar Alghadeer Yan Ting Zhao Yan Ting Zhao Yan Ting Zhao Devon Duron Ehnes Devon Duron Ehnes Devon Duron Ehnes Vivian N. Vo Vivian N. Vo Vivian N. Vo Sydney Y. Kim Sydney Y. Kim Druthi Jithendra Druthi Jithendra Ashish Phal Ashish Phal Ashish Phal Natasha I. Edman Natasha I. Edman Natasha I. Edman Natasha I. Edman Thomas Schlichthaerle Thomas Schlichthaerle David Baker David Baker David Baker Jessica E. Young Jessica E. Young Julie Mathieu Julie Mathieu Hannele Ruohola-Baker Hannele Ruohola-Baker Hannele Ruohola-Baker Hannele Ruohola-Baker Hannele Ruohola-Baker |
| author_sort | Sesha Hanson-Drury |
| collection | DOAJ |
| description | Over 90% of the U.S. adult population suffers from tooth structure loss due to caries. Most of the mineralized tooth structure is composed of dentin, a material produced and mineralized by ectomesenchyme derived cells known as odontoblasts. Clinicians, scientists, and the general public share the desire to regenerate this missing tooth structure. To bioengineer missing dentin, increased understanding of human tooth development is required. Here we interrogate at the single cell level the signaling interactions that guide human odontoblast and ameloblast development and which determine incisor or molar tooth germ type identity. During human odontoblast development, computational analysis predicts that early FGF and BMP activation followed by later HH signaling is crucial. Here we generate a differentiation protocol based on this sci-RNA-seq analysis to produce mature hiPSC derived odontoblasts in vitro (iOB). Further, we elucidate the critical role of FGF signaling in odontoblast maturation and its biomineralization capacity using the de novo designed FGFR1/2c isoform specific minibinder scaffolded as a C6 oligomer that acts as a pathway agonist. Using computational tools, we show on a molecular level how human molar development is delayed compared to incisors. We reveal that enamel knot development is guided by FGF and WNT in incisors and BMP and ROBO in the molars, and that incisor and molar ameloblast development is guided by FGF, EGF and BMP signaling, with tooth type specific intensity of signaling interactions. Dental ectomesenchyme derived cells are the primary source of signaling ligands responsible for both enamel knot and ameloblast development. |
| first_indexed | 2024-03-11T12:20:13Z |
| format | Article |
| id | doaj.art-d5ec883d1daf42f1a753762ef71e295b |
| institution | Directory Open Access Journal |
| issn | 2673-4915 |
| language | English |
| last_indexed | 2024-03-11T12:20:13Z |
| publishDate | 2023-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Dental Medicine |
| spelling | doaj.art-d5ec883d1daf42f1a753762ef71e295b2023-11-07T03:11:38ZengFrontiers Media S.A.Frontiers in Dental Medicine2673-49152023-07-01410.3389/fdmed.2023.12095031209503Single cell RNA sequencing reveals human tooth type identity and guides in vitro hiPSC derived odontoblast differentiation (iOB)Sesha Hanson-Drury0Sesha Hanson-Drury1Sesha Hanson-Drury2Anjali P. Patni3Anjali P. Patni4Anjali P. Patni5Anjali P. Patni6Deborah L. Lee7Deborah L. Lee8Deborah L. Lee9Ammar Alghadeer10Ammar Alghadeer11Ammar Alghadeer12Ammar Alghadeer13Yan Ting Zhao14Yan Ting Zhao15Yan Ting Zhao16Devon Duron Ehnes17Devon Duron Ehnes18Devon Duron Ehnes19Vivian N. Vo20Vivian N. Vo21Vivian N. Vo22Sydney Y. Kim23Sydney Y. Kim24Druthi Jithendra25Druthi Jithendra26Ashish Phal27Ashish Phal28Ashish Phal29Natasha I. Edman30Natasha I. Edman31Natasha I. Edman32Natasha I. Edman33Thomas Schlichthaerle34Thomas Schlichthaerle35David Baker36David Baker37David Baker38Jessica E. Young39Jessica E. Young40Julie Mathieu41Julie Mathieu42Hannele Ruohola-Baker43Hannele Ruohola-Baker44Hannele Ruohola-Baker45Hannele Ruohola-Baker46Hannele Ruohola-Baker47Department of Oral Health Sciences, School of Dentistry, University of Washington, Seattle, WA, United StatesDepartment of Biochemistry, School of Medicine, University of Washington, Seattle, WA, United StatesInstitute for Stem Cell and Regenerative Medicine, School of Medicine, University of Washington, Seattle, WA, United StatesDepartment of Oral Health Sciences, School of Dentistry, University of Washington, Seattle, WA, United StatesDepartment of Biochemistry, School of Medicine, University of Washington, Seattle, WA, United StatesInstitute for Stem Cell and Regenerative Medicine, School of Medicine, University of Washington, Seattle, WA, United StatesDepartment of Genetic Engineering, SRM Institute of Science and Technology, Chennai, IndiaDepartment of Oral Health Sciences, School of Dentistry, University of Washington, Seattle, WA, United StatesDepartment of Biochemistry, School of Medicine, University of Washington, Seattle, WA, United StatesInstitute for Stem Cell and Regenerative Medicine, School of Medicine, University of Washington, Seattle, WA, United StatesDepartment of Oral Health Sciences, School of Dentistry, University of Washington, Seattle, WA, United StatesDepartment of Biochemistry, School of Medicine, University of Washington, Seattle, WA, United StatesInstitute for Stem Cell and Regenerative Medicine, School of Medicine, University of Washington, Seattle, WA, United StatesDepartment of Biomedical Dental Sciences, College of Dentistry, Imam Abdulrahman bin Faisal University, Dammam, Saudi ArabiaDepartment of Oral Health Sciences, School of Dentistry, University of Washington, Seattle, WA, United StatesDepartment of Biochemistry, School of Medicine, University of Washington, Seattle, WA, United StatesInstitute for Stem Cell and Regenerative Medicine, School of Medicine, University of Washington, Seattle, WA, United StatesDepartment of Oral Health Sciences, School of Dentistry, University of Washington, Seattle, WA, United StatesDepartment of Biochemistry, School of Medicine, University of Washington, Seattle, WA, United StatesInstitute for Stem Cell and Regenerative Medicine, School of Medicine, University of Washington, Seattle, WA, United StatesDepartment of Biochemistry, School of Medicine, University of Washington, Seattle, WA, United StatesInstitute for Stem Cell and Regenerative Medicine, School of Medicine, University of Washington, Seattle, WA, United StatesDepartment of Biology, University of Washington, Seattle, WA, United StatesDepartment of Oral Health Sciences, School of Dentistry, University of Washington, Seattle, WA, United StatesInstitute for Stem Cell and Regenerative Medicine, School of Medicine, University of Washington, Seattle, WA, United StatesInstitute for Stem Cell and Regenerative Medicine, School of Medicine, University of Washington, Seattle, WA, United StatesDepartment of Biotechnology, SRM Institute of Science and Technology, Chennai, IndiaDepartment of Biochemistry, School of Medicine, University of Washington, Seattle, WA, United StatesInstitute for Stem Cell and Regenerative Medicine, School of Medicine, University of Washington, Seattle, WA, United StatesDepartment of Bioengineering, University of Washington, Seattle, WA, United StatesDepartment of Biochemistry, School of Medicine, University of Washington, Seattle, WA, United StatesInstitute for Protein Design, University of Washington, Seattle, WA, United States0Molecular and Cellular Biology Graduate Program, University of Washington, Seattle, WA, United States1Medical Scientist Training Program, University of Washington, Seattle, WA, United StatesDepartment of Biochemistry, School of Medicine, University of Washington, Seattle, WA, United StatesInstitute for Protein Design, University of Washington, Seattle, WA, United StatesDepartment of Biochemistry, School of Medicine, University of Washington, Seattle, WA, United StatesInstitute for Stem Cell and Regenerative Medicine, School of Medicine, University of Washington, Seattle, WA, United StatesInstitute for Protein Design, University of Washington, Seattle, WA, United StatesInstitute for Stem Cell and Regenerative Medicine, School of Medicine, University of Washington, Seattle, WA, United States2Department of Laboratory Medicine and Pathology, University of Washington, Seattle, WA, United StatesInstitute for Stem Cell and Regenerative Medicine, School of Medicine, University of Washington, Seattle, WA, United States3Department of Comparative Medicine, School of Medicine, University of Washington, Seattle, WA, United StatesDepartment of Oral Health Sciences, School of Dentistry, University of Washington, Seattle, WA, United StatesDepartment of Biochemistry, School of Medicine, University of Washington, Seattle, WA, United StatesInstitute for Stem Cell and Regenerative Medicine, School of Medicine, University of Washington, Seattle, WA, United StatesDepartment of Biology, University of Washington, Seattle, WA, United StatesDepartment of Bioengineering, University of Washington, Seattle, WA, United StatesOver 90% of the U.S. adult population suffers from tooth structure loss due to caries. Most of the mineralized tooth structure is composed of dentin, a material produced and mineralized by ectomesenchyme derived cells known as odontoblasts. Clinicians, scientists, and the general public share the desire to regenerate this missing tooth structure. To bioengineer missing dentin, increased understanding of human tooth development is required. Here we interrogate at the single cell level the signaling interactions that guide human odontoblast and ameloblast development and which determine incisor or molar tooth germ type identity. During human odontoblast development, computational analysis predicts that early FGF and BMP activation followed by later HH signaling is crucial. Here we generate a differentiation protocol based on this sci-RNA-seq analysis to produce mature hiPSC derived odontoblasts in vitro (iOB). Further, we elucidate the critical role of FGF signaling in odontoblast maturation and its biomineralization capacity using the de novo designed FGFR1/2c isoform specific minibinder scaffolded as a C6 oligomer that acts as a pathway agonist. Using computational tools, we show on a molecular level how human molar development is delayed compared to incisors. We reveal that enamel knot development is guided by FGF and WNT in incisors and BMP and ROBO in the molars, and that incisor and molar ameloblast development is guided by FGF, EGF and BMP signaling, with tooth type specific intensity of signaling interactions. Dental ectomesenchyme derived cells are the primary source of signaling ligands responsible for both enamel knot and ameloblast development.https://www.frontiersin.org/articles/10.3389/fdmed.2023.1209503/fullodontoblastsingle cell RNA sequencingenamel knotcell signalingde novo designed mini protein bindershuman tooth |
| spellingShingle | Sesha Hanson-Drury Sesha Hanson-Drury Sesha Hanson-Drury Anjali P. Patni Anjali P. Patni Anjali P. Patni Anjali P. Patni Deborah L. Lee Deborah L. Lee Deborah L. Lee Ammar Alghadeer Ammar Alghadeer Ammar Alghadeer Ammar Alghadeer Yan Ting Zhao Yan Ting Zhao Yan Ting Zhao Devon Duron Ehnes Devon Duron Ehnes Devon Duron Ehnes Vivian N. Vo Vivian N. Vo Vivian N. Vo Sydney Y. Kim Sydney Y. Kim Druthi Jithendra Druthi Jithendra Ashish Phal Ashish Phal Ashish Phal Natasha I. Edman Natasha I. Edman Natasha I. Edman Natasha I. Edman Thomas Schlichthaerle Thomas Schlichthaerle David Baker David Baker David Baker Jessica E. Young Jessica E. Young Julie Mathieu Julie Mathieu Hannele Ruohola-Baker Hannele Ruohola-Baker Hannele Ruohola-Baker Hannele Ruohola-Baker Hannele Ruohola-Baker Single cell RNA sequencing reveals human tooth type identity and guides in vitro hiPSC derived odontoblast differentiation (iOB) Frontiers in Dental Medicine odontoblast single cell RNA sequencing enamel knot cell signaling de novo designed mini protein binders human tooth |
| title | Single cell RNA sequencing reveals human tooth type identity and guides in vitro hiPSC derived odontoblast differentiation (iOB) |
| title_full | Single cell RNA sequencing reveals human tooth type identity and guides in vitro hiPSC derived odontoblast differentiation (iOB) |
| title_fullStr | Single cell RNA sequencing reveals human tooth type identity and guides in vitro hiPSC derived odontoblast differentiation (iOB) |
| title_full_unstemmed | Single cell RNA sequencing reveals human tooth type identity and guides in vitro hiPSC derived odontoblast differentiation (iOB) |
| title_short | Single cell RNA sequencing reveals human tooth type identity and guides in vitro hiPSC derived odontoblast differentiation (iOB) |
| title_sort | single cell rna sequencing reveals human tooth type identity and guides in vitro hipsc derived odontoblast differentiation iob |
| topic | odontoblast single cell RNA sequencing enamel knot cell signaling de novo designed mini protein binders human tooth |
| url | https://www.frontiersin.org/articles/10.3389/fdmed.2023.1209503/full |
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