Original Report

Genotype–Phenotype Correlation of Tracheal Cartilaginous Sleeves and Fgfr2 Mutations in Mice

Austin S. Lam MD

orcid.org/0000-0002-2297-6376

Department of Otolaryngology – Head & Neck Surgery, University of Washington School of Medicine, Seattle, Washington, U.S.A.

Division of Pediatric Otolaryngology – Head & Neck Surgery, Seattle Children's Hospital, Seattle, Washington, U.S.A.

Seattle Children's Research Institute, Center for Developmental Biology and Regenerative Medicine, Seattle, Washington, U.S.A.

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Carrie C. Liu MD, MPH,

Carrie C. Liu MD, MPH

orcid.org/0000-0002-9262-2792

Department of Otolaryngology – Head & Neck Surgery, University of Washington School of Medicine, Seattle, Washington, U.S.A.

Division of Pediatric Otolaryngology – Head & Neck Surgery, Seattle Children's Hospital, Seattle, Washington, U.S.A.

Current address: Divisions of Otolaryngology – Head and Neck Surgery, and Pediatric Surgery, Department of Surgery, University of Calgary, Calgary, Alberta, Canada

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Gail H. Deutsch MD,

Gail H. Deutsch MD

Department of Pathology, University of Washington School of Medicine, Seattle, Washington, U.S.A.

Department of Pathology, Seattle Children's Hospital, Seattle, Washington, U.S.A.

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Joshua Rivera BS,

Joshua Rivera BS

Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York City, New York, U.S.A.

Current address: Center for Personalized Cancer Therapy, University of Massachusetts, Boston, Massachusetts, U.S.A.

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Jonathan A. Perkins DO,

Jonathan A. Perkins DO

orcid.org/0000-0003-0181-3997

Department of Otolaryngology – Head & Neck Surgery, University of Washington School of Medicine, Seattle, Washington, U.S.A.

Division of Pediatric Otolaryngology – Head & Neck Surgery, Seattle Children's Hospital, Seattle, Washington, U.S.A.

Craniofacial Center, Seattle Children's Hospital, Seattle, Washington, U.S.A.

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Greg Holmes PhD,

Greg Holmes PhD

Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York City, New York, U.S.A.

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Ethylin W. Jabs MD,

Ethylin W. Jabs MD

Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York City, New York, U.S.A.

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Michael L. Cunningham MD, PhD,

Michael L. Cunningham MD, PhD

Seattle Children's Research Institute, Center for Developmental Biology and Regenerative Medicine, Seattle, Washington, U.S.A.

Craniofacial Center, Seattle Children's Hospital, Seattle, Washington, U.S.A.

Department of Pediatrics, University of Washington School of Medicine, Seattle, Washington, U.S.A.

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John P. Dahl MD, PhD, MBA,

Corresponding Author

John P. Dahl MD, PhD, MBA

[email protected]

Department of Otolaryngology – Head & Neck Surgery, University of Washington School of Medicine, Seattle, Washington, U.S.A.

Division of Pediatric Otolaryngology – Head & Neck Surgery, Seattle Children's Hospital, Seattle, Washington, U.S.A.

Craniofacial Center, Seattle Children's Hospital, Seattle, Washington, U.S.A.

Send correspondence to John P. Dahl, MD, PhD, MBA, Seattle Children's Hospital, 4800 Sand Point Way NE, Seattle, WA 98105. E-mail: [email protected]

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Austin S. Lam MD,

Austin S. Lam MD

orcid.org/0000-0002-2297-6376

Department of Otolaryngology – Head & Neck Surgery, University of Washington School of Medicine, Seattle, Washington, U.S.A.

Division of Pediatric Otolaryngology – Head & Neck Surgery, Seattle Children's Hospital, Seattle, Washington, U.S.A.

Seattle Children's Research Institute, Center for Developmental Biology and Regenerative Medicine, Seattle, Washington, U.S.A.

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Carrie C. Liu MD, MPH,

Carrie C. Liu MD, MPH

orcid.org/0000-0002-9262-2792

Department of Otolaryngology – Head & Neck Surgery, University of Washington School of Medicine, Seattle, Washington, U.S.A.

Division of Pediatric Otolaryngology – Head & Neck Surgery, Seattle Children's Hospital, Seattle, Washington, U.S.A.

Current address: Divisions of Otolaryngology – Head and Neck Surgery, and Pediatric Surgery, Department of Surgery, University of Calgary, Calgary, Alberta, Canada

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Gail H. Deutsch MD,

Gail H. Deutsch MD

Department of Pathology, University of Washington School of Medicine, Seattle, Washington, U.S.A.

Department of Pathology, Seattle Children's Hospital, Seattle, Washington, U.S.A.

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Joshua Rivera BS,

Joshua Rivera BS

Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York City, New York, U.S.A.

Current address: Center for Personalized Cancer Therapy, University of Massachusetts, Boston, Massachusetts, U.S.A.

Search for more papers by this author

Jonathan A. Perkins DO,

Jonathan A. Perkins DO

orcid.org/0000-0003-0181-3997

Department of Otolaryngology – Head & Neck Surgery, University of Washington School of Medicine, Seattle, Washington, U.S.A.

Division of Pediatric Otolaryngology – Head & Neck Surgery, Seattle Children's Hospital, Seattle, Washington, U.S.A.

Craniofacial Center, Seattle Children's Hospital, Seattle, Washington, U.S.A.

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Greg Holmes PhD,

Greg Holmes PhD

Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York City, New York, U.S.A.

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Ethylin W. Jabs MD,

Ethylin W. Jabs MD

Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York City, New York, U.S.A.

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Michael L. Cunningham MD, PhD,

Michael L. Cunningham MD, PhD

Seattle Children's Research Institute, Center for Developmental Biology and Regenerative Medicine, Seattle, Washington, U.S.A.

Craniofacial Center, Seattle Children's Hospital, Seattle, Washington, U.S.A.

Department of Pediatrics, University of Washington School of Medicine, Seattle, Washington, U.S.A.

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John P. Dahl MD, PhD, MBA,

Corresponding Author

John P. Dahl MD, PhD, MBA

[email protected]

Department of Otolaryngology – Head & Neck Surgery, University of Washington School of Medicine, Seattle, Washington, U.S.A.

Division of Pediatric Otolaryngology – Head & Neck Surgery, Seattle Children's Hospital, Seattle, Washington, U.S.A.

Craniofacial Center, Seattle Children's Hospital, Seattle, Washington, U.S.A.

Send correspondence to John P. Dahl, MD, PhD, MBA, Seattle Children's Hospital, 4800 Sand Point Way NE, Seattle, WA 98105. E-mail: [email protected]

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First published: 04 September 2020

https://doi.org/10.1002/lary.29060

Citations: 9

Editor's Note: This Manuscript was accepted for publication on August 10, 2020.

Supported by the National Institute on Deafness and Other Communication Disorders (NIDCD)/ National Institutes of Health (NIH) grant T32DC000018 (a.s.l.); and The National Institute of Dental and Craniofacial Research (NIDCR)/NIH grants R01 DE022988 and NIH/Eunice Kennedy Shriver National Institute of Child Health and Human Development P01 HD078233 (e.w.j., g.h., j.r.). The authors have no other funding, financial relationships, or conflicts of interest to disclose.

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Abstract

Objectives

To characterize tracheal cartilage morphology in mouse models of fibroblast growth factor receptor (Fgfr2)-related craniosynostosis syndromes. To establish relationships between specific Fgfr2 mutations and tracheal cartilaginous sleeve (TCS) phenotypes in these mouse models.

Methods

Postnatal day 0 knock-in mouse lines with disease-specific genetic variations in the Fgfr2 gene (Fgfr2^C342Y/C342Y, Fgfr2^C342Y/+, Fgfr2^+/Y394C, Fgfr2^+/S252W, and Fgfr2^+/P253R) as well as line-specific controls were utilized. Tracheal cartilage morphology as measured by gross analyses, microcomputed tomography (μCT), and histopathology were compared using Chi-squared and single-factor analysis of variance statistical tests.

Results

A greater proportion of rings per trachea were abnormal in Fgfr2^C342Y/+ tracheas (63%) than Fgfr2^+/S252W (17%), Fgfr2^+/P253R (17%), Fgfr2^+/Y394C (12%), and controls (10%) (P < .001 for each vs. Fgfr2^C342Y/+). TCS segments were found only in Fgfr2^C342Y/C342Y (100%) and Fgfr2^C342Y/+ (72%) tracheas. Cricoid and first-tracheal ring fusion was noted in all Fgfr2^C342Y/C342Y and 94% of Fgfr2^C342Y/+ samples. The Fgfr2^C342Y/C342Y and Fgfr2^C342Y/+ groups were found to have greater areas and volumes of cartilage than other lines on gross analysis and μCT. Histologic analyses confirmed TCS among the Fgfr2^C342Y/C342Y and Fgfr2^C342Y/+ groups, without appreciable differences in cartilage morphology, cell size, or density; no histologic differences were observed among other Fgfr2 lines compared to controls.

Conclusion

This study found TCS phenotypes only in the Fgfr2^C342Y mouse lines. These lines also had increased tracheal cartilage compared to other mutant lines and controls. These data support further study of the Fgfr2 mouse lines and the investigation of other Fgfr2 variants to better understand their role in tracheal development and TCS formation.

Level of Evidence

NA Laryngoscope, 131:E1349–E1356, 2021

Supporting Information

Filename

Description

lary29060-sup-0001-Supinfo.docxWord 2007 document , 15 KB

Supplemental Methods. Supplemental Methods Document.

Table S1. Whole-mount morphology data, alcian blue staining measures, microcomputed-tomography and histology data for Fgfr2^C342Y groups and their line-specific controls. n – number of samples per group. SD – standard deviation.

Table S2. Whole-mount morphology data, alcian blue staining measures, microcomputed-tomography and histology data for the Fgfr2^Y394C group and their line-specific controls. n – number of samples per group. SD – standard deviation.

Table S3. Whole-mount morphology data, alcian blue staining measures, microcomputed-tomography and histology data for the Fgfr2^S252W group and their line-specific controls. n – number of samples per group. SD – standard deviation.

Table S4. Whole-mount morphology data, alcian blue staining measures, microcomputed-tomography and histology data for the Fgfr2^P253R group and their line-specific controls. n – number of samples per group. SD – standard deviation.

Table S5. Whole-mount morphology data, alcian blue staining measures, microcomputed-tomography and histology data between Fgfr2 gene variants and pooled controls. n – number of samples per group. SD – standard deviation.

lary29060-sup-0002-Tables.docxWord 2007 document , 50.3 KB

Appendix S2. Tables.

Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

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Citing Literature

Volume131, Issue4

April 2021

Pages E1349-E1356

Genotype–Phenotype Correlation of Tracheal Cartilaginous Sleeves and Fgfr2 Mutations in Mice

Abstract

Objectives

Methods

Results

Conclusion

Level of Evidence

Supporting Information

BIBLIOGRAPHY

Citing Literature

References

Information

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Genotype–Phenotype Correlation of Tracheal Cartilaginous Sleeves and Fgfr2 Mutations in Mice

Abstract

Objectives

Methods

Results

Conclusion

Level of Evidence

Supporting Information

BIBLIOGRAPHY

Citing Literature

References

Related

Information