RFX3 Modulation of FOXJ1 regulation of cilia genes in the human airway epithelium

Lukas Didon, Rachel K. Zwick, Ion Wa Chao, Matthew S. Walters, Rui Wang, Neil R. Hackett, Ronald Crystal

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Background: Ciliated cells play a central role in cleansing the airways of inhaled contaminants. They are derived from basal cells that include the airway stem/progenitor cells. In animal models, the transcription factor FOXJ1 has been shown to induce differentiation to the ciliated cell lineage, and the RFX transcription factor-family has been shown to be necessary for, but not sufficient to induce, correct cilia development.Methods: To test the hypothesis that FOXJ1 and RFX3 cooperatively induce expression of ciliated genes in the differentiation process of basal progenitor cells toward a ciliated cell linage in the human airway epithelium, primary human airway basal cells were assessed under conditions of in vitro differentiation induced by plasmid-mediated gene transfer of FOXJ1 and/or RFX3. TaqMan PCR was used to quantify mRNA levels of basal, secretory, and cilia-associated genes.Results: Basal cells, when cultured in air-liquid interface, differentiated into a ciliated epithelium, expressing FOXJ1 and RFX3. Transfection of FOXJ1 into resting basal cells activated promoters and induced expression of ciliated cell genes as well as both FOXJ1 and RFX3, but not basal cell genes. Transfection of RFX3 induced expression of RFX3 but not FOXJ1, nor the expression of cilia-related genes. The combination of FOXJ1 + RFX3 enhanced ciliated gene promoter activity and mRNA expression beyond that due to FOXJ1 alone. Corroborating immunoprecipitation studies demonstrated an interaction between FOXJ1 and RFX3.Conclusion: FOXJ1 is an important regulator of cilia gene expression during ciliated cell differentiation, with RFX3 as a transcriptional co-activator to FOXJ1, helping to induce the expression of cilia genes in the process of ciliated cell differentiation of basal/progenitor cells.

Original languageEnglish
Article number70
JournalRespiratory Research
Volume14
Issue number1
DOIs
Publication statusPublished - 3 Jul 2013
Externally publishedYes

Fingerprint

Cilia
Epithelium
Genes
Stem Cells
Gene Expression
Transfection
Cell Differentiation
Messenger RNA
Cell Lineage
Regulator Genes
Immunoprecipitation
Cultured Cells
Plasmids
Transcription Factors
Animal Models
Air
Polymerase Chain Reaction

Keywords

  • Basal cell
  • Ciliated cell differentiation
  • FOXJ1
  • Human
  • Lung epithelium
  • RFX3

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine

Cite this

Didon, L., Zwick, R. K., Chao, I. W., Walters, M. S., Wang, R., Hackett, N. R., & Crystal, R. (2013). RFX3 Modulation of FOXJ1 regulation of cilia genes in the human airway epithelium. Respiratory Research, 14(1), [70]. https://doi.org/10.1186/1465-9921-14-70

RFX3 Modulation of FOXJ1 regulation of cilia genes in the human airway epithelium. / Didon, Lukas; Zwick, Rachel K.; Chao, Ion Wa; Walters, Matthew S.; Wang, Rui; Hackett, Neil R.; Crystal, Ronald.

In: Respiratory Research, Vol. 14, No. 1, 70, 03.07.2013.

Research output: Contribution to journalArticle

Didon, Lukas ; Zwick, Rachel K. ; Chao, Ion Wa ; Walters, Matthew S. ; Wang, Rui ; Hackett, Neil R. ; Crystal, Ronald. / RFX3 Modulation of FOXJ1 regulation of cilia genes in the human airway epithelium. In: Respiratory Research. 2013 ; Vol. 14, No. 1.
@article{7c09144c1fd74784bfe6fcb571fe2833,
title = "RFX3 Modulation of FOXJ1 regulation of cilia genes in the human airway epithelium",
abstract = "Background: Ciliated cells play a central role in cleansing the airways of inhaled contaminants. They are derived from basal cells that include the airway stem/progenitor cells. In animal models, the transcription factor FOXJ1 has been shown to induce differentiation to the ciliated cell lineage, and the RFX transcription factor-family has been shown to be necessary for, but not sufficient to induce, correct cilia development.Methods: To test the hypothesis that FOXJ1 and RFX3 cooperatively induce expression of ciliated genes in the differentiation process of basal progenitor cells toward a ciliated cell linage in the human airway epithelium, primary human airway basal cells were assessed under conditions of in vitro differentiation induced by plasmid-mediated gene transfer of FOXJ1 and/or RFX3. TaqMan PCR was used to quantify mRNA levels of basal, secretory, and cilia-associated genes.Results: Basal cells, when cultured in air-liquid interface, differentiated into a ciliated epithelium, expressing FOXJ1 and RFX3. Transfection of FOXJ1 into resting basal cells activated promoters and induced expression of ciliated cell genes as well as both FOXJ1 and RFX3, but not basal cell genes. Transfection of RFX3 induced expression of RFX3 but not FOXJ1, nor the expression of cilia-related genes. The combination of FOXJ1 + RFX3 enhanced ciliated gene promoter activity and mRNA expression beyond that due to FOXJ1 alone. Corroborating immunoprecipitation studies demonstrated an interaction between FOXJ1 and RFX3.Conclusion: FOXJ1 is an important regulator of cilia gene expression during ciliated cell differentiation, with RFX3 as a transcriptional co-activator to FOXJ1, helping to induce the expression of cilia genes in the process of ciliated cell differentiation of basal/progenitor cells.",
keywords = "Basal cell, Ciliated cell differentiation, FOXJ1, Human, Lung epithelium, RFX3",
author = "Lukas Didon and Zwick, {Rachel K.} and Chao, {Ion Wa} and Walters, {Matthew S.} and Rui Wang and Hackett, {Neil R.} and Ronald Crystal",
year = "2013",
month = "7",
day = "3",
doi = "10.1186/1465-9921-14-70",
language = "English",
volume = "14",
journal = "Respiratory Research",
issn = "1465-9921",
publisher = "BioMed Central",
number = "1",

}

TY - JOUR

T1 - RFX3 Modulation of FOXJ1 regulation of cilia genes in the human airway epithelium

AU - Didon, Lukas

AU - Zwick, Rachel K.

AU - Chao, Ion Wa

AU - Walters, Matthew S.

AU - Wang, Rui

AU - Hackett, Neil R.

AU - Crystal, Ronald

PY - 2013/7/3

Y1 - 2013/7/3

N2 - Background: Ciliated cells play a central role in cleansing the airways of inhaled contaminants. They are derived from basal cells that include the airway stem/progenitor cells. In animal models, the transcription factor FOXJ1 has been shown to induce differentiation to the ciliated cell lineage, and the RFX transcription factor-family has been shown to be necessary for, but not sufficient to induce, correct cilia development.Methods: To test the hypothesis that FOXJ1 and RFX3 cooperatively induce expression of ciliated genes in the differentiation process of basal progenitor cells toward a ciliated cell linage in the human airway epithelium, primary human airway basal cells were assessed under conditions of in vitro differentiation induced by plasmid-mediated gene transfer of FOXJ1 and/or RFX3. TaqMan PCR was used to quantify mRNA levels of basal, secretory, and cilia-associated genes.Results: Basal cells, when cultured in air-liquid interface, differentiated into a ciliated epithelium, expressing FOXJ1 and RFX3. Transfection of FOXJ1 into resting basal cells activated promoters and induced expression of ciliated cell genes as well as both FOXJ1 and RFX3, but not basal cell genes. Transfection of RFX3 induced expression of RFX3 but not FOXJ1, nor the expression of cilia-related genes. The combination of FOXJ1 + RFX3 enhanced ciliated gene promoter activity and mRNA expression beyond that due to FOXJ1 alone. Corroborating immunoprecipitation studies demonstrated an interaction between FOXJ1 and RFX3.Conclusion: FOXJ1 is an important regulator of cilia gene expression during ciliated cell differentiation, with RFX3 as a transcriptional co-activator to FOXJ1, helping to induce the expression of cilia genes in the process of ciliated cell differentiation of basal/progenitor cells.

AB - Background: Ciliated cells play a central role in cleansing the airways of inhaled contaminants. They are derived from basal cells that include the airway stem/progenitor cells. In animal models, the transcription factor FOXJ1 has been shown to induce differentiation to the ciliated cell lineage, and the RFX transcription factor-family has been shown to be necessary for, but not sufficient to induce, correct cilia development.Methods: To test the hypothesis that FOXJ1 and RFX3 cooperatively induce expression of ciliated genes in the differentiation process of basal progenitor cells toward a ciliated cell linage in the human airway epithelium, primary human airway basal cells were assessed under conditions of in vitro differentiation induced by plasmid-mediated gene transfer of FOXJ1 and/or RFX3. TaqMan PCR was used to quantify mRNA levels of basal, secretory, and cilia-associated genes.Results: Basal cells, when cultured in air-liquid interface, differentiated into a ciliated epithelium, expressing FOXJ1 and RFX3. Transfection of FOXJ1 into resting basal cells activated promoters and induced expression of ciliated cell genes as well as both FOXJ1 and RFX3, but not basal cell genes. Transfection of RFX3 induced expression of RFX3 but not FOXJ1, nor the expression of cilia-related genes. The combination of FOXJ1 + RFX3 enhanced ciliated gene promoter activity and mRNA expression beyond that due to FOXJ1 alone. Corroborating immunoprecipitation studies demonstrated an interaction between FOXJ1 and RFX3.Conclusion: FOXJ1 is an important regulator of cilia gene expression during ciliated cell differentiation, with RFX3 as a transcriptional co-activator to FOXJ1, helping to induce the expression of cilia genes in the process of ciliated cell differentiation of basal/progenitor cells.

KW - Basal cell

KW - Ciliated cell differentiation

KW - FOXJ1

KW - Human

KW - Lung epithelium

KW - RFX3

UR - http://www.scopus.com/inward/record.url?scp=84879831717&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84879831717&partnerID=8YFLogxK

U2 - 10.1186/1465-9921-14-70

DO - 10.1186/1465-9921-14-70

M3 - Article

VL - 14

JO - Respiratory Research

JF - Respiratory Research

SN - 1465-9921

IS - 1

M1 - 70

ER -