Pax9 is required for cardiovascular development and interacts with Tbx1 in the pharyngeal endoderm to control 4th pharyngeal arch artery morphogenesis

Helen M. Phillips; Catherine A.  Stothard; Wasay M. Shaikh Qureshi; Anastasia L.  Kousa; J. Alberto  Briones-Leon; Ramada R.  Khasawneh; Chloe  O'Loughlin; Rachel  Sanders; Silvia Mazzotta; Rebecca Dodds; Kerstin  Seidel; Timothy C Bates; Mitsushiro Nakatomi; Simon J.  Cockell; Jurgen E. Schneider; Timothy J. Mohun; Rene' Maehr; Ralf Kist; Heiko Peters; Simon D. Bamforth

doi:10.1242/dev.177618

Pax9 is required for cardiovascular development and interacts with Tbx1 in the pharyngeal endoderm to control 4th pharyngeal arch artery morphogenesis

Helen M. Phillips
, Catherine A. Stothard
, Wasay M. Shaikh Qureshi
, Anastasia L. Kousa
, J. Alberto Briones-Leon
, Ramada R. Khasawneh
, Chloe O'Loughlin
, Rachel Sanders
, Silvia Mazzotta
, Rebecca Dodds
, Kerstin Seidel
, Timothy C Bates
, Mitsushiro Nakatomi
, Simon J. Cockell
, Jurgen E. Schneider
, Timothy J. Mohun
, Rene' Maehr
, Ralf Kist
, Heiko Peters
, Simon D. Bamforth^* (Corresponding Author)

^*Corresponding author for this work

Medical Sciences

Newcastle University
Memorial Sloan-Kettering Cancer Center
University of Massachusetts
University of Leeds
The Francis Crick Institute

Research output: Contribution to journal › Article › peer-review

33 Citations (Scopus)

6 Downloads (Pure)

Abstract

Developmental defects affecting the heart and aortic arch arteries are a significant phenotype observed in individuals with 22q11 deletion syndrome and are caused by a microdeletion on chromosome 22q11. TBX1, one of the deleted genes, is expressed throughout the pharyngeal arches and is considered a key gene, when mutated, for the arch artery defects. Pax9 is expressed in the pharyngeal endoderm and is downregulated in Tbx1 mutant mice. We show here that Pax9-deficient mice are born with complex cardiovascular malformations that affect the outflow tract and aortic arch arteries with failure of the 3rd and 4th pharyngeal arch arteries to form correctly. Transcriptome analysis indicated that Pax9 and Tbx1 may function together, and mice double heterozygous for Tbx1/Pax9 presented with a significantly increased incidence of interrupted aortic arch when compared with Tbx1 heterozygous mice. Using a novel Pax9Cre allele, we demonstrated that the site of this Tbx1-Pax9 genetic interaction is the pharyngeal endoderm, therefore revealing that a Tbx1-Pax9-controlled signalling mechanism emanating from the pharyngeal endoderm is required for crucial tissue interactions during normal morphogenesis of the pharyngeal arch artery system.

Original language	English
Article number	dev177618
Number of pages	16
Journal	Development
Volume	146
Issue number	18
Early online date	23 Sept 2019
DOIs	https://doi.org/10.1242/dev.177618
Publication status	Published - Sept 2019

Bibliographical note

We thank Jessica Addison, Kathleen Allinson and Divya Venkatesh for technical
assistance, Sushma Grellscheid for facilitating the RNA-seq experiment, and
Nicoletta Bobola for critically reading the manuscript. Tbx1 +/– mice were obtained
from Robert Kelly and Virginia Papaioannou. We acknowledge the Newcastle
University Flow Cytometry Core Facility (FCCF) for assistance with the generation of
flow cytometry data.

Data Availability Statement

The RNA-seq data have been deposited in GEO under accession number
GSE128087.

Funding

This work was funded by a British Heart Foundation Intermediate Basic Science Research Fellowship (FS/08/016/24741 to S.D.B.), a British Heart Foundation project grant (PG/16/39/32115 to S.D.B.), a British Heart Foundation PhD Non- Clinical PhD Studentship (FS/16/8/31984 to S.D.B.) and a grant from the Newcastle upon Tyne Hospitals NHS Foundation Trust (BH120404 to S.D.B.). J.A.B.-L. received a PhD Scholarship from the Consejo Nacional de Ciencia y Tecnologı́a (CONACyT). R.R.K. received a PhD Scholarship from Yarmouk University, Jordan.

Funders	Funder number
British Heart Foundation	FS/08/016/24741, PG/16/39/32115, FS/16/8/31984
Newcastle upon Tyne Hospitals NHS Foundation Trust	BH120404

Keywords

Pharyngeal endoderm
Arch artery development
Tbx1
Pax9
Neural crest
22q11 deletion syndrome

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Cite this

Phillips, H. M., Stothard, C. A., Qureshi, W. M. S., Kousa, A. L., Briones-Leon, J. A., Khasawneh, R. R., O'Loughlin, C., Sanders, R., Mazzotta, S., Dodds, R., Seidel, K., Bates, T. C., Nakatomi, M., Cockell, S. J., Schneider, J. E., Mohun, T. J., Maehr, R., Kist, R., Peters, H., & Bamforth, S. D. (2019). Pax9 is required for cardiovascular development and interacts with Tbx1 in the pharyngeal endoderm to control 4th pharyngeal arch artery morphogenesis. Development, 146(18), Article dev177618. https://doi.org/10.1242/dev.177618

Phillips, HM, Stothard, CA, Qureshi, WMS, Kousa, AL, Briones-Leon, JA, Khasawneh, RR, O'Loughlin, C, Sanders, R, Mazzotta, S, Dodds, R, Seidel, K, Bates, TC, Nakatomi, M, Cockell, SJ, Schneider, JE, Mohun, TJ, Maehr, R, Kist, R, Peters, H & Bamforth, SD 2019, 'Pax9 is required for cardiovascular development and interacts with Tbx1 in the pharyngeal endoderm to control 4th pharyngeal arch artery morphogenesis', Development, vol. 146, no. 18, dev177618. https://doi.org/10.1242/dev.177618

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title = "Pax9 is required for cardiovascular development and interacts with Tbx1 in the pharyngeal endoderm to control 4th pharyngeal arch artery morphogenesis",

abstract = "Developmental defects affecting the heart and aortic arch arteries are a significant phenotype observed in individuals with 22q11 deletion syndrome and are caused by a microdeletion on chromosome 22q11. TBX1, one of the deleted genes, is expressed throughout the pharyngeal arches and is considered a key gene, when mutated, for the arch artery defects. Pax9 is expressed in the pharyngeal endoderm and is downregulated in Tbx1 mutant mice. We show here that Pax9-deficient mice are born with complex cardiovascular malformations that affect the outflow tract and aortic arch arteries with failure of the 3rd and 4th pharyngeal arch arteries to form correctly. Transcriptome analysis indicated that Pax9 and Tbx1 may function together, and mice double heterozygous for Tbx1/Pax9 presented with a significantly increased incidence of interrupted aortic arch when compared with Tbx1 heterozygous mice. Using a novel Pax9Cre allele, we demonstrated that the site of this Tbx1-Pax9 genetic interaction is the pharyngeal endoderm, therefore revealing that a Tbx1-Pax9-controlled signalling mechanism emanating from the pharyngeal endoderm is required for crucial tissue interactions during normal morphogenesis of the pharyngeal arch artery system.",

keywords = "Pharyngeal endoderm, Arch artery development, Tbx1, Pax9, Neural crest, 22q11 deletion syndrome",

author = "Phillips, \{Helen M.\} and Stothard, \{Catherine A.\} and Qureshi, \{Wasay M. Shaikh\} and Kousa, \{Anastasia L.\} and Briones-Leon, \{J. Alberto\} and Khasawneh, \{Ramada R.\} and Chloe O'Loughlin and Rachel Sanders and Silvia Mazzotta and Rebecca Dodds and Kerstin Seidel and Bates, \{Timothy C\} and Mitsushiro Nakatomi and Cockell, \{Simon J.\} and Schneider, \{Jurgen E.\} and Mohun, \{Timothy J.\} and Rene' Maehr and Ralf Kist and Heiko Peters and Bamforth, \{Simon D.\}",

note = "We thank Jessica Addison, Kathleen Allinson and Divya Venkatesh for technical assistance, Sushma Grellscheid for facilitating the RNA-seq experiment, and Nicoletta Bobola for critically reading the manuscript. Tbx1 +/{\textendash} mice were obtained from Robert Kelly and Virginia Papaioannou. We acknowledge the Newcastle University Flow Cytometry Core Facility (FCCF) for assistance with the generation of flow cytometry data.",

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doi = "10.1242/dev.177618",

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T1 - Pax9 is required for cardiovascular development and interacts with Tbx1 in the pharyngeal endoderm to control 4th pharyngeal arch artery morphogenesis

AU - Phillips, Helen M.

AU - Stothard, Catherine A.

AU - Qureshi, Wasay M. Shaikh

AU - Kousa, Anastasia L.

AU - Briones-Leon, J. Alberto

AU - Khasawneh, Ramada R.

AU - O'Loughlin, Chloe

AU - Sanders, Rachel

AU - Mazzotta, Silvia

AU - Dodds, Rebecca

AU - Seidel, Kerstin

AU - Bates, Timothy C

AU - Nakatomi, Mitsushiro

AU - Cockell, Simon J.

AU - Schneider, Jurgen E.

AU - Mohun, Timothy J.

AU - Maehr, Rene'

AU - Kist, Ralf

AU - Peters, Heiko

AU - Bamforth, Simon D.

N1 - We thank Jessica Addison, Kathleen Allinson and Divya Venkatesh for technical assistance, Sushma Grellscheid for facilitating the RNA-seq experiment, and Nicoletta Bobola for critically reading the manuscript. Tbx1 +/– mice were obtained from Robert Kelly and Virginia Papaioannou. We acknowledge the Newcastle University Flow Cytometry Core Facility (FCCF) for assistance with the generation of flow cytometry data.

PY - 2019/9

Y1 - 2019/9

N2 - Developmental defects affecting the heart and aortic arch arteries are a significant phenotype observed in individuals with 22q11 deletion syndrome and are caused by a microdeletion on chromosome 22q11. TBX1, one of the deleted genes, is expressed throughout the pharyngeal arches and is considered a key gene, when mutated, for the arch artery defects. Pax9 is expressed in the pharyngeal endoderm and is downregulated in Tbx1 mutant mice. We show here that Pax9-deficient mice are born with complex cardiovascular malformations that affect the outflow tract and aortic arch arteries with failure of the 3rd and 4th pharyngeal arch arteries to form correctly. Transcriptome analysis indicated that Pax9 and Tbx1 may function together, and mice double heterozygous for Tbx1/Pax9 presented with a significantly increased incidence of interrupted aortic arch when compared with Tbx1 heterozygous mice. Using a novel Pax9Cre allele, we demonstrated that the site of this Tbx1-Pax9 genetic interaction is the pharyngeal endoderm, therefore revealing that a Tbx1-Pax9-controlled signalling mechanism emanating from the pharyngeal endoderm is required for crucial tissue interactions during normal morphogenesis of the pharyngeal arch artery system.

AB - Developmental defects affecting the heart and aortic arch arteries are a significant phenotype observed in individuals with 22q11 deletion syndrome and are caused by a microdeletion on chromosome 22q11. TBX1, one of the deleted genes, is expressed throughout the pharyngeal arches and is considered a key gene, when mutated, for the arch artery defects. Pax9 is expressed in the pharyngeal endoderm and is downregulated in Tbx1 mutant mice. We show here that Pax9-deficient mice are born with complex cardiovascular malformations that affect the outflow tract and aortic arch arteries with failure of the 3rd and 4th pharyngeal arch arteries to form correctly. Transcriptome analysis indicated that Pax9 and Tbx1 may function together, and mice double heterozygous for Tbx1/Pax9 presented with a significantly increased incidence of interrupted aortic arch when compared with Tbx1 heterozygous mice. Using a novel Pax9Cre allele, we demonstrated that the site of this Tbx1-Pax9 genetic interaction is the pharyngeal endoderm, therefore revealing that a Tbx1-Pax9-controlled signalling mechanism emanating from the pharyngeal endoderm is required for crucial tissue interactions during normal morphogenesis of the pharyngeal arch artery system.

KW - Pharyngeal endoderm

KW - Arch artery development

KW - Tbx1

KW - Pax9

KW - Neural crest

KW - 22q11 deletion syndrome

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U2 - 10.1242/dev.177618

DO - 10.1242/dev.177618

M3 - Article

SN - 0950-1991

VL - 146

JO - Development

JF - Development

IS - 18

M1 - dev177618

ER -

Pax9 is required for cardiovascular development and interacts with Tbx1 in the pharyngeal endoderm to control 4th pharyngeal arch artery morphogenesis

Abstract

Bibliographical note

Data Availability Statement

Funding

Keywords

Access to Document

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