C11orf70 Mutations Disrupting the Intraflagellar Transport-Dependent Assembly of Multiple Axonemal Dyneins Cause Primary Ciliary Dyskinesia

Mahmoud R. Fassad, Amelia Shoemark, Pierrick le Borgne, France Koll, Mitali Patel, Mellisa Dixon, Jane Hayward, Charlotte Richardson, Emily Frost, Lucy Jenkins, Thomas Cullup, Eddie M. K. Chung, Michel Lemullois, Anne Aubusson-Fleury, Claire Hogg, David R. Mitchell, Anne-Marie Tassin, Hannah M. Mitchison (Lead / Corresponding author)

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Abstract

Primary ciliary dyskinesia (PCD) is a genetically and phenotypically heterogeneous disorder characterized by destructive respiratory disease and laterality abnormalities due to randomized left-right body asymmetry. PCD is mostly caused by mutations affecting the core axoneme structure of motile cilia that is essential for movement. Genes that cause PCD when mutated include a group that encode proteins essential for the assembly of the ciliary dynein motors and the active transport process that delivers them from their cytoplasmic assembly site into the axoneme. We screened a cohort of affected individuals for disease-causing mutations using a targeted next generation sequencing panel and identified two unrelated families (three affected children) with mutations in the uncharacterized C11orf70 gene (official gene name CFAP300). The affected children share a consistent PCD phenotype from early life with laterality defects and immotile respiratory cilia displaying combined loss of inner and outer dynein arms (IDA+ODA). Phylogenetic analysis shows C11orf70 is highly conserved, distributed across species similarly to proteins involved in the intraflagellar transport (IFT)-dependant assembly of axonemal dyneins. Paramecium C11orf70 RNAi knockdown led to combined loss of ciliary IDA+ODA with reduced cilia beating and swim velocity. Tagged C11orf70 in Paramecium and Chlamydomonas localizes mainly in the cytoplasm with a small amount in the ciliary component. IFT139/TTC21B (IFT-A protein) and FLA10 (IFT kinesin) depletion experiments show that its transport within cilia is IFT dependent. During ciliogenesis, C11orf70 accumulates at the ciliary tips in a similar distribution to the IFT-B protein IFT46. In summary, C11orf70 is essential for assembly of dynein arms and C11orf70 mutations cause defective cilia motility and PCD.

Original languageEnglish
Pages (from-to)956-972
Number of pages17
JournalAmerican Journal of Human Genetics
Volume102
Issue number5
DOIs
Publication statusPublished - 3 May 2018

Fingerprint

Axonemal Dyneins
Kartagener Syndrome
Cilia
Dyneins
Mutation
Paramecium
Axoneme
Carrier Proteins
Genes
Chlamydomonas
Kinesin
Active Biological Transport
RNA Interference
Names
Cytoplasm
Proteins
Phenotype

Keywords

  • primary ciliary dyskinesia
  • Kartagener syndrome
  • mutation
  • intraflagellar transport
  • dynein
  • Paramecium

Cite this

Fassad, Mahmoud R. ; Shoemark, Amelia ; le Borgne, Pierrick ; Koll, France ; Patel, Mitali ; Dixon, Mellisa ; Hayward, Jane ; Richardson, Charlotte ; Frost, Emily ; Jenkins, Lucy ; Cullup, Thomas ; Chung, Eddie M. K. ; Lemullois, Michel ; Aubusson-Fleury, Anne ; Hogg, Claire ; Mitchell, David R. ; Tassin, Anne-Marie ; Mitchison, Hannah M. / C11orf70 Mutations Disrupting the Intraflagellar Transport-Dependent Assembly of Multiple Axonemal Dyneins Cause Primary Ciliary Dyskinesia. In: American Journal of Human Genetics. 2018 ; Vol. 102, No. 5. pp. 956-972.
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abstract = "Primary ciliary dyskinesia (PCD) is a genetically and phenotypically heterogeneous disorder characterized by destructive respiratory disease and laterality abnormalities due to randomized left-right body asymmetry. PCD is mostly caused by mutations affecting the core axoneme structure of motile cilia that is essential for movement. Genes that cause PCD when mutated include a group that encode proteins essential for the assembly of the ciliary dynein motors and the active transport process that delivers them from their cytoplasmic assembly site into the axoneme. We screened a cohort of affected individuals for disease-causing mutations using a targeted next generation sequencing panel and identified two unrelated families (three affected children) with mutations in the uncharacterized C11orf70 gene (official gene name CFAP300). The affected children share a consistent PCD phenotype from early life with laterality defects and immotile respiratory cilia displaying combined loss of inner and outer dynein arms (IDA+ODA). Phylogenetic analysis shows C11orf70 is highly conserved, distributed across species similarly to proteins involved in the intraflagellar transport (IFT)-dependant assembly of axonemal dyneins. Paramecium C11orf70 RNAi knockdown led to combined loss of ciliary IDA+ODA with reduced cilia beating and swim velocity. Tagged C11orf70 in Paramecium and Chlamydomonas localizes mainly in the cytoplasm with a small amount in the ciliary component. IFT139/TTC21B (IFT-A protein) and FLA10 (IFT kinesin) depletion experiments show that its transport within cilia is IFT dependent. During ciliogenesis, C11orf70 accumulates at the ciliary tips in a similar distribution to the IFT-B protein IFT46. In summary, C11orf70 is essential for assembly of dynein arms and C11orf70 mutations cause defective cilia motility and PCD.",
keywords = "primary ciliary dyskinesia, Kartagener syndrome, mutation, intraflagellar transport, dynein, Paramecium",
author = "Fassad, {Mahmoud R.} and Amelia Shoemark and {le Borgne}, Pierrick and France Koll and Mitali Patel and Mellisa Dixon and Jane Hayward and Charlotte Richardson and Emily Frost and Lucy Jenkins and Thomas Cullup and Chung, {Eddie M. K.} and Michel Lemullois and Anne Aubusson-Fleury and Claire Hogg and Mitchell, {David R.} and Anne-Marie Tassin and Mitchison, {Hannah M.}",
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Fassad, MR, Shoemark, A, le Borgne, P, Koll, F, Patel, M, Dixon, M, Hayward, J, Richardson, C, Frost, E, Jenkins, L, Cullup, T, Chung, EMK, Lemullois, M, Aubusson-Fleury, A, Hogg, C, Mitchell, DR, Tassin, A-M & Mitchison, HM 2018, 'C11orf70 Mutations Disrupting the Intraflagellar Transport-Dependent Assembly of Multiple Axonemal Dyneins Cause Primary Ciliary Dyskinesia', American Journal of Human Genetics, vol. 102, no. 5, pp. 956-972. https://doi.org/10.1016/j.ajhg.2018.03.024

C11orf70 Mutations Disrupting the Intraflagellar Transport-Dependent Assembly of Multiple Axonemal Dyneins Cause Primary Ciliary Dyskinesia. / Fassad, Mahmoud R.; Shoemark, Amelia; le Borgne, Pierrick; Koll, France; Patel, Mitali; Dixon, Mellisa; Hayward, Jane; Richardson, Charlotte; Frost, Emily; Jenkins, Lucy; Cullup, Thomas; Chung, Eddie M. K.; Lemullois, Michel; Aubusson-Fleury, Anne; Hogg, Claire; Mitchell, David R.; Tassin, Anne-Marie; Mitchison, Hannah M. (Lead / Corresponding author).

In: American Journal of Human Genetics, Vol. 102, No. 5, 03.05.2018, p. 956-972.

Research output: Contribution to journalArticle

TY - JOUR

T1 - C11orf70 Mutations Disrupting the Intraflagellar Transport-Dependent Assembly of Multiple Axonemal Dyneins Cause Primary Ciliary Dyskinesia

AU - Fassad, Mahmoud R.

AU - Shoemark, Amelia

AU - le Borgne, Pierrick

AU - Koll, France

AU - Patel, Mitali

AU - Dixon, Mellisa

AU - Hayward, Jane

AU - Richardson, Charlotte

AU - Frost, Emily

AU - Jenkins, Lucy

AU - Cullup, Thomas

AU - Chung, Eddie M. K.

AU - Lemullois, Michel

AU - Aubusson-Fleury, Anne

AU - Hogg, Claire

AU - Mitchell, David R.

AU - Tassin, Anne-Marie

AU - Mitchison, Hannah M.

N1 - Funding for this study was provided by Action Medical Research (GN2101; H.M.M.) and Great Ormond Street Children’s Charity grant (V4515; H.M.M.) and Leadership awards (V1299, V2217; H.M.M.). M.R.F. is supported by the British Council Newton-Mosharafa Fund and the Ministry of Higher Education in Egypt. We acknowledge support from the NIHR Biomedical Research Centre at Great Ormond Street Hospital for Children NHS Foundation Trust and University College London (Doctoral Trainee Support Award; M.R.F.). Work by A.S. is independent research funded by a postdoctoral research fellowship from the National Institute of Health Research and Health Education England.

PY - 2018/5/3

Y1 - 2018/5/3

N2 - Primary ciliary dyskinesia (PCD) is a genetically and phenotypically heterogeneous disorder characterized by destructive respiratory disease and laterality abnormalities due to randomized left-right body asymmetry. PCD is mostly caused by mutations affecting the core axoneme structure of motile cilia that is essential for movement. Genes that cause PCD when mutated include a group that encode proteins essential for the assembly of the ciliary dynein motors and the active transport process that delivers them from their cytoplasmic assembly site into the axoneme. We screened a cohort of affected individuals for disease-causing mutations using a targeted next generation sequencing panel and identified two unrelated families (three affected children) with mutations in the uncharacterized C11orf70 gene (official gene name CFAP300). The affected children share a consistent PCD phenotype from early life with laterality defects and immotile respiratory cilia displaying combined loss of inner and outer dynein arms (IDA+ODA). Phylogenetic analysis shows C11orf70 is highly conserved, distributed across species similarly to proteins involved in the intraflagellar transport (IFT)-dependant assembly of axonemal dyneins. Paramecium C11orf70 RNAi knockdown led to combined loss of ciliary IDA+ODA with reduced cilia beating and swim velocity. Tagged C11orf70 in Paramecium and Chlamydomonas localizes mainly in the cytoplasm with a small amount in the ciliary component. IFT139/TTC21B (IFT-A protein) and FLA10 (IFT kinesin) depletion experiments show that its transport within cilia is IFT dependent. During ciliogenesis, C11orf70 accumulates at the ciliary tips in a similar distribution to the IFT-B protein IFT46. In summary, C11orf70 is essential for assembly of dynein arms and C11orf70 mutations cause defective cilia motility and PCD.

AB - Primary ciliary dyskinesia (PCD) is a genetically and phenotypically heterogeneous disorder characterized by destructive respiratory disease and laterality abnormalities due to randomized left-right body asymmetry. PCD is mostly caused by mutations affecting the core axoneme structure of motile cilia that is essential for movement. Genes that cause PCD when mutated include a group that encode proteins essential for the assembly of the ciliary dynein motors and the active transport process that delivers them from their cytoplasmic assembly site into the axoneme. We screened a cohort of affected individuals for disease-causing mutations using a targeted next generation sequencing panel and identified two unrelated families (three affected children) with mutations in the uncharacterized C11orf70 gene (official gene name CFAP300). The affected children share a consistent PCD phenotype from early life with laterality defects and immotile respiratory cilia displaying combined loss of inner and outer dynein arms (IDA+ODA). Phylogenetic analysis shows C11orf70 is highly conserved, distributed across species similarly to proteins involved in the intraflagellar transport (IFT)-dependant assembly of axonemal dyneins. Paramecium C11orf70 RNAi knockdown led to combined loss of ciliary IDA+ODA with reduced cilia beating and swim velocity. Tagged C11orf70 in Paramecium and Chlamydomonas localizes mainly in the cytoplasm with a small amount in the ciliary component. IFT139/TTC21B (IFT-A protein) and FLA10 (IFT kinesin) depletion experiments show that its transport within cilia is IFT dependent. During ciliogenesis, C11orf70 accumulates at the ciliary tips in a similar distribution to the IFT-B protein IFT46. In summary, C11orf70 is essential for assembly of dynein arms and C11orf70 mutations cause defective cilia motility and PCD.

KW - primary ciliary dyskinesia

KW - Kartagener syndrome

KW - mutation

KW - intraflagellar transport

KW - dynein

KW - Paramecium

U2 - 10.1016/j.ajhg.2018.03.024

DO - 10.1016/j.ajhg.2018.03.024

M3 - Article

VL - 102

SP - 956

EP - 972

JO - American Journal of Human Genetics

JF - American Journal of Human Genetics

SN - 0002-9297

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