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Functional analysis of novel desert hedgehog gene variants improves the clinical interpretation of genomic data and provides a more accurate diagnosis for patients with 46, XY differences of sex development

BACKGROUND: Desert hedgehog (DHH) gene variants are known to cause 46, XY differences/disorders of sex development (DSD). We have identified six patients with 46, XY DSD with seven novel DHH gene variants. Many of these variants were classified as variants of uncertain significance due to their hete...

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Autores principales: Ayers, Katie, van den Bergen, Jocelyn, Robevska, Gorjana, Listyasari, Nurin, Raza, Jamal, Atta, Irum, Riedl, Stefan, Rothacker, Karen, Choong, Catherine, Faradz, Sultana M H, Sinclair, Andrew
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BMJ Publishing Group 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6591740/
https://www.ncbi.nlm.nih.gov/pubmed/31018998
http://dx.doi.org/10.1136/jmedgenet-2018-105893
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author Ayers, Katie
van den Bergen, Jocelyn
Robevska, Gorjana
Listyasari, Nurin
Raza, Jamal
Atta, Irum
Riedl, Stefan
Rothacker, Karen
Choong, Catherine
Faradz, Sultana M H
Sinclair, Andrew
author_facet Ayers, Katie
van den Bergen, Jocelyn
Robevska, Gorjana
Listyasari, Nurin
Raza, Jamal
Atta, Irum
Riedl, Stefan
Rothacker, Karen
Choong, Catherine
Faradz, Sultana M H
Sinclair, Andrew
author_sort Ayers, Katie
collection PubMed
description BACKGROUND: Desert hedgehog (DHH) gene variants are known to cause 46, XY differences/disorders of sex development (DSD). We have identified six patients with 46, XY DSD with seven novel DHH gene variants. Many of these variants were classified as variants of uncertain significance due to their heterozygosity or associated milder phenotype. To assess variant pathogenicity and to refine the spectrum of DSDs associated with this gene, we have carried out the first reported functional testing of DHH gene variant activity. METHODS: A cell co-culture method was used to assess DHH variant induction of Hedgehog signalling in cultured Leydig cells. Protein expression and subcellular localisation were also assessed for DHH variants using western blot and immunofluorescence. RESULTS: Our co-culture method provided a robust read-out of DHH gene variant activity, which correlated closely with patient phenotype severity. While biallelic DHH variants from patients with gonadal dysgenesis showed significant loss of activity, variants found as heterozygous in patients with milder phenotypes had no loss of activity when tested with a wild type allele. Taking these functional results into account improved clinical interpretation. CONCLUSION: Our findings suggest heterozygous DHH gene variants are unlikely to cause DSD, reaffirming that DHH is an autosomal recessive cause of 46, XY gonadal dysgenesis. Functional characterisation of novel DHH variants improves variant interpretation, leading to greater confidence in patient reporting and clinical management.
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spelling pubmed-65917402019-07-11 Functional analysis of novel desert hedgehog gene variants improves the clinical interpretation of genomic data and provides a more accurate diagnosis for patients with 46, XY differences of sex development Ayers, Katie van den Bergen, Jocelyn Robevska, Gorjana Listyasari, Nurin Raza, Jamal Atta, Irum Riedl, Stefan Rothacker, Karen Choong, Catherine Faradz, Sultana M H Sinclair, Andrew J Med Genet Diagnostics BACKGROUND: Desert hedgehog (DHH) gene variants are known to cause 46, XY differences/disorders of sex development (DSD). We have identified six patients with 46, XY DSD with seven novel DHH gene variants. Many of these variants were classified as variants of uncertain significance due to their heterozygosity or associated milder phenotype. To assess variant pathogenicity and to refine the spectrum of DSDs associated with this gene, we have carried out the first reported functional testing of DHH gene variant activity. METHODS: A cell co-culture method was used to assess DHH variant induction of Hedgehog signalling in cultured Leydig cells. Protein expression and subcellular localisation were also assessed for DHH variants using western blot and immunofluorescence. RESULTS: Our co-culture method provided a robust read-out of DHH gene variant activity, which correlated closely with patient phenotype severity. While biallelic DHH variants from patients with gonadal dysgenesis showed significant loss of activity, variants found as heterozygous in patients with milder phenotypes had no loss of activity when tested with a wild type allele. Taking these functional results into account improved clinical interpretation. CONCLUSION: Our findings suggest heterozygous DHH gene variants are unlikely to cause DSD, reaffirming that DHH is an autosomal recessive cause of 46, XY gonadal dysgenesis. Functional characterisation of novel DHH variants improves variant interpretation, leading to greater confidence in patient reporting and clinical management. BMJ Publishing Group 2019-07 2019-04-24 /pmc/articles/PMC6591740/ /pubmed/31018998 http://dx.doi.org/10.1136/jmedgenet-2018-105893 Text en © Author(s) (or their employer(s)) 2019. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ. This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/.
spellingShingle Diagnostics
Ayers, Katie
van den Bergen, Jocelyn
Robevska, Gorjana
Listyasari, Nurin
Raza, Jamal
Atta, Irum
Riedl, Stefan
Rothacker, Karen
Choong, Catherine
Faradz, Sultana M H
Sinclair, Andrew
Functional analysis of novel desert hedgehog gene variants improves the clinical interpretation of genomic data and provides a more accurate diagnosis for patients with 46, XY differences of sex development
title Functional analysis of novel desert hedgehog gene variants improves the clinical interpretation of genomic data and provides a more accurate diagnosis for patients with 46, XY differences of sex development
title_full Functional analysis of novel desert hedgehog gene variants improves the clinical interpretation of genomic data and provides a more accurate diagnosis for patients with 46, XY differences of sex development
title_fullStr Functional analysis of novel desert hedgehog gene variants improves the clinical interpretation of genomic data and provides a more accurate diagnosis for patients with 46, XY differences of sex development
title_full_unstemmed Functional analysis of novel desert hedgehog gene variants improves the clinical interpretation of genomic data and provides a more accurate diagnosis for patients with 46, XY differences of sex development
title_short Functional analysis of novel desert hedgehog gene variants improves the clinical interpretation of genomic data and provides a more accurate diagnosis for patients with 46, XY differences of sex development
title_sort functional analysis of novel desert hedgehog gene variants improves the clinical interpretation of genomic data and provides a more accurate diagnosis for patients with 46, xy differences of sex development
topic Diagnostics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6591740/
https://www.ncbi.nlm.nih.gov/pubmed/31018998
http://dx.doi.org/10.1136/jmedgenet-2018-105893
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