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Copy number variation in patients with disorders of sex development due to 46,XY gonadal dysgenesis.

White S, Ohnesorg T, Notini A, Roeszler K, Hewitt J, Daggag H, Smith C, Turbitt E, Gustin S, van den Bergen J, Miles D, Western P, Arboleda V, Schumacher V, Gordon L, Bell K, Bengtsson H, Speed T, Hutson J, Warne G, Harley V, Koopman P, Vilain E, Sinclair A - PLoS ONE (2011)

Bottom Line: Functional analysis of potential SRY binding sites within this deleted region identified five putative enhancers, suggesting that sequences additional to the known SRY-binding TES enhancer influence human testis-specific SOX9 expression.Thirdly, we identified a small deletion immediately downstream of GATA4, supporting a role for GATA4 in gonad development in humans.These CNV analyses give new insights into the pathways involved in human gonad development and dysfunction, and suggest that rearrangements of non-coding sequences disturbing gene regulation may account for significant proportion of DSD cases.

View Article: PubMed Central - PubMed

Affiliation: Murdoch Childrens Research Institute, Royal Children's Hospital, Melbourne, Victoria, Australia.

ABSTRACT
Disorders of sex development (DSD), ranging in severity from mild genital abnormalities to complete sex reversal, represent a major concern for patients and their families. DSD are often due to disruption of the genetic programs that regulate gonad development. Although some genes have been identified in these developmental pathways, the causative mutations have not been identified in more than 50% 46,XY DSD cases. We used the Affymetrix Genome-Wide Human SNP Array 6.0 to analyse copy number variation in 23 individuals with unexplained 46,XY DSD due to gonadal dysgenesis (GD). Here we describe three discrete changes in copy number that are the likely cause of the GD. Firstly, we identified a large duplication on the X chromosome that included DAX1 (NR0B1). Secondly, we identified a rearrangement that appears to affect a novel gonad-specific regulatory region in a known testis gene, SOX9. Surprisingly this patient lacked any signs of campomelic dysplasia, suggesting that the deletion affected expression of SOX9 only in the gonad. Functional analysis of potential SRY binding sites within this deleted region identified five putative enhancers, suggesting that sequences additional to the known SRY-binding TES enhancer influence human testis-specific SOX9 expression. Thirdly, we identified a small deletion immediately downstream of GATA4, supporting a role for GATA4 in gonad development in humans. These CNV analyses give new insights into the pathways involved in human gonad development and dysfunction, and suggest that rearrangements of non-coding sequences disturbing gene regulation may account for significant proportion of DSD cases.

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Expression analysis of Camk1d and Dnajc15 in developing mouse gonads.Expression analysis was performed on cDNA from sorted GFP- (somatic) cells derived from male and female mouse gonads, embryonic days 12.5–15.5. The data is normalised such that the lowest expression is 1.0, and represent mean values ±SEM from three independent experiments. Shown here are the expression patterns for a) Camk1d and b) Dnajc15. Comparisons between male and female expression for Camk1d and Dnajc15 were performed with a 2-tailed t-test. ** p<0.005; * p<0.05.
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pone-0017793-g004: Expression analysis of Camk1d and Dnajc15 in developing mouse gonads.Expression analysis was performed on cDNA from sorted GFP- (somatic) cells derived from male and female mouse gonads, embryonic days 12.5–15.5. The data is normalised such that the lowest expression is 1.0, and represent mean values ±SEM from three independent experiments. Shown here are the expression patterns for a) Camk1d and b) Dnajc15. Comparisons between male and female expression for Camk1d and Dnajc15 were performed with a 2-tailed t-test. ** p<0.005; * p<0.05.

Mentions: Several previously unreported CNVs were identified that affected the coding region of genes not currently associated with gonad development. The genes affected were GPR83, ACBD7, CAMK1D, OLAH, NEIL2 and DNAJC15 (table S3). To assess the potential of these genes to be involved in DSD, expression analysis of the orthologous mouse genes was performed on cDNA from purified somatic cells isolated from male and female mouse gonads at the time of sex differentiation. Two genes (Dnajc15 and Camk1d) showed sexually dimorphic expression in the somatic cells (Figure 4). Dnajc15 was expressed more highly in the female gonad than the male (p<0.005 at E13.5, E14.5, E15.5). Conversely, Camk1d expression was significantly higher in the developing male gonad when compared to the female gonad (p<0.05 at E13.5, p<0.005 at E15.5).


Copy number variation in patients with disorders of sex development due to 46,XY gonadal dysgenesis.

White S, Ohnesorg T, Notini A, Roeszler K, Hewitt J, Daggag H, Smith C, Turbitt E, Gustin S, van den Bergen J, Miles D, Western P, Arboleda V, Schumacher V, Gordon L, Bell K, Bengtsson H, Speed T, Hutson J, Warne G, Harley V, Koopman P, Vilain E, Sinclair A - PLoS ONE (2011)

Expression analysis of Camk1d and Dnajc15 in developing mouse gonads.Expression analysis was performed on cDNA from sorted GFP- (somatic) cells derived from male and female mouse gonads, embryonic days 12.5–15.5. The data is normalised such that the lowest expression is 1.0, and represent mean values ±SEM from three independent experiments. Shown here are the expression patterns for a) Camk1d and b) Dnajc15. Comparisons between male and female expression for Camk1d and Dnajc15 were performed with a 2-tailed t-test. ** p<0.005; * p<0.05.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3049794&req=5

pone-0017793-g004: Expression analysis of Camk1d and Dnajc15 in developing mouse gonads.Expression analysis was performed on cDNA from sorted GFP- (somatic) cells derived from male and female mouse gonads, embryonic days 12.5–15.5. The data is normalised such that the lowest expression is 1.0, and represent mean values ±SEM from three independent experiments. Shown here are the expression patterns for a) Camk1d and b) Dnajc15. Comparisons between male and female expression for Camk1d and Dnajc15 were performed with a 2-tailed t-test. ** p<0.005; * p<0.05.
Mentions: Several previously unreported CNVs were identified that affected the coding region of genes not currently associated with gonad development. The genes affected were GPR83, ACBD7, CAMK1D, OLAH, NEIL2 and DNAJC15 (table S3). To assess the potential of these genes to be involved in DSD, expression analysis of the orthologous mouse genes was performed on cDNA from purified somatic cells isolated from male and female mouse gonads at the time of sex differentiation. Two genes (Dnajc15 and Camk1d) showed sexually dimorphic expression in the somatic cells (Figure 4). Dnajc15 was expressed more highly in the female gonad than the male (p<0.005 at E13.5, E14.5, E15.5). Conversely, Camk1d expression was significantly higher in the developing male gonad when compared to the female gonad (p<0.05 at E13.5, p<0.005 at E15.5).

Bottom Line: Functional analysis of potential SRY binding sites within this deleted region identified five putative enhancers, suggesting that sequences additional to the known SRY-binding TES enhancer influence human testis-specific SOX9 expression.Thirdly, we identified a small deletion immediately downstream of GATA4, supporting a role for GATA4 in gonad development in humans.These CNV analyses give new insights into the pathways involved in human gonad development and dysfunction, and suggest that rearrangements of non-coding sequences disturbing gene regulation may account for significant proportion of DSD cases.

View Article: PubMed Central - PubMed

Affiliation: Murdoch Childrens Research Institute, Royal Children's Hospital, Melbourne, Victoria, Australia.

ABSTRACT
Disorders of sex development (DSD), ranging in severity from mild genital abnormalities to complete sex reversal, represent a major concern for patients and their families. DSD are often due to disruption of the genetic programs that regulate gonad development. Although some genes have been identified in these developmental pathways, the causative mutations have not been identified in more than 50% 46,XY DSD cases. We used the Affymetrix Genome-Wide Human SNP Array 6.0 to analyse copy number variation in 23 individuals with unexplained 46,XY DSD due to gonadal dysgenesis (GD). Here we describe three discrete changes in copy number that are the likely cause of the GD. Firstly, we identified a large duplication on the X chromosome that included DAX1 (NR0B1). Secondly, we identified a rearrangement that appears to affect a novel gonad-specific regulatory region in a known testis gene, SOX9. Surprisingly this patient lacked any signs of campomelic dysplasia, suggesting that the deletion affected expression of SOX9 only in the gonad. Functional analysis of potential SRY binding sites within this deleted region identified five putative enhancers, suggesting that sequences additional to the known SRY-binding TES enhancer influence human testis-specific SOX9 expression. Thirdly, we identified a small deletion immediately downstream of GATA4, supporting a role for GATA4 in gonad development in humans. These CNV analyses give new insights into the pathways involved in human gonad development and dysfunction, and suggest that rearrangements of non-coding sequences disturbing gene regulation may account for significant proportion of DSD cases.

Show MeSH
Related in: MedlinePlus