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Disorders of sex development expose transcriptional autonomy of genetic sex and androgen-programmed hormonal sex in human blood leukocytes.

Holterhus PM, Bebermeier JH, Werner R, Demeter J, Richter-Unruh A, Cario G, Appari M, Siebert R, Riepe F, Brooks JD, Hiort O - BMC Genomics (2009)

Bottom Line: Gender appears to be determined by independent programs controlled by the sex-chromosomes and by androgen-dependent programming during embryonic development.However, a significantly larger gene set in the PBMC only reflected the degree of external genital masculinization independent of the sex chromosomes and independent of concurrent post-natal sex steroid hormone levels.A significant fraction of gene expression differences between males and females in the human appears to have its roots in early embryogenesis and is not only caused by sex chromosomes but also by long-term sex-specific hormonal programming due to presence or absence of androgen during the time of external genital masculinization.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pediatrics, Christian Albrechts University of Kiel, Germany. holterhus@pediatrics.uni-kiel.de

ABSTRACT

Background: Gender appears to be determined by independent programs controlled by the sex-chromosomes and by androgen-dependent programming during embryonic development. To enable experimental dissection of these components in the human, we performed genome-wide profiling of the transcriptomes of peripheral blood mononuclear cells (PBMC) in patients with rare defined "disorders of sex development" (DSD, e.g., 46, XY-females due to defective androgen biosynthesis) compared to normal 46, XY-males and 46, XX-females.

Results: A discrete set of transcripts was directly correlated with XY or XX genotypes in all individuals independent of male or female phenotype of the external genitalia. However, a significantly larger gene set in the PBMC only reflected the degree of external genital masculinization independent of the sex chromosomes and independent of concurrent post-natal sex steroid hormone levels. Consequently, the architecture of the transcriptional PBMC-"sexes" was either male, female or even "intersex" with a discordant alignment of the DSD individuals' genetic and hormonal sex signatures.

Conclusion: A significant fraction of gene expression differences between males and females in the human appears to have its roots in early embryogenesis and is not only caused by sex chromosomes but also by long-term sex-specific hormonal programming due to presence or absence of androgen during the time of external genital masculinization. Genetic sex and the androgen milieu during embryonic development might therefore independently modulate functional traits, phenotype and diseases associated with male or female gender as well as with DSD conditions.

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Hierarchical clustering analysis of 34 microarray experiments on PBMC total RNA derived from 9 normal males, 10 normal females, one 46,XY-male with CAH and 14 individuals with DSD [see additional files 2, 3, 4, 5]. (A) Transcript levels of 157 transcripts identified by SAM as differing significantly between PBMC from 9 normal males and 10 normal females. Transcripts are grouped by hierarchical cluster analysis and displayed in rows while experiments are displayed in columns. Expression values per gene are centered by the mean log2 Cy5/Cy3 normalized ratio across the arrays. Increasing blue intensity in the heat map corresponds to higher relative transcript levels compared to the mean expression level across all 34 array experiments. Increasing yellow intensity corresponds to relatively decreased transcript levels compared to the mean. Dark grey corresponds to missing data. Examples taken from these gene clusters are marked by color within the gene tree on the left of the heat map. The red gene tree corresponds to the enlarged gene cluster D, the blue gene tree corresponds to F, the pink gene tree corresponds to G, and the green gene tree corresponds to E, respectively. Gene symbols of the named transcripts are shown on the right. (B) Enlarged cluster dendrogram of the PBMC samples demonstrating the degree of relatedness (Pearson correlation) between the expression patterns of the 157 transcripts. The length of the arms of the dendrogram reflects the degree of correlation between experiments. Samples are color coded to reflect the degree of external genital virilization according to Prader that had been applied to both XX and XY individuals to enable comparability of genital phenotypes independent of the sex chromosomes. The dark grey bar below the experiment cluster represents individuals with an XX karyotype, light grey represents an XY karyotype. The major subdivison of the individuals corresponds strictly to the karyotype. The second level of subdivision in each of the major arms reflects mostly the phenotype of external genital virilization independent of the karyotype. (C) Schematic representation of the Prader stages of external genital virilization. (D) X-chromosome gene cluster (E) Y-chromosome gene cluster (F) Genes with predominantly higher transcript levels in the phenotypic females and the normal females independent of the karyotype (G) Genes with higher transcript levels in the strongly virilized individuals and the normal males independent of the karyotype.
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Figure 1: Hierarchical clustering analysis of 34 microarray experiments on PBMC total RNA derived from 9 normal males, 10 normal females, one 46,XY-male with CAH and 14 individuals with DSD [see additional files 2, 3, 4, 5]. (A) Transcript levels of 157 transcripts identified by SAM as differing significantly between PBMC from 9 normal males and 10 normal females. Transcripts are grouped by hierarchical cluster analysis and displayed in rows while experiments are displayed in columns. Expression values per gene are centered by the mean log2 Cy5/Cy3 normalized ratio across the arrays. Increasing blue intensity in the heat map corresponds to higher relative transcript levels compared to the mean expression level across all 34 array experiments. Increasing yellow intensity corresponds to relatively decreased transcript levels compared to the mean. Dark grey corresponds to missing data. Examples taken from these gene clusters are marked by color within the gene tree on the left of the heat map. The red gene tree corresponds to the enlarged gene cluster D, the blue gene tree corresponds to F, the pink gene tree corresponds to G, and the green gene tree corresponds to E, respectively. Gene symbols of the named transcripts are shown on the right. (B) Enlarged cluster dendrogram of the PBMC samples demonstrating the degree of relatedness (Pearson correlation) between the expression patterns of the 157 transcripts. The length of the arms of the dendrogram reflects the degree of correlation between experiments. Samples are color coded to reflect the degree of external genital virilization according to Prader that had been applied to both XX and XY individuals to enable comparability of genital phenotypes independent of the sex chromosomes. The dark grey bar below the experiment cluster represents individuals with an XX karyotype, light grey represents an XY karyotype. The major subdivison of the individuals corresponds strictly to the karyotype. The second level of subdivision in each of the major arms reflects mostly the phenotype of external genital virilization independent of the karyotype. (C) Schematic representation of the Prader stages of external genital virilization. (D) X-chromosome gene cluster (E) Y-chromosome gene cluster (F) Genes with predominantly higher transcript levels in the phenotypic females and the normal females independent of the karyotype (G) Genes with higher transcript levels in the strongly virilized individuals and the normal males independent of the karyotype.

Mentions: Transcript profiles for 9 normal male and 10 normal female controls were measured using spotted cDNA microarrays with more than 44,000 cDNA elements representing 26,000 unique genes. The Significance Analysis of Microarrays (SAM) procedure [20] was used to identify 157 sex-specific transcripts (121 unique named genes) (false discovery rate < 0.09): 136 transcripts with higher expression in males (86.6%) and 21 (13.4%) with higher expression in females. Gene expression across these 157 sex-dimorphic transcripts was analyzed across all 19 controls and 14 DSD-patients. The DSD patients were comprised of ten 46,XY subjects whose genitalia ranged from normal female to Prader 4, three 46,XX subjects with masculinized genitalia from high prenatal androgenic steroid levels due to congenital adrenal hyperplasia (CAH, 21-hydroxlase deficiency) and one Prader 4 45,X0/46,XY subject. In addition, one normally masculinized 46,XY male with CAH was investigated (Additional file 1 and Figure 1A, B, C, additional files 2, 3, 4, 5).


Disorders of sex development expose transcriptional autonomy of genetic sex and androgen-programmed hormonal sex in human blood leukocytes.

Holterhus PM, Bebermeier JH, Werner R, Demeter J, Richter-Unruh A, Cario G, Appari M, Siebert R, Riepe F, Brooks JD, Hiort O - BMC Genomics (2009)

Hierarchical clustering analysis of 34 microarray experiments on PBMC total RNA derived from 9 normal males, 10 normal females, one 46,XY-male with CAH and 14 individuals with DSD [see additional files 2, 3, 4, 5]. (A) Transcript levels of 157 transcripts identified by SAM as differing significantly between PBMC from 9 normal males and 10 normal females. Transcripts are grouped by hierarchical cluster analysis and displayed in rows while experiments are displayed in columns. Expression values per gene are centered by the mean log2 Cy5/Cy3 normalized ratio across the arrays. Increasing blue intensity in the heat map corresponds to higher relative transcript levels compared to the mean expression level across all 34 array experiments. Increasing yellow intensity corresponds to relatively decreased transcript levels compared to the mean. Dark grey corresponds to missing data. Examples taken from these gene clusters are marked by color within the gene tree on the left of the heat map. The red gene tree corresponds to the enlarged gene cluster D, the blue gene tree corresponds to F, the pink gene tree corresponds to G, and the green gene tree corresponds to E, respectively. Gene symbols of the named transcripts are shown on the right. (B) Enlarged cluster dendrogram of the PBMC samples demonstrating the degree of relatedness (Pearson correlation) between the expression patterns of the 157 transcripts. The length of the arms of the dendrogram reflects the degree of correlation between experiments. Samples are color coded to reflect the degree of external genital virilization according to Prader that had been applied to both XX and XY individuals to enable comparability of genital phenotypes independent of the sex chromosomes. The dark grey bar below the experiment cluster represents individuals with an XX karyotype, light grey represents an XY karyotype. The major subdivison of the individuals corresponds strictly to the karyotype. The second level of subdivision in each of the major arms reflects mostly the phenotype of external genital virilization independent of the karyotype. (C) Schematic representation of the Prader stages of external genital virilization. (D) X-chromosome gene cluster (E) Y-chromosome gene cluster (F) Genes with predominantly higher transcript levels in the phenotypic females and the normal females independent of the karyotype (G) Genes with higher transcript levels in the strongly virilized individuals and the normal males independent of the karyotype.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC2713997&req=5

Figure 1: Hierarchical clustering analysis of 34 microarray experiments on PBMC total RNA derived from 9 normal males, 10 normal females, one 46,XY-male with CAH and 14 individuals with DSD [see additional files 2, 3, 4, 5]. (A) Transcript levels of 157 transcripts identified by SAM as differing significantly between PBMC from 9 normal males and 10 normal females. Transcripts are grouped by hierarchical cluster analysis and displayed in rows while experiments are displayed in columns. Expression values per gene are centered by the mean log2 Cy5/Cy3 normalized ratio across the arrays. Increasing blue intensity in the heat map corresponds to higher relative transcript levels compared to the mean expression level across all 34 array experiments. Increasing yellow intensity corresponds to relatively decreased transcript levels compared to the mean. Dark grey corresponds to missing data. Examples taken from these gene clusters are marked by color within the gene tree on the left of the heat map. The red gene tree corresponds to the enlarged gene cluster D, the blue gene tree corresponds to F, the pink gene tree corresponds to G, and the green gene tree corresponds to E, respectively. Gene symbols of the named transcripts are shown on the right. (B) Enlarged cluster dendrogram of the PBMC samples demonstrating the degree of relatedness (Pearson correlation) between the expression patterns of the 157 transcripts. The length of the arms of the dendrogram reflects the degree of correlation between experiments. Samples are color coded to reflect the degree of external genital virilization according to Prader that had been applied to both XX and XY individuals to enable comparability of genital phenotypes independent of the sex chromosomes. The dark grey bar below the experiment cluster represents individuals with an XX karyotype, light grey represents an XY karyotype. The major subdivison of the individuals corresponds strictly to the karyotype. The second level of subdivision in each of the major arms reflects mostly the phenotype of external genital virilization independent of the karyotype. (C) Schematic representation of the Prader stages of external genital virilization. (D) X-chromosome gene cluster (E) Y-chromosome gene cluster (F) Genes with predominantly higher transcript levels in the phenotypic females and the normal females independent of the karyotype (G) Genes with higher transcript levels in the strongly virilized individuals and the normal males independent of the karyotype.
Mentions: Transcript profiles for 9 normal male and 10 normal female controls were measured using spotted cDNA microarrays with more than 44,000 cDNA elements representing 26,000 unique genes. The Significance Analysis of Microarrays (SAM) procedure [20] was used to identify 157 sex-specific transcripts (121 unique named genes) (false discovery rate < 0.09): 136 transcripts with higher expression in males (86.6%) and 21 (13.4%) with higher expression in females. Gene expression across these 157 sex-dimorphic transcripts was analyzed across all 19 controls and 14 DSD-patients. The DSD patients were comprised of ten 46,XY subjects whose genitalia ranged from normal female to Prader 4, three 46,XX subjects with masculinized genitalia from high prenatal androgenic steroid levels due to congenital adrenal hyperplasia (CAH, 21-hydroxlase deficiency) and one Prader 4 45,X0/46,XY subject. In addition, one normally masculinized 46,XY male with CAH was investigated (Additional file 1 and Figure 1A, B, C, additional files 2, 3, 4, 5).

Bottom Line: Gender appears to be determined by independent programs controlled by the sex-chromosomes and by androgen-dependent programming during embryonic development.However, a significantly larger gene set in the PBMC only reflected the degree of external genital masculinization independent of the sex chromosomes and independent of concurrent post-natal sex steroid hormone levels.A significant fraction of gene expression differences between males and females in the human appears to have its roots in early embryogenesis and is not only caused by sex chromosomes but also by long-term sex-specific hormonal programming due to presence or absence of androgen during the time of external genital masculinization.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pediatrics, Christian Albrechts University of Kiel, Germany. holterhus@pediatrics.uni-kiel.de

ABSTRACT

Background: Gender appears to be determined by independent programs controlled by the sex-chromosomes and by androgen-dependent programming during embryonic development. To enable experimental dissection of these components in the human, we performed genome-wide profiling of the transcriptomes of peripheral blood mononuclear cells (PBMC) in patients with rare defined "disorders of sex development" (DSD, e.g., 46, XY-females due to defective androgen biosynthesis) compared to normal 46, XY-males and 46, XX-females.

Results: A discrete set of transcripts was directly correlated with XY or XX genotypes in all individuals independent of male or female phenotype of the external genitalia. However, a significantly larger gene set in the PBMC only reflected the degree of external genital masculinization independent of the sex chromosomes and independent of concurrent post-natal sex steroid hormone levels. Consequently, the architecture of the transcriptional PBMC-"sexes" was either male, female or even "intersex" with a discordant alignment of the DSD individuals' genetic and hormonal sex signatures.

Conclusion: A significant fraction of gene expression differences between males and females in the human appears to have its roots in early embryogenesis and is not only caused by sex chromosomes but also by long-term sex-specific hormonal programming due to presence or absence of androgen during the time of external genital masculinization. Genetic sex and the androgen milieu during embryonic development might therefore independently modulate functional traits, phenotype and diseases associated with male or female gender as well as with DSD conditions.

Show MeSH
Related in: MedlinePlus