Intrinsic androgen-dependent gene expression patterns revealed by comparison of genital fibroblasts from normal males and individuals with complete and partial androgen insensitivity syndrome

<p>Abstract</p> <p>Background</p> <p>To better understand the molecular programs of normal and abnormal genital development, clear-cut definition of androgen-dependent gene expression patterns, without the influence of genotype (46, XX vs. 46, XY), is warranted. Previously, we have identified global gene expression profiles in genital-derived fibroblasts that differ between 46, XY males and 46, XY females with complete androgen insensitivity syndrome (CAIS) due to inactivating mutations of the androgen receptor (AR). While these differences could be due to cell autonomous changes in gene expression induced by androgen programming, recent work suggests they could also be influenced by the location from which the fibroblasts were harvested (topology). To minimize the influence of topology, we compared gene expression patterns of fibroblasts derived from identical urogenital anlagen: the scrotum in normally virilized 46, XY males and the labia majora from completely feminized 46, XY individuals with CAIS.</p> <p>Results</p> <p>612 transcripts representing 440 unique genes differed significantly in expression levels between scrotum and CAIS labia majora, suggesting the effects of androgen programming. While some genes coincided with those we had identified previously (TBX3, IGFBP5, EGFR, CSPG2), a significant number did not, implying that topology had influenced gene expression in our previous experiments. Supervised clustering of gene expression data derived from a large set of fibroblast cultures from individuals with partial AIS revealed that the new, topology controlled data set better classified the specimens.</p> <p>Conclusion</p> <p>Inactivating mutations of the AR, in themselves, appear to induce lasting changes in gene expression in cultured fibroblasts, independent of topology and genotype. Genes identified are likely to be relevant candidates to decipher androgen-dependent normal and abnormal genital development.</p

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

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

Intrinsic androgen-dependent gene expression patterns revealed by comparison of genital fibroblasts from normal males and individuals with complete and partial androgen insensitivity syndrome-1

<p><b>Copyright information:</b></p><p>Taken from "Intrinsic androgen-dependent gene expression patterns revealed by comparison of genital fibroblasts from normal males and individuals with complete and partial androgen insensitivity syndrome"</p><p>http://www.biomedcentral.com/1471-2164/8/376</p><p>BMC Genomics 2007;8():376-376.</p><p>Published online 18 Oct 2007</p><p>PMCID:PMC2212662.</p><p></p>interpretable data across the experiments whose expression levels were at least 2-fold different from the mean expression across all samples in at least 5 microarrays. (A) The cluster dendrogram demonstrating the degree of relatedness (Pearson correlation) between the expression patterns of the 259 genes in the cultured fibroblast samples. The length of the arms of the dendrogram reflects the degree of correlation between the samples. Samples are color coded to reflect the localization of the biopsy and the degree of external genital virilization according to a grading scheme developed by Sinnecker et al. [10]. The grey bar below indicates whether a sample was derived from dataset 1, 2 or 3. ''L.n.d.'' signifies that the biopsy localization was not accurately documented. Italics indicate a sample with a 46, XX karyotype. (B) Schematic depiction of the external genitalia phenotype of the cases from which the fibroblast cultures were derived using color coding that corresponds to the degree of genital ambiguity and the location of the biopsy. Color coding corresponds to the bar below the dendrogram in (A). (C) Cluster of AR-dependent transcripts that are highly expressed in the left ''male'' major branch of the cluster that are expressed at significantly lower levels in the ''female'' branch of the cluster on the right. TBX3, previously reported in ulnar mammary syndrome and IGF2, previously reported as being down-regulated in CAIS [20] are shown in this cluster. (D) Cluster of AR-dependent transcripts that are expressed at significantly lower levels in the lefthand ''male'' branch of the cluster and at higher levels in the righthand ''female'' branch. These include many extracellular matrix genes such as proteoglycan testican, versican, and fibrillin 1. Detailed data on figure 2 is available in additional files , , ,

Intrinsic androgen-dependent gene expression patterns revealed by comparison of genital fibroblasts from normal males and individuals with complete and partial androgen insensitivity syndrome-3

<p><b>Copyright information:</b></p><p>Taken from "Intrinsic androgen-dependent gene expression patterns revealed by comparison of genital fibroblasts from normal males and individuals with complete and partial androgen insensitivity syndrome"</p><p>http://www.biomedcentral.com/1471-2164/8/376</p><p>BMC Genomics 2007;8():376-376.</p><p>Published online 18 Oct 2007</p><p>PMCID:PMC2212662.</p><p></p>normal scrotal cell lines. Semi-quantitative RT-PCR was performed on four samples of normal male scrotal fibroblasts (S4, S5, S8, S9), as well as on four samples of labia majora derived from CAIS patients (ARD402, ARD411, ARD682, ARD1097). Differences in expression levels between different cell lines were calculated according to the ΔΔ-CT method [19]. The y-axis represents the ratios of expression levels of CAIS labia majora fibroblasts divided by those from scrotal fibroblasts

Intrinsic androgen-dependent gene expression patterns revealed by comparison of genital fibroblasts from normal males and individuals with complete and partial androgen insensitivity syndrome-2

<p><b>Copyright information:</b></p><p>Taken from "Intrinsic androgen-dependent gene expression patterns revealed by comparison of genital fibroblasts from normal males and individuals with complete and partial androgen insensitivity syndrome"</p><p>http://www.biomedcentral.com/1471-2164/8/376</p><p>BMC Genomics 2007;8():376-376.</p><p>Published online 18 Oct 2007</p><p>PMCID:PMC2212662.</p><p></p>on (B). In each of the two clusters A and B, the microarray experiments used to generate the underlying list of significant genes by SAM were removed before clustering resulting 57 remaining experiments in (A) and 58 in (B), respectively. The color code is the same as in Fig. 2. Based on the previous gene set, several highly virilized AIS 2 patients and normal scrotal fibroblasts were incorrectly classified female

Intrinsic androgen-dependent gene expression patterns revealed by comparison of genital fibroblasts from normal males and individuals with complete and partial androgen insensitivity syndrome-4

<p><b>Copyright information:</b></p><p>Taken from "Intrinsic androgen-dependent gene expression patterns revealed by comparison of genital fibroblasts from normal males and individuals with complete and partial androgen insensitivity syndrome"</p><p>http://www.biomedcentral.com/1471-2164/8/376</p><p>BMC Genomics 2007;8():376-376.</p><p>Published online 18 Oct 2007</p><p>PMCID:PMC2212662.</p><p></p>ual transcripts are grouped by hierarchical cluster analysis and are displayed in rows while experiments are represented in columns. Expression values per gene are centered by the mean logred/green normalized ratio. Increasing red intensity corresponds to higher relative transcript levels compared to the mean expression level across all 15 array experiments. Increasing green intensity corresponds to relatively decreased transcript levels compared to the mean. On the right side, examples of individual genes (gene symbols according to S.O.U.R.C.E. []) discussed in the paper or falling into the biological processes and cellular pathways detected by PANTHER are displayed. Detailed data on figure 1 is available in additional files , , ,

Intrinsic androgen-dependent gene expression patterns revealed by comparison of genital fibroblasts from normal males and individuals with complete and partial androgen insensitivity syndrome-0

<p><b>Copyright information:</b></p><p>Taken from "Intrinsic androgen-dependent gene expression patterns revealed by comparison of genital fibroblasts from normal males and individuals with complete and partial androgen insensitivity syndrome"</p><p>http://www.biomedcentral.com/1471-2164/8/376</p><p>BMC Genomics 2007;8():376-376.</p><p>Published online 18 Oct 2007</p><p>PMCID:PMC2212662.</p><p></p>ual transcripts are grouped by hierarchical cluster analysis and are displayed in rows while experiments are represented in columns. Expression values per gene are centered by the mean logred/green normalized ratio. Increasing red intensity corresponds to higher relative transcript levels compared to the mean expression level across all 15 array experiments. Increasing green intensity corresponds to relatively decreased transcript levels compared to the mean. On the right side, examples of individual genes (gene symbols according to S.O.U.R.C.E. []) discussed in the paper or falling into the biological processes and cellular pathways detected by PANTHER are displayed. Detailed data on figure 1 is available in additional files , , ,