Deletion Porr Filmer - Deletion Sex

Deletion Porr Filmer - Deletion Sex

Dashboard
Publications
Account settings
Log out

Journal List

Eur J Hum Genet

v.20(3); 2012 Mar

PMC3283189

Eur J Hum Genet. 2012 Mar; 20(3): 348–351.
Published online 2011 Nov 9. doi: 10.1038/ejhg.2011.204
Find articles by Leendert Looijenga
1 Murdoch Childrens Research Institute, Royal Children's Hospital, Melbourne, Victoria, Australia
2 Monash Institute of Medical Research, Melbourne, Victoria, Australia
3 University of Melbourne, Melbourne, Victoria, Australia
4 Erasmus MC-University Medical Center, Rotterdam, The Netherlands
5 Institute for Molecular Bioscience, The University of Queensland, Brisbane, Queensland, Australia
6 Prince Henry's Institute, Melbourne, Victoria, Australia
7 University Hospital Ghent, Ghent, Belgium
* Monash Institute of Medical Research, 27-31 Wright Street, Clayton Victoria 3168, Australia. Tel +61 3 9902 4812; Fax +61 3 9594 7114; E-mail: ude.hsanom@etihw.nafets
Received 2011 May 12; Revised 2011 Sep 19; Accepted 2011 Sep 29.
Copyright © 2012 Macmillan Publishers Limited
This article has been cited by other articles in PMC.
Keywords: disorders of sex development, copy number, WWOX, gonad, microarrays
Wilhelm D, Palmer S, Koopman P. Sex determination and gonadal development in mammals. Physiol Rev. 2007; 87 :1–28. [ PubMed ] [ Google Scholar ] Lee PA, Houk CP, Ahmed SF, Hughes IA. Consensus statement on management of intersex disorders. International Consensus Conference on Intersex. Pediatrics. 2006; 118 :e488–e500. [ PubMed ] [ Google Scholar ] White SJ, Ohnesorg T, Notini A, et al. Copy number variation in patients with disorders of sex development due to 46,XY gonadal dysgenesis. PLoS One. 2011; 6 :e17793. [ PMC free article ] [ PubMed ] [ Google Scholar ] Sutton E, Hughes J, White SJ, et al. Identification of SOX3 as an XX male sex reversal gene in mice and humans. J Clin Invest. 2011; 121 :328–341. [ PMC free article ] [ PubMed ] [ Google Scholar ] White SJ, Breuning MH, den Dunnen JT. Detecting copy number changes in genomic DNA: MAPH and MLPA. Methods Cell Biol. 2004; 75 :751–768. [ PubMed ] [ Google Scholar ] Hersmus R, Kalfa N, de Leeuw B, et al. FOXL2 and SOX9 as parameters of female and male gonadal differentiation in patients with various forms of disorders of sex development (DSD) J Pathol. 2008; 215 :31–38. [ PubMed ] [ Google Scholar ] Looijenga LH, Stoop H, de Leeuw HP, et al. POU5F1 (OCT3/4) identifies cells with pluripotent potential in human germ cell tumors. Cancer Res. 2003; 63 :2244–2250. [ PubMed ] [ Google Scholar ] Stoop H, Honecker F, van de Geijn GJ, et al. Stem cell factor as a novel diagnostic marker for early malignant germ cells. J Pathol. 2008; 216 :43–54. [ PubMed ] [ Google Scholar ] Cools M, Wolffenbuttel KP, Drop SL, Oosterhuis JW, Looijenga LH. Gonadal development and tumor formation at the crossroads of male and female sex determination. Sex Dev. 2011; 5 :167–180. [ PubMed ] [ Google Scholar ] Cools M, Drop SL, Wolffenbuttel KP, Oosterhuis JW, Looijenga LH. Germ cell tumors in the intersex gonad: old paths, new directions, moving frontiers. Endocr Rev. 2006; 27 :468–484. [ PubMed ] [ Google Scholar ] Notini AJ, Craig JM, White SJ. Copy number variation and mosaicism. Cytogenet Genome Res. 2008; 123 :270–277. [ PubMed ] [ Google Scholar ] Paige AJ, Taylor KJ, Taylor C, et al. WWOX: a candidate tumor suppressor gene involved in multiple tumor types. Proc Natl Acad Sci USA. 2001; 98 :11417–11422. [ PMC free article ] [ PubMed ] [ Google Scholar ] Aqeilan RI, Trapasso F, Hussain S, et al. Targeted deletion of Wwox reveals a tumor suppressor function. Proc Natl Acad Sci USA. 2007; 104 :3949–3954. [ PMC free article ] [ PubMed ] [ Google Scholar ] Bednarek AK, Laflin KJ, Daniel RL, Liao Q, Hawkins KA, Aldaz CM. WWOX, a novel WW domain-containing protein mapping to human chromosome 16q23.3–24.1, a region frequently affected in breast cancer. Cancer Res. 2000; 60 :2140–2145. [ PubMed ] [ Google Scholar ] Del Mare S, Salah Z, Aqeilan RI. WWOX: its genomics, partners, and functions. J Cell Biochem. 2009; 108 :737–745. [ PubMed ] [ Google Scholar ] Aqeilan RI, Palamarchuk A, Weigel RJ, Herrero JJ, Pekarsky Y, Croce CM. Physical and functional interactions between the Wwox tumor suppressor protein and the AP-2gamma transcription factor. Cancer Res. 2004; 64 :8256–8261. [ PubMed ] [ Google Scholar ] Aqeilan RI, Pekarsky Y, Herrero JJ, et al. Functional association between Wwox tumor suppressor protein and p73, a p53 homolog. Proc Natl Acad Sci USA. 2004; 101 :4401–4406. [ PMC free article ] [ PubMed ] [ Google Scholar ] Aqeilan RI, HaganJP, de Bruin A, et al. Targeted ablation of the WW domain-containing oxidoreductase tumor suppressor leads to impaired steroidogenesis. Endocrinology. 2009; 150 :1530–1535. [ PMC free article ] [ PubMed ] [ Google Scholar ] Ludes-Meyers JH, Kil H, Nunez MI, et al. WWOX hypomorphic mice display a higher incidence of B-cell lymphomas and develop testicular atrophy. Genes Chromosomes Cancer. 2007; 46 :1129–1136. [ PMC free article ] [ PubMed ] [ Google Scholar ] Bouteille N, Driouch K, Hage PE, et al. Inhibition of the Wnt/beta-catenin pathway by the WWOX tumor suppressor protein. Oncogene. 2009; 28 :2569–2580. [ PubMed ] [ Google Scholar ] Maatouk DM, DiNapoli L, Alvers A, Parker KL, Taketo MM, Capel B. Stabilization of beta-catenin in XY gonads causes male-to-female sex-reversal. Hum Mol Genet. 2008; 17 :2949–2955. [ PMC free article ] [ PubMed ] [ Google Scholar ] Nunez MI, Ludes-Meyers J, Aldaz CM. WWOX protein expression in normal human tissues. J Mol Histol. 2006; 37 :115–125. [ PMC free article ] [ PubMed ] [ Google Scholar ]
Articles from European Journal of Human Genetics are provided here courtesy of Nature Publishing Group
Wilhelm D, Palmer S, Koopman P. Sex determination and gonadal development in mammals. Physiol Rev. 2007; 87 :1–28. [ PubMed ] [ Google Scholar ] [ Ref list ]
Lee PA, Houk CP, Ahmed SF, Hughes IA. Consensus statement on management of intersex disorders. International Consensus Conference on Intersex. Pediatrics. 2006; 118 :e488–e500. [ PubMed ] [ Google Scholar ] [ Ref list ]
White SJ, Ohnesorg T, Notini A, et al. Copy number variation in patients with disorders of sex development due to 46,XY gonadal dysgenesis. PLoS One. 2011; 6 :e17793. [ PMC free article ] [ PubMed ] [ Google Scholar ] [ Ref list ]
Sutton E, Hughes J, White SJ, et al. Identification of SOX3 as an XX male sex reversal gene in mice and humans. J Clin Invest. 2011; 121 :328–341. [ PMC free article ] [ PubMed ] [ Google Scholar ] [ Ref list ]
White SJ, Breuning MH, den Dunnen JT. Detecting copy number changes in genomic DNA: MAPH and MLPA. Methods Cell Biol. 2004; 75 :751–768. [ PubMed ] [ Google Scholar ] [ Ref list ]
Hersmus R, Kalfa N, de Leeuw B, et al. FOXL2 and SOX9 as parameters of female and male gonadal differentiation in patients with various forms of disorders of sex development (DSD) J Pathol. 2008; 215 :31–38. [ PubMed ] [ Google Scholar ] [ Ref list ]
Looijenga LH, Stoop H, de Leeuw HP, et al. POU5F1 (OCT3/4) identifies cells with pluripotent potential in human germ cell tumors. Cancer Res. 2003; 63 :2244–2250. [ PubMed ] [ Google Scholar ] [ Ref list ]
Stoop H, Honecker F, van de Geijn GJ, et al. Stem cell factor as a novel diagnostic marker for early malignant germ cells. J Pathol. 2008; 216 :43–54. [ PubMed ] [ Google Scholar ] [ Ref list ]
Cools M, Wolffenbuttel KP, Drop SL, Oosterhuis JW, Looijenga LH. Gonadal development and tumor formation at the crossroads of male and female sex determination. Sex Dev. 2011; 5 :167–180. [ PubMed ] [ Google Scholar ] [ Ref list ]
Cools M, Drop SL, Wolffenbuttel KP, Oosterhuis JW, Looijenga LH. Germ cell tumors in the intersex gonad: old paths, new directions, moving frontiers. Endocr Rev. 2006; 27 :468–484. [ PubMed ] [ Google Scholar ] [ Ref list ]
Notini AJ, Craig JM, White SJ. Copy number variation and mosaicism. Cytogenet Genome Res. 2008; 123 :270–277. [ PubMed ] [ Google Scholar ] [ Ref list ]
Paige AJ, Taylor KJ, Taylor C, et al. WWOX: a candidate tumor suppressor gene involved in multiple tumor types. Proc Natl Acad Sci USA. 2001; 98 :11417–11422. [ PMC free article ] [ PubMed ] [ Google Scholar ] [ Ref list ]
Aqeilan RI, Trapasso F, Hussain S, et al. Targeted deletion of Wwox reveals a tumor suppressor function. Proc Natl Acad Sci USA. 2007; 104 :3949–3954. [ PMC free article ] [ PubMed ] [ Google Scholar ] [ Ref list ]
Bednarek AK, Laflin KJ, Daniel RL, Liao Q, Hawkins KA, Aldaz CM. WWOX, a novel WW domain-containing protein mapping to human chromosome 16q23.3–24.1, a region frequently affected in breast cancer. Cancer Res. 2000; 60 :2140–2145. [ PubMed ] [ Google Scholar ] [ Ref list ]
Del Mare S, Salah Z, Aqeilan RI. WWOX: its genomics, partners, and functions. J Cell Biochem. 2009; 108 :737–745. [ PubMed ] [ Google Scholar ] [ Ref list ]
Aqeilan RI, Palamarchuk A, Weigel RJ, Herrero JJ, Pekarsky Y, Croce CM. Physical and functional interactions between the Wwox tumor suppressor protein and the AP-2gamma transcription factor. Cancer Res. 2004; 64 :8256–8261. [ PubMed ] [ Google Scholar ] [ Ref list ]
Aqeilan RI, Pekarsky Y, Herrero JJ, et al. Functional association between Wwox tumor suppressor protein and p73, a p53 homolog. Proc Natl Acad Sci USA. 2004; 101 :4401–4406. [ PMC free article ] [ PubMed ] [ Google Scholar ] [ Ref list ]
Aqeilan RI, HaganJP, de Bruin A, et al. Targeted ablation of the WW domain-containing oxidoreductase tumor suppressor leads to impaired steroidogenesis. Endocrinology. 2009; 150 :1530–1535. [ PMC free article ] [ PubMed ] [ Google Scholar ] [ Ref list ]
Ludes-Meyers JH, Kil H, Nunez MI, et al. WWOX hypomorphic mice display a higher incidence of B-cell lymphomas and develop testicular atrophy. Genes Chromosomes Cancer. 2007; 46 :1129–1136. [ PMC free article ] [ PubMed ] [ Google Scholar ] [ Ref list ]
Bouteille N, Driouch K, Hage PE, et al. Inhibition of the Wnt/beta-catenin pathway by the WWOX tumor suppressor protein. Oncogene. 2009; 28 :2569–2580. [ PubMed ] [ Google Scholar ] [ Ref list ]
Maatouk DM, DiNapoli L, Alvers A, Parker KL, Taketo MM, Capel B. Stabilization of beta-catenin in XY gonads causes male-to-female sex-reversal. Hum Mol Genet. 2008; 17 :2949–2955. [ PMC free article ] [ PubMed ] [ Google Scholar ] [ Ref list ]
Nunez MI, Ludes-Meyers J, Aldaz CM. WWOX protein expression in normal human tissues. J Mol Histol. 2006; 37 :115–125. [ PMC free article ] [ PubMed ] [ Google Scholar ] [ Ref list ]

Format:

AMA

APA

MLA

NLM

NLM

NIH

HHS

USA.gov

External link. Please review our privacy policy .
The new PMC design is here!
Learn more
about navigating our updated article layout. The
PMC legacy view
will also be available for a limited time.

An official website of the United States government

The .gov means it’s official.

Federal government websites often end in .gov or .mil. Before
sharing sensitive information, make sure you’re on a federal
government site.

The site is secure.

The https:// ensures that you are connecting to the
official website and that any information you provide is encrypted
and transmitted securely.

1 Murdoch Childrens Research Institute, Royal Children's Hospital, Melbourne, Victoria, Australia
2 Monash Institute of Medical Research, Melbourne, Victoria, Australia
1 Murdoch Childrens Research Institute, Royal Children's Hospital, Melbourne, Victoria, Australia
3 University of Melbourne, Melbourne, Victoria, Australia
1 Murdoch Childrens Research Institute, Royal Children's Hospital, Melbourne, Victoria, Australia
4 Erasmus MC-University Medical Center, Rotterdam, The Netherlands
4 Erasmus MC-University Medical Center, Rotterdam, The Netherlands
4 Erasmus MC-University Medical Center, Rotterdam, The Netherlands
5 Institute for Molecular Bioscience, The University of Queensland, Brisbane, Queensland, Australia
6 Prince Henry's Institute, Melbourne, Victoria, Australia
7 University Hospital Ghent, Ghent, Belgium
4 Erasmus MC-University Medical Center, Rotterdam, The Netherlands
1 Murdoch Childrens Research Institute, Royal Children's Hospital, Melbourne, Victoria, Australia
3 University of Melbourne, Melbourne, Victoria, Australia
Disorders of sex development (DSD) are congenital conditions where chromosomal, gonad or genital development is atypical. In a significant proportion of 46,XY DSD cases it is not possible to identify a causative mutation, making genetic counseling difficult and potentially hindering optimal treatment. Here, we describe the analysis of a 46,XY DSD patient that presented at birth with ambiguous genitalia. Histological analysis of the surgically removed gonads showed bilateral undifferentiated gonadal tissue and immature testis, both containing malignant germ cells. We screened genomic DNA from this patient for deletions and duplications using an Illumina whole-genome SNP microarray. This analysis revealed a heterozygous deletion within the WWOX gene on chromosome 16, removing exons 6–8. Analysis of parental DNA showed that the deletion was inherited from the mother. cDNA analysis confirmed that the deletion maintained the reading frame, with exon 5 being spliced directly onto exon 9. This deletion is the first description of a germline rearrangement affecting the coding sequence of WWOX in humans. Previously described Wwox knockout mouse models showed gonadal abnormalities, supporting a role for WWOX in human gonad development.
In the early stages of human embryogenesis the developing gonads are bipotent, being capable of forming either testes or ovaries. In males the expression of the Y chromosomal SRY gene initiates testis development, whereas ovarian development in principle occurs only in the absence of SRY (reviewed in Wilhelm et al 1 ). Following the establishment of sex-specific expression of key regulatory genes in the gonad, gonadal differentiation results in development of the external genitalia. As a result there are two main stages in gonad formation, and disruption of either can lead to disorders of sex development (DSD). DSD are surprisingly common, with ambiguous genitalia estimated to occur with an incidence of 1 in 4500 live births. 2
A number of genes important in the regulation of sex determination have been identified, yet in as many as 70% of 46,XY DSD cases no genetic cause has been identified. We have previously demonstrated the power of whole-genome copy number analysis with high-density microarrays to identify causative mutations in DSD. 3 , 4 Here, we describe the use of this approach to identify a multi-exon heterozygous deletion in the WWOX gene of a 46,XY DSD patient.
Genomic DNA was hybridized onto an Illumina 610-Quad microarray at the Australian Genome Research Facility (Melbourne, Australia) following the manufacturer's instructions. Data were analyzed using Genome Studio data analysis software (Illumina).
Deletion screening of the WWOX gene was performed with MLPA. Probe design, the MLPA reaction and data analysis were performed as described previously. 5
RNA was extracted from lymphocytes obtained from the index case using standard procedures, with cDNA generated using random hexamers and the Transcriptor High Fidelity cDNA synthesis kit (Roche, Mannheim, Germany) according to the manufacturer's instructions. The PCR amplification across the deletion used the following primers; F-5′-CGAAACCGCCAAGTCTTTT-3′, R-5′-CGTCTCTTCGCTCTGAGCTT-3′, and was run under the following conditions:
1 cycle: 60 s 95°C; 35 cycles: 30 s 95°C; 30 s 58°C; 60 s 72°C; 1 cycle: 20 min 72°C.
Sanger sequencing was conducted at the Department of Pathology, University of Melbourne, Australia.
Research on human tissue samples was performed according to the Code for Proper Secondary Use of Human Tissue in the Netherlands, as developed by the Dutch Federation of Medical Scientific Societies (FMWV) version 2002 and approved by an Institutional Review Board (MEC 02.981). Immunohistochemical detection of formalin-fixed paraffin-embedded tissue was performed for SOX9 and FOXL2, 6 OCT3/4 7 and KITL, 8 as described previously.
Physical examination of the index patient at the age of 10 days revealed unfused labioscrotal folds, impalpable gonads, clitoral hypertrophy 20 mm in size and a perineal urogenital sinus. Genitography demonstrated the presence of a vagina and underdeveloped uterus. Chromosomal analysis showed a 46,XY karyotype with no visible aberrations. Sequence analysis of the SRY and NR5A1 genes did not reveal any variants, and MLPA analysis with a commercially available kit (MLPA P185-B1) containing probes targeted at the WNT4, SRY, NR0B1, SOX9 and NR5A1 genes did not show any deletions or duplications.
Small dysgenic gonads were present in the abdomen, and pathological analysis following complete removal at 2 years of age showed that they contained edematous infantile testicular parenchyma ( Figure 1 ). The epididymis was completely separated from the rete testis on both sides, and tubular epithelium was identified at the left side. The left gonad consisted of centrally located edematous testicular tissue, positive for immunohistochemical detection of SOX9 (indicative for Sertoli cells) and negative for FOXL2 (a granulosa cell marker). A gradual transition toward undifferentiated gonadal tissue, containing both SOX9 and FOXL2-positive cells at the upper and lower poles were identified. Undifferentiated gonadal tissue is a gonadal pattern found specifically in patients with gonadal dysgenesis and is typically characterized by the combined expression of FOXL2 and SOX9 (usually with a preponderance of FOXL2), suggesting limited differentiation of the supportive cell lineage into pre-granulosa and pre-Sertoli cells 9 . Immature OCT3/4-positive germ cells were also found, either dispersed in ovarian-like stroma or organized together with Sertoli/granulosa cells in cord-like structures, reminiscent of sex cords. These structures have been recognized as the precursor lesion for gonadoblastoma 10 .
Representative histological and immunohistochemical findings for the left gonad: ( a ) total overview of the gonad histology (H&E); ( b ) higher magnification ( × 2.5 magnification, indicated by square in ( a ); immunohistochemical detection of ( c ) SOX9, positive in the Sertoli cells; ( d ) FOXL2, positive in the granulosa cells; ( e ) OCT3/4; ( f ) TSPY; ( g ) KITL, all positive in the transformed germ cells. All immunohistochemical images are
Tara Pink Porr Filmer - Tara Pink Sex
Elektriker Porr Filmer - Elektriker Sex
Läsa Porr Filmer - Läsa Sex

Deletion Porr Filmer - Deletion Sex

Report Page