Collagen Content in Skin and Internal Organs of the Tight Skin Mouse: An Animal Model of Scleroderma.

The Tight Skin mouse is a genetically induced animal model of tissue fibrosis caused by a large in-frame mutation in the gene encoding fibrillin-1 (Fbn-1). We examined the influence of gender on the collagen content of tissues in C57BL/6J wild type (+/+) and mutant Tight Skin (Tsk/+) mice employing hydroxyproline assays. Tissue sections were stained with Masson\u27s trichrome to identify collagen in situ. Adult Tsk/+ mice skin contains ~15% more collagen, on average, than skin from +/+ mice of the same gender. The heart of Tsk/+ males had significantly more collagen than that of +/+ males. No significant gender differences were found in lungs and kidney collagen content. Overall, the collagen content of Tsk/+ males and +/+ males was higher than that of their Tsk/+ and +/+ female counterparts, respectively. Our data confirm increased deposition of collagen in skin and hearts of Tsk/+ mice; however, the effects of the Tsk mutation on collagen content are both tissue specific and gender specific. These results indicate that comparative studies of collagen content between normal and Tsk/+ mice skin and internal organs must take into account gender differences caused by expression of the androgen receptor

A tandem duplication within the fibrillin 1 gene is associated with the mouse tight skin mutation.

Mice carrying the Tight skin (Tsk) mutation have thickened skin and visceral fibrosis resulting from an accumulation of extracellular matrix molecules. These and other connective tissue abnormalities have made Tskl + mice models for scleroderma, hereditary emphysema, and myocardial hypertrophy. Previously we localized Tsk to mouse chromosome 2 in a region syntenic with human chromosome 15. The microfibrillar glycoprotein gene, fibrillin 1 (FBN1), on human chromosome 15q, provided a candidate for the Tsk mutation. We now demonstrate that the Tsk chromosome harbors a 30- to 40-kb genomic duplication within the Fbn1 gene that results in a larger than normal in-frame Fbn1 transcript. These findings provide hypotheses to explain some of the phenotypic characteristics of Tskl + mice and the lethality of Tsk/Tsk embryos

Collagen Content in Skin and Internal Organs of the Tight Skin Mouse: An Animal Model of Scleroderma

The Tight Skin mouse is a genetically induced animal model of tissue fibrosis caused by a large in-frame mutation in the gene encoding fibrillin-1 (Fbn-1). We examined the influence of gender on the collagen content of tissues in C57BL/6J wild type (+/+) and mutant Tight Skin (Tsk/+) mice employing hydroxyproline assays. Tissue sections were stained with Masson's trichrome to identify collagen in situ. Adult Tsk/+ mice skin contains ∼15% more collagen, on average, than skin from +/+ mice of the same gender. The heart of Tsk/+ males had significantly more collagen than that of +/+ males. No significant gender differences were found in lungs and kidney collagen content. Overall, the collagen content of Tsk/+ males and +/+ males was higher than that of their Tsk/+ and +/+ female counterparts, respectively. Our data confirm increased deposition of collagen in skin and hearts of Tsk/+ mice; however, the effects of the Tsk mutation on collagen content are both tissue specific and gender specific. These results indicate that comparative studies of collagen content between normal and Tsk/+ mice skin and internal organs must take into account gender differences caused by expression of the androgen receptor

Histology and CHD5 expression in xenografts derived from NLF and IMR5 cells transfected with either CHD5 sense or CHD5 antisense constructs

) Histology of NLF and IMR5 xenograft tumors (see ) after 5 weeks of growth. NLF--AS tumors were composed of undifferentiated cells with scant cytoplasm (top left), whereas NLF- tumors showed areas of necrosis () and differentiation (; top right). Cells in the IMR5--AS tumors were undifferentiated (bottom left), whereas cells in the IMR5- tumors had a more elongated appearance (bottom right). Bar = 20 μm. ) Relative expression of CHD5, normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH), was determined by real-time reverse transcription–polymerase chain reaction in parental NLF and IMR5 cell lines, as well as the CHD5 sense and CHD5 antisense transfected lines used for the xenograft experiments. The normalized values indicated by each bar graph represent the mean of three measurements. Replicate measurements were within 10% of the mean for each bar shown. ) Expression of CHD5 protein, as detected by immunoblotting, is shown for the NLF and IMR5 parental lines and the corresponding sense- and antisense-transfected cells used in these experiments.<p><b>Copyright information:</b></p><p>Taken from ", a Tumor Suppressor Gene Deleted From 1p36.31 in Neuroblastomas"</p><p></p><p>JNCI Journal of the National Cancer Institute 2008;100(13):940-949.</p><p>Published online 2 Jul 2008</p><p>PMCID:PMC2483574.</p><p></p

Effect of altered CHD5 expression on clonogenicity in neuroblastoma cell lines

Plasmids containing CHD5 or antisense CHD5 (CHD5-AS) were stably transfected into NLF and IMR5 neuroblastoma cell lines, both of which have hemizygous 1p deletion and amplification, and into SK-N-SH and SK-N-FI neuroblastoma cell lines, which have neither. Transfected cells were plated on soft agar, and colonies were counted 3 weeks later. ) NLF cells. CHD5 vs CHD5-AS, < .001. ) IMR5 cells. CHD5 vs CHD5-AS, = .001. ) SK-N-SH cells. CHD5 vs CHD5-AS, = .16. ) SK-N-FI cells. CHD5 vs CHD5-AS, = .10. Means and 95% confidence intervals () are shown. values (two-sided) were calculated using the Student test. Data are representative of three independent experiments performed in quadruplicate.<p><b>Copyright information:</b></p><p>Taken from ", a Tumor Suppressor Gene Deleted From 1p36.31 in Neuroblastomas"</p><p></p><p>JNCI Journal of the National Cancer Institute 2008;100(13):940-949.</p><p>Published online 2 Jul 2008</p><p>PMCID:PMC2483574.</p><p></p

Methylation of the CHD5 promoter and 5′ coding region in four neuroblastoma cell lines

–) CHD5 promoter methylation in NLF () and IMR5 () cells, both of which have hemizygous 1p deletions and virtually no CHD5 expression. –) CHD5 promoter methylation in SK-N-SH () and SK-N-FI cells (), which lack 1p deletion and have low expression of CHD5. Methylation at a given GC dinucleotide, as determined by methylation-specific sequencing, is shown as a percentage of sequences analyzed (10 per site). Methylation was determined as the number of clones with methylation of the given region divided by 10 and is expressed as a percentage.<p><b>Copyright information:</b></p><p>Taken from ", a Tumor Suppressor Gene Deleted From 1p36.31 in Neuroblastomas"</p><p></p><p>JNCI Journal of the National Cancer Institute 2008;100(13):940-949.</p><p>Published online 2 Jul 2008</p><p>PMCID:PMC2483574.</p><p></p

Association of CHD5 expression with risk factors and outcome in primary neuroblastomas

) Normalized CHD5 expression in 101 primary neuroblastomas stratified based on the presence (n = 26) or absence (n = 75) of 1p deletion (two-sample test, < .001). ) Association of normalized CHD5 expression with the risk group (low, intermediate, high, and ultrahigh) as defined above and in (). Briefly, for this study, low-risk patients were defined as infants (<p><b>Copyright information:</b></p><p>Taken from ", a Tumor Suppressor Gene Deleted From 1p36.31 in Neuroblastomas"</p><p></p><p>JNCI Journal of the National Cancer Institute 2008;100(13):940-949.</p><p>Published online 2 Jul 2008</p><p>PMCID:PMC2483574.</p><p></p

Genomic landscape of pediatric adrenocortical tumors

Pediatric adrenocortical carcinoma is a rare malignancy with poor prognosis. Here we analyze 37 adrenocortical tumors (ACTs) by whole genome, whole exome and/or transcriptome sequencing. Most cases (91%) show loss of heterozygosity (LOH) of chromosome 11p, with uniform selection against the maternal chromosome. IGF2 on chromosome 11p is overexpressed in 100% of the tumors. TP53 mutations and chromosome 17 LOH with selection against wild-type TP53 are observed in 28 ACTs (76%). Chromosomes 11p and 17 undergo copy-neutral LOH early during tumorigenesis, suggesting tumor-driver events. Additional genetic alterations include recurrent somatic mutations in ATRX and CTNNB1 and integration of human herpesvirus-6 in chromosome 11p. A dismal outcome is predicted by concomitant TP53 and ATRX mutations and associated genomic abnormalities, including massive structural variations and frequent background mutations. Collectively, these findings demonstrate the nature, timing and potential prognostic significance of key genetic alterations in pediatric ACT and outline a hypothetical model of pediatric adrenocortical tumorigenesis