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A porcine model of osteosarcoma.

Saalfrank A, Janssen KP, Ravon M, Flisikowski K, Eser S, Steiger K, Flisikowska T, Müller-Fliedner P, Schulze É, Brönner C, Gnann A, Kappe E, Böhm B, Schade B, Certa U, Saur D, Esposito I, Kind A, Schnieke A - Oncogenesis (2016)

Bottom Line: Transformed porcine MSCs exhibit genomic instability, with complex karyotypes, and develop into sarcomas on transplantation into immune-deficient mice.Unlike in Trp53 mutant mice, osteosarcoma developed in the long bones and skull, closely recapitulating the human disease.These animals thus promise a model for juvenile osteosarcoma, a relatively uncommon but devastating disease.

View Article: PubMed Central - PubMed

Affiliation: Chair of Livestock Biotechnology, Technische Universität München, Freising, Germany.

ABSTRACT
We previously produced pigs with a latent oncogenic TP53 mutation. Humans with TP53 germline mutations are predisposed to a wide spectrum of early-onset cancers, predominantly breast, brain, adrenal gland cancer, soft tissue sarcomas and osteosarcomas. Loss of p53 function has been observed in >50% of human cancers. Here we demonstrate that porcine mesenchymal stem cells (MSCs) convert to a transformed phenotype after activation of latent oncogenic TP53(R167H) and KRAS(G12D), and overexpression of MYC promotes tumorigenesis. The process mimics key molecular aspects of human sarcomagenesis. Transformed porcine MSCs exhibit genomic instability, with complex karyotypes, and develop into sarcomas on transplantation into immune-deficient mice. In pigs, heterozygous knockout of TP53 was sufficient for spontaneous osteosarcoma development in older animals, whereas homozygous TP53 knockout resulted in multiple large osteosarcomas in 7-8-month-old animals. This is the first report that engineered mutation of an endogenous tumour-suppressor gene leads to invasive cancer in pigs. Unlike in Trp53 mutant mice, osteosarcoma developed in the long bones and skull, closely recapitulating the human disease. These animals thus promise a model for juvenile osteosarcoma, a relatively uncommon but devastating disease.

No MeSH data available.


Related in: MedlinePlus

Expression profiles of cancer-related genes in transformed MSCs and sarcoma-derived tumour cells. (a) Differential gene expression heat maps relative to wild-type MSCs. Genes are grouped by functional categories, as indicated. The key at bottom right indicates fold-change values (log2 scale) represented as a colour gradient from blue (downregulation) to red (upregulation). See also Supplementary Table S2. (b) Sequence analysis of KRAS RT–PCR products amplified from exon 1 to exon 4. Cell types as shown. Codon 12 is indicated by a red box. RT–PCR products carrying the GAT sequence (G12D mutation) are the predominant species in poSARCO cells. (c) PoSARCO cells express higher levels of active GTP-bound Ras proteins (21 kDa) than the parental MSC-PKCM cells.
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fig2: Expression profiles of cancer-related genes in transformed MSCs and sarcoma-derived tumour cells. (a) Differential gene expression heat maps relative to wild-type MSCs. Genes are grouped by functional categories, as indicated. The key at bottom right indicates fold-change values (log2 scale) represented as a colour gradient from blue (downregulation) to red (upregulation). See also Supplementary Table S2. (b) Sequence analysis of KRAS RT–PCR products amplified from exon 1 to exon 4. Cell types as shown. Codon 12 is indicated by a red box. RT–PCR products carrying the GAT sequence (G12D mutation) are the predominant species in poSARCO cells. (c) PoSARCO cells express higher levels of active GTP-bound Ras proteins (21 kDa) than the parental MSC-PKCM cells.

Mentions: Microarray analysis was carried out to identify transcriptional changes associated with the stages of porcine MSC transformation. Figure 2a and Supplementary Table S1 show expression profiles for six groups of genes associated with cellular transformation: cellular transformation, telomere maintenance, cell cycle control, p53 target genes, apoptosis, and chromosomal instability (CIN).


A porcine model of osteosarcoma.

Saalfrank A, Janssen KP, Ravon M, Flisikowski K, Eser S, Steiger K, Flisikowska T, Müller-Fliedner P, Schulze É, Brönner C, Gnann A, Kappe E, Böhm B, Schade B, Certa U, Saur D, Esposito I, Kind A, Schnieke A - Oncogenesis (2016)

Expression profiles of cancer-related genes in transformed MSCs and sarcoma-derived tumour cells. (a) Differential gene expression heat maps relative to wild-type MSCs. Genes are grouped by functional categories, as indicated. The key at bottom right indicates fold-change values (log2 scale) represented as a colour gradient from blue (downregulation) to red (upregulation). See also Supplementary Table S2. (b) Sequence analysis of KRAS RT–PCR products amplified from exon 1 to exon 4. Cell types as shown. Codon 12 is indicated by a red box. RT–PCR products carrying the GAT sequence (G12D mutation) are the predominant species in poSARCO cells. (c) PoSARCO cells express higher levels of active GTP-bound Ras proteins (21 kDa) than the parental MSC-PKCM cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4815050&req=5

fig2: Expression profiles of cancer-related genes in transformed MSCs and sarcoma-derived tumour cells. (a) Differential gene expression heat maps relative to wild-type MSCs. Genes are grouped by functional categories, as indicated. The key at bottom right indicates fold-change values (log2 scale) represented as a colour gradient from blue (downregulation) to red (upregulation). See also Supplementary Table S2. (b) Sequence analysis of KRAS RT–PCR products amplified from exon 1 to exon 4. Cell types as shown. Codon 12 is indicated by a red box. RT–PCR products carrying the GAT sequence (G12D mutation) are the predominant species in poSARCO cells. (c) PoSARCO cells express higher levels of active GTP-bound Ras proteins (21 kDa) than the parental MSC-PKCM cells.
Mentions: Microarray analysis was carried out to identify transcriptional changes associated with the stages of porcine MSC transformation. Figure 2a and Supplementary Table S1 show expression profiles for six groups of genes associated with cellular transformation: cellular transformation, telomere maintenance, cell cycle control, p53 target genes, apoptosis, and chromosomal instability (CIN).

Bottom Line: Transformed porcine MSCs exhibit genomic instability, with complex karyotypes, and develop into sarcomas on transplantation into immune-deficient mice.Unlike in Trp53 mutant mice, osteosarcoma developed in the long bones and skull, closely recapitulating the human disease.These animals thus promise a model for juvenile osteosarcoma, a relatively uncommon but devastating disease.

View Article: PubMed Central - PubMed

Affiliation: Chair of Livestock Biotechnology, Technische Universität München, Freising, Germany.

ABSTRACT
We previously produced pigs with a latent oncogenic TP53 mutation. Humans with TP53 germline mutations are predisposed to a wide spectrum of early-onset cancers, predominantly breast, brain, adrenal gland cancer, soft tissue sarcomas and osteosarcomas. Loss of p53 function has been observed in >50% of human cancers. Here we demonstrate that porcine mesenchymal stem cells (MSCs) convert to a transformed phenotype after activation of latent oncogenic TP53(R167H) and KRAS(G12D), and overexpression of MYC promotes tumorigenesis. The process mimics key molecular aspects of human sarcomagenesis. Transformed porcine MSCs exhibit genomic instability, with complex karyotypes, and develop into sarcomas on transplantation into immune-deficient mice. In pigs, heterozygous knockout of TP53 was sufficient for spontaneous osteosarcoma development in older animals, whereas homozygous TP53 knockout resulted in multiple large osteosarcomas in 7-8-month-old animals. This is the first report that engineered mutation of an endogenous tumour-suppressor gene leads to invasive cancer in pigs. Unlike in Trp53 mutant mice, osteosarcoma developed in the long bones and skull, closely recapitulating the human disease. These animals thus promise a model for juvenile osteosarcoma, a relatively uncommon but devastating disease.

No MeSH data available.


Related in: MedlinePlus