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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

Transformed MSCs and osteosarcoma-derived cells show increased resistance to radiation. (a) Survival of cells after 137Cs irradiation after 2 weeks of culture. Left: Colonies stained with crystal violet; cell types and doses as indicated. Right: Mean survival±s.d., n=4. No significant differences were observed at 10 Gy irradiation. However, at 2 Gy transformed MSC showed increased viability compared with WT-MSCs: MSC-P (P=0.0184), MSCK-PKC (P=0.0500). MSC-PKCM cells did not attain significance (P=0.1558). Results were similar at the intermediate dose 6 Gy: MSC-P (P=0.0011); MSCK-PKC (P=0.0081); MSC-PKCM (P=0.0874). (b) Osteosarcoma cells show increased radioresistance. Cell survival in response to irradiation was assessed after 2 weeks of cell culture, as above. Cells were derived from a tumour on the left tibia of homozygous TP53 knockout pig (animal ID: 242) compared with cells from a tumour-free bone of the same animal. Tumour cells showed increased clonal growth after irradiation.
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fig6: Transformed MSCs and osteosarcoma-derived cells show increased resistance to radiation. (a) Survival of cells after 137Cs irradiation after 2 weeks of culture. Left: Colonies stained with crystal violet; cell types and doses as indicated. Right: Mean survival±s.d., n=4. No significant differences were observed at 10 Gy irradiation. However, at 2 Gy transformed MSC showed increased viability compared with WT-MSCs: MSC-P (P=0.0184), MSCK-PKC (P=0.0500). MSC-PKCM cells did not attain significance (P=0.1558). Results were similar at the intermediate dose 6 Gy: MSC-P (P=0.0011); MSCK-PKC (P=0.0081); MSC-PKCM (P=0.0874). (b) Osteosarcoma cells show increased radioresistance. Cell survival in response to irradiation was assessed after 2 weeks of cell culture, as above. Cells were derived from a tumour on the left tibia of homozygous TP53 knockout pig (animal ID: 242) compared with cells from a tumour-free bone of the same animal. Tumour cells showed increased clonal growth after irradiation.

Mentions: Several mutant p53 isoforms confer resistance to radiation.33 p53-deficient MSCs or MSCs expressing mutant p53 show increased resistance to 137Cs irradiation (Figure 6a). Similarly, cells derived from an osteosarcoma on the left tibia of homozygous TP53 knockout pig 242 showed increased radioresistance compared with control cells derived from a tumour-free bone of the same animal (Figure 6b). Tumour-derived and normal cell isolates were characterised as positive for osteocalcin, osteonectin, type 1 collagen, alkaline phosphatase and vimentin and negative for EpCam.


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)

Transformed MSCs and osteosarcoma-derived cells show increased resistance to radiation. (a) Survival of cells after 137Cs irradiation after 2 weeks of culture. Left: Colonies stained with crystal violet; cell types and doses as indicated. Right: Mean survival±s.d., n=4. No significant differences were observed at 10 Gy irradiation. However, at 2 Gy transformed MSC showed increased viability compared with WT-MSCs: MSC-P (P=0.0184), MSCK-PKC (P=0.0500). MSC-PKCM cells did not attain significance (P=0.1558). Results were similar at the intermediate dose 6 Gy: MSC-P (P=0.0011); MSCK-PKC (P=0.0081); MSC-PKCM (P=0.0874). (b) Osteosarcoma cells show increased radioresistance. Cell survival in response to irradiation was assessed after 2 weeks of cell culture, as above. Cells were derived from a tumour on the left tibia of homozygous TP53 knockout pig (animal ID: 242) compared with cells from a tumour-free bone of the same animal. Tumour cells showed increased clonal growth after irradiation.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig6: Transformed MSCs and osteosarcoma-derived cells show increased resistance to radiation. (a) Survival of cells after 137Cs irradiation after 2 weeks of culture. Left: Colonies stained with crystal violet; cell types and doses as indicated. Right: Mean survival±s.d., n=4. No significant differences were observed at 10 Gy irradiation. However, at 2 Gy transformed MSC showed increased viability compared with WT-MSCs: MSC-P (P=0.0184), MSCK-PKC (P=0.0500). MSC-PKCM cells did not attain significance (P=0.1558). Results were similar at the intermediate dose 6 Gy: MSC-P (P=0.0011); MSCK-PKC (P=0.0081); MSC-PKCM (P=0.0874). (b) Osteosarcoma cells show increased radioresistance. Cell survival in response to irradiation was assessed after 2 weeks of cell culture, as above. Cells were derived from a tumour on the left tibia of homozygous TP53 knockout pig (animal ID: 242) compared with cells from a tumour-free bone of the same animal. Tumour cells showed increased clonal growth after irradiation.
Mentions: Several mutant p53 isoforms confer resistance to radiation.33 p53-deficient MSCs or MSCs expressing mutant p53 show increased resistance to 137Cs irradiation (Figure 6a). Similarly, cells derived from an osteosarcoma on the left tibia of homozygous TP53 knockout pig 242 showed increased radioresistance compared with control cells derived from a tumour-free bone of the same animal (Figure 6b). Tumour-derived and normal cell isolates were characterised as positive for osteocalcin, osteonectin, type 1 collagen, alkaline phosphatase and vimentin and negative for EpCam.

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