A genetically inducible porcine model of intestinal cancer

Morten M. Callesen; Sigrid S. Árnadóttir; Iben Lyskjær; Mai Britt W. Ørntoft; Søren Høyer; Frederik Dagnæs-Hansen; Ying Liu; Rong Li; Henrik Callesen; Mads H. Rasmussen; Martin F. Berthelsen; Martin K. Thomsen; Pawel J. Schweiger; Kim B. Jensen; Søren Laurberg; Torben F. Ørntoft; Jannik E. Elverløv-Jakobsen; Claus L. Andersen

doi:10.1002/1878-0261.12136

A genetically inducible porcine model of intestinal cancer

Morten M. Callesen, Sigrid S. Árnadóttir, Iben Lyskjær, Mai Britt W. Ørntoft, Søren Høyer, Frederik Dagnæs-Hansen, Ying Liu, Rong Li, Henrik Callesen, Mads H. Rasmussen, Martin F. Berthelsen, Martin K. Thomsen, Pawel J. Schweiger, Kim B. Jensen, Søren Laurberg, Torben F. Ørntoft, Jannik E. Elverløv-Jakobsen, Claus L. Andersen^*

^*Corresponding author for this work

13 Citations (Scopus)

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Abstract

Transgenic porcine cancer models bring novel possibilities for research. Their physical similarities with humans enable the use of surgical procedures and treatment approaches used for patients, which facilitates clinical translation. Here, we aimed to develop an inducible oncopig model of intestinal cancer. Transgenic (TG) minipigs were generated using somatic cell nuclear transfer by handmade cloning. The pigs encode two TG cassettes: (a) an Flp recombinase-inducible oncogene cassette containing KRAS-G12D, cMYC, SV40LT - which inhibits p53 - and pRB and (b) a 4-hydroxytamoxifen (4-OHT)-inducible Flp recombinase activator cassette controlled by the intestinal epithelium-specific villin promoter. Thirteen viable transgenic minipigs were born. The ability of 4-OHT to activate the oncogene cassette was confirmed in vitro in TG colonic organoids and ex vivo in tissue biopsies obtained by colonoscopy. In order to provide proof of principle that the oncogene cassette could also successfully be activated in vivo, three pigs were perorally treated with 400 mg tamoxifen for 2 × 5 days. After two months, one pig developed a duodenal neuroendocrine carcinoma with a lymph node metastasis. Molecular analysis of the carcinoma and metastasis confirmed activation of the oncogene cassette. No tumor formation was observed in untreated TG pigs or in the remaining two treated pigs. The latter indicates that tamoxifen delivery can probably be improved. In summary, we have generated a novel inducible oncopig model of intestinal cancer, which has the ability to form metastatic disease already two months after induction. The model may be helpful in bridging the gap between basic research and clinical usage. It opens new venues for longitudinal studies of tumor development and evolution, for preclinical assessment of new anticancer regimens, for pharmacology and toxicology assessments, as well as for studies into biological mechanisms of tumor formation and metastasis.

Original language	English
Journal	Molecular Oncology
Volume	11
Issue number	11
Pages (from-to)	1616–1629
Number of pages	14
ISSN	1574-7891
DOIs	https://doi.org/10.1002/1878-0261.12136
Publication status	Published - Nov 2017

Keywords

Inducible intestinal cancer
Oncopig
Tissue-specific activation
Transgenic porcine model

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Callesen, M. M., Árnadóttir, S. S., Lyskjær, I., Ørntoft, M. B. W., Høyer, S., Dagnæs-Hansen, F., Liu, Y., Li, R., Callesen, H., Rasmussen, M. H., Berthelsen, M. F., Thomsen, M. K., Schweiger, P. J., Jensen, K. B., Laurberg, S., Ørntoft, T. F., Elverløv-Jakobsen, J. E., & Andersen, C. L. (2017). A genetically inducible porcine model of intestinal cancer. Molecular Oncology, 11(11), 1616–1629. https://doi.org/10.1002/1878-0261.12136

Callesen, MM, Árnadóttir, SS, Lyskjær, I, Ørntoft, MBW, Høyer, S, Dagnæs-Hansen, F, Liu, Y, Li, R, Callesen, H, Rasmussen, MH, Berthelsen, MF, Thomsen, MK, Schweiger, PJ , Jensen, KB, Laurberg, S, Ørntoft, TF, Elverløv-Jakobsen, JE & Andersen, CL 2017, 'A genetically inducible porcine model of intestinal cancer', Molecular Oncology, vol. 11, no. 11, pp. 1616–1629. https://doi.org/10.1002/1878-0261.12136

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title = "A genetically inducible porcine model of intestinal cancer",

abstract = "Transgenic porcine cancer models bring novel possibilities for research. Their physical similarities with humans enable the use of surgical procedures and treatment approaches used for patients, which facilitates clinical translation. Here, we aimed to develop an inducible oncopig model of intestinal cancer. Transgenic (TG) minipigs were generated using somatic cell nuclear transfer by handmade cloning. The pigs encode two TG cassettes: (a) an Flp recombinase-inducible oncogene cassette containing KRAS-G12D, cMYC, SV40LT - which inhibits p53 - and pRB and (b) a 4-hydroxytamoxifen (4-OHT)-inducible Flp recombinase activator cassette controlled by the intestinal epithelium-specific villin promoter. Thirteen viable transgenic minipigs were born. The ability of 4-OHT to activate the oncogene cassette was confirmed in vitro in TG colonic organoids and ex vivo in tissue biopsies obtained by colonoscopy. In order to provide proof of principle that the oncogene cassette could also successfully be activated in vivo, three pigs were perorally treated with 400 mg tamoxifen for 2 × 5 days. After two months, one pig developed a duodenal neuroendocrine carcinoma with a lymph node metastasis. Molecular analysis of the carcinoma and metastasis confirmed activation of the oncogene cassette. No tumor formation was observed in untreated TG pigs or in the remaining two treated pigs. The latter indicates that tamoxifen delivery can probably be improved. In summary, we have generated a novel inducible oncopig model of intestinal cancer, which has the ability to form metastatic disease already two months after induction. The model may be helpful in bridging the gap between basic research and clinical usage. It opens new venues for longitudinal studies of tumor development and evolution, for preclinical assessment of new anticancer regimens, for pharmacology and toxicology assessments, as well as for studies into biological mechanisms of tumor formation and metastasis.",

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doi = "10.1002/1878-0261.12136",

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volume = "11",

pages = "1616–1629",

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T1 - A genetically inducible porcine model of intestinal cancer

AU - Callesen, Morten M.

AU - Árnadóttir, Sigrid S.

AU - Lyskjær, Iben

AU - Ørntoft, Mai Britt W.

AU - Høyer, Søren

AU - Dagnæs-Hansen, Frederik

AU - Liu, Ying

AU - Li, Rong

AU - Callesen, Henrik

AU - Rasmussen, Mads H.

AU - Berthelsen, Martin F.

AU - Thomsen, Martin K.

AU - Schweiger, Pawel J.

AU - Jensen, Kim B.

AU - Laurberg, Søren

AU - Ørntoft, Torben F.

AU - Elverløv-Jakobsen, Jannik E.

AU - Andersen, Claus L.

PY - 2017/11

Y1 - 2017/11

N2 - Transgenic porcine cancer models bring novel possibilities for research. Their physical similarities with humans enable the use of surgical procedures and treatment approaches used for patients, which facilitates clinical translation. Here, we aimed to develop an inducible oncopig model of intestinal cancer. Transgenic (TG) minipigs were generated using somatic cell nuclear transfer by handmade cloning. The pigs encode two TG cassettes: (a) an Flp recombinase-inducible oncogene cassette containing KRAS-G12D, cMYC, SV40LT - which inhibits p53 - and pRB and (b) a 4-hydroxytamoxifen (4-OHT)-inducible Flp recombinase activator cassette controlled by the intestinal epithelium-specific villin promoter. Thirteen viable transgenic minipigs were born. The ability of 4-OHT to activate the oncogene cassette was confirmed in vitro in TG colonic organoids and ex vivo in tissue biopsies obtained by colonoscopy. In order to provide proof of principle that the oncogene cassette could also successfully be activated in vivo, three pigs were perorally treated with 400 mg tamoxifen for 2 × 5 days. After two months, one pig developed a duodenal neuroendocrine carcinoma with a lymph node metastasis. Molecular analysis of the carcinoma and metastasis confirmed activation of the oncogene cassette. No tumor formation was observed in untreated TG pigs or in the remaining two treated pigs. The latter indicates that tamoxifen delivery can probably be improved. In summary, we have generated a novel inducible oncopig model of intestinal cancer, which has the ability to form metastatic disease already two months after induction. The model may be helpful in bridging the gap between basic research and clinical usage. It opens new venues for longitudinal studies of tumor development and evolution, for preclinical assessment of new anticancer regimens, for pharmacology and toxicology assessments, as well as for studies into biological mechanisms of tumor formation and metastasis.

AB - Transgenic porcine cancer models bring novel possibilities for research. Their physical similarities with humans enable the use of surgical procedures and treatment approaches used for patients, which facilitates clinical translation. Here, we aimed to develop an inducible oncopig model of intestinal cancer. Transgenic (TG) minipigs were generated using somatic cell nuclear transfer by handmade cloning. The pigs encode two TG cassettes: (a) an Flp recombinase-inducible oncogene cassette containing KRAS-G12D, cMYC, SV40LT - which inhibits p53 - and pRB and (b) a 4-hydroxytamoxifen (4-OHT)-inducible Flp recombinase activator cassette controlled by the intestinal epithelium-specific villin promoter. Thirteen viable transgenic minipigs were born. The ability of 4-OHT to activate the oncogene cassette was confirmed in vitro in TG colonic organoids and ex vivo in tissue biopsies obtained by colonoscopy. In order to provide proof of principle that the oncogene cassette could also successfully be activated in vivo, three pigs were perorally treated with 400 mg tamoxifen for 2 × 5 days. After two months, one pig developed a duodenal neuroendocrine carcinoma with a lymph node metastasis. Molecular analysis of the carcinoma and metastasis confirmed activation of the oncogene cassette. No tumor formation was observed in untreated TG pigs or in the remaining two treated pigs. The latter indicates that tamoxifen delivery can probably be improved. In summary, we have generated a novel inducible oncopig model of intestinal cancer, which has the ability to form metastatic disease already two months after induction. The model may be helpful in bridging the gap between basic research and clinical usage. It opens new venues for longitudinal studies of tumor development and evolution, for preclinical assessment of new anticancer regimens, for pharmacology and toxicology assessments, as well as for studies into biological mechanisms of tumor formation and metastasis.

KW - Inducible intestinal cancer

KW - Oncopig

KW - Tissue-specific activation

KW - Transgenic porcine model

U2 - 10.1002/1878-0261.12136

DO - 10.1002/1878-0261.12136

M3 - Journal article

C2 - 28881081

AN - SCOPUS:85031092360

SN - 1574-7891

VL - 11

SP - 1616

EP - 1629

JO - Molecular Oncology

JF - Molecular Oncology

IS - 11

ER -

A genetically inducible porcine model of intestinal cancer

Abstract

Keywords

UN SDGs

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