A new mouse model to study the role of ectopic Nanos3 expression in cancer
Abstract Background NANOS3 is a gene conserved throughout evolution. Despite the quite low conservation of Nanos sequences between different organisms and even between Nanos paralogs, their role in germ cell development is remarkably universal. Human Nanos3 expression is normally restricted to the g...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2019-06-01
|
| Series: | BMC Cancer |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12885-019-5807-x |
| _version_ | 1819002722841526272 |
|---|---|
| author | Vanessa Andries Evi De Keuckelaere Katrien Staes Tino Hochepied Joachim Taminau Kelly Lemeire Philippe Birembaut Geert Berx Frans van Roy |
| author_facet | Vanessa Andries Evi De Keuckelaere Katrien Staes Tino Hochepied Joachim Taminau Kelly Lemeire Philippe Birembaut Geert Berx Frans van Roy |
| author_sort | Vanessa Andries |
| collection | DOAJ |
| description | Abstract Background NANOS3 is a gene conserved throughout evolution. Despite the quite low conservation of Nanos sequences between different organisms and even between Nanos paralogs, their role in germ cell development is remarkably universal. Human Nanos3 expression is normally restricted to the gonads and the brain. However, ectopic activation of this gene has been detected in various human cancers. Until now, Nanos3 and other Nanos proteins have been studied almost exclusively in germ cell development. Methods Transgenic mice were generated by targeted insertion of a human Nanos3 cDNA into the ROSA26 locus. The transgene could be spatiotemporally induced by Cre recombinase activity removing an upstream floxed STOP cassette. A lung tumor model with ectopic Nanos3 expression was based on the lung-specific activation of the reverse tetracycline transactivator gene, in combination with a tetO-CMV promoter controlling Cre expression. When doxycycline was provided to the mice, Cre was activated leading to deletion of TP53 alleles and activation of both oncogenic KRasG12D and Nanos3. Appropriate controls were foreseen. Tumors and tumor-derived cell cultures were analyzed in various ways. Results We describe the successful generation of Nanos3LSL/− and Nanos3LSL/LSL mice in which an exogenous human NANOS3 gene can be activated in vivo upon Cre expression. These mice, in combination with different conditional and doxycycline-inducible Cre lines, allow the study of the role of ectopic Nanos3 expression in several cancer types. The Nanos3LSL mice were crossed with a non-small cell lung cancer (NSCLC) mouse model based on conditional expression of oncogenic KRas and homozygous loss of p53. This experiment demonstrated that ectopic expression of Nanos3 in the lungs has a significant negative effect on survival. Enhanced bronchiolar dysplasia was observed when Nanos3-expressing NSCLC mice were compared with control NSCLC mice. An allograft experiment, performed with cell cultures derived from primary lung tumors of control and Nanos3-expressing NSCLC mice, revealed lymph node metastasis in mice injected with Nanos3-expressing NSCLC cells. Conclusions A new mouse model was generated allowing examination of Nanos3-associated pathways and investigation of the influence of ectopic Nanos3 expression in various cancer types. This model might identify Nanos3 as an interesting target in cancer therapeutics. |
| first_indexed | 2024-12-20T23:09:38Z |
| format | Article |
| id | doaj.art-033078d3a4a34ff59eb98f36de05bc24 |
| institution | Directory Open Access Journal |
| issn | 1471-2407 |
| language | English |
| last_indexed | 2024-12-20T23:09:38Z |
| publishDate | 2019-06-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Cancer |
| spelling | doaj.art-033078d3a4a34ff59eb98f36de05bc242022-12-21T19:23:45ZengBMCBMC Cancer1471-24072019-06-0119111710.1186/s12885-019-5807-xA new mouse model to study the role of ectopic Nanos3 expression in cancerVanessa Andries0Evi De Keuckelaere1Katrien Staes2Tino Hochepied3Joachim Taminau4Kelly Lemeire5Philippe Birembaut6Geert Berx7Frans van Roy8VIB-UGent Center for Inflammation Research (IRC)VIB-UGent Center for Inflammation Research (IRC)VIB-UGent Center for Inflammation Research (IRC)VIB-UGent Center for Inflammation Research (IRC)Department of Biomedical Molecular Biology, Ghent UniversityVIB-UGent Center for Inflammation Research (IRC)INSERM UMRS 1250, Department of Biopathology, CHU Maison-Blanche, University Hospital of Reims & University of Reims Champagne-ArdenneDepartment of Biomedical Molecular Biology, Ghent UniversityVIB-UGent Center for Inflammation Research (IRC)Abstract Background NANOS3 is a gene conserved throughout evolution. Despite the quite low conservation of Nanos sequences between different organisms and even between Nanos paralogs, their role in germ cell development is remarkably universal. Human Nanos3 expression is normally restricted to the gonads and the brain. However, ectopic activation of this gene has been detected in various human cancers. Until now, Nanos3 and other Nanos proteins have been studied almost exclusively in germ cell development. Methods Transgenic mice were generated by targeted insertion of a human Nanos3 cDNA into the ROSA26 locus. The transgene could be spatiotemporally induced by Cre recombinase activity removing an upstream floxed STOP cassette. A lung tumor model with ectopic Nanos3 expression was based on the lung-specific activation of the reverse tetracycline transactivator gene, in combination with a tetO-CMV promoter controlling Cre expression. When doxycycline was provided to the mice, Cre was activated leading to deletion of TP53 alleles and activation of both oncogenic KRasG12D and Nanos3. Appropriate controls were foreseen. Tumors and tumor-derived cell cultures were analyzed in various ways. Results We describe the successful generation of Nanos3LSL/− and Nanos3LSL/LSL mice in which an exogenous human NANOS3 gene can be activated in vivo upon Cre expression. These mice, in combination with different conditional and doxycycline-inducible Cre lines, allow the study of the role of ectopic Nanos3 expression in several cancer types. The Nanos3LSL mice were crossed with a non-small cell lung cancer (NSCLC) mouse model based on conditional expression of oncogenic KRas and homozygous loss of p53. This experiment demonstrated that ectopic expression of Nanos3 in the lungs has a significant negative effect on survival. Enhanced bronchiolar dysplasia was observed when Nanos3-expressing NSCLC mice were compared with control NSCLC mice. An allograft experiment, performed with cell cultures derived from primary lung tumors of control and Nanos3-expressing NSCLC mice, revealed lymph node metastasis in mice injected with Nanos3-expressing NSCLC cells. Conclusions A new mouse model was generated allowing examination of Nanos3-associated pathways and investigation of the influence of ectopic Nanos3 expression in various cancer types. This model might identify Nanos3 as an interesting target in cancer therapeutics.http://link.springer.com/article/10.1186/s12885-019-5807-xNANOSCre-loxPROSA26 locusMouse embryogenesisLung cancer model |
| spellingShingle | Vanessa Andries Evi De Keuckelaere Katrien Staes Tino Hochepied Joachim Taminau Kelly Lemeire Philippe Birembaut Geert Berx Frans van Roy A new mouse model to study the role of ectopic Nanos3 expression in cancer BMC Cancer NANOS Cre-loxP ROSA26 locus Mouse embryogenesis Lung cancer model |
| title | A new mouse model to study the role of ectopic Nanos3 expression in cancer |
| title_full | A new mouse model to study the role of ectopic Nanos3 expression in cancer |
| title_fullStr | A new mouse model to study the role of ectopic Nanos3 expression in cancer |
| title_full_unstemmed | A new mouse model to study the role of ectopic Nanos3 expression in cancer |
| title_short | A new mouse model to study the role of ectopic Nanos3 expression in cancer |
| title_sort | new mouse model to study the role of ectopic nanos3 expression in cancer |
| topic | NANOS Cre-loxP ROSA26 locus Mouse embryogenesis Lung cancer model |
| url | http://link.springer.com/article/10.1186/s12885-019-5807-x |
| work_keys_str_mv | AT vanessaandries anewmousemodeltostudytheroleofectopicnanos3expressionincancer AT evidekeuckelaere anewmousemodeltostudytheroleofectopicnanos3expressionincancer AT katrienstaes anewmousemodeltostudytheroleofectopicnanos3expressionincancer AT tinohochepied anewmousemodeltostudytheroleofectopicnanos3expressionincancer AT joachimtaminau anewmousemodeltostudytheroleofectopicnanos3expressionincancer AT kellylemeire anewmousemodeltostudytheroleofectopicnanos3expressionincancer AT philippebirembaut anewmousemodeltostudytheroleofectopicnanos3expressionincancer AT geertberx anewmousemodeltostudytheroleofectopicnanos3expressionincancer AT fransvanroy anewmousemodeltostudytheroleofectopicnanos3expressionincancer AT vanessaandries newmousemodeltostudytheroleofectopicnanos3expressionincancer AT evidekeuckelaere newmousemodeltostudytheroleofectopicnanos3expressionincancer AT katrienstaes newmousemodeltostudytheroleofectopicnanos3expressionincancer AT tinohochepied newmousemodeltostudytheroleofectopicnanos3expressionincancer AT joachimtaminau newmousemodeltostudytheroleofectopicnanos3expressionincancer AT kellylemeire newmousemodeltostudytheroleofectopicnanos3expressionincancer AT philippebirembaut newmousemodeltostudytheroleofectopicnanos3expressionincancer AT geertberx newmousemodeltostudytheroleofectopicnanos3expressionincancer AT fransvanroy newmousemodeltostudytheroleofectopicnanos3expressionincancer |