Mouse Models of Lung Cancer...therapy,and patient management,the cure rate for advanced non–small-cell lung cancer (NSCLC) remains dismal. A comprehensive understanding of the genetic

Mouse Models of Lung CancerAmit Dutt1,2 and Kwok-Kin Wong2,3

Abstract Human lung cancer is responsible for f30% of all cancer deaths worldwide with >160,000deaths in the United States alone annually. Recent advances in the identification of novel muta-tions relevant to lung cancer from a myriad of genomic studies might translate into meaningfuldiagnostic and therapeutic progress. Towards this end, a genetic model animal system that canvalidate the oncogenic roles of these mutations in vivo would facilitate the understanding ofthe pathogenesis of lung cancer as well as provide ideal preclinical models for targeted therapytesting.The mouse is a promising model system, as complex human genetic traits causal to lungcancer, from inherited polymorphisms to somatic mutations, can be recapitulated in its genomevia genetic manipulation.We present here a brief overview of the existing mouse models of lungcancers and the challenges and opportunities for building the next generation of lung cancermouse models.

Lung cancer is the leading cause of cancer-related mortality forboth men and women in the United States and in the world(1). Despite advances in cytotoxic drug development, radio-therapy, and patient management, the cure rate for advancednon–small-cell lung cancer (NSCLC) remains dismal. Acomprehensive understanding of the genetic alterationsinvolved in the initiation and progression of human lungcancer would facilitate progress in clinical treatment and earlydiagnosis. The recent genome-scale effort to identify lungcancer–relevant genes has generated many candidate genes ofpotential therapeutic relevance. Success with a systematicgenome resequencing approach, which has identified muta-tions in the EGFR kinase domain in human lung cancer andshowed their correlation with patient response to the kinaseinhibitors gefitinib and erlotinib (2–6), has revolutionizedtargeted therapy in lung cancer. Similarly, complementarygenome-wide analytic approaches such as comparative or arraygenomic hybridization to detect copy number alteration (7, 8),as well as single-nucleotide polymorphism array analyses (9),have proved highly productive in discovering novel lungcancer–specific genomic alterations. In vitro studies have begunto validate some of these gene candidates of potential clinical

relevance to lung cancer. In particular, the recently identifiedsomatic mutations in EGFR, BRAF , and PI3K kinases have beenshown to possess transforming activity. However, althoughthese in vitro cell culture studies are informative, they cannotfully recapitulate the complex nature of de novo tumorigenesisor in vivo responses to therapy. Thus, the development ofanimal models harboring these mutations will likely yieldadditional insights into the underlying pathophysiologicperturbations. With such models in hand, it will also becomepossible to generate preclinical models to yield histopathologicand molecular surrogates for a specific mutation-directedactivity in a mature tumor, and thus better evaluate the efficacyand specificity of emerging agents directed against such lesions.

The ability to easily manipulate the mouse genome, alongwith the anatomic and physiologic similarity of the mouse tohumans, makes the mouse an ideal system to model humandiseases. Different generations of mouse models have evolvedemploying diverse innovative strategies to model lung cancer assummarized in Fig. 1. Although they have all been informativeand further propel our understanding of human lung cancer,they still do not fully recapitulate the complexities of humanlung cancer. The existing models and the strategies for thedevelopment of better mouse models will be discussed in thisreview (for additional review, see ref. 10). Although we haveattempted to be as comprehensive as possible, our list of mousemodels is likely to be incomplete.

Evolving Mouse Models for Human Lung Cancer

Classic transgenic models for ectopic expression. The devel-opment of methods for manipulating the mouse genome,combined with the sequencing of the mouse genome, hasadvanced our ability to generate new mouse models of humandiseases. Some of the earliest lung cancer mouse models werebased on the introduction of a transgene under the control ofheterologous promoters. To generate a transgenic mouse, alinearized plasmid vector carrying a transgene is integrated intothe genome of the fertilized oocytes following microinjection

Authors’Affiliations: 1Broad Institute of Harvard and Massachusetts Institute ofTechnology, Cambridge, Massachusetts; 2Department of Medical Oncology, Dana-Farber Cancer Institute; and 3Department of Medicine, Brigham andWomen’sHospital, Boston, MassachusettsReceived 2/21/06; revised 5/3/06; accepted 5/22/06.Grant support: Swiss National Science Foundation Fellowship, no. PBZHB-106297 (A. Dutt); NIH grant K08AG 2400401, the Sidney Kimmel Foundation forCancer Research, and the Joan Scarangello Foundation to Conquer Lung Cancerand the Flight Attendant Medical Research Institute (K.K.Wong).Presented at theThird Cambridge Conference on Novel Agents in theTreatment ofLung Cancer: Advances in EGFR-Targeted Agents, September 23-24, 2005 inCambridge, Massachusetts.Requests for reprints: Kwok-KinWong, Department of Medical Oncology, LoweCenter for Thoracic Oncology, Dana-Farber Cancer Institute, Harvard MedicalSchool, 44 Binney Street, D810, Boston, MA 02115. Phone: 617-632-6084;E-mail: [email protected].

F2006 American Association for Cancer Research.doi:10.1158/1078-0432.CCR-06-0414

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and later implanted into pseudopregnant mothers. In additionto constitutively expressed promoters such as keratin, geneshave also been targeted to specific subsets of lung epithelialcells using alveolar type II cell–specific surfactant protein C(SP-C) promoter (11), the nonciliated secretory cell–specificClara cell secretory protein (CCSP) promoter (12, 13), andcalcitonin gene – related peptide (CGRP) promoter (14).Examples of transgenic mice that model lung cancer includemodels with expression of human papillomavirus-16 E6/E7under the keratin-5 promoter, which developed lung adeno-carcinoma (15). Similarly, simian virus large T antigen (Tag),targeted with SP-C (16, 17) or CCSP (18, 19) promoter,resulted in adenocarcinoma of alveolar type II cells and distalClara cells of the lung, respectively. Furthermore, SP-C specificexpression of c-myc , epidermal growth factor (EGF; ref. 20),c-met-related tyrosine kinase, Ron (21), Raf-1 (22), anddominant-negative p53 (23) transgenes developed bronchio-loalveolar adenocarcinomas.

These transgenic mice are ideal for overexpression of themodeled oncogenes, a phenomenon that is frequently observeddue to amplification or epigenetic modification in humancancer. However, a major biological caveat to this strategy isthat heterologous promoters might not recapitulate the trueexpression pattern of the gene during development, adult life,and/or tumorigenesis. Further, oncogenes in such transgenicmice are constitutively expressed and cannot be regulated toassess the role of the transgene in tumor maintenance after theinitiation and establishment of the tumor.Classic knockout/knock-in models. In a knockout technolo-

gy, a tumor suppressor gene in embryonic stem cells (24, 25)is inactivated or replaced by site-specific homologous recom-bination between the chromosomal DNA and recombinantDNA flanked by homologous DNA sequences (the targetingvector). Recombinant DNA could code either for an antibioticmarker or for any intervening sequence that interrupts thecoding region of the target gene to knock out or to knock ina novel sequence. The embryonic stem cells with the modifiedgenome are injected into a pre-implantation embryo (blasto-coele of 3.5-day-old embryo). The incorporation of themodified embryonic stem cell into the murine germ lineof the chimeric mouse gives rise to genetically modifiedloci, enabling comparative studies with wild-type micelittermates.

The knockout strategy allows preferential deletion of selectivetumor suppressor genes implicated in lung cancer. In particular,targeting genes deleted early during human lung tumordevelopment, such as the nonessential genes on chromosome3p, FHIT and VHL , is likely to closely recapitulate a cancerphenotype. For example, 44% of Fhit-Vhl doubly deficient micewere found to develop spontaneous lung adenocarcinomas(26). However, this strategy cannot be used to study essentialtumor suppressor genes like Rb-1 or WT-1 , which areembryonic lethal if deleted (27, 28). On the other hand, othertumor suppressor knockout mutant animals that are notembryonic lethal, such as p53, p16, p19, or p16/p19 mutantmice, either do not develop lung adenocarcinomas or succumbto other tumors like lymphomas and sarcomas early on, thusprecluding the development of lung cancer. Given the complexgenetic alterations in human lung cancer, it is unlikely thatdeletion of a single tumor suppressor gene is sufficient tophenocopy human lung cancer in the mouse.

The knock-in approach, in contrast, has been applied toactivate oncogenes. In one study, one allele of mouse K-ras wasreplaced with an oncogenic K-rasG12D allele, which could beexpressed only if somatic recombination occurred between thetargeted allele and the wild-type allele (29). This strategybypasses potential alterations in development due to the earlyexpression of the mutant allele. Subsequently, following aspontaneous somatic recombination event, the endogenouspromoter gets juxtaposed to express the mutant allele and, thus,physiologic levels of expression are obtained. Mice with thetargeted K-ras mutation developed pulmonary adenocarcino-mas because of the sporadic activation of the mutant allele.However, these mice also developed other tumor types inaddition to lung adenocarcinomas.

Conditional bitransgenic tetracycline-inducible mouse models. Inlung cancer, most oncogenic activating mutations are somaticwhereas the classic transgenic mouse carries transgenes ofwhich the expression is limited by the inherent temporal-spatialexpression pattern of the chosen heterologous promoter. Thenext generation of transgenic mice was designed to overcomethis drawback by conditionally regulating the expression of agene in a specific somatic tissue, providing leverage toresearchers in generating inducible mouse tumor models. Thebinary transgenic conditional model is an inducible model thatoffers to overcome basic deficiencies of the classic model bysequentially splitting temporal-spatial regulation into individ-ual events (30, 31). Based on the principle of gene activatorprotein–mediated positive regulation of the lac operon inbacteria, a gene of interest is cloned downstream to a regulatorelement that requires the cooperation of at least two differentmolecules to initiate transcription, a regulator protein and itscognate binding partner (32). The expression of the regulatorprotein under a tissue-specific promoter defines the spatialregulation of the gene of interest whereas the temporalregulation is achieved by exogenously supplementing thebinding partner.

One of the classic examples of a bitransgenic induciblesystem that has been frequently exploited is the reversetetracycline transactivator (rtTA) inducible system. In thissystem, the bitransgenic mouse harbors two different trans-genes. One transgene, the rtTA gene linked to a tissue specificpromoter, drives the expression of the rtTA protein in a specificcell type. In the presence of tetracycline/doxycycline, the rtTAprotein activates the expression of the second transgene, thetargeted gene linked to the tetracycline-responsive promoterelement (Tet-Op ; refs. 33, 34), as shown in Fig. 2. Thisbitransgenic regulatory system has been successfully employedto generate different inducible models of lung cancer. Forexample, bitransgenic CCSP-rtTA/Tet-Op-FGF-7 mice (35) andCCSP-rtTA/Tet-Op-K-Ras4G12D mice (36) developed epithelialcell hyperplasia, adenomatous hyperplasia, and eventuallyadenocarcinomas after administration of doxycycline. Moreinterestingly, these tumors regressed following withdrawal ofdoxycycline, showing that the continual expression of thesegenes is necessary for tumor maintenance. Similar experimentsdone in a Tp53- or Rb1-deficient background resulted in aneven faster tumor onset. Thus, by combining the inducibletransgenic system with the traditional knockout model, thecomplex mutant mice showed that the expression of oncogenicK-ras in combination with tumor suppressor deficiency issynergistic in lung tumorigenesis.

MouseModels of Lung Cancer

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This bitransgenic system provides a high level of tight,inducible, and reversible gene expression in the appropriatetissue in vivo to recapitulate the somatic mutation at a definedstage of life in an otherwise normal mouse. The most strikingfeature of this system is its ability to switch off oncogeneexpression following the withdrawal of doxycycline, whichallows the assessment of the role of these alleles in themaintenance of established lung tumors. However, thesemurine lung tumor models develop pulmonary adenocarcino-mas with limited metastatic potential, albeit with a strikinghistopathologic similarity to human adenocarcinomas. Finally,considering the high complexity of lung cancer genetics, thisregulatory system is an approach that likely needs to becombined with other conditional genetic systems to fullyrecapitulate human lung cancer.Conditional bitransgenic mouse models involving Cre/loxP

system. As it is becoming increasingly clear that mammalian geneexpression is regulated by multiple complex mechanisms, thegeneration of conditional knockout models using tissue-specificexpression of recombinases of the Cre/loxP and Flp/FRT systemshas revolutionized lung cancer modeling, whereby mutation at aspecific locus can be directed to occur strictly in a set ofdifferentiated cells while the genome of the adjacent cells remainsunaltered (37–39). The versatility of this robust system makes itan efficient tool with dual applicability to generating conditionalalleles of both tumor suppressor genes and oncogenes.

A target region to be deleted in the gene locus can be markedfor deletion by signal sequences of loxP or FRT that areidentified by their site-specific recombinase Cre or Flp,respectively. The expression of the recombinase leads to theprecise removal of the stretch of DNA between the recombinasesignal sequences. To generate a conditional null, hypomorphicmutation or a reduction-of-function allele of a tumor suppres-sor gene, an appropriate sequence of a coding or noncodingregion is flanked by a loxP or FRT site (commonly referred asthe region being floxed). Similarly, as a complementaryapproach, to conditionally activate an oncogene, one couldflox the region that prevents its expression. To add anadditional level of maneuverability to regulate the expressionof the target gene, the recombinase is expressed under a tissue-specific or inducible promoter like the tetracycline-induciblesystem (40), Protamine-Cre (41), and CMV-Cre (42).

An alternative approach to Cre-mediated recombination inthe lung is to employ engineered viruses (such as adenovirus)via nasal instillation (43). This strategy allows closer recapit-ulation of lung cancer events by localized and timely regulationof target gene expression in a relatively small number of cells.As a proof of principle, adenoviral-mediated expression ofmutant alleles of oncogenic K-RasG12D or K-RasG12V inbronchioloalveolar cells produced multiple adenomas andadenocarcinomas, revealing the precise role of RAS oncogenesin the development of human lung cancer (43, 44). And more

Fig. 1. Evolving mouse models of human lung cancer.




recently, Olive et al. and Lang et al. independently generatedconditional knock-in mice with dominant-negative p53 mod-ified with two point mutations, p53R270H (45) and p53R172H(45, 46), at regions analogous to human p53 found to bemutated in patients with Li-Fraumeni syndrome. Followingadenoviral-mediated activation of the mutant allele, 19% ofp53R270H/+ mice developed lung adenocarcinomas withmalignant characteristics and metastasis that were found to besimilar to human lung adenocarcinoma. However, one ofthe major downsides of this gene activation system is that thevariability in delivery and infection with the virus adds to theheterogeneity of results and experimental inconsistency.Compound conditional mouse models. A natural extension of

the new transgenic technology has yielded a newer generationof compound conditional mouse models that integrate variedfeatures from the above models. The conditional knock-in andknockout genetic systems, for example, allow the ability tosimultaneously activate and/or delete multiple genes in atemporal- and tissue-specific manner. In a recent report,compound mouse mutant models with conditional activationof a K-ras transgene (Kras2), along with conditional inactivationof the alleles of Rb or p53 by Cre/loxP system, showedinvolvement of specific factors in regulating non–small-celllung cancer and small-cell lung cancer (SCLC). The activation ofKRas2 could induce hyperplasia that developed into non–small-cell lung cancer. Development of small-cell lung cancerinstead required inactivation of two tumor suppressor genes,Ink4A/Arf and Tp53 (47). Similarly, a mouse carrying condi-tional alleles of both mutant K-ras LSL-K-rasG12D allele and

mutant p53 LSL-p53R270H allele has been generated thatdeveloped aggressive lung cancer with induction of stromaldesmoplasia, local invasion, and distant metastases. Thesecompound mutant mouse models more faithfully recapitulatethe causal genetic phenotypic correlation for human lung cancer(48). Another compound mouse with conditional mutations inthe K-ras proto-oncogene and different alleles of p53 tumorsuppresser gene (a contact mutant, a structural mutant, or a nullallele) has recently been reported. It recapitulates several aspectsof advanced human pulmonary adenocarcinoma with develop-ment of tumor growth and progression, along with inductionof a desmoplastic response and metastasis (49).

Complex Genetics of Lung Cancer NecessitatesComplex Mouse Models

The current era of genetically engineered compoundconditional mice has generated models that more methodi-cally recapitulate the sporadic forms of cancers in human.These new model systems create an opportunity to evaluatehow different combinations of mutations would affect theinitiation and progression of lung cancer. In addition, thesenovel mouse models would allow for the dissection of howdifferent combinations of oncogene or tumor suppressorgenes might differentially affect the responses of the lungtumors to novel targeted therapeutics. It is interesting thatwith the exception of one elegant small-cell lung cancermouse model (50), all the other mouse lung cancer models todate only develop adenocarcinoma. Presently, there is no





genetic mouse model of squamous cell carcinoma of the lung.A better understanding of the cell of origin that give rise tolung squamous cell carcinoma and identification of uniquegene alterations that are specific to lung squamous cellcarcinoma might help the squamous cell carcinoma mousemodel development.

The recently discovered EGFR, BRAF , and PI3K mutationsinvolved in human lung cancer are yet to be exploited withappropriate murine models to determine their role in lungtumorigenesis. Murine lung cancer models with inducibleexpression of these mutations in the lung are in the processof being generated in several laboratories. In combination withthe existing oncogene and tumor suppressor mutant alleles thatare relevant to lung cancer, these novel alleles will help furtherour understanding of human lung cancer pathogenesis and aidin the development of novel therapeutics. Lastly, with the rapidadvances of lung cancer genome analyses, many additionalnovel mutations will no doubt be uncovered. Mouse modelingwill play a role in the elucidation and validation of these newpotential therapeutic targets.

Open Discussion

Dr. Thomas Lynch: What do you think the ultimatesignificance of this EGFRvIII mutation is going to be in lungcancer biology?

Dr. Wong: We need to screen additional adenocarcinomasto see if these mutations are only found in squamous cellcarcinomas. That would then be a very interesting question asto why this mutation is only in squamous cell carcinomasversus the kinase mutations, which are predominantly found inadenocarcinomas. That may suggest that there are differentbiological forces driving it.

Dr. Lynch: Question for Dr. Meyerson or Dr. Haber: Haveeither of your labs looked for the vIII mutation in other groupsof lung cancer patients?

Dr. Daniel Haber: We have. We looked by RT-PCR atgefitinib responders, nonresponders, and in broad samples. Wefound it in brain tumors but not otherwise.

Dr. Matthew Meyerson: We have found it. We know theyare erbB3 mutations because we see novel genomic DNArearrangements that we can’t imagine would be artifacts.

Dr. Wong: Just to go back to the question of whetherthere is any clinical relevance to the vIII mutation, Dr.Meyerson and I have been talking about what patientpopulations to look at, among patients who initiallyresponded to erlotinib or gefitinib. One interesting popula-tion would be the BR.21 trial in which there was the 4% to5% response rate in the nonadenocarcinoma group. Onemight think that some of those patients might harbor theEGFRvIII mutation.

Dr. Glenwood Goss: Ian Lorimer, who works in myinstitution, was one of the original investigators on the vIIImutation [Clin Cancer Res 1995;1:859–64]. So we can do it inour lab; we will look at the mutation.

Dr. Pasi Janne: If you look at the vIII mutations in mice inthe adenocarcinomas versus the kinase mutations, do they lookhistologically the same or different?

Dr. Wong: They do look histologically the same. We havesome preliminary evidence in our laboratory suggesting thatadditional tumor suppressor mutations can actually transforman adenocarcinoma into a squamous cell carcinoma, at least inmouse.

Dr. Bruce Johnson: Do you see histologic evidence ofapoptosis in the treated mice? Also, in your controls versus yourexperimental group, is it actually regression or is it a growthdelay?

Dr. Wong: It is actually regression. When you treat the micewith erlotinib, after 2 days of treatment there is strikingapoptosis. After a week, there is a big hole with evidence ofscarring where the tumor should be. So there is activeremodeling taking place where the tumor was.

Dr. John Heymach: These tumors were reversible when youtook away the doxycycline—was it with just one of the alleles ormore? It is intriguing and reminiscent of the work by LyndaChin and Ron DePinho [Nature 1999;400:468–72] that driv-ing Ras you get tumors, but take it off and those tumors regress.Do you see any cytogenetic abnormalities in your tumors yet,

Fig. 2. Example of a conditionalbitransgenic inducible mouse using thetetracycline regulatory system.




and do you think that it is going to be reversible until you startto get additional cytogenetic abnormalities accumulating?

Dr. Wong: With the Ras experiments that you describe inthe lab of Ron DePinho and Lynda Chin, when they look atthose cell lines there are not a lot of cytogenetic abnormalities.We are doing those kinds of experiments now, trying to cultureout these primary tumors. We find it very difficult to do at themoment, unless we put p53 deficiency in these tumors; then itbecomes a lot easier. We are going to be looking at genomicinstability in these tumors.

Dr. Jeffrey Settleman: I have a general philosophicalquestion about mouse models in lung cancer. These aremodels where in a few months, you have a single gene-drivenevent that results in a rapidly growing tumor. How does thatcompare to what is happening in human lung cancer patientswhere maybe it is 30 years that a tumor is developing? Whatdo you think will ultimately be the value of drug studies inthat setting?

Dr. Wong: Obviously, the human lung tumors will havemany more mutations. They are a lot more complex, withaneuploidy and multiple chromosomal aberrations. In themouse model, you don’t have any of that when it is just drivenby a single oncogene. With the other tumor suppressor allelesthat we have, we can try to recapitulate the human condition byintroducing p53 deficiency, pTEN deficiency, or telomeredysfunction into this model. I think the proper number ofalleles that you can put into a mouse and start rebuilding whatyou see in people as tumor progression we may be able torecapitulate the condition. Presently, this is a very simplisticmodel and we are going to try to build on top of that.

Dr. Heymach: One of the values of doing mouse modelsas opposed to in vitro studies is that, in mouse models, youhave the capability of understanding interactions betweendifferent cellular compartments and using that as a drugscreening strategy. If the only question is how well does atarget get inhibited, in vitro studies allow you to do muchhigher throughput studies. I have seen in vivo data thatmatch what is seen in vitro, but I haven’t seen anythingthat gives us qualitatively new information. When you arelooking for specific targets and asking biochemical andpathway questions, I think those studies can be better donein cell lines.

Dr. Paul Bunn: In my opinion, an orthotopic model withhuman tumors is a much better model because it has all thegenetic changes. Having a single genetic change and then tryingto do drug testing in that model is almost ludicrous. You canlearn about that one gene but it is not going to help you withhuman therapy.

Dr. Johnson: Let me respond to that. If a genomic studyhas identified some mutated gene that is important in bothinitiating and driving the tumor, then perhaps you can learnsomething about its role as a necessary but not sufficient step.To find out if, by inhibiting that particular genetic lesion, thereis an impact on tumor growth is a worthwhile initial step.Then following up with orthotopic models or whatever youthink is important to actually verify its relevance in humans isa second step.

Dr. Bunn: I don’t think providing scientific value is thequestion. Is it a good way to screen for drugs? I don’t think it is.But I think you learn something.

References1. Jemal A,Tiwari RC, MurrayT, et al. Cancer statistics,2004. CACancerJClin 2004;54:8^29.

2. Greulich H, ChenTH, FengW, et al. Oncogenic trans-formation by inhibitor-sensitive and -resistant EGFRmutants. PLoSMed 2005;2:e313.

3. LynchTJ, Bell DW, Sordella R, et al. Activatingmuta-tions in the epidermal growth factor receptor underly-ing responsiveness of non-small-cell lung cancer togefitinib. NEngl JMed 2004;350:2129^39.

4. LynchTJ, Adjei AA, Bunn PA, Jr., et al. Novel agentsin the treatment of lung cancer: conference summarystatement. Clin Cancer Res 2004;10:4199^204s.

5. Paez JG, Janne PA, Lee JC, et al. EGFR mutations inlung cancer: correlation with clinical response to gefi-tinib therapy. Science 2004;304:1497^500.

6. PaoW, Miller VA, Kris MG. ‘Targeting’ the epidermalgrowth factor receptor tyrosine kinase with gefitinib(Iressa) in non-small cell lung cancer (NSCLC). SeminCancer Biol 2004;14:33^40.

7. Kallioniemi A, Kallioniemi OP, Sudar D, et al. Com-parative genomic hybridization for molecular cyto-genetic analysis of solid tumors. Science 1992;258:818^21.

8. Lucito R, HealyJ, AlexanderJ, et al. Representationaloligonucleotide microarray analysis: a high-resolutionmethod to detect genome copy number variation.Genome Res 2003;13:2291^305.

9. Zhao X, Li C, Paez JG, et al. An integrated view ofcopy number and allelic alterations in the cancer ge-nome using single nucleotide polymorphism arrays.Cancer Res 2004;64:3060^71.

10. Meuwissen R, Berns A. Mouse models for humanlung cancer. Genes Dev 2005;19:643^64.

11. Glasser SW, KorfhagenTR,Wert SE, et al. Geneticelement from human surfactant protein SP-C geneconfers bronchiolar-alveolar cell specificity in trans-genic mice. AmJPhysiol 1991;261:L349^56.

12. Stripp BR, Sawaya PL, Luse DS, et al. cis-actingelements that confer lung epithelial cell expression ofthe CC10 gene. JBiol Chem1992;267:14703^12.

13. Ray MK, Magdaleno SW, Finegold MJ, DeMayoFJ. cis-acting elements involved in the regulationof mouse Clara cell-specific 10-kDa protein gene.In vitro and in vivo analysis. J Biol Chem 1995;270:2689^94.

14. SundayME, Haley KJ, Emanuel RL, et al. Fetal alve-olar epithelial cells contain [D-Ala(2)]-deltorphin I-likeimmunoreactivity: y- and A-opiate receptors mediateopposite effects in developing lung. Am JRespir CellMol Biol 2001;25:447^56.

15. Carraresi L,Tripodi SA, Mulder LC, et al. Thymic hy-perplasia and lung carcinomas in a line of mice trans-genic for keratin 5-driven HPV16 E6/E7 oncogenes.Oncogene 2001;20:8148^53.

16.Wikenheiser KA, Clark JC, Linnoila RI, StahlmanMT, Whitsett JA. Simian virus 40 large T antigen di-rected by transcriptional elements of the humansurfactant protein C gene produces pulmonaryadenocarcinomas in transgenic mice. Cancer Res1992;52:5342^52.

17.Wikenheiser KA,WhitsettJA.Tumor progression andcellular differentiation of pulmonary adenocarcinomasin SV40 largeTantigen transgenic mice. Am J RespirCell Mol Biol 1997;16:713^23.

18. DeMayo FJ, Finegold MJ, HansenTN, Stanley LA,Smith B, Bullock DW. Expression of SV40 Tantigenunder control of rabbit uteroglobin promoter in trans-genic mice. AmJPhysiol 1991;261:L70^6.

19. Magdaleno SM, Wang G, Mireles VL, Ray MK,Finegold MJ, DeMayo FJ. Cyclin-dependent kinaseinhibitor expression in pulmonary Clara cells trans-formed with SV40 largeTantigen in transgenic mice.Cell Growth Differ1997;8:145^55.

20. Ehrhardt A, Bartels T, Geick A, Klocke R, Paul D,

Halter R. Development of pulmonary bronchiolo-alve-olar adenocarcinomas in transgenicmice overexpress-ing murine c-myc and epidermal growth factor inalveolar type II pneumocytes. Br J Cancer 2001;84:813^8.

21. ChenYQ, ZhouYQ, Fisher JH,Wang MH. Targetedexpression of the receptor tyrosine kinase RON in dis-tal lung epithelial cells results in multiple tumor forma-tion: oncogenic potential of RON in vivo. Oncogene2002;21:6382^6.

22. Kerkhoff E, Fedorov LM, Siefken R, Walter AO,PapadopoulosT, Rapp UR. Lung-targeted expressionof the c-Raf-1 kinase in transgenic mice exposes anovel oncogenic character of the wild-type protein.Cell Growth Differ 2000;11:185^90.

23.Morris GF, Hoyle GW, AthasGB, et al. Lung-specificexpression in mice of a dominant negative mutantform of the p53 tumor suppressor protein. J La StateMed Soc1998;150:179^85.

24. Melton DW. Gene targeting in the mouse. BioEs-says16:633^8.

25. Capecchi MR. The new mouse genetics: alteringthe genome by gene targeting. Trends Genet 1989;5:70^6.

26. Zanesi N, Mancini R, Sevignani C, et al. Lung can-cer susceptibility in Fhit-deficient mice is increasedby Vhl haploinsufficiency. Cancer Res 2005;65:6576^82.

27. JacksT, Fazeli A, Schmitt EM, Bronson RT, GoodellMA,Weinberg RA. Effects of an Rb mutation in themouse. Nature1992;359:295^300.

28. KreidbergJA, Sariola H, LoringJM, et al.WT-1is re-quired for early kidney development. Cell 1993;74:679^91.

29. Johnson L, Mercer K, Greenbaum D, et al. Somaticactivation of the K-ras oncogene causes early onsetlung cancer in mice. Nature 2001;410:1111^6.





30. Lewandoski M. Conditional control of gene expres-sion in the mouse. Nat Rev Genet 2001;2:743^55.

31. DeMayo FJ,Tsai SY. Targeted gene regulation andgene ablation. Trends Endocrinol Metab 2001;12:348^53.

32. ZhouY, Zhang X, Ebright RH. Identification of theactivating region of catabolite gene activator protein(CAP): isolation and characterization of mutants ofCAP specifically defective in transcription activation.Proc Natl Acad Sci US A1993;90:6081^5.

33. Gossen M, Freundlieb S, Bender G, Muller G,HillenW, Bujard H. Transcriptional activation by tetra-cyclines in mammalian cells. Science 1995;268:1766^9.

34. Furth PA, St Onge L, Boger H, et al. Temporal con-trol of gene expression in transgenic mice by a tetracy-cline-responsive promoter. Proc Natl Acad Sci U S A1994;91:9302^6.

35. Tichelaar JW, Lu W, Whitsett JA. Conditionalexpressionof fibroblast growth factor-7 in the developingandmature lung. JBiol Chem 2000;275:11858^64.

36. Fisher GH,Wellen SL, Klimstra D, et al. Inductionand apoptotic regression of lung adenocarcinomas byregulation of a K-Ras transgene in the presence andabsence of tumor suppressor genes. Genes Dev2001;15:3249^62.

37. Gu H, ZouYR, Rajewsky K. Independent control ofimmunoglobulin switch recombination at individualswitch regions evidenced through Cre-loxP-mediatedgene targeting. Cell 1993;73:1155^64.

38. KuhnR,SchwenkF,AguetM,RajewskyK. Induciblegene targeting inmice. Science1995;269:1427^9.

39. Kilby NJ, Snaith MR, Murray JA. Site-specificrecombinases: tools for genome engineering. TrendsGenet 1993;9:413^21.

40. Perl AK,Wert SE, Nagy A, Lobe CG,Whitsett JA.Early restriction of peripheral and proximal cell line-ages during formation of the lung. Proc Natl Acad SciUS A 2002;99:10482^7.

41. O’Gorman S, Dagenais NA, Qian M, Marchuk Y.Protamine-Cre recombinase transgenes efficiently re-combine target sequences in the male germ line ofmice, but not in embryonic stem cells. Proc Natl AcadSci US A1997;94:14602^7.

42. Schwenk F, Baron U, Rajewsky K. A cre-transgenicmouse strain for the ubiquitous deletion of loxP-flanked gene segments including deletion in germcells. Nucleic Acids Res1995;23:5080^1.

43. Jackson EL,Willis N, Mercer K, et al. Analysis oflung tumor initiation and progression using condition-al expression of oncogenic K-ras. Genes Dev 2001;15:3243^8.

44. Guerra C, Mijimolle N, Dhawahir A, et al. Tumor in-duction by an endogenous K-ras oncogene is highlydependent on cellular context. Cancer Cell 2003;4:111^20.

45. Olive KP,Tuveson DA, Ruhe ZC, et al. Mutant p53gain of function in two mouse models of Li-Fraumenisyndrome. Cell 2004;119:847^60.

46. Lang GA, IwakumaT, SuhYA, et al. Gain of functionof a p53 hotspot mutation in a mouse model of Li-Fraumeni syndrome. Cell 2004;119:861^72.

47.WangY, Zhang Z, Lubet R,You M. Tobacco smoke-induced lung tumorigenesis in mutant A/J mice withalterations in K-ras, p53, or Ink4a/Arf. Oncogene2005;24:3042^9.

48.Tuveson DA, ShawAT,Willis NA, et al. Endogenousoncogenic K-ras(G12D) stimulates proliferation andwidespread neoplastic and developmental defects.Cancer Cell 2004;4:357^87.

49. Jackson EL, Olive KP,Tuveson DA, et al. The differ-ential effects of mutant p53 alleles on advancedmurine lung cancer. Cancer Res 2005;65:10280^8.

50. Meuwissen R, Linn SC, Linnoila RI, ZevenhovenJ, Mooi WJ, Berns A. Induction of small cell lungcancer by somatic inactivation of both Trp53 andRb1 in a conditional mouse model. Cancer Cell2003;4:181^9.




2006;12:4396s-4402s. Clin Cancer Res Amit Dutt and Kwok-Kin Wong Mouse Models of Lung Cancer

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