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JNCI
Vol. 101, Issue 1
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January 7, 2009
 NEWS
resistance to gemcitabine, thestandard therapy for pancreaticcancer, and sensitivity to mito-mycin C, a rarely used treat-ment. The patient who firstfailed gemcitabine therapy  received mitomycin C and hasbeen in remission for morethan 2 years. The trial’s principal inves-tigator, Manuel Hidalgo, M.D., Ph.D., stressed thattumorgrafts, at present, canbe of only limited use forindividualizing patient treat-ment because of the time andresources necessary to createtumorgraft banks. Many patients die even before theirmouse models are ready fordrug testing. “All we’re trying to achievehere is proof of concept, that thereis some prediction,” Hidalgo said. Tumorgrafts will be used mainly to screennew drug candidates for activity and todiscover biomarkers that can predict drugeffects, he said. For example, an April
Cancer Research
report by Hidalgo’s groupdescribed using pancreatic tumorgrafts toderive a 25-gene signature that success-fully predicted mouse sensitivity to epi-dermal growth factor receptor inhibitors. A human trial prospectively testing theepidermal growth factor receptor bio-markers is pending.Patient selection using such biomarkersis widely viewed as the future of cancertherapy, although few prospective bio-marker trials have taken place. “If we really  want to make progress, that’s the way togo,” Hidalgo said. And for biomarker discovery, tumor-grafts are much easier to work with thanprimary tumor tissue because of the abun-dance and renewability of the tumormaterial. “You can go back and you can
From Human to Mouse and Back: “Tumorgraft”Models Surge in Popularity
By Ken Garber
M
ouse xenograft mod-els of cancer, under-standably, have aterrible reputation. Althoughresearchers and companiesroutinely use these humantumors in mice for preclinicaldrug testing, individual modelspoorly predict how drugs willact in the clinic. Retrospectivere views published by theNational Cancer Institute in2001 and the National CancerInstitute of Canada in 2003came to the same conclusion:Drugs that work against cancerin xenograft mice rarely work in people with the same tumor, with the exception of lungand possibly ovarian cancer.“There’s this mantra: ‘Xenografts don’tpredict for human effects,” said PeterHoughton, Ph.D., a cancer researcher atthe St. Jude Children’s Research Hospitalin Memphis, Tenn.But not all xenograft models poorly predict drug effects. In direct transfer, or“explant,” xenografts, recently dubbed“tumorgrafts,” tumors taken from patientsare chopped into fragments slightly smaller than a pencil eraser and implanteddirectly into immunodeficient, or “nude,”mice. (Standard xenografts use perma-nent cell lines, not primary tumors, as thesource of tumor material.) In several ret-rospective studies, drug effects in suchmice closely mirror human effects whendifferences in drug activity between spe-cies are taken into account. “They work extremely well, as long as you factor inthe drug systemic exposure,” Houghtonsaid.Now, these direct transfer xenografts,first used in the 1970s, are surging in popu-larity, as research groups and drug compa-nies apply new molecular techniques to
New drug testing in mouse models by the NCI-supported Pediatric Preclinical TestingProgram. The 3-year-old program, which has already sent several drugs into clinical trials,relies heavily on explant xenograft models.
these mice to begin creating personalizedcancer treatments.
Of Mice and Men
Explant models are, to be sure, more tediousto create. Human tumors must be freshly implanted “[and] they’re hard to grow,”said David Sidransky, M.D., a cancerresearcher at Johns Hopkins Hospital inBaltimore, at the 2008 American Associationfor Cancer Research (AACR) conferenceon molecular diagnostics in cancer thera-peutic development. But “once growing,they’re very predictive of response.” Tumorgrafts (Sidransky’s preferredterm) are already being used in clinical set-tings. At Johns Hopkins, researchers takeresected pancreatic tumors from newly diagnosed patients, implant them into mul-tiple nude mice, and then test 10 approvedand experimental treatments against thesepersonal tumorgraft banks. Results are usedto guide therapy when the patients relapse(
 J Natl Cancer Inst 
2007;99:1057.) At the AACR meeting, Sidransky described onecase in which the mouse model predicted
 
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repeat and repeat and repeat if you think  you’ve found something that’s real, asopposed to patient samples where you getbasically one shot at doing something,”said Houghton.
Power To Predict
 The drug industry, in the last few years,has recognized the potential utility of tumorgrafts, and drug candidate screeningusing tumorgraft models is now becomingroutine. “Some of the large pharmaceuticalcompanies have established very large pan-els of [tumorgrafted mice] in the particularcancer types that they’re interested in … tosee how their drugs are working,”Houghton said. For example, at the AACRmeeting, Frederic de Sauvage, Ph.D., of Genentech in South San Francisco, Calif.,described using such models to decipherhow experimental inhibitors of the hedge-hog signaling path- way target tumorconnective tissue.How strong isthe evidence thattumorgrafts canaccurately predictactual human response to drugs, giventhat standard xenografts have done sopoorly? Heinz-Herbert Fiebig, M.D.,Ph.D., a medical oncologist and cancerresearcher in Freiburg, Germany, hasbeen creating direct transfer xenograftmodels since the late 1970s and retro-spectively testing their predictive powerfor human treatments. Fiebig has reportedconsistently good results. For example, in2004 he reported in the
 European Journal of Cancer 
that his xenografts correctly predicted response in 90% of patients (19of 21 tumors) and resistance in 97% (57of 59). The failure of standard cell linede-rived xenografts to predict, said Fiebig, iscaused by changes in culture. Tumor cellsin culture tend to undergo selective pres-sure to become less differentiated andmore homogeneous, so that the tumorsthey give rise to in mice no longer faith-fully reflect the original tumor. Mosttumorgrafts, on the other hand, closely mirror their parent human tumors micro-scopically, he said. And Houghton’s grouphas shown that they change minimally onthe molecular level, when taking intoaccount the absence of surrounding humanconnective tissue, or stroma. “They havemaintained the [gene] expression profilesand genomic profiles very, very accu-rately,” Houghton said. Tumorgrafts, though, have their ownproblems, including the high transplanta-tion failure rate and the labor required formultiple transfers, or “passages,” frommouse to mouse (needed to propagateeach human tumor in multiple mice). Theoverall “take rate” varies from 40% to60% for nonsmall-cell lung cancer, coloncancer, and melanomas down to 12% 20% for breast cancer and just 3% forprostate cancer, said Fiebig. So tumor-grafts cost much more than conventional xenografts.Both Houghtonand Fiebig stress theimportance of limit-ing the number of mouse-to-mouse pas-sages to prevent thetumors in the mousehost from gradually mutating away fromthe parent tumors. Frozen tumors make upa permanent repository that allows unlim-ited replenishment. “As a measure of pre-caution, we go back to the frozen masterstock after 1012 passages” to generate newmouse tumors, said Fiebig.
Mining the Model
 The main knock against tumorgrafts is theneed for more prospective trials showingthat they predict drug activity in humans.Several such tumorgraft validation studiesare ongoing or planned. But for many researchers, the evidence is already con- vincing, and use of the models is growing.In 2005, Houghton’s group reported aclinical trial in pediatric neuroblastoma with the drug topotecan, guided by indi- vidual drug level adjustments and dosingschedules tested in a direct transfer mouse xenograft model. “The response rates inthe clinic were exactly as predicted fromthe modelsa 60% response rate,”Houghton said.On the basis of those results, the NCI isfunding a multicenter drug developmentinitiative, the Pediatric Preclinical TestingProgram, which uses 60 mouse tumormodels, with most of the tumors takendirectly from patients rather than from cellculture. “Basically the drugs that work inhuman cancers, childhood cancers, work inthe mice,” said Houghton. “Now we’reusing those models to identify new drugs, which are being fast-tracked into pediatrictrials.”Similar work is under way at Fiebig’scompany, Oncotest GmbH in Freiburg,Germany, which now has a bank of morethan 200 tumors established directly frompatients. Oncotest has also derived morethan 40 cell lines from these tumors, whichare used for biochemical and molecularstudies. U.S. drug companies especially have shown “a hugeinterest” in these toolsfor drug development,said Fiebig, beginning4 years ago when geneexpression chips werefirst used to character-ize these tumors on amolecular level.Personalized ther-apy, based on such gene expression signa-tures, is the ultimate goal. Fiebig hasidentified signatures in his models thatpredict response to 14 cytotoxic drugs andtwo targeted agents, bevacizumab andcetuximab. Overall response rates for bothtargeted drugs and most of the cytotoxics were close to those of past clinical results.Fiebig is planning to validate these signa-tures further using samples from clinicaltrials that have been completed and for which the outcomes are known, and even-tually in prospective clinical trials.In 2006, Sidransky and Hidalgo alsocofounded a company, BioMerk, to com-mercialize the tumorgraft platform. Thefollowing year, Champions Biotechnology in Arlington, Va., acquired BioMerk,later naming Sidransky chairman andHidalgo chief scientific officer.Champions, in collaboration with South Texas Accelerated Research Therapeutics,a San Antonio company, is establishing
Peter Houghton, Ph.D.
“There’s this mantra:‘Xenografts don’t predict for human effects.’ ” 
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