FluorescenceVisualization Detection of Field Alterations in Tumor Margins of Oral Cancer Patients CatherineF.Poh, 1,2,3 LeweiZhang, 1 DonW.Anderson, 5 J.ScottDurham, 5 P.MicheleWilliams, 1,3 RobertW.Priddy, 1 KenW.Berean, 6 SamsonNg, 1 OliviaL.Tseng, 7 CalumMacAulay, 4 andMiriamP.Rosin 2,7 Abstract Purpose: Geneticallyalteredcellscouldbecomewidespreadacrosstheepitheliumofpatients withoralcancer,ofteninclinicallyandhistologicallynormaltissue,andcontributetorecurrent disease.Molecularapproacheshavebeguntoyieldinformationoncancer/riskfields;tissueoptics couldfurtherextendourunderstandingofalterationtophenotypeasaresultofmolecularchange. Experimental Design: Weusedasimplehand-helddeviceintheoperatingroomtodirectly visualizesubclinicalfieldchangesaroundoralcancers,documentingalterationtofluorescence. Atotalof122oralmucosabiopsieswereobtainedfrom20surgicalspecimenswitheachbiopsy beingassessedforlocation,fluorescencevisualization(FV)status,histology,andlossof heterozygosity(LOH;10markersonthreeregions:3p14,9p21,and17p13). Results: AlltumorsshowedFVloss(FVL).For19ofthe20tumors,thelossextendedinat leastonedirectionbeyondtheclinicallyvisibletumor,withtheextensionvaryingfrom4to 25mm.Thirty-twoof36FVLbiopsiesshowedhistologicchange(including7squamouscell carcinoma/carcinomas in situ,10severedysplasias,and15mild/moderatedysplasias)compared with1ofthe66FVretained(FVR)biopsies.Molecularanalysisonmarginswithlow-gradeorno dysplasiashowedasignificantassociationofLOHinFVLbiopsies,withLOHat3pand/or9p (previouslyassociatedwithlocaltumorrecurrence)presentin12of19FVLbiopsiescompared with3of13FVRbiopsies(P =0.04). Conclusions: Thesedatahave,forthefirsttime,shownthatdirectFVcanidentifysubclinical high-riskfieldswithcancerousandprecancerouschangesintheoperatingroomsetting. In 1953, Slaughter published a hallmark article in which he emphasized the importance of examining the field surrounding oral cancers for both risk assessment and management of this disease (1). There has been extensive research in this area since then, more recently, using molecular technology. It is becoming increasingly apparent that genetically altered cells could become widespread across the epithelium of patients with oral cancer, into clinically and histologically normal tissue, and that these cells could drive the process of field cancerization (2, 3). In recognition of this, surgeons try to remove oral squamous cell carcinomas (SCC) with a significant width of surrounding normal-looking oral mucosa, if anato- mically allowed. However, the occult disease varies in size and a wealth of evidence suggests that it frequently extends beyond the tumor clearance. This extension may be responsible for the high rate of recurrence of carcinomas at the primary site (f10-30% of cases; refs. 4–9). There is a pressing need to develop new approaches that can be easily used in clinical prac- tice to facilitate the detection of these clinically occult fields. One such new approach may involve the use of tissue optics. The association of cancer development with the loss of normal tissue autofluorescence has been reported for a number of tissues and organs (10 – 15). More recently, visual aids using optical methods to detect such loss have been shown to reveal premalignant and malignant lesions that are not detected by unaided eyes (16 – 18). We have reported the development of a simple hand-held device that facilitated the detection of autofluorescence loss in both visible and occult high-risk oral lesions through direct fluorescence visualization (FV; refs. 17, 18). The interaction of light with tissue has generally been found to highlight changes in the structure and metabolic activity of the areas optically sampled. Specifically, the loss of autofluorescence is believed to reflect a complex mixture of alterations to intrinsic tissue fluorophore distribution, such as the breakdown of the collagen matrix and a decrease in flavin adenine dinucleotide concentration due to tissue remodeling and increased metabolism associated with neo- plastic development. Correspondingly, structural changes in tissue morphology associated with neoplastic development Imaging, Diagnosis, Prognosis Authors’Affiliations: 1 FacultyofDentistry,UniversityofBritishColumbia, Departmentsof 2 CancerControlResearch, 3 OralOncology,and 4 CancerImaging, BritishColumbiaCancerAgency,Departmentsof 5 Surgery(Otolaryngology)and 6 Pathology,VancouverGeneralHospital,Vancouver,BritishColumbia,Canada,and 7 SchoolofKinesiology,SimonFraserUniversity,Burnaby,BritishColumbia,Canada Received5/30/06;revised7/25/06;accepted8/10/06. Grant support: R01DE13124andR01DE17013fromtheNationalInstituteof DentalandCraniofacialResearchandMOP-77663fromtheCanadianInstitutes ofHealthResearch,andaClinicianScientistAwardfromtheCanadianInstitutesof HealthResearch(C.F.Poh). Thecostsofpublicationofthisarticleweredefrayedinpartbythepaymentofpage charges.Thisarticlemustthereforebeherebymarked advertisement inaccordance with18U.S.C.Section1734solelytoindicatethisfact. Requestsforreprints: MiriamP.Rosin,BCOralCancerPreventionProgram,BC CancerAgency/CancerControlResearchCentre,675West10thAvenue,Vancouver, BritishColumbia,CanadaV5Z1L3.Phone:604-675-8078;Fax:604-675-8180; E-mail:Miriam__Rosin@shaw.ca. F 2006AmericanAssociationforCancerResearch. doi:10.1158/1078-0432.CCR-06-1317 www.aacrjournals.org ClinCancerRes2006;12(22)November15,2006 6716 Downloaded from http://aacrjournals.org/clincancerres/article-pdf/12/22/6716/1921523/6716.pdf by guest on 17 June 2022