InacollaborativeresearchprojectthatIinitiatedbetweenthelabsofProf.PaulSondelandProf.Paul Harariduringmyresidencytraining,weexploredanovelapproachtoaugmentinganti-tumorimmuneresponse bycombiningtwoestablishedcancertreatments,radiationtherapy(RT)andtumor-specificantibodies(mAbs). Insingle-tumormurinemodelsofmelanoma,neuroblastoma,andheadandnecksquamouscellcarcinomawe observedacooperativeanti-tumorinteractionbetweenlocalRTandintratumoral(IT)injectionoftumor-specific mAbresulting,inpart,fromenhancedantibody-dependentcell-mediatedcytotoxicity.Tofurtheraugmentthis response,weinvestigatedcombinedRTandIT-immunocytokine(IC),afusion-proteinlinkingtumor-specific mAbtoIL2.Withthisweobservedagreateranti-tumorimmuneresponseresultingincompleteregressionof established(~5weekpostengraftment)tumorsinmostanimalsandamemoryTcellresponsethatrejected re-challengewithsimilartumorcells.ThisdemonstratedthatcombinedRT+IT-ICcanelicitapotent?in situ?tumorvaccineresponse.Tcellcheckpointblockadeisbecomingastandardofoncologiccarein certaincancersettingsandwethereforetestedthebenefitofaddinglocalRTandIT-ICtosystemiccheckpoint blockadeinourmurinemelanomamodel.Inmicebearinglargeprimarytumors(~7weekpostengraftment)or microscopicdistantsitesofdisease(IVinjectedonthedayofRT),thetriple-combinationofprimarytumorRT followedbyIT-ICinjectionofthistumorandsystemicTcellcheckpointblockadewithanti-cytotoxicT- lymphocyteantigen-4(CTLA-4)antibodyimprovedanimalsurvivalcomparedtocombinationsofanytwoof thesethreeinterventions.However,inthepresenceofestablished(~3-weekpostengraftment)distantsitesof diseasewehavenowobservedakeylimitationtoourinsituvaccinationstrategy.Inthepresenceofanun- radiatedsecondmacroscopictumor,thecombinationofprimarytumorRT+primarytumorIT-IC injectionisnomoreeffectivethanprimarytumorRTalone.Thissuggeststhatanuntreated,distant secondtumormaysuppressthegenerationofananti-tumorimmuneresponseintheprimarytumorfollowing RT+IT-IC.Wedescribethisas?concomitantimmunetolerance?(CIT). HereIhypothesizethat1)insituvaccinationmaybeachievedusinglocalRTandITinjectionoftumor- specificmAbwithIL2,2)regulatoryTcell(Tregs)harboredinanestablished,macroscopic,distanttumorsite mayexertCIT,and3)deliveringexternalbeamRT(EBRT)andIT-ICtoamouse?sprimarytumor,together withIV131I-NM404,amoleculartargetedradiotherapy(MTRT),willeliminateCITandenableaneffectiveinsitu vaccineresponsetoeradicatealltumorsinmicewithprimaryanddistantsitesofdisease. Iproposetotestthesehypothesesinmyindependentresearchlabusingestablishedtechniquesand animalmodelsthatIhaveusedtogeneraterobustpreliminarydata.Ihavesecuredthesupportofsix collaboratorswhoseexpertconsultationand/ormaterialassistancewillhelpensurethesuccessofthis researcheffort.Thisproposaldoesnotoverlapwithpriororongoingprojectsinthelabsofmyresearch mentors,whohaveexpressedtheircompletesupportformycareerdevelopment,myintellectual independence,andmypursuitoftheseresearchobjectivesintheindependentlabthatIwilllaunchinOctober. IhavepreviouslyappliedforthisDP5award(forRM-15-006)and,incomparisontothatpriorapplication, thecurrentproposalincludesmultiplerevisionsguidedbyreviewerfeedback.Thisapplicationalsoreflectsthe intervalpublicationofanadditionalfirst-authororiginalresearchmanuscriptthatisdirectlyrelatedtothe proposedresearch.Inanefforttoenhancethelevelofinnovationinthisproposal,Ihavecompletelyrevised Aim2and3.
Aim2 (previousAim2and3)nowspecificallyevaluatesthecriticalmechanisticunderpinningsof tumor-specificCITusingtransgenicmurinemodelstodeciphertheroleofTregsinthisnever-before-reported phenomenon.MeanwhileinAim3,IbuildfromourpreliminaryobservationthatCITcanbeovercomeby deliveringRTtothedistantsecondtumorsiteandhereIproposeanaltogetheruniqueapproachto circumventingCITbycombininganinsitutumorvaccinestrategywithasystemicmoleculartargeted radiotherapy(131I-NM404),whichmaysafelydeliverRTtoallsitesofmetastaticdiseasewithoutunduetoxicity orsystemicimmunesuppression.Inadditiontothesechangesspecificallyaimedatenhancingtheinnovation ofthisapplication,Ihavealsoincorporatedadditional,moreclinicallyrelevant,spontaneousandinducible transgenictumormodels(Aim1.3andthereafter).Thiswasdoneinanefforttofurtherstrengthenourrobust experimentalapproach.Tofurtherclarifythisapproach,Ihavealsomoreexplicitlyincorporatedastatistical planinthediscussionofeachsub-aim.WhilemypriorDP5proposalwasscoredquitefavorablyandadvanced tothefinalroundofinterviews,itismyhopethatwiththeserevisionsthecurrentapplicationmaywarrant funding.Thisawardwouldclearlyprovideatremendousaccelerationtomyindependentearlycareer developmentandthepaceatwhichmylabwillbeabletopursuetheexperimentsoutlinedherein.

Public Health Relevance

Weproposetoinvestigateanewapproachtocancertreatment,inwhichwedeliverradiationtoasingle tumorsiteandtheninjectthatsitewithanantibodythatspecificallybindstotumorcells.Ourpreliminarydata suggestthatthiscombinedtreatmentmaytriggerapatient?simmunesystemtorecognizeandattackthe targetedtumor.Thisapproachmayenhancetheresponsetoexistingcancerimmunotherapiesandthereby improvetreatmentofthemanycancersforwhichtumor-specificantibodiesalreadyexist.

Agency
National Institute of Health (NIH)
Institute
Office of The Director, National Institutes of Health (OD)
Type
Early Independence Award (DP5)
Project #
5DP5OD024576-03
Application #
9772144
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Miller, Becky
Project Start
2017-09-01
Project End
2022-08-31
Budget Start
2019-09-01
Budget End
2020-08-31
Support Year
3
Fiscal Year
2019
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Radiation-Diagnostic/Oncology
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Morris, Zachary S; Guy, Emily I; Werner, Lauryn R et al. (2018) Tumor-Specific Inhibition of In Situ Vaccination by Distant Untreated Tumor Sites. Cancer Immunol Res 6:825-834
Werner, Lauryn R; Kler, Jasdeep S; Gressett, Monica M et al. (2017) Transcriptional-mediated effects of radiation on the expression of immune susceptibility markers in melanoma. Radiother Oncol 124:418-426