Repository at Hanyang UniversityGRADUATE SCHOOL OF BIOMEDICAL SCIENCE AND ENGINEERING[S](의생명공학전문대학원)BIOMEDICAL SCIENCE(의생명과학과)Theses (Ph.D.)
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CRISPR-baseddevelopmentandscreeningofdeubiquitinasesforthesensitizationofcancercellstosmallmoleculetherapies
- Title
- CRISPR-baseddevelopmentandscreeningofdeubiquitinasesforthesensitizationofcancercellstosmallmoleculetherapies
- Author
- 아룬판디안찬드라세카란
- Alternative Author(s)
- 아룬판디안찬드라세카란
- Advisor(s)
- SureshRamakrishna
- Issue Date
- 2021.2
- Publisher
- 한양대학교
- Degree
- Doctor
- Abstract
- Despiteadvancesincancertherapies,cancerisstillcausingmillionsofdeathsworldwide.Thestandardchemotherapeutictreatmentssuchaslowmolecularweightmoleculesfortreatingvariouscancershavenotbeensuccessful.Further,incrementinthedrugconcentrationisnotpossibleasthedosageisalreadyreachedthemaximumtolerablelevels.Raiseindosagelevelsmaycausedamagestovariousnormaltissuesororgans.Currentdevelopmentsincludetargeted,patient-specificorprecisiontherapiesarealternativeapproachesbuttheoutcomesareverybleak. Deubiquitinatingenzymes(DUBs)areafamilyofproteasesthatremoveubiquitinfromproteinsundergoingdegradation.DUBsplayseveralfunctionalrolesotherthandeubiquitinationsuchasconferringresistancetocancertherapiesandmediatestumorprogression.ConventionalscreeningmethodsforDUBshavelimitations.Aloss-of-functionstudybasedontheknockoutofDUBgenesinmammaliancellscanprovideanexcellentmodelforexploringDUBfunction.Here,IusedCRISPR-Cas9toperformgenome-scaleknockoutoftheentiresetofgenesencodingDUBsandthensystematicallyscreenedforDUBsthatinvolvedinthesmallmoleculeresistancemechanism. Toprovidetheproof-of-conceptthatoverexpressionofDUBsimpedessmallmolecule-basedtherapies,IexploitedourDUBknockoutlibrarytoscreenforprobableDUBcandidate(s)thatinvolveinresistancetoYM155,ananticancercompound.IhaveidentifiedUSP32asapotentialmolecularmarkerforYM155resistancebynegativelyregulatingSLC35F2,acarrierforYM155.IshowedthatUSP32andSLC35F2haveanegativecorrelationfromapanelofcancercelllinestested.CRISPR-mediatedgenerationofUSP32knockoutinMCF7andBT474cellspromotesYM155-drivenDNAdamagebyupregulatingSLC35F2.Importantly,USP32depletionenhancedYM155-mediatedsuppressionofcancerprogressioninvitroandinvivo. IalsoreportedthatourCRISPR-Cas9DUBknockoutlibraryapproachtoidentifyaregulatorforCdc25AtosensitizeATRinhibitors-basedcancertherapies.IfoundUSP3asdeubiquitinaseofCdc25A.DepletionofUSP3reducestheproteinlevelofCdc25AandsensitizesthecancercellstotheATRinhibitors,AZ20andcaffeine. Together,applyingmystrategytootherpotentialsmallmoleculesscheduledforcancertherapymayincreaseitsclinicalefficacy,especiallyduringpostclinicalstages.Ibelievethatthisapproachwillbehelpfultounderstandandovercometheresistanceproblemsexhibitedbyseveralsmallmoleculesandmayrewritetheirtherapeutics.
- URI
- http://hanyang.dcollection.net/common/orgView/200000485415https://repository.hanyang.ac.kr/handle/20.500.11754/188241
- Appears in Collections:
- GRADUATE SCHOOL OF BIOMEDICAL SCIENCE AND ENGINEERING[S](의생명공학전문대학원) > BIOMEDICAL SCIENCE(의생명과학과) > Theses (Ph.D.)
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