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Final PresentationFinal Presentation
Anonymous student MK and NMAnonymous student MK and NM
Image credits: Left, courtesy of Torsten Witmann, used with permission. Right, Donald Bliss and Sriram Subramaniam, National Library of Medicine, NIH.
OUR MISSIONOUR MISSIONCANCERCANCER
METASTASISMETASTASIS
COMMON DISEASE, KILLS 500,000 A YEARCOMMON DISEASE, KILLS 500,000 A YEAR ABNORMAL CELL GROWTH AND DIVISIONABNORMAL CELL GROWTH AND DIVISIONCAN ORIGINATE IN DIFFERENT ORGANSCAN ORIGINATE IN DIFFERENT ORGANS
SPREAD FROM ORIGINAL TUMORSPREAD FROM ORIGINAL TUMORTRAVELS THROUGH BLOOD/LYMPHTRAVELS THROUGH BLOOD/LYMPHMAIN CAUSE OF DEATHMAIN CAUSE OF DEATH
OUR MISSIONOUR MISSIONDIAGNOSTIC IMAGING TOOLSDIAGNOSTIC IMAGING TOOLSBETTER UNDERSTANDING OF METASTASISBETTER UNDERSTANDING OF METASTASISPATTERNS OF MOVEMENT, TIMINGPATTERNS OF MOVEMENT, TIMING
LEAD TO DEVELOPMENTS IN TREATMENTSLEAD TO DEVELOPMENTS IN TREATMENTSCOULD BE USED FOR KILLING CANCER CELLSCOULD BE USED FOR KILLING CANCER CELLS
A SOLUTIONA SOLUTION
IMPACTIMPACT
SCIENCE BEHIND ITSCIENCE BEHIND ITSIGNALSSIGNALS OVEREXPRESSION OF ENZYMESOVEREXPRESSION OF ENZYMES
COXCOX--2: PREVENTS APOPTOSIS2: PREVENTS APOPTOSISMMPMMP--1: BREAKS BASEMENT MEMBRANE1: BREAKS BASEMENT MEMBRANE
Courtesy of Larry Marnett, Ph.D. Used with permission. http://www.mc.vanderbilt.edu/lens/article/?id=49&pg=999
Image: http://www.rcsb.org/
SCIENCE BEHIND ITSCIENCE BEHIND ITDETECTIONDETECTION RIBOZYMES CLEAVE MRNARIBOZYMES CLEAVE MRNA
PRODUCTS OF REACTIONSPRODUCTS OF REACTIONSBINDING ENDS LUCIFERASE INHIBITIONBINDING ENDS LUCIFERASE INHIBITIONEXPRESSION LEADS TO LIGHTEXPRESSION LEADS TO LIGHT
Image removed due copyright restrictions. Fluorescent imaged mouse, from http://www.caliperls.com/tech/ optical-imaging/image-gallery/oncology-angiogenesis-models.htm
HOW IT WORKSHOW IT WORKSTREATMENTTREATMENT REMOVE PATIENT CELLSREMOVE PATIENT CELLS
ADD PLASMIDS/CHANGE DNAADD PLASMIDS/CHANGE DNADELIVER MODIFIED TDELIVER MODIFIED T--CELLSCELLSADD DOSE OF LUCIFERINADD DOSE OF LUCIFERINTAKE IMAGES WITH CCD CAMERATAKE IMAGES WITH CCD CAMERA
Image removed due copyright restrictions. Photo of CCD imaging system, http://www.caliperls.com/ products/contract-research/in-vivo/optical-imaging-studies.htm
HOW IT WORKSHOW IT WORKSSYSTEM OVERVIEWSYSTEM OVERVIEW
ENDOCYTOSISENDOCYTOSIS
DIFFUSIONDIFFUSIONBINDINGBINDING
EXPRESSIONEXPRESSIONLIGHTLIGHT
HOW IT WORKSHOW IT WORKSTIMING DIAGRAMTIMING DIAGRAM
COLLAGEN RECEPTORCOLLAGEN RECEPTOR
INHIBITORINHIBITOR
LIGHT PRODUCERLIGHT PRODUCER
““DEBRISDEBRIS””
PROSTANOIDSPROSTANOIDS
SS
SSSS
AA
AAAA
HOW IT WORKSHOW IT WORKSDEVICE OVERVIEWDEVICE OVERVIEW
INHIBITORINHIBITORCOLLAGENCOLLAGEN RECEPTORRECEPTOR
LIGHT PRODUCERLIGHT PRODUCER
BREAKING IT DOWNBREAKING IT DOWNCOLLAGEN RECEPTORCOLLAGEN RECEPTOR
COLLAGEN BINDSCOLLAGEN BINDS ENDOCYTOSIS OFENDOCYTOSIS OF COLLAGENCOLLAGEN
ENDO180ENDO180
BREAKING IT DOWNBREAKING IT DOWNCOLLAGEN RECEPTORCOLLAGEN RECEPTOR
GFPGFPMCMVMCMV
ENDO180ENDO180
STRONG PROMOTERSTRONG PROMOTERENDO180 RECEPTOR GENEENDO180 RECEPTOR GENEGFP FOR TESTING/DEBUGGFP FOR TESTING/DEBUG
PARTSPARTS
BREAKING IT DOWNBREAKING IT DOWNINHIBITORINHIBITOR
INHIBITION ENDSINHIBITION ENDSCLEAVAGE OF MRNACLEAVAGE OF MRNA
RIBOZYMERIBOZYME
BREAKING IT DOWNBREAKING IT DOWNINHIBITORINHIBITOR
RIBOZYMERIBOZYME
PROMOTER DEPENDENT ON LUCIFERASEPROMOTER DEPENDENT ON LUCIFERASERIBOZYME GENERIBOZYME GENEDOUBLE APTAMER LOOPDOUBLE APTAMER LOOP
PARTSPARTS
APTAMERSAPTAMERS
BREAKING IT DOWNBREAKING IT DOWNLIGHT PRODUCERLIGHT PRODUCER
OXIDIZESOXIDIZES PIGMENTPIGMENT
LIGHTLIGHT
LUCIFERINLUCIFERIN OO22
LUCIFERASELUCIFERASE
BREAKING IT DOWNBREAKING IT DOWNLIGHT PRODUCERLIGHT PRODUCERPARTSPARTS
STRONGEST PROMOTER, COULD VARYSTRONGEST PROMOTER, COULD VARYLUCIFERASE GENELUCIFERASE GENEGFP FOR TESTING/DEBUGGFP FOR TESTING/DEBUG
H2CMVH2CMVSV40SV40 LUCIFERASELUCIFERASE GFPGFP
POTENTIAL PROBLEMSPOTENTIAL PROBLEMS
CHANGESCHANGES DIFFERENT RECEPTORSDIFFERENT RECEPTORSLONGER DETECTION TIMELONGER DETECTION TIMEOTHER SIGNAL BESIDES LIGHTOTHER SIGNAL BESIDES LIGHT
UNKNOWNSUNKNOWNS DEGREE OF VISIBILITYDEGREE OF VISIBILITYCOLLAGEN DEGRADATIONCOLLAGEN DEGRADATIONIMMUNE RESPONSEIMMUNE RESPONSE
MAKING IT WORKMAKING IT WORK
DEBUGGINGDEBUGGING PRESENCE OF LUCIFERINPRESENCE OF LUCIFERINLUCIFERASE REACTIONSLUCIFERASE REACTIONSUSE OF GFP IN VITROUSE OF GFP IN VITROFREEZEFREEZE--FRACTURE METHODFRACTURE METHOD
TESTINGTESTING PROMOTER STRENGTHPROMOTER STRENGTHININ--VITRO BINDING TO RIBOZYMEVITRO BINDING TO RIBOZYMEENDOCYTOSIS EXPERIMENTSENDOCYTOSIS EXPERIMENTSEXPRESSION OF RECEPTOR/LUCIFERASEEXPRESSION OF RECEPTOR/LUCIFERASE
Image removed due copyright restrictions. D-Luciferin Firefly vial from Caliper Life Sciences (http://www.caliperls.com)
MAKING IT WORKMAKING IT WORK
A PLANA PLAN RECEPTOR EVOLUTION FOR SPECIFICITYRECEPTOR EVOLUTION FOR SPECIFICITYSLOW DEGRADATIONSLOW DEGRADATIONAPTAMER DEVELOPMENTAPTAMER DEVELOPMENTIN VITRO BINDING/REACTIONSIN VITRO BINDING/REACTIONSLUCIFERIN ADDITION AND EFFECTSLUCIFERIN ADDITION AND EFFECTSIN VIVO TESTING/TRIALSIN VIVO TESTING/TRIALS
Image removed due to copyright restrictions.“Firefly Luciferase antibody for ICC/IF (Rat)”by Mal Niladri.http://www.abcam.co.jp/index.html?pageconfig=reviews&intAbID=21176&intAbReviewID=5843
QUESTIONS REMAININGQUESTIONS REMAINING
SAFETYSAFETY POSSIBILITY OF REJECTIONPOSSIBILITY OF REJECTIONLUCIFERIN EFFECTSLUCIFERIN EFFECTSTAXING ON BODYTAXING ON BODYDEGRADATION DELAYDEGRADATION DELAY
O O H H N N
HN H
O O NH
H2N + NH2 Figure by MIT OpenCourseWare.
QUESTIONS REMAININGQUESTIONS REMAINING
SECURITYSECURITY DESIGNED TO SURVIVE IN HUMANDESIGNED TO SURVIVE IN HUMANPOSSIBLE TOXIN DELIVERYPOSSIBLE TOXIN DELIVERYSHOULD NOT MAKE PUBLICSHOULD NOT MAKE PUBLICTT--CELL ENGINEERED TO DIECELL ENGINEERED TO DIE
Electron microscope image removed due to copyright restrictions.
image source: http://www.ncc.go.jp/Courtesy of the National Cancer Center (Japan). Used with permission.
WOULD IT SELLWOULD IT SELLSUNK COSTSSUNK COSTS
TT--CELL HARVESTING: ~$700CELL HARVESTING: ~$700DNA TRANSFECTION: ~$300DNA TRANSFECTION: ~$300LUCIFERIN DOSE: $400 PER GRAMLUCIFERIN DOSE: $400 PER GRAM
FIXED COSTSFIXED COSTS
CCD CAMERA: ~$3,000CCD CAMERA: ~$3,000RIBOZYME SEQUENCING: $100+RIBOZYME SEQUENCING: $100+RECEPTOR PLASMID ANDRECEPTOR PLASMID ANDLUCIFERASE PLASMID: $1000+LUCIFERASE PLASMID: $1000+
WOULD IT SELLWOULD IT SELLEFFECTIVENESSEFFECTIVENESS
COMPETITIONCOMPETITION
GRADIENT OF LIGHTGRADIENT OF LIGHTDYNAMIC OVER A PERIOD OF TIMEDYNAMIC OVER A PERIOD OF TIMESCALE AND 3D CLARITYSCALE AND 3D CLARITY
STATIC TESTSSTATIC TESTSCTC/TMEM BLOOD TESTSCTC/TMEM BLOOD TESTSLYMPH NODE SCREENINGLYMPH NODE SCREENING
OUR PROBLEMOUR PROBLEM
OUR SOLUTIONOUR SOLUTION
IN SUMMARYIN SUMMARYCANCER DEATHS DUE TO METASTASISCANCER DEATHS DUE TO METASTASISLACK OF KNOWLEDGELACK OF KNOWLEDGENO ACCURATE, DETAILED TESTINGNO ACCURATE, DETAILED TESTING
ACTIVE MAPPING OF METASTASISACTIVE MAPPING OF METASTASISSHOWS PROBLEM AREASSHOWS PROBLEM AREASGIVES SENSE OF HOW/WHEN IT TRAVELSGIVES SENSE OF HOW/WHEN IT TRAVELS
FOR THEIR HELP!FOR THEIR HELP!
THANKS TOTHANKS TO……NATALIE KULDELLNATALIE KULDELLMENTORMENTOR ““RARA”” (anonymous)(anonymous)DREW ENDYDREW ENDYROGER KAMMROGER KAMM
AGI STACHOWIAKAGI STACHOWIAKCHRISTINA SMOLKECHRISTINA SMOLKECHRIS ANDERSONCHRIS ANDERSON
MIT OpenCourseWarehttp://ocw.mit.edu
20.020 Introduction to Biological Engineering Design Spring 2009
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