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

PUTTING IT TOGETHERPUTTING IT TOGETHERREVIEW OF SYSTEMREVIEW OF SYSTEM

LIGHTLIGHT

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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