WELCOME TO OUR MONTHLY
BIOMEDICAL ENGINEERING
SCIENTIFIC INTEREST GROUP
SEMINAR SERIES AND TODAY WE HAVE
Dr. PAULA HAMMOND FROM MIT
SHE IS THE
INSTITUTE PROFESSOR AT THE KOCH
INSTITUTE FOR INTEGRATIVE CANCER
RESEARCH AND THE HEAD OF THE
DEPARTMENT OF CHEMICAL
ENGINEERING AT MIT.
Dr. HAMMOND'S ACHIEVEMENTSES
ARE NUMEROUS BEING NAMED THE
NATIONAL SCIENCE ENGINEERING AND
MEDICINE AND SHE'S CURRENTLY
SERVING ON PRESIDENT JOE BIDEN'S
COUNCIL ADVISERS ON SCIENCE AND
TECHNOLOGY AND HER TALK TODAY
WILL BE ON ELECTRO STATIC
ASSEMBLY OF LAYER BY LAYER IN
BIO MEDICINE.
TAKE IT AWAY.
>> THANK YOU SO MUCH.
I'M EXCITED TO HAVE A CHANCE TO
TALK TO THIS GROUP TODAY.
WE HAVE BEEN FOCUSING ON THE USE
OF THE ELECTRO STATIC ASSEMBLY
METHODS TO GENERATE SYSTEMS THAT
CAN BE USED FOR DRUG DELIVERY
AND HOPEFULLY, WE CAN TALK IN AND
HAVE DISCUSSION.
ONE IS NANO PARTICLES FOR
DELIVERY TO TUMORS BUT THE OTHER
IS CHARGED POLYMERIC SYSTEMS FOR
DELIVERY OF ARTHRITIS.
SO, OUR WORK THAT IS FOCUSED ON
CANCER, TAKES ADVANTAGE OF A
TECHNIQUE THAT WE'VE BEEN USING
FOR SEVERAL YEARS IN OUR LAB
WHICH INVOLVES THAL TERNE
NATURING POSITIVELY AND NEGATIVE
LOW CHARGED POLYMERIC SPECIES TO
CREATE CO'S I HAVE NANO SCALE IN
NATURE AND WE FOUND THAT WE CAN
ACTUALLY APPLY THESE SYSTEMS TO
A RANGE OF DIFFERENT SURFACES
FROM THE SURFACES OF DRESSINGS
FOR WOUND HEALING AND TO
APPLYING THEM ON SURFACES THAT
CAN HELP PROMOTE THE GENERATION
OF BALM.
HOWEVER, IN THIS CASE, WE'VE
BEEN TAKING LYPOSOME S&P LGA
THAT CAN BE LOADED WITH A
CHEMOTHERAPY DRUG IN THE CORE
AND USING THOSE PARTICLES AS
SURFACES THAT CAN ESSENTIALLY
SUPPORT THE GENERATION OF MULTI
LAYERS.
AND IN THESE LAYERS, WE
INCORPORATE A POSITIVELY-CHARGED
POLYMER ON TOP OF THE NEGATIVELY
CHARGED NANO PARTICLE AND THEN
WE CAN INCORPORATE A NEWCLEIC
AND THE HEAVILY CHARGED ACID IS
GOING TO HAVE A NICE PROTECTIVE
LAYER ON TOP OF IT, WHICH IS
ANOTHER POSITIVELY-CHARGED
LAYER.
SO NOW WE'VE COMPACTED OUR
NEUCLEIC ACID INTO A THIN FOAM.
OUR INITIAL GOAL IN THIS WORK
WAS TO CREATE COMBINATION
SYSTEMS AND THE IDEA WAS TO
ADDRESS CANCERS THAT HAVE A
NUMBER OF DIFFERENT GENETIC
MODIFICATIONS THAT WHEN SILENCES
WOULD ALLOW THE CHEMOTHERAPY TO
BE MUCH MORE EFFECTIVE.
SO IN OTHER WORDS, SILENCING THE
TUMOR DEFENSE MECHANISMS THAT
ARE PREVALENT IN A NUMBER OF
TUMOR TYPES AND I'LL GIVE ONE
EXAMPLE OF THAT.
HOWEVER, WE WERE MORE AND MORE
INTERESTED IN THIS FINAL LAYER,
WHICH IS THE LAST LAYER WE PUT
DOWN, THIS IS A NEGATIVELY
CHARGED OUTER LAYER.
WE NEED THE NEGATIVE CHARGE SO
THE NOW PARTICLE IS POSITIVITY
CHARGED AND GET OXYGENIZED AND
GET ELIMINATED OR CLEARED ON THE
BLOOD STREAM AND SO THIS IS PART
OF THE REASON FOR THIS OUTER
LAYER AND WE TAKE AND WE CAN USE
POLYMERS THAT HAVE HUGE AMOUNTS
OF HYDRATION AND ASSOCIATIONED
WATER THAT PROVIDES US WITH A
PENALTY FOR DISRUPTION OF THAT
HYDRATION LAYER IF PROTEINS DO
ABSORB ON THE SURFACE AND
PROVIDE A LAYER THAT CAN EXIST
FOR MORE TIMEFRAMES THAN WE
MIGHT TYPICALLY EXPECT FROM A
NEUTRAL NANOPARTICLE.
THIS SYSTEM A LOUSE A LARGE
AMOUNT OF MODULARITY SO WE HAVE
LAYERED LYPOSOMES WHICH ONLY
INCREASE IN SIZE BY 10 OR 15
NANO MEETERS IN THICKNESS AND WE
CAN LAYER PARTICLES, GOLD
NANOPARTICLES AND ANY SORT OF
METAL NANO PARTICLE AND THIS CAN
BE USED FOR IMAGING.
THESE ARE CENTERING AND WE JUST
ONE NEGATIVELY CHARGED FOLLOWED
BY ONE POSITIVELY CHARGED BY ONE
NEGATIVE LEON THE PARTICLES AND
WE CAN SEE INTERESTING RESULTS.
FOR EXAMPLE, HERE WE'RE LOOKING
AT A POLY ELECTRO LIGHT BUY
LAYER WHICH CONSISTS OF POLY LIE
SING FOLLOWING BY HYDRO LONIC
ACID.
IF WE CONSTRUCT THIS ON TO A
NANO PARTICLE AT ONE CONDITION
WE CAN SEE THAT AT THAT PH, 4.7,
WE HAVE A VERY STRONGLY
NEGATIVELY CHARGED NANO PARTICLE
AND THIS IS ABLE TO EXHIBIT NON
HALF LIES AND IS ABLE TO
ESSENTIALLY EXIST IN BLOOD
STREAM FOR SOME TIME.
HOWEVER, WHEN THE PH IS
DECREASED SLIGHTLY, WE GET
SWELLING OF THE BUY LAYER
BECAUSE WE'RE BEGIN TO GO
PROTEASOME NATURE THE ACIDS ON
THE BACKBONE AND WE ARE ERASING
THE ICONIC CROSS LINKS TO HOLD
THAT TOGETHER AND THAT LEADS TO
SWELLING AND THAT SWELLING CAN
HELP FACILITATE THINGS AS SIRNA
OR OTHER MOLECULES THAT MIGHT BE
INCAPSULATED UNDER THESE LAYERS.
WHAT WE CAN ALSO SEE IS A
SIGNIFICANT SHIFT IN THE CHARGE
OF THE NANO PARTICLE AS WE GO
DOWN IN PH.
EVEN AT A MODERATE SHIFT, WE CAN
GET CHANGE IN THE ZETA POTENTIAL
GIVING US A NEUTRAL NANO
PARTICLE OR IN SOME CASES EVEN A
SLIGHT POSITIVE CHARGE.
IT GOES FROM STEALTHY MODE TO A
STICKY MODE JUST THROUGH A MICRO
ENVIRONMENT SHIFT AND THAT MEANS
FOR THE NANO PARTICLES IN THIS
FASHION WE CAN HAVE A SYSTEM
THAT IS VERY STABLE AND DOESN'T
INTERACT VERY MUCH WITH CELLS
WHEN IT'S IN THE BLOOD STREAM
AND IT GETS NO A TIME OR AND IT
GETS INTO THE TUMOR COMPARTMENT
AND WE CAN SEE OFTEN THE SHIFT
IN THE PH DUE TO HYPOXIA IN THAT
TUMOR LEADING TO THIS CHANGE IN
THE NET CHARGE AND ESSENTIAL LOW
THIS NANO PARTICLE IS SA IS A STICKIER
NANO PARTICLE AND ONE CELLS
INTERACT WITH AND WE SEE NON
SPOKE UPTAKE.
NON-SPECIFIC UPTAKE ASIDE, THIS
IS AN INTERESTING MECHANISM, WE
HAVE FOUND THAT THIS OUTER LAYER
CAN BE VERY INTERESTING FOR A
SPECIFIC INCOMPETENTER ACTIONS
AND AND IT'S KNOWN TO BUY THE
RECEPTORS AND IN A LARGE NUMBER
OF THE SOLID EPITHELIAL TYPE
TUMORS WE SEE HIGH OVER
EXPRESSION OF THE RECEPTOR AND
FOR THAT REASON WE CAN SEE VERY
STRONG UPTAKE OF OUR NANO
PARTICLE SHOWN IN RED BY TUMOR
CELLS AND WE HAVE SEEN THIS WITH
TRIPLE NEGATIVE BREAST CANCER,
OVARIAN CANCER AND SOME FORMS OF
GLEGLIOBLASTOMA AS WELL AS HIGH
GRADE OVARIAN CANCER.
HERE YOU CAN SEE A SECTION OF
TUMOR TISSUE THAT WE STAINED
FOLLOWING AN IB INJECTION AND
THIS IS A FLANK TUMOR AND WE CAN
SEE VERY NICE KIND OF LAYER OF
CELLS.
HOWEVER, WE CAN ALSO SEE THESE
TUMOR CELLS THAT OVER PRESS CD44
LABELED IN GREEN AND THE
PARTICLES PENETRATE THE LAYER
AND GET INTO THE TUMOR ITSELF.
I MENTIONED NON SMALL CELL
CANCER AND WE HAVE A GREAT
COLLABORATORS HERE AT.
>> Mitch: AND THE COKE INSTITUTE
AND ONE OF OUR COLLABORATORS IS
TYLER JACKS AND HE PROVIDED US
WITH A MODEL OF CELLS THAT ARE
ESSENTIALLY EXHIBITING TWO
PHOTOGRAPHIC MUTATIONS IN NON
SMALL HUNG CANCER WHICH MAKES IT
PARTICULARLY AGGRESSIVE AND MORE
RESISTANT TO TREATMENT.
THESE TWO GENETIC MUTATIONS ARE
THE PRESENCE OF KRAS, ACTUALLY
ENABLES MUCH MORE AGGRESSIVE AND
NET A STATIC TYPE AND IT'S
UNDRUGABLE WITH SMALL MOLECULES.
AND WE ALSO LOOK AT THE LOSS OF
THE P53 GENE.
THE P53 FUNCTION, WHICH
ESSENTIALLY IS A GUARDIAN
FUNCTION WHICH MANAGING DNA
DAMAGE AND CAUSES A POP TOE SIS
WHEN DNA DAMAGE GOES TOO FAR IN
THESE CELLS, IS ABSENT IN THE
NUMBER OF TUMORS OF INTEREST
HERE.
WE LOOK TO REPLACE THE LOSS OF
P53 OR THE SINGLE POINT MUTATION
OF IT, WITH A MICRO RNA WHICH IS
KNOWN TOE REPLACE SOME OF THOSE
AFFECT OR FUNCTIONS THAT P53
PROVIDED.
WE GET THE MONITORING BACK AND
THAT ENABLES CELL APOPTOSIS.
SO IN THIS CASE, WE LOOKED AT A
COMBINATION THERAPY AND WE USE
SIS PLATEN, WE DESIGN OUR
LYPOSOME SO IT HAS NEGATIVELY
CHARGED LIPIDS AND WE CAN BUILD
OUR MULTI-LAYER AND HERE WE
ABSORB A POLY BECAUSE WE FOUND
THAT IT ACTUALLY FACILITATES
ESCAPE VERY NICELY IN THESE
SYSTEMS.
WE THEN ABSORBED A COMBINATION
OF KRAS SIRNA SO WE SILENCED THE
SIRNA AND KRAS WITH AN SIRNA MOL
ACTUAL AND THESE ARE ROUGHLY THE
SAME SIZE, NEGATIVELY CHARGED 20
TO 21 BASE PAIR MOLECULES THAT
ABSORB NICELY INTO OUR LAYER.
WE TOP OFF WITH HIGH LOR ONIC
ACID WITH CD44 TARGETING.
WHEN WE LOOK A LOT THE RELEASE
OF THE COMPONENTS OF THE
THEORIES IN OUR NANO PARTICLE,
WE CAN SEE THE SIRNA, IN THE
LAYERS IS RELEASED MORE RAPIDLY
THAN THE SIS-PLATEN INCORPORATED
IN THE CORE AND WE GET A TWO TO
THROW TIMES MORE RAPID RELEASE
OF THE SRNA ALLOWING US TO
ESSENTIALLY REPROGRAM THE TUMOR
CELLS THAT ARE PRESENT BEFORE
THE SISPLATEN REACHES ITS PEAK
ACCUMULATION IN THE CELL.
WE LOOKED AT WHERE THESE NANO
PARTICLES GO IN HEALTHY MICE AND
IN MICE THAT EXHIBIT LUNG CANCER
AND WE CAN SEE THE USUAL
ESSENTIALLY ACCUMULATION AND THE
EXCRETION THROUGH THE LIVER THAT
WE SEE WITH NANO PARTICLES BUT
THIS IS A COMMON THEME IN
NANOMEDICINE AND OBSERVED
BETWEEN THE 12 AND 24 HOUR
POINT.
THIS IS A 48-EIGHT HOUR POINT.
HOWEVER, WE SEE NO ACCUMULATION
IN LUNGS IN HEALTHY MICE AND A
VERY SIGNIFICANT ACCUMULATION IN
THE LUNGS OF TUMORRED MICE.
40 TO 50 FOLD MEASUREMENTS
DIFFERENCE.
AND WHAT WE CAN SEE IS
ESSENTIALLY THAT THE TUMOR ARE
VERY HEAVILY STAINED WHEN WE
LOOK FOR CD44.
NOW, WHEN WE TAKE OUR NANO
PARTICLE AND INCORPORATE THE
DRUGS, ESSENTIALLY, THE PLATEN
COMBINED WITH THE SIRNA AND
MICRO RNA COMBINATION, WE CAN
LOOK AT WHAT HAPPENS IN MICE AND
IN THIS CASE, WE ESSENTIALLY ARE
LOOKING AT THE KP MOUSE MODEL
DEVELOPED BY THE TYPIER LAB.
WHAT WE CAN FIND IS THAT FOR
CONTROL WE SEE A NUMBER OF THESE
TUMORS AND WE HAVE A LARGE TUMOR
MASSES THAT ARE GROWING AND WHEN
WE LOOK AT JUST RNA, OR JUST
SISPLATIN ALONE, WE SEE A
DECEASE BUT NOT SIGNIFICANT IN
THESE ANIMALS AND THIS IS
BECAUSE THIS IS A FAIRLY RESIST
APARTMENT FORM OF THE CANCER.
WE CAN SEE HERE, IN SURVIVAL,
THE RNA DOESN'T DO MUCH ON ITS
OWN.
SIS PLAT INIS IMPROVED COMPARED
TO THE CONTROL.
OTHER OF OUR WORK, OTHER AREAS
OF WORK, HAVE LOOKED AT
INHIBITOR COMBINATIONS.
WE BEGAN TO REALLY FOCUS ON
OVARIAN CANCER.
OUR INTEREST IN THIS IS TRYING
TO ADDRESS SOME OF THE REAL
DIFFICULTIES IN ESSENTIALLY
TARGETING OVARIAN CANCER CELLS
EFFECTIVELY AND DELIVERING
THERAPIES THAT WILL ENABLE THE
GREATER SURVIVAL AND WE'LL TALK
ABOUT THAT IN A MINUTE.
BUT TO DO THAT EFFECTIVELY, WE
WANTED TO LOOK AT THE RANGE OF
OUTER LAYERS THAT WE COULD
EXPERIMENT WITH.
LOOK AT AND UNDERSTAND IF WE CAN
HAVE SYSTEMS THAT HAVE A NATIVE
AFFINITY FOR OVARIAN CANCER
CELLS.
SO WE START BI STARTED BY MAKING A SMALL
LIBRARY.
THIS IS A WORK OF MY FORMER
STUDENT, SANTEE KOREA WHO
DESIGNED A NANOPARTICLE LIBRARY
WHERE WE LOOKED AT A RANGE OF
DIFFERENT NEGATIVE LOW CHARGED
POLYMER, INCLUDING HIGHER ACID
AND SULPHATED SURFACE CHEM IS
TREES AND WE JUST STARTED TO
ESSENTIALLY USE ROBOTICS TO
GENERATE THESE NANO PARTICLES
AND TEST THEM.
THIS ALLOWED US TO ASK A SIMPLE
QUESTION, ARE THERE SYSTEMS THAT
HAVE A HIGHER AFFINITY FOR HIGH
GRADE OVARIAN CANCER CELLS
COMPARED TO HEALTHY CELLS.
SO, IN THIS WORK, WE ACTUALLY
LOOKED AT 10 DIFFERENT OVARIAN
CANCER CELL LINES AS WELL AS
SEVEN HEALTHY CELL LINES AND WE
FOUND THAT THERE WERE SYSTEMS
WHICH GAVE US A HIGH AMOUNT OF
NANO PARTICLE ASSOCIATION USING
CELL SORTING.
SO, FROM THOSE EXPERIMENTS, WE
NARROWED OUR STUDIES TO HIGH LER
ON I CAN ACID AND WE RECOGNIZED
THAT WAS GOING TO BE AN
IMPORTANT OUTER LAYER AND WE
ALSO FOUND THAT POLY LGLUTAMIC
ACID AND POLY-L-ASPARTIC SHOWED
A ASSOCIATION OF CELLS THAN OUR
HIGH ALLURE ON I CAN ACID.
WE LOOKED CLOSELY AT WHAT WAS
GOING ON AND WE FOUND, WITH HIGH
LURE ON I CAN ACID WE CAN SEE
THESE NANO PARTICLES DO GET THIS IS
NORTH WE ANTICIPATE FROM THE
CD44.
WITH POLY LGLUTAMIC ACID, THEY
ASSOCIATED ON THE CELLS' OUTER
SURFACE.
SO WE HAVE SURFACE MEMBRANE
ASSOCIATION.
OVER LONG PERIODS OF TIME, VERY
LITTLE UPTAKE IS OBSERVED.
THIS IS INTERESTING TO US AND WE
DIDN'T QUITE UNDERSTAND IF
THERE'S A SPECIFIC RECEPTORS
ASSOCIATED WITH IT.
POLY LASPARTIC ACID WE SEE
GRADUAL COMMUNICATION ON THE
SURFACE MEMBRANE AND WE SEE
THERE'S UPTAKE AND THAT UPTAKE,
UNLIKE THE UPTAKE OF HA,
DECORATED NANO PARTICLES IS
CAVOLAR SO DIFFERENT PROTEINS
ARE INVOLVED IN THE UPTAKE OF
THESE NANO PARTICLES.
WE'RE CONTINUING TO TRY TO
UNDERSTAND THESE INTERACTIONS.
IN THE MEANTIME, WE FOUND THAT
THESE COULD BE ADVANTAGEOUS TO
OUR APPROACH.
HYALURONIC ACID IS INTERESTING
FOR DELIVERING DRUGS THAT NEED
TO GET INSIDE.
THAT INCLUDES SIRNA AND
DNA-DAMAGING DRUGS.
HOWEVER, THE SURFACE BOUND
ARRANGEMENT HERE, ALLOWS US TO
THINK ABOUT NANO PARTICLES THAT
CAN DELIVER SOMETHING THAT WE
DON'T WANT TO HAVE IN THE
TRAFFICKING MECHANISM BECAUSE WE
DON'T WANT TO SEE DEGRADATION.
THEREFORE, PROTEIN-RELATED DRUGS
ARE GOING TO BE INTERESTING AND
IN PARTICULAR, IF WE WANT TO
RELEASE OUR DRUG TO NEIGHBORING
CELLS, THIS WOULD BE A NICE WAY
OF DOING IT.
WHILE STILL HAVING HIGH
ACCUMULATION IN THE TUMOR.
I SHOULD MENTION, WE'RE THINKING
ABOUT THESE MECHANISMS THAT SOME
WORK THAT HAS BEEN GOING ON IN
OUR LAB, HAS USED PRISM TO LOOK
AT RAPID SCREENS TO UNDERSTAND
NANO PARTICLE INTERACTIONS WITH
SPECIFIC CELLS TYPES AND LOOK AT
WHICH GENES AND IT ALSO LET'S US
ASSOCIATE WHICH GENES ARE BEING
EXPRESSED UNDER CERTAIN UPTAKE
CONDITIONS AND IN THAT WORK, WE
HE DID FIND IN DEED THAT POLY L
ACID PARTICLES ACTUALLY THE
UPTAKE IN THOSE CASES TEND TO BE
ASSOCIATED WITH ENRICHMENTS IN
GENE NETWORKS ASSOCIATED WITH
INTER CELLULAR UPTAKE WHEREAS
POLY LGLUTANIC ACID WAS ENRICHED
IN CELL SURFACE BINDING
INTERACTIONS IN THAT WORK SO
WE'RE SEEING A CORRELATION AND
WE'RE HOPING TO UNDERSTAND THE
SYSTEMS BOYOLOGY OF THOSE
SYSTEMS.
HOWEVER, IN THE MEANTIME, WE
WANT TO TAKE ADVANTAGE OF THIS
POLY LGLUTANIC ACID'S TENDENCY
TO ASSOCIATE WITH THE OUTSIDE OF
OVARIAN CANCER CELLS AND IN
COLLABORATION, WE GUN TO ASK THE
QUESTION CAN WE DELIVER A
CYTOKINE THAT CAN ACTIVATE THE
IMMUNE SYSTEM IN OVARIAN CANCER.
NOW, THIS IS GENERAL IDEA IS
INTERESTING TO US BECAUSE
OVARIAN CANCER TENDS TO BE A
RELATIVELY COLD TUMOR.
ESPECIALLY THE ADVANCED OVARIAN
CANCER THAT LEADS TO AT LEAST
70% RECURRENCE.
WE CAN ACTUALLY ADDRESS SOME OF
THIS RECURRENCE IF WE CAN
STIMULATE A VERY EFFECTIVE
IMMUNE RESPONSE AGAINST THE
TUMOR.
HOWEVER, THERE ARE TYPICALLY
SMALLER NUMBERS OF IMMUNE CELLS
PRESENT IN THESE TUMORS.
ONE WAY OF CHANGING THAT IS TO
INTRODUCE SOMETHING THAT CAN
ACTUALLY STIMULATE THE IMMUNE
ENVIRONMENT AND AN APPROACH IS
TO USE IL12 OR SIMILAR CYTOKINE
THAT CAN ACTUALLY TRIGGER SOME
OF THESE IMMUNE PATHWAYS.
IL-12 IS INTERESTING, BECAUSE IT
LEADS TO THE INTER FEWER AN
GAMMA AND IT LEADS TO SIGNALING
THAT CAN ESSENTIALLY ACTIVATE
CD4, POSITIVE AND CD8 POSITIVE T
CELLS AND WHEN WE ARE ABLE TO
ACTIVATE THESE
ANTIGEN-PRESENTING CELLS.
WHEN WE'RE DOING THIS IN THE
TUMOR ENVIRONMENT, WE CAN GET
ANTIGEN-SPECIFIC RESPONSES TO
THE TUMOR.
SO, THIS IS THE GENERAL IDEA.
THE PROBLEM WITH IL-12 AND THE
REASON WHY WE'RE NOT USING IT
BROAD LOW, ALTHOUGH THERE HAVE
BEEN ATTEMPTS IN THE PAST, IS
BECAUSE IT'S VERY POTENT AND
CYTOKINES HAS BEEN NON TO HAVE A
NUMBER OF OFF-TARGET EFFECTS
BEAUCE IT'S SUCH A POTENT
STIMULATER OF INTERFERON KAMA
AND OF COURSE SYSTEMIC
STIMULATION OF INTER FEWER ON
LEFT A LARGE SCALE HOST EFFECT
SO WE'RE TRYING TO CONTROL WHERE
IL-12 IS EXPOSED.
IN OUR CASE, WE THINK THIS IS AN
ABSOLUTE OPPORTUNITY TO
INTRODUCE IP INJECTION OF OUR
NANO PARTICLES WHICH THEN
ASSOCIATE WITH THESE TUMOR CELLS
AND THE QUESTION IS CAN WE USE
OUR NANOPARTICLE TO DO THAT?
WE LOOKED AT A NANOPARTICLE
DESIGNS THAT BEGINS WITH A LIP
OSEM BUT WE USE THE TAG THAT
DARYL'S LAB HAD BEEN USING TO
ATTACH THESE PROTEINS.
WE HAVE A SCIL-12 AND IT'S
ATTACHED.
WE CAN LAYER ON TOP OF THAT,
POLY-L-ARGINE AND A POLIAN TON
AND IT WAS ON -- WE DID
COMPARISONS WITH HYALURONIC
ACID.
WE USED UNDERSTANDING THE
RELEASE KEN ET TICKS OF THIS
SYSTEM BECAUSE THE IL-12 DOES
GET TO BE PRE IN THE SYSTEMS
THAT I'VE JUST DESCRIBED AND IN
THIS STUDY, WE LABELED
ESSENTIALLY FRET DYES FOR THE
IL12 AND PLR TO SEE WHEN THESE
SEPARATE FROM THE NANOPARTICLE
AND WE CAN SEE THAT THE PLR IS
ESSENTIALLY ESCAPING FIRST AND
THEN WE SEE THE IL-12 ESCAPING
SHORT LOSHORTLY AFTERWARDS.
ONE OF THE MODELS IS THAT
POTENTIALLY THE BUY LAYER GETS
DIS RESULTED OR DESTABILIZED AND
IT'S RELEASED WITH THE OUTER
LAYER AND THE IL-12 RELEASED
AFTERWARD.
ANOTHER IS THERE'S JUST
SIGNIFICANT SWELLING THAT CAN
OCCUR WHICH ALLOWS THE IL-12 TO
RELEASE.
AND THE QUESTION OF WHETHER IT
CAN WORK IN OVARIAN CANCER, WAS
REALLY IMPORTANT TO US.
WE DID SOME EARLY STUDIES
LOOKING AT PROSTATE CANCER
MODELS AND OTHER MODEL THAT'S
WERE MORE COMMON AND MORE
RESPONSIVE TO IMMUNOTHERAPY.
OVARIAN CANCER WAS THE ULTIMATE
GOAL AND IN THIS WORK, WE USED A
MODEL AND A FAIRLY AGGRESSIVE
ONE KNOWN AS HM-1 SOLVE IN THIS
MODEL, WE CAN TRACK WHERE
NANOPARTICLE GOES.
WE LET THAT INTRAVENOUS
INJECTION AND WHAT WE FOUND WAS
THAT THERE'S ACCUMULATION IN
BOTH CASES.
IV GIVES US A NICE ACCUMULATION
FOR INTERVENE YUS AND IT'S
SOMEWHERE BETWEEN 8% AND 9%
ACCUMULATION.
HOWEVER, THE INJECTION GIVES US
SOMETHING CLOSER TO AN 85%
ASSOCIATION OF THE NANO PART CAL
OF THE TUMOR AND WE THINK THIS
IS THE ROUTE THAT EX SIGHTS US
MOST, ALTHOUGH INTRAVENOUS IS
SOMETHING THAT WE STUDY
ALONGSIDE THIS WORK.
SO, GOING WITH THIS IP INJECTION
APPROACH, WE LOCKED AT
ESSENTIALLY THE INTRODUCTION
WITH IP INJECTIONS OF REASONABLY
HIGH DOSE OF THE IL-12 AND WE
LOOKED FIRST AT TOXICITY AND YOU
CAN SEE THE WEIGHT OF ANIMALS
AND HERE WE'RE LOOKING AT
ESSENTIALLY THE DROP THAT WE
EXPECT WHEN WE HAVE FREE IL-12
COMPARED TO THE CONTROL ANIMALS
WHICH IS VERY SIGNIFICANT.
THE ARROWS ALSO INDICATE LOSS OF
LIFE.
WE LOSE SOME ANIMALS DUE TO THE
TOXICITY, HOWEVER, WITH OUR
IL-12 NANOPARTICLE, LAYERED WITH
THE POLY GLUTAMIC ACID, WE SEE
WE'RE ABLE TO PRESERVE THE
WEIGHTS OF ANIMALS AND
ESSENTIALLY RESCUE THAT
TOXICITY.
WITHOUT THE LAYERS ON THE
NANOPARTICLE WE STILL SEE THAT
THERE IS A NUMBER OF -- A
SIGNIFICANT LOSS.
SO, WE ARE EXCITED ABOUT THE
FACT THAT THE LAYERS ARE ABLE TO
PROTECT AND ALSO TO TARGET.
WHEN WE LOOKED AT EFFICACY IN
HM1 TUMORS, WE FOUND THERE WAS
SIGNIFICANT ENHANCEMENT IN
SURVIVAL IN THE IL-1 L-12PLE SYSTEM
COMPARED TO OUR UNTARGETED
SYSTEM.
IT GOT US VERY EXCITED.
WE'RE EXCITED ABOUT THE FACT WE
WERE SEEING ABOUT A 30% SURVIVAL
RATE WE LOOKED AT SERUM
CYTOKINES IN THESE ANIMALS AND
WE WERE PARTICULARLY INTERESTED
IN WHETHER WE WERE GOING TO SEE
ANY SORT OF ACCUMULATION OF
INTER FEWER AN GAMMA AFTER
INJECTION AND WE SEE THAT THE
INTER FEWER ON GAMMA IS LOWER
COMPARED TO THE FREE IL-12.
IN THE HEALTHY MICE AND IN
TUMOR-BEARING MICE, WE SEE IT'S
EVEN MORE SIGNIFICANT.
OF COURSE, WE DID DO A MUCH MORE
DETAILED LOOK AT WHICH IMMUNE
CELLS ARE PRESENT AFTER OUR
TREATMENTS.
THIS IS JUST A LITTLE BIT OF
THAT DATA.
HERE WE'RE LOOKING AT
ESSENTIALLY, THE PRESENCE OF
IMMUNE CELLS IN KT CELLS AND K
CELLS AND C8 POSITIVE CELLS IN
TUMORS AND WE'RE COMPARING THEM
AGAINST THE PRE IL-12 WHICH WE
ALSO ANTICIPATE SHOULD PRESENT
THESE IMMUNE CELLS AN CELLS.
IN ALL OF OUR CASES, NANO 12
NANOPARTICLES CAN DO AS WELL AS
THE FREE IL12 IF NOT BETTER.
WHEN WE LOOK ELSEWHERE OUTSIDE
OF THE TUMOR, WE SEE THAT
THERE'S A RELATIVELY LOW
PRESENCE IN A SIGH TEASE AND IN
DISPLAYING.
WE DON'T SEE THERE'S AN
ENHANCEMENT AND THESE OTHER
AREAS SO WE LOCKED AT HM1 MODEL
BEFORE.
I DIDN'T REALLY DESCRIBE IT IN
MUCH DETAIL BUT THIS MODEL HAS
MODERATE T-CELL PRESENCE AND IT
HAS A HIGH AMOUNT OF
INFILTRATION OF GRANULAR SITES
AND IT'S RESPONSIVE TO DNA
DAMAGING AGENTS BUT IT'S
RESISTANT TO PD1 AND PDL-1
BLOCKADES.
WE LOOKED AT THE KPCA MODEL.
THIS IS A COLD TUMOR.
WITH MODERATE AMOUNTSES OF T
REGULATORY CELLS SO IT'S
RESPONSIBLE TO -- RESPONSIVE TO
CHECKPOINT INHIBITORS.
SO, THIS ON THE LEFT IS THE DATA
YOU SAW BEFORE FOR THE HM1
MODEL.
WHEN WE LOOK AT THE KPCA MODEL
THE NANOPARTICLE IS MORE
RESPONSIVE THAN IT WAS IN THE
HM1.
THIS IS TRUE ALSO FOR THE FRO
IL12 BUT WE'RE SEEING THIS
EFFICACY ENHANCEMENT IS
CONSISTENT ACROSS MODELS.
WE CONTINUE TO DO THIS WORK, WE
HAVE RECENT DATA THAT I HAVEN'T
BEEN ABLE TO INCORPORATE IN THIS
TALK YET BUT I HOPE I CAN TALK
TO YOU ABOUT IT LATER.
WHICH INCLUDES A DIFFERENT MEANS
OF ATTACHING THE IL12 TO THE
NANOPARTICLE WHICH SEEMS TO BE
GIVING US A VERY SIGNIFICANT
ENHANCEMENT IN THAT SURVIVAL
NUMBER.
SO WE'RE LOOKING AT THAT AND
WE'RE ALSO TRYING TO UNDERSTAND
THE MECHANISM FOR THAT AND
HOPEFULLY WE CAN BRING MORE TO
THE TABLE WITH RESPECT TO HOW TO
CONTROL THE CYTOKYNK FOR MORE
EXTENDED TIME PERIODS.
BEFORE MOVING ON, I SHOULD
MENTION WE HAVE LOOKED AT OTHER
KINDS OF CANCERS.
THIS IS AN EXAMPLE OF WORK IN
WHICH WE TARGETED LYMPHOMA USING
AN SIRNA AGAINST BCL.
AND IN THIS CASE, WE'RE LOOKING
AT AN ANTI BODY ATTACHMENT TO
OUR OUTER LAYER.
THE OUTER LAYER WAS HYALURONIC
ACID BUT WE ADDED AN ANTI BODY
WHICH BINDS TO THE CD-20
RECEPTORS AND IN THIS WORK, WE
FOUND THAT WE WERE ABLE TO GET
MORE EFFECTIVE TARGETING AND I
DON'T HAVE TIME TO GO OVER THE
DATA WITH THE COMBINATION SYSTEM
AND WE WERE ABLE TO SEE GREATER
SURVIVALABILITY IN THIS LYMPHOMA
MODEL JUST WITH THE APPLICATION
OF THIS SIBCL2 AND HERE WE CAN
SEE THE COMBINATION GIVES US A
MUCH LOWER AMOUNT OF CANCER
SHOWN AS THIS STRONGLY
LUMINESCENT THAT YOU CAN SEE IN
THE ANIMALS HERE.
WE'VE ALSO LOOKED AT THE IMPACT
OF THE MECHANIC AT PROPERTIES OF
THESE NANO CHEM KELLS INSPIRED
BY WORK AS OTHERS SUCH AS DEBRA
AND ESPECIALLY WORK THERE WAS
FOUND THAT ELASTICITY OF
NANOPARTICLES CAN INFLUENCE
BLOOD CIRCULATION TIME AND HALF
LIFE AND AND HERE WE THOUGHT
BECAUSE OUR NANOPARTICLE IS VERY
FLEXIBLE, WE CAN APPLY IT TO
BOTH SOFT AND RIDGE ENTER
NANOPARTICLES AND USE THAT CORE
TO DETERMINE THE TRAFFICKING AND
NATURE OF TRAFFICKING FOR THE
NANOPARTICLE.
WE USE LYPO SEEM LINE LIPOSOMES AND
BECAUSE WE'RE USING FULLY UNSAT
RAIDED LIPIDS THE CHOLESTEROL TO
BE USED TO ESSENTIALLY TAKE US
FROM A MORE CRYSTALLY KIND OF
NANOPARTICLE WHICH IS RIDGED TO
A SOFTER AND MORE FLUENT-LIKE
LIPID LAYER AND WE CAN LAYER BY
LAYER ON TOP OF BOTH OF THOSE
AND GET NEARLY THE SAME RESULTS,
SHOWN BY THE BLUE AND GREEN BARS
IN TERMS OF THE CHARGE AND THE
SIZE.
THE CRYO EM IMAGES ARE
CONSISTENT WITH THAT AND HERE
WE'RE BASICALLY LOOKING AT
BEFORE AND AFTER WE INTRODUCE
THE LAYERS ON OUR RIDGED AND OUR
SOFT NANOPARTICLES.
WE DO FIND THAT THERE IS AN
ENHANCEMENT IN HALF LIFE IN THIS
SYSTEM.
WHEN WE LOOK AT THE SOFTER
NANOPARTICLES WITH THE SAME
OUTER LAYER.
AND VERY BRIEFLY, WE FOUND THAT
THE SOFTER LBO NANOPARTICLES
HAVE HIGHER TUMOR ACCUMULATION
AS SHOWN HERE ON THE RIGHT AS
WELL AS FROM THE IMAGES HERE
WHERE THE TOP ARE THE RIDGED AND
THE BOTTOM ARE THE SOFT AND
WE'RE LOOKING FOR THE SIGNAL OF
THE TUMOR.
CY7.
WHEN WE LOOK CROSS SECTIONS, OF
THE TUMORS, IN THESE CASES AND
LOOKED AT THE PENETRATION AND
HERE WE'RE LOOKING AT OVARIAN
TUMORS, WE SEE THAT THERE'S A
SIGNIFICANT ENHANCEMENT OF
PRESENCE OF THE NANO PART CAL
WHEN WE HAVE THE SOFT LBL
COMPARED TO THE RIDGED
NANOPARTICLE AND WE'VE SEEN THE
RIDGED ONES THAT WE HAVE SURFACE
ACCUMULATION BUT NOT DEEP
PENETRATION.
SO, IN SUMMARY, WE HAVE FOUND
THAT WE CAN USE THIS OUTER LAYER
CHEMISTRY AS A WAY TO TARGET OUR
NANOPARTICLES TO TUMORS.
WE CAN ACTUALLY USE THE NATURE
OF THAT OUTER LAYER AS A WAY TO
ESSENTIALLY ADAPT HOW THE NANO
PART CAL TRAFFICKING IN CELLS
AND IT CAN IMPACT THE HALF LIFE
TO THE DEGREE OF TUMOR AND SO WE
HAVE WE ARE LEARNING HOW TO USE
TO ESSENTIALLY ENHANCE THE
PRESENCE OF THESE NANOPARTICLES
AND THE NATURE OF THEIR RELEASE
AND USE THEM AS A WAY TO TARGET
THE DELIVERY OF PROTEINS AS WELL
AS SIRNA AND CHEMOTHERAPY DRUGS.
WITH THAT, I'LL TRANSITION AND
TRY TO BE BRIEFER WITH OUR TOPIC
ON OSTEO ARTHRITIS.
IN THIS CASE, WE'RE ABLE TO
TREAT TRAUMA INDUCED AS TEE OWE
ARTHRITIS SO THIS IS OFTEN
CALLED POST TRAUMATIC OSTEO
ARTHRITIS OR PTOA AND IT HAPPENS
WHEN THERE'S A BIT OF DAMAGE TO
THE CARTILAGE PERHAPS DUE TO A
WORK INCIDENT AND SOME SORT OF
DAMAGE OR TRAUMA A THIS IS
SOMETHING THAT IS OF GREAT
INTEREST TO THE DOD FOR EXAMPLE,
AND FOR SOLDIERS AND MILITARY
PERSONNEL BUT IS OF INTEREST OF
ALSO TO ATHLETES AND TO ANYONE
WHO HAS SIGNIFICANT PHYSICAL
ELEMENTS TO THEIR WORK.
WHEN THIS TEAR OR DAMAGE
HAPPENS, OVER TIME, THERE'S
INFLAMMATION WHICH LEADS TO PAIN
AND WE'VE LEARNED HOW TO HANDLE
THE INPRACTICE MISSION WITH
STEROIDS AND DO A GOODS JOB OR
ORAL NON STEROID ANTI
INFLAMMATORY DRUGS BUT IT'S NOT
REALLY ELIMINATING OR TREATING
THE PROBLEM, IT'S TREATING THE
PAIN AND THE INFLAMMATION.
TYPICALLY WHAT HAPPENS IS THAT
OVER TIME, THIS INFLAMMATION
OCCURS AND THERE'S ENOUGH DAMAGE
THERE'S A NEED FOR A FULL JOINT
REPLACEMENT.
THERE'S NOT A DISEASE MODIFYING
DRUGS WHICH ESSENTIALLY REVERSES
SOME OF THESE PATHWAYS TOWARDS
ESSENTIALLY PROGRESSIVE
INFLAMMATION AND THE REASON FOR
THIS IS NOT BECAUSE WE DON'T
UNDERSTAND THE KINDS OF
THERAPIES THAT MIGHT WORK, THERE
ARE PROTECTORS AND STIMULATE THE
CARTILAGE SITES THAT RECITE
WITHIN CARTILAGE TO PRODUCE MORE
CARTILAGE AND THERE ARE DRUGS
WHICH CAN REUSE THE TENDENCY
TOWARDS INFLAMMATION HOWEVER, TO
MANAGE THE DELIVERY OF PRETEENS
IN THE JOINT AND ONE OF THE
PRIMARY ONES IS THAT THERE'S A
LOT OF RAPID CLEARANCE THAT
TAKES PLACE WITHIN THE JOINT AND
SO IF YOU INTRODUCE A FREE
PROTEIN, INTO THE JOINT, EVEN IF
IT HAS ANNA TRACTION TO THE
CARTILAGE, IT WILL RESIDE THERE
FOR A SHORT TIME PERIOD BECAUSE
THERE'S A TENDENCY FOR THERE TO
BE A CLEARENS THROUGH THE VESSEL
AND THE VENUALS.
THE OTHER ISSUE IS CARTILAGE
ITSELF IS A VERY SENSE
NEGATIVELY CHARGED POLYMER AND
HERE WE SHOW THE PRO KROLL
GLYCANS PRESENT WHICH HAVE
HEAVILY BRANCHED AND CHARGED
THESE ARE ESSENTIALLY NEGATIVE
LOW CHARGED POLY SACKRIDES AND
WE HAVE A PROBLEM THAT ANYTHING
THAT IS LARGER IS GOING TO
ESSENTIALLY BIND TO THE TOP AND
WON'T PENETRATE, EVEN IF IT'S
POSITIVELY CHARGED.
IF IT'S SMALLER, A SMALL
MOLECULE HAS A CHANCE OF GETTING
IN AND SPENDING TIME WITHIN THIS
MATRIX AND REACHING THE SITES
THAT RECITE IN THEM BUT THERE
ARE NO BLOOD VESSELS TO HELP US,
THEY'RE A VASCULAR TISSUES SO
WE'VE BEEN INTERESTING IN
DESIGNING NANO SYSTEMS THAT CAN
HELP DELIVER PROTEIN DRUGS AND
OTHER KINDS OF DRUGS DIRECTLY TO
AND IT'S RELEVANT WHEN WE HAVE A
DRUG THAT NEEDS TO INTERACT
DIRECTLY ON THE CELLS INCLUDING
GROW FACTORS THAT CAN STIMULATE
THE GENERATION CARTILAGE.
WE NEED SMALL ENOUGH TO DEAL
WITH 15 NANO METER OR SMALLER
MESH SIZE THIS MATRIX PRESENTS.
IT NEEDS TO HAVE STRONG POSITIVE
CHARGE IT CAN COMBINE SO IT
CAN'T BE POSITIVE LOW CHARGED,
IT ONLY BINDS TO THE SURFACE BUT
DOESN'T MAKE ITS WAY OVER TIME
PERIODS THAT ARE REASONABLE INTO
THE CARTILAGE.
SO WE DECIDED TO WORK WITH
DENDROMERS BECAUSE THEY'RE VERY
NICE, BEAUTIFULLY BRANCHED
POLYMERIC ARCHITECTURES THAT ARE
HEAVILY CONTROLLED.
THEIR SIZE, DEPENDING ON THE
GENERATION, FOR EACH BRANCHING
EVENT, REPRESENTS A GENERATION,
DETERMINES THE SIZE, GIVES US A
NICE RANGE AND WE CAN PURCHASE
DENDRIMERS IN THE SIZE RANGE OF
FOUR TO SEVEN NANO MEETERS AND
THEY CAN PRESENT ANYWHERE FROM
64 TO 256 THREE GROUPS AND ALL
OF THESE GROUPS PROVIDE AN
OPPORTUNITY FOR POSITIVE CHARGE
SO THAT'S ONE OF THE REASONS
WE'RE INTERESTED IN THIS MOT
MOLECULE.
SIZE AND CHARGE ARE WHERE WE
WANT TO BE.
ON TOP OF THAT, THEY PROVIDE PRE
CHEMICAL HANDALS THAT ALLOW US
TO ATTACH OUR CARGO PROTEIN TO
THE SURFACE THE DENDRIMER.
WITH THAT SAID, ALL OF THIS
CHARGE CAN BE PROBLEMATIC.
BY ITSELF IT'S TOXIC TO CELLS
BECAUSE IT CAN ACTUALLY BREAK
THROUGH A NUMBER OF CELL
MEMBRANES AND IT CAN INTRODUCE
TOXICITY SO WE WANT TO CONTROL
THE CHARGE.
WE WANT TO TITRATE IT.
IF IT'S TOO STICKY, IT WILL GET
STUCK AT THE CARTILAGE SURFACE
SO WE WANT TO MANAGE THAT
STICKINESS ISSUE AS WELL.
AND ALLOW THIS NANOPARTICLE TO
INTERACT WITH THE MATRIX THAT
HAVE AN INTERACTION THAT ALLOWS
BINDING AND DEBINDING SO THAT IT
CAN MAKE ITS WAY INTO THE
CARTILAGE.
ONE WAY OF DOING THIS IS
INTRODUCING POLY ETHYLENE GLY
CON, A HYDRATED SHARP POLYMER
CHAIN WE CAN ADD BECAUSE IT'S
NEUTRAL AND IT HAS A LARGE
AMOUNT OF WATER ASSOCIATION AND
IT'S A VERY FLEXIBLE MOLECULE.
IT'S BEEN KNOWN TO SHIEL KNOWN TO SHIELD A
CHARGE.
IT SHIELDS SOME OF THE CHARGE
THAT IS PRESENT ON THESE
DENDRIMER MOLECULES AND OF
COURSE, EACH REACTION WE HAVE
DOES ELIMINATE ONE OF THE
PRIMARY AMINES AND WE ARE A
LOWER BASIS TEE OF THE AMEEN.
LET'S LOOK AT THE BIG PICTURE.
WE HAVE SOME DESIGN PARAMETER
SPACE.
FOR THE PANAM DENDRIMER, WE
LOOKED AT TWO DIFFERENT
GENERATIONS.
ONE SMALLER AND ONE LARGER.
AND WE ALWAYS ATTACHED IGF,
WHICH IS THE PROTEIN DRUG OF
CHOICE FOR OUR STUDY AND CAN
STIMULATE THE GENERATION OF
MATRIX MOLECULES BY THE CON
DEGREE SITES SO WE ATTACHED THAT
USING A COMMON CON -- WE USE THE
DYE TO TRACK WHAT IS GOING ON.
THE PEG CHAIN BECAME ONE OF OUR
BIGGEST MODIFICATIONS AND WE
LOOKED AT DIFFERENT LENGTHS AS
WELL AS DIFFERENT DENSITIES OF
THE PEG ITSELF.
SO YOU CAN IMAGINE VISUALLY A
LIBRARY IN WHICH WE ARE
INCREASING THE SIZE OF THE
POLYMER CHAINS THAT WE ATTACH AS
WELL AS THE DENSITY OF THE
CHAINS THAT WE PUT DOWN AND OF
COURSE, THE LESS THAT WE HAVE
PRESENT, THE HIGHER THE NET
CHARGE OF THE MOLECULE.
WE THEN LOOK AT HOW DEEPLY THE
NANOMATERIALS PENETRATE INTO
CARTILAGE AS WELL AS THE NET
AMOUNT THAT IS TAKEN UP OR THE
UPTAKE BY USING BOTH CARTILAGE
DISKS TO ESSENTIALLY LOOK AT
THESE LABELED NANOMATERIALS.
IN DOING THIS WORK, WE MEASURED
NOT ONLY THAT PERCENT UPTAKE OR
NET UPTAKE INTO THE CARTILAGE,
WE ALSO LOOKED AT CELL
VIABILITY.
ESSENTIALLY, WE'RE INTERESTED IN
SYSTEMS WHICH GAVE US A VERY
HIGH PERCENT VIABILITY OF CELLS
CLOSE TO 100% IN DOING SO, WE
COULD SEE A TREND WHICH
INCREASING PEG A LAYING OF OUR
DEN DEMER LED TO ESSENTIALLY
HIGH UPTAKE UP TO A CERTAIN
POINT AND THEN WE REACHED A
POINT OF PEG WHICH THE UPTAKE
DROPS SIGNIFICANTLY THE INVERSE
BEHAVIOR FOR VIABILITY IN WHICH
WE SEE THAT VIABILITY IS VERY
LOW UP TO A CERTAIN POINT AND WE
SEE VIABILITY IS PRESERVED AND
SO, WE'RE LOOKING FOR A SWEET
SPOT WHERE WE MAINTAIN VIABILITY
WHILE HAVING HIGH UPTAKE.
AND WE FOUND THAT THAT SWEET
SPOT INTENDED TO EXIST SOMEWHERE
BETWEEN THE POINT WHERE WE HAVE
THESE PEG CHAINS PACKING RIGHT
NEXT TO EACH OTHER IF WE CAN
IMAGINE THAT THEY'RE FLEXIBLE
AND THEY'RE SWITCHING AROUND AND
THEY'RE RANDOM WALK ON THE DEN
DRUM SURFACE THEY'RE BEGINNING
TO TOUCH THAT WE SEE THAT WE ARE
ENTERING A WINDOW OF GOOD
VIABILITY AND WE CONTINUE IN
THAT WINDOW OF GOOD VIABILITY
AND DROP IN UPTAKE WHEN WE GET
BEYOND ABOUT TWO TIMES THAT
DENSITY.
NOW, THAT PREVIOUS WORK WAS DONE
BY MY FORMER GRADUATE STUDENT
BRETT, GUYGER AND NOW BRANDON
JOHNSTON IN OUR LAB IS LOOKING
AT THE SPECIFIC INTERACTIONS
THESE PEG CHAINS HAVE WHEN THEY
ARE ON THIS ATTACHED SURFACE AND
THIS INCLUDES HYDROGEN PONDING
WHICH CONTRIBUTES TO THE
SHIELDER, OF THE PEG CHAIN AND
AS WELL AS THE UMBRELLA EFFECT
OF ESSENTIALLY THIS PEG CHAIN
SURROUNDING THE CHARGE.
AND HE WAS ABLE TO DERIVE A WAY
OF CORRELATING THE AMOUNT OF
SORT OF FREELY AVAILABLE OR
ACCESSIBLE PRIMARY MEANS THAT
WILL CONTRIBUTE TO THE CHARGE
BEHAVIOR OF THE SYSTEM AS
OPPOSED TO THE ONES THAT ARE
ESSENTIALLY HIDDEN BY PEG AND HE
DID THIS BY USING A COMPETITIVE
SCREENING ASSAY IN WHICH YOU
LOOK AT HOW MUCH SALT IS
REQUIRED BEFORE YOU GET COMPLETE
DISRUPTION OF THE DENDRIMER FROM
THE CARTILAGE.
MATRIX.
I DON'T HAVE TIME TO GO INTO THE
DETAILS OF THIS APPROACH.
IT'S A BEAUTIFUL APPROACH.
THAT ALLOWED US TO CORRELATE THE
NUMBER OF FREE AMINES THAT PRO
CHARGE VERSUS THOSE THAT ARE
OFFERED UP BY PEG.
THAT ALLOWED US TO MAKE MORE
DIRECT COMPARISONS BETWEEN OUR
DIFFERENT FORMULATIONS AND LOOK
FOR TRENDS.
SO, THIS IS A LITTLE BIT MORE
ABOUT HOW THIS WORK WAS DONE AND
I'LL SKIP AHEAD A LITTLE BIT
JUST TO EXPLAIN THAT IN BEING
ABLE TO MAKE THESE CORRELATIONS,
WE CAN PLOT SOMETHING LIKE THE
NUMBER OF ACCESSIBLE CHARGED
AMINES ON ONE POINT, ON ONE AXIS
AGAINST THE TOTAL NUMBER OF
GROUPS THAT WERE MODIFIED AND WE
CAN SEE THAT AS WE INCREASE THE
PEG CHAIN LENGTH, WE CAN SEE A
TREND IN WHICH WE'RE GETTING
LESS AND LESS ACCESSIBLE CHARGE
ON THESE MOLECULES SO WE CAN
ACTUALLY TUNE THAT WITH A GREAT
DEAL OF ACCURACY.
WE'VE DONE THIS FOR TWO
DIFFERENT GENERATIONS,
GENERATION 4 AND 6 AND WE CAN
MAKE CORRELATIONS WITH RESPECT
TO THE LENGTH OF THE PEG CHAIN
AND THE AMOUNT OF SCREENING WE
GET.
THIS ALLOWS US TO LOOK AT THE
BALANCE OF UPTAKE AND HOW LONG
THAT DENDRIMER RESIDES IN THE
MATRIX AS WELL AS THE AB OPTION
ABSORPTION KEN ET TICKS AND WE
HAVE DOPE PENETRATION AND THE
TIME IN THE CARTILAGE.
WE LOOK AT HOW WE MEASURE UPTAKE
OVER 24 HOURS SO NOW WE'RE
LOOKING AT A TIMED UPTAKE AND WE
CAN SEE THERE ARE SYSTEMS WHICH
GIVE US SIGNIFICANTLY MORE
UPTAKE AND THIS IS THE PEG 4
SYSTEM AND EVEN THOUGH WE HAVE
THE SAME NUMBER OF ACCESSIBLE
CHARGED AMINES IN THIS
COMPARISON.
THIS IS BECAUSE FASTER UPTAKE
HAPPENS WITH THE SHORTER PEG
CHAINS.
THAT'S PART OF THE STORY.
WE CAN GET VERY RAPID UPTAKE
WITH THE SHORTER PEG CHAINS
GIVING THE SAME AMOUNT OF
SHIELDING.
WHEN THE PEG CHAIN LENGTH GETS
LONGER, THEY CONTRIBUTE TO THE
NATURE OF THE CONJIGATE AND IF
WE WAIT LONGER TIME PERIODS,
EVEN THOUGH IT'S FASTER FOR THE
SHORTER CHAINS, OVER LONGER TIME
PERIODS THE UPTAKE IS GREATER
WHEN WE HAVE LONGER CHAINS.
IF WE LOOK AT LONG TIME PERIODS,
THE LONGER CHAINS, ALTHOUGH THEY
TAKE LONGER TO GET IN, MORE OF
THEM GET INTO THE MIDDLE
THICKNESS OF THE CARTILAGE OVER
A PERIOD OF SEVERAL DAYS.
AND WE THINK ABOUT THESE
INJECTIONS, WE'RE ABLE TO THEN
MONITOR HOW LONG WE HAVE TO GET
INTO THE CARTILAGE, THAT TERMS
WHICH ONE OF THESE FORMULATIONS
WE GO WITH.
NOW WE TOOK ONE OF OUR OPTIMIZED
SYSTEMS AND LOOKED AT
ESSENTIALLY THE DEN DEGREE MER
IGF-1 AND WE'RE LOOKING AT 45%
PEG
WE'RE LOOKING AT THE JOINT OF
THE ANIMAL, FOR IGF ALONE WHICH
IS LABELS, WE CAN SEE A
SIGNIFICANT ENHANCEMENT IN THE
TIME IN THE CARTILAGE.
WE SEE VERY RAPID CLEARANCE FOR
THE IGF ONE ALONE.
IF WE MEASURE IT, THE CLEARANCE
IS IN LESS THAN A DAY WE'VE LOST
ALMOST ALL OF IT.
WHEREAS THE HALF LIFE FOR OUR
IGF IS 4.2 DAYS.
WHEN WE LOOK AT THE TIME AT
THERAPEUTIC EFFICACY, PRE IGF
GIVES US A FEW DAVIS EFFICACY IN
TERMS OF THERAPEUTIC PRESENCE AS
OPPOSED TO 30 DAYS THAT WE CAN
SEE IN THE PANAM AND FINALLY,
HERE WE CAN SEE FROM BLUE
STAINING, HERE THAT WE HAVE
INJURY MODEL.
HERE WE'VE REMOVED OUR CAUSED
DAMAGE TO PART OF THE CARTILAGE,
AND WE LOOK AT THE RECOVERY OF
THIS DARK BLUE STAIN OVER TIME
WHEN WE INJECT WITH IGF-1 ALONE
WE CAN SEE THAT WE'RE NOT
GETTING AS MUCH REGENERATION
HOWEVER, WE CAN SEE SIGNIFICANT
REGENERATION OF THE CARTILAGE
AND IT LOOKS ALMOST LIKE THE
CONTROL.
NOW WE'RE LOOKING AT WAYS WHERE
WE CAN MAKE THESE SYSTEMS MORE
RESPONSIVE, BY HAVING THEM
RESPOND TO A VERY INFLAMMATORY
ENVIRONMENT USING MMP RESPONSIVE
SYSTEMS THAT MAKE THE SYSTEM
MORE CHARGED AND WE HAVE BEGUN
TO LOOK AT WAYS IN WHICH WE CAN
GENERATE THESEN TIME SENSITIVE
FORM LEASE WITH THE SAME LEVEL
OF CONTROL THAT WE WERE ABLE TO
GENERATE THE CHARGE PEG SYSTEMS.
SO I'LL STOP HERE AND I TALKED
ABOUT ELECTROSTATIC ASSEMBLY AND
HOW WE APPLY IT TO NANOMATERIALS
AND USE IT TO GET MULTI-DRUG
RELEASE TARGETED TISSUE
CAPABILITIES AND WE HAVE
TARGETING ELEMENTS TO OUR SYSTEM
HOW WE CAN CONTROL TRAFFICKING,
BOTH WITH NOT THE TISSUE AND
WITH THE CELL AND HOW WE CAN
LEVERAGE THIS TO TARGET
DIFFERENT CELL TYPES AND ADDRESS
DISEASE SO I'D LIKE TO
ACKNOWLEDGE THE STUDENTS AND
POSTDOCS WHO CONTRIBUTED TO THIS
WORK WHOSE NAMES ARE HERE BOTH
ON THE NANOPARTICLE SIDE AND THE
DENDRIMER SIDE FOR OUR AS TRIO
ARTHRITIS.
COLLABORATIONORS, FOR THESE
PROJECTS, DARYL IRVIN ANDAL
INAND THE FUNDING WHICH INCLUDES
NIH, NIBIB AND NIH-NCI WE HAVE
FUNDING FROM THE DLD AND IN
PARTICULAR FOR THE A OSTEO
ARTHRITIS AND WE FOCUS ON
OVARIAN CANCER AND I'D LIKE TO
WELCOME ANY QUESTIONS WITH THE
TIME WE HAVE LEFT.
THANK YOU.
>> THANK YOU, THERE ARE MORE
QUESTIONS COMING IN AND WE'LL
HAVE TIME TO GET TO WE CAN GIVE
A FEW.
SO, FOR THE HA INCORPORATING
PARTICLES, WHAT MOLECULAR WEIGHT
HA IS AT?
BECAUSE, YOU KNOW, DEPENDING ON
THE MOLECULAR WEIGHT, IT WAS
FIFTH DESIGN ELEMENT?
>> VERY GOOD QUESTION.
WHEN WE FIRST STARTED THIS WORK,
WE WERE WORKING WITH
INTERMEDIATE MOLECULAR WEIGHTS,
20 TO 50,000, THIS RANGE, AND
MORE RECENTLY, MY STUDENTS HAVE
BEGUN TO LOOK AT WHETHER WE CAN
SEE DIFFERENCES WITH MOLECULAR
WEIGHT WHEN WE ABSORB ON THE
NANOPARTICLE AND WE KNOW THAT WE
ARE ESSENTIALLY ENGAGING THE
CD-44 RECEPTOR AND IT'S NOT
CLEAR WHETHER THE MOLECULAR
WEIGHT IS GOING TO MAKE A
DIFFERENCE IN TERMS OF TLR
RECEPTORS AND UP WITH OF THE
THINGS THAT I SHOULD MENTION IS
THAT WHEN WE ABSORB THESE
POLYMER CHAINS, THEY'RE
EXHIBITING TAIL-LIKE BEHAVIOR SO
IT'S AS IF WE HAVE FRAGMENTS
WAVING ABOUT AS OPPOSED TO
HAVING LONG CHAINS THAT ARE
TETHERED AT ONE END.
>> IT'S VERY INTERESTING.
>> THINK OF A INCH WORM ON A
SURFACE AS OPPOSED TO A SINGLE
TEETHER.
>> WAS THERE INTERNALIZED BY A
CELL WHAT HAPPENS TO THEM NEXT.
HO HOW ARE THEY DEGRADED AND HOW
DOES THAT FACTOR INTO YOUR
DESIGN?
>> GREAT QUESTION.
WHEN WE WORK WITH HYPO ZOMAL
CHARACTERS, WE HAVE THE CONTENT
OF THE CELL THAT UNDERGOES IT'S
OWN SORT OF TRACKING AND WE HAVE
THESE POLYMERS WHICH TEND TO BE
POLY STEP SIDES SO WE THINK THEY
UNDERGO A MORE TYPICAL CLEAR NOT
AND EARLY ON, WE DID STUDIES AND
WE CAN SEE SOME OF THESE
POLYMERS EXCRETED WITH POLY L
LICING AND POLY L.
WITH THE POLY L GLUTAMIC
STUDIES, WE EXPECT SIMILAR
RESULTS AND THE LGA DOES
BREAKDOWN SO WHEN WE USE THE
PLGA CORE, WE KNOW THAT ALSO
BREAKS DOWN AND OWN FAIRLY
REASONABLE BLEE SHORT TIMEFRAMES
BECAUSE IN NANOPARTICLE FORM,
THEY BREAKDOWN MUCH MORE RAPIDLY
THAN THEY WOULD IF THEY WERE IN
A MACRO SCOPIC FORM.
>> GREAT.
>> AND SO, ARE YOU FAMILIAR WITH
A PAPER THAT CAME OUT ONE OR TWO
YEARS GO ABOUT A DORE THRESHOLD
OF NANOPARTICLE DELIVERY AND
IT'S OVER DELIVER IS THE DOSE OF
NANOPARTICLES SOMETHING THAT YOU
CONSIDER YOUR SYSTEMIC DELIVERY
AND DO YOU OBSERVE THAT
PHENOMENON?
>> I REMEMBER HEARING ABOUT THAT
WORK AND I INTEND TO TAKE A
DEEPER DIVE INTO IT.
WE HAVE NOT THOUGHT ABOUT
FLOODING.
YOU CAN FLOOD THE SYSTEM WITH
NANOPARTICLES AND INTRODUCE THE
TRULY THEY ARE.
WHEN WE WORK WITH THE MODELS WE
WORK WITH, WE'RE REALLY WORKING
TO GET HIGH DOSE OF OUR
NANOPARTICLE IN AND SOMETIMES
EVEN THE NUMBER OF INJECTIONS
CAN CONSTRAIN HOW WE SET UP OUR
EXPERIMENT.
SO, WE HAVEN'T TRIED THAT AND IT
WOULD PROBABLY REQUIRED SOME
WORK WITH OUR CURRENT ANIMAL
INFORMED IT WOULD BE I. I'M MUCH
IS THEY STICK TO THE TUMOR THEN
LET'S SAY TRYING TO TRICK
SYSTEMIC DELIVERY INTO HELPING
US ALTHOUGH, I'M DEFINITELY
READY TO TRY TRICKS THAT WORK.
>> IT'S TO GET THEM TO
ACCUMULATE, RIGHT?
>> ABSOLUTELY.
ABSOLUTELY.
AND I THINK IF YOU ARE ARE
WORKING WITH SYSTEMS THAT ARE
POTENT ENOUGH AND SYSTEMIC
INJECTIONS 8% INJECTED DOSE IS
GOING TO GET YOU THERE AND YOU
DON'T HAVE ANY SIDE EFFECTS
THEN, YOU ARE ALREADY IN A SWEET
SPOT COMPARED TO PRE DRUG.
>> GREAT.
WELL, THAT IS ABOUT TIME.
THANK YOU AGAIN FOR YOUR TALK.
I REALLY ENJOYED IT AND I KNOW A
LOT OF FOLKS HERE ENJOYED IT AS
WELL AND THANK YOU AGAIN AND IT
WAS GREAT HAVING YOU TODAY.
IT WAS GREAT TO HAVE A CHANCE TO
PRESENT.
THANK YOU, EVERYONE, VERY MUCH
AND I HOPE TO MEET YOU IN THE
FUTURE.
>> OF COURSE.
>> AND EVERYONE ELSE IN THE
BIOMEDICAL ENGINEERING SIG,
WE MEET AGAIN NEXT MONTH AND
WE'LL SEE YOU ALL THEN.
THANK YOU.
>> BYE-BYE.