GOOD AFTERNOON, AND WELCOME
TO THE NIH'S WEDNESDAY AFTERNOON
LECTURE SERIES.
IT'S MY HONOR TO BE HERE FOR THE
FIRST TIME AND LOOKING FORWARD
TO THE LECTURE.
IT HONORS THE LEGACY OF MARGARET
PITTMAN ESTABLISHED IN 1994 IT'S
GIVEN BY A RESEARCHER WHO HAS
THE EXCELLENCE AND DRIVE THAT
MADE MARGARET PITTMAN A LEADER.
IT'S MY PLEASURE TOE INTRODUCE
DR. CAROLYN BERTOZZI WHO WILL
COME OUT IN A MINUTE.
A FORMER COLLEAGUE OF MINE FROM
STANFORD.
SHE'S AN INVESTIGATOR AT THE
HOWARD HUGHES MEDICAL AND FROM
CHEM-H AND BASS PROFESSOR OF
CHEMISTRY AND SYSTEMS BIOLOGY
AND RADIOLOGY AT STANFORD.
BEFORE I TELL YOU HER BACKGROUND
I SHOULD LET YOU KNOW IF YOU'RE
ATTENDING FOR CME CREDIT THE
CODE IS 57692 AND IF YOU'RE
WATCHING ONLINE AND WOULD TO
SUBMIT QUESTIONS CLICK ON THE
SEND LIVE FEEDBACK BUTTON
ANYTIME DURING THE LECTURE.
WE'RE HONORED TO HAVE
DR. BERTOZZI A CHEMICAL
BIOLOGIST AND KNOWN HER RESEARCH
INTERESTS THAT SPAN CHEMISTRY
AND BIOLOGY AND WORKS ON CELL
SURFACE SUGARS IMPORTANT TO
HEALTH AND DISEASE AND HOW TO
USE BIO CHEMICAL CHEMISTRY AND
COINED THE THERM FOR
BIOORTHOGONAL QUICK CHEMICAL
REACTIONS WITH SYSTEMS AND LOOKS
AT GLYCOSYLATION ASSOCIATED WITH
CANCER AND INFLAMMATION AND USES
IT TO DEVELOP NEW DIAGNOSTIC AND
THERAPEUTIC APPROACHES IN THE
AAREA OF IMMUNO ONCOLOGY AND
RECEIVED THE NOBEL PRIZE IN
CHEMISTRY.
DR. BERTOZZI WAS AWARDED AN
AWARD AT AGE 33 AND THE FIRST
WOMAN TO RECEIVE THE LEMELSON
M.I.T. PRIZE AND HAS MANY OTHER
HONORS AND POSITIONS.
HER RESEARCH IS REFLECTED IN
ABOUT 440 PUBLICATIONS ACROSS
THE SPECTRUM OF CHEMISTRY AND
BIOLOGY JOURNALS AND MENTORED 81
FORMER Ph.D. STUDENTS AND 58
FORMER POSTDOCTORAL FELLOWS AND
COMPLETED HER DEGREE FROM
CHEMISTRY IN HARVARD IN 1988.
AFTER BEING IN THE FIELD OF
CELLULAR IMMUNOLOGY SHE JOINED
THE U.C. BERKELEY FACULTY IN
1996 AND IN 2015 JOINED THE
FACULTY IN STANFORD UNIVERSITY
AT CHEM-H WHICH STANDS FOR
MEDICAL CHEMICAL MEDICINE
APPLIED TO HEALTH.
JOIN ME IN GIVING A WARM WELCOME
TO DR. BERTOZZI.
>> THANK YOU.
HELLO.
HELLO?
THANK YOU SO MUCH FOR COMING
TODAY.
THANK YOU, DR. BHATTACHARYA FOR
THE VERY KIND INVITATION AND FOR
HOSTING ME TODAY.
I'VE ALREADY LEARNED SO MUCH
JUST FROM MY FIRST MORNING HERE
INTERACTING WITH THE SCIENTISTS
AND KIND OF FINDING OUT WHAT'S
AT THE LEADING EDGE OF SCIENCE
WHICH OFTEN COMES RIGHT OUT OF
THE NIH.
YOU GUYS ARE AN AMAZING GROUP OF
PHYSICIANS AND SCIENTISTS AND
STAFF.
YOU DO SO MUCH TO SERVE YOUR
COUNTRY.
I'M ETERNALLY GRATEFUL FOR THE
SUPPORT I'VE HAD FROM THE
NATIONAL INSTITUTES OF HEALTH
AND I'LL TALK ABOUT WORK FUNDED
BY THE NIH STARTING WHEN I WAS A
BRAND NEW ASSISTANT PROFESSOR AT
THE TIME THAT WAS AT U.C.
BERKELEY SO THIS TAKES ME BACK
TO THE LATE 1990s BUT HOPEFULLY
BY STARTING IN THE EARLY DAYS,
YOU'LL GET TO UNDERSTAND THE
PROGRESSION OF A PROGRAM THAT
STARTED VERY MUCH AS A BASIC
SCIENCE PROJECT BUT EVENTUALLY
EVOLVED IN A MORE TRANSLATIONAL
DIRECTION TO THE POINT WHERE
THERE ARE NOW HUMAN THERAPEUTICS
AND CLINICAL TRIALS THAT
LEVERAGE SOME OF THE CHEMISTRIES
I'LL TALK ABOUT TODAY.
BEFORE I GET INTO ANY OF THE
DETAILS, I WOULD ALSO LIKE TO
SORT OF SHARE THE HONOR OF
HELPING YOU CELEBRATE THE LIFE
AND CAREER AND THE IMPACT OF
DR. PITMAN.
I READ A BIT ABOUT HER LAST
NIGHT.
SHE CLEARLY WAS AN INCREDIBLY
INFLUENTIAL FIGURE AND HAD A
HUGE IMPACT IN HER ACADEMIC
POSITIONS AS WELL AS THE NIH.
IT'S AN EXTRA HONOR FOR ME TO BE
ON THE STAGE RECOGNIZING HER
CONTRIBUTIONS.
SO WITH THAT, LET'S TALK A
LITTLE BIT ABOUT CHEMISTRY.
I ALWAYS LIKE TO GET A SHOW OF
STANDS SO I UNDERSTAND MY
AUDIE
AUDIENCE.
HOW MANY PEOPLE HERE WOULD
CONSIDER THEMSELVES TO BE A
CHEMIST?
ALL RIGHT.
HOW MANY PEOPLE HERE WOULD
CONSIDER THEMSELVES TO BE A NOT
CHEMIST?
IT'S ACTUALLY QUITE A FEW.
THERE'S NOTHING WRONG WITH THAT.
NOTHING TO BE ASHAMED OF
WHATSOEVER.
THAT'S A JOKE FOR CHEMISTS.
REGARDLESS OF WHETHER YOU'RE A
CHEMIST OR BIOLOGIST OR ANOTHER
TYPE OF SCIENTIST YOU'LL
PROBABLY RECOGNIZE THIS GADGET.
WHEN YOU TAKE ORGANIC CHEMISTRY
LAB COURSES AND I THINK THE
LASER POINTER IS SUPPOSED TO
WORK BUT SEEMS TO BE SILENT.
THERE IT IS.
WHEN YOU TAKE AN ORGANIC
CHEMISTRY CLASS, WHICH I TEACH
AT STANFORD AND YOU'RE IN THE
LABORATORY AND DOING A CHEMICAL
REACTION, YOU'LL BE USING A
GADGET LIKE THIS ONE.
SO THIS IS A ROUND BOTTOM FLASK.
HIGHLY ENGINEERED GLASSWARE AND
THE LOGIC OF THE GLASSWARE IS IT
ALLOWS YOU TO CONTROL EVERY
PARAMETER OF THE REACTION.
SO YOU AS THE CHEMIST OR THE
CHEMISTRY STUDENT, YOU CONTROL
EXACTLY WHAT GOES INTO THIS
FLASK, THE CONCENTRATIONS OF THE
REAGENTS, THE SOLVENT YOU
DISSOLVE THE REAGENTS IN WHICH
MATTERS WHEN YOU DO ORGANIC
CHEMISTRY AND EVEN CONTROL THE
ATMOSPHERE ABOVE THE LIQUID.
SOMETIMES YOU'LL PURGE IT TO GET
RID OF THE MOISTURE FROM THE
ATMOSPHERE.
YOU EVEN CONTROL THE RATE AT
WHICH THE STORING OCCURS AND
CONTROL THE TEMPERATURE WITH
THIS DOWN MERE AND ALL OF THIS
RIGID CONTROL ALLOWS YOU TO FORM
ALL KINDS OF BONDS USING ALL
KINDS OF REAGENTS AND WE HAVE
GIANT TELEPHONE BOOKS FULL OF
LISTS OF ORGANIC REACTIONS YOU
CAN DO IF YOU'RE DOING THEM IN
THIS KIND OF SETTING.
BACK IN THE 1990s IN THE EARLY
'90s AS A POSTDOCTORAL FELLOW IN
AN IMMUNOLOGY LAB LEARNING ABOUT
THE MYSTERIES OF BIOLOGY THAT
WERE DIFFICULT TO ANSWER AT THE
TIME AND UNDERSTAND I STARTED TO
BECOME INTERESTED IN THE IDEA OF
DOING CHEMISTRY IN LESS
CONTROLLED ENVIRONMENTS AND
SPECIFICALLY TO DO CHEMISTRY IN
THE ENVIRONMENT OF LIVING
SYSTEMS.
AT THE TIME I WAS THINKING ABOUT
DOING CHEMISTRY ON LIVING CELLS
IN A DISH OR IN THE BODY OF
MODEL ORGANISMS IN THE
LABORATORY BUT YOU DON'T HAVE TO
EXTRAPOLATE TOO FAR TO START
THINKING ABOUT WHAT IF YOU CAN
DO CHEMICAL REACTIONS TO FORM
NEW BONDS, NEW FORMS OF MATTER
INSIDE THE HUMAN BODY AND IF YOU
COULD DO THAT, WHAT
OPPORTUNITIES MIGHT OPEN UP IN
THE WORLD OF THERAPEUTICS
DEVELOPMENT OR DIAGNOSTICS
TECHNOLOGIES.
SO IT WAS THAT KIND OF THINKING
THAT ULTIMATELY LED TO THE
DEVELOPMENT TO THIS NEW BRAND OF
CHEMISTRY WE CALL BIOORTHOGONAL
CHEMISTRY WHICH IS A BIT OF A
MOUTHFUL.
PEOPLE OFTEN ASK WHAT'S THAT
MEAN AND THIS IS NOT IN THE
WEBSTER AS A DICTIONARY BUT SOME
DAY MIGHT BE.
IT MEANS CHEMISTRY BETWEEN
REAGENTS THAT NEITHER INTERACT
WITH NOR INTERFERE WITH A
BIOLOGICAL SETTING.
SO THEY'RE LITERALLY ORTHOGONAL
TO BIOLOGY.
I RECRUITED A WHOLE BRAND NEW
GROUP OF STUDENTS WHEN I STARTED
MY FIRST FACULTY APPOINTMENT IN
BERKELEY AND TAKING NOTATIONS ON
SPACE AND THERE'S CHEMISTRY FROM
THE ELEMENTS ON THE PERIOD TRICK
TABLE.
BUT ONLY A SMALL PORTION OF
THOSE CHEMISTRIES OCCUR IN
BIOLOGY.
AS VAST AS THAT BIOLOGICAL
CHEMICAL SPACE IS AND YOU COULD
JUST TAKE INVENTORY OF YOUR
BODY, ALL THE CHEMICALS IN YOUR
BODY ARE IN THIS GALAXY OF
BIOLOGICAL REACTIVITY SPACE BUT
IN THE UNIVERSE WE THOUGHT THERE
COULD BE A GALAXY OF CHEMICALS
THAT DON'T EXIST PLANET EARTH
EARTH THAT ARE NOT IN THE HUMAN
BODY MAYBE THEY'RE CHEMICALS
INVENTED BY CHEMISTS NATURE
HASN'T ARISEN YET.
IF WE COULD IDENTIFY REACTIONS
AMONGST THE BIOORTHOGONAL
CHEMICALS THOSE ARE WHAT WE CAN
DO WITHOUT INTERFERENCE IN THE
HUMAN BODY IN PARTICULAR.
THAT WAS THE NOTION.
AS ABSTRACT AS IT SOUNDS I WAS
WORKING IN THE IMMUNOLOGY LAB
AND MY GOAL WAS TO STUDY CHANGES
IN THE PATTERN OF CELL SURFACE
GLYCOSYLATION.
SPECIFICALLY THE SITE OF
INFLAMMATION.
THIS IS MY OPPORTUNITY TO GIVE A
TEXTBOOK DRAWING HOW PEOPLE
DEPICT THE SURFACE OF CELLS AND
THE PATTERNS OF GLYCANS ON THE
CELL SURFACE.
MOST WILL KNOW CELLS ARE BOUNDED
BY A PLASMA MEMBRANE.
ANCHOR READY MOLECULES LIKE GLUC
YO LIPIDS MOST ARE GLYCOPROTEINS
AND GLYCOBIOLOGY IS AN AREA I
DEVELOPED AN INTEREST IN AS A
GRAD STUDENT BUT BLOSSOMED IN MY
POSTDOC YEARS.
IT TURNS OUT THE STRUCTURES OF
THESE GLYCANS, WHICH BY THE WAY,
THE GLYCOSCIENTISTS USE AS A
SYMBOLIC LEXICON TO DESCRIBE
THEM AND MADE OF SIMPLE SUGAR
BUILDING BLOCKS ON THE TABLE.
EACH BUILDING BLOCK HAS A SHAPE
AND COLOR THAT'S DISCREET AND
YOU CAN SEE FROM THE CARTOONS
THEY GET LINKED IN PATTERNS AND
THE TOTALITY OF ALL THESE
GLYCANS ON THE SURFACE OF THE
CELL WE CALL THAT THE
GLYCOCALYX.
I WAS CLEAR THE IS GLYCOCALYX
CHANGES AND THERE WERE SHIFTS IN
THE PATTERN OF IT IN THE SETTING
OF CANCER BUT IT WAS REALLY HARD
TO CHARACTERIZE THESE CHANGES
AND NO ONE REALLY HAD ANY CLUE
ABOUT THE FUNCTIONAL
SIGNIFICANCE SO THIS WAS EARLY
DAYS IN THE FIELD OF CANCER
GLYCO SCIENCE.
IF THEY'RE CHANGING MAYBE YOU
CAN DETECT A TUMOR IN THE HUMAN
BODY BY CT OR PET IMAGING IF YOU
HAD A WAY TO LIGHT UP THE SUGARS
WHICH WOULD DIFFERENTIATE THE
HEALTHY CELLS FROM CANCEROUS
CELLS.
THERE WAS ONE PARTICULAR
PHENOTYPE, A GLYCOPHENOTYPE AS
WE WOULD SAY VERY WIDE SUPPRESS
AMONGST CANCERS AND IT'S THE
OVERPRODUCTION OF GLYCANS THAT
TERMINATE WITH A SIMPLE
MONOSACCHARIDE UNIT CALLED
SIALIC ACID.
THIS IS THE CHEAT SHEET FOR THE
SUGARS AND SYMBOLS.
THE ACID IS THIS PINK DIAMOND.
AND HEALTHY CELLS WILL HAVE A
CERTAIN POWDER OF GLYCANS WILL
AND I USE MET FOAPHORS AND I WO
CALL THAT A GARDEN AND THAT WELL
MANICURED GARDEN OVERGROWS AND
IT BECOMES A TROPICAL JUNGLE OF
SIALIC ACID AND CALLED IT A
GLY
GLYCOCAYLX AND IF WE CAN LIGHT
UP UP THAT MAY BE A WAY TO
DIFFERENTIATE IT FROM THE TUMOR
TISSUE WHICH IS A POWERFUL
CAPABILITY TRYING TO TREAT OR
MONITOR OR DIAGNOSE CANCERS.
BACK IN THE EARLY 1990s WE DID
NOT HAVE A WAY OF IMAGING
SUGARS.
THERE WAS ONE OF THE TOOL TO
LIGHT THEM UP IN LIVING SYSTEMS.
AS IT HAPPENS, BY THE WAY,
SORRY, I HAVE AN A LITTLE
SHAMELESS PLUG, I'M GIVING A
TALK TOMORROW ALSO AT NCATS IN
ROCKVILLE AND IN THAT TALK I'LL
TELL YOU WHY IT HAPPENS SO DON'T
ASK ME THAT NOW IF YOU PLAN TO
GO TO THE TALK TOMORROW BECAUSE
YOU'LL SPOIL IT.
IN THIS TALK I'M TRYING TO TALK
ABOUT IMAGING AND WHY BUT COME
TOMORROW IF YOU WANT TO KNOW
WHY.
SO I WENT TO A CONFERENCE I WAS
STILL A POSTDOC WORKING ON MY
JOB APPLICATIONS AND LANDED IN A
CONFERENCE IN SOUTH HAMPTON,
ENGLAND.
IT WAS A BOUTIQUE MIDDLEWEIGHT
OFF THE BEATEN TRACK AND MED
DR. REUTER AND HE HAD WORK
UNPUBLISHED THAT FOUND THE
PATHWAY HAS PRECURSORS AND THE
SUGAR KNOWN AS THIS PRECURSOR IS
A NORMAL PRECURSOR FOR SIALIC
ACID.
THERE'S SIX ACIDS THAT TRANSFORM
IT AND A COUPLE MORE GET THAT
ATTACHED TO THE GLYCANS ON THE
CELL SURFACE.
THEY COVERED IT IN OIL TO
STRETCH THE CARBON CHAIN AND AS
LONG AS THE MODIFICATIONS WERE
NOT TOO EXTRAVAGANT, ALL THE
ENZYMES IN THE PATHWAY WOULD
STAY FINE.
FREE PASS.
YOU FEED THE CELL WITH THIS AND
DUMP IT IN THE MEDIA AND WAIT A
COUPLE HOURS AND THE
CORRESPONDING ALTERED SIALIC
ACID WITH THE SAME SIDE CHAIN
APPEARS ON THE CELL SURFACE.
WHEN I SAW THE TALK AN IDEA
AROSE.
WHAT IF YOU CAN EMBED WITHIN THE
SUGAR SOME KIND OF A CHEMICAL
HANDLE.
SOMETHING INERT AND WOULD NOT
INTERFERE OR INTERACT WITH THE
CELL.
LET THAT CHEMICAL HANDLE MAKE IT
ITS WAY TO THE CELL SURFACE
SIALIC ACID WITH AN IMAGING
PROBE.
I CAME BACK FROM THE CONFERENCE
AND SAT AND WROTE MY
APPLICATIONS AND THIS IS A
GRAPHIC FROM A JOB APPLICATION.
YOU WRITE A PROPOSAL WHEN YOU
APPLY FOR FACULTY JOBS.
BY THE WAY, THIS GRAPHIC WAS
VERY STATE OF THE ART CIRCA
1994.
NOT SO MUCH NOW.
BUT IN THAT PROPOSAL, I WROTE,
OKAY, WHAT WE'RE GOING TO DO IS
DEVELOP A GLYCAN IMAGING
TECHNOLOGY WHERE THE FIRST STEP
WOULD BE TO FEED CELLS A
CHEMICALLY ALTERED SUGAR THAT
MAKES ITS WAY THROUGH BIO
SYNTHESIS THROUGH THE GLYCAN AND
WHEN THAT'S DONE WE'LL HAD A
PROMOLECULE AND THE IDEA IS A
CHEMICAL GROUP LET'S CALL IT X
FOR NOW ON THE SUGAR AND
COMPLEMENTARY GROUP CALL IT Y ON
THE PROBE AND THEY'D FIND EACH
OTHER WITHIN THE LIVING SYSTEM
ON MAYBE LIVE CELLS OR ORGANISMS
AND A CHEMICAL REACTION WOULD
OCCUR THAT BASICALLY ATTACHES
THE PROBE TO THE SUGAR AND NOW
YOU CAN VISUALIZE THE SUGAR.
THAT WAS THE IDEA.
SO I GOT THE JOB AT BERKELEY AND
I REALIZ REALIZED IMMEDIATELY WE
TO INVENT AND IT WOULD REQUIRE
BIOORTHOGONALLY AND WE HAD TO
INVENT WHAT WOULD BE USEFUL FOR
AN EXPERIMENT LIKE THIS ONE.
THERE I WAS THIS IS U.C.
BERKELEY AND ME AT THE SITE IF
YOU DON'T RECOGNIZE ME AND I
APPLIED FOR MY FIRST GRANT AT
NIH AND AFTER A COUPLE FALSE
STARTS IT WAS FUNDED BY THROUGH
THE NATIONAL INSTITUTE OF
MEDICAL SCIENCES AND THAT'S WHEN
I LAUNCHED THIS SEARCH FOR
BIOORTHOGONAL CHEMICAL REACTION.
SO THANK YOU, TO THE NIH.
THIS WAS IN REACTION TO A FAMOUS
CHEMIST IN THE EARLY 1800s TO
REACTED TO THIS GROUP WHERE
THERE'S THREE NITROGENS IN A ROW
WITH THIS CHEMICAL GROUP WHICH A
REAGENT.
THERE'S A CHEMISTRY BY WHICH A
PERSON CAN REDUCE THEM AND
THAT'S THE REDUCTION.
HOW MANY HERE HAVE DONE THIS
TYPE OF REDUCTION?
I KNEW IT.
IT'S AN OLDIE BUT GOODIE AND I
HAD DONE THIS MYSELF AS A GRAD
STUDENT BUT IT OCCURRED TO ME A
SLIGHT MODIFICATION OF THE
STAUDINGER REDUCTION MAY JOIN
THEM TOGETHER AND THAT'S WHAT
THIS IS.
THIS IS A CHEMISTRY YOU CAN DO
ON CELLS, NO PROBLEM, THERE'S NO
HARM TO THE CELL, NO
INTERFERENCE WITH ANY OF THE
BIOLOGICAL CHEMICALS, AZIDES AND
PHOSPHINES WERE MADE BY HUMANS.
THEY'RE NOT NATURAL CHEMICAL
GROUPS.
THIS CHEMISTRY WAS SO WELL
TOLERATED IN BIOLOGICAL SETTINGS
WE COULD EVEN PERFORM THIS IN
THE BODY OF A MOUSE AND WE
PUBLISHED -- THIS IS THE WORK OF
TWO FORMER STUDENTS A PROFESSOR
NOW AT U.C. IRVINE, ANOTHER AT
BODEN COLLEGE AND SHOWED YOU CAN
INJECT A PRECURSOR OF THE SIALIC
ACID WITH THE AZIDE AND THE
SUGAR WOULD GET METABOLIZED IN
THE BODY OF THE MOUSE.
A COUPLE DAYS LATER YOU'D FIND
THE GLYCANS AND INTRODUCE YOUR
PHOSPHINE PROBE.
IT'S HERE AND THAT WILL REACT
WITH THE AZIDE AND THE REST IS
JUST A PROBE.
SOMETHING YOU CAN FIND AND QUANT
TIGHT ON THE CELLS AFTER THE
REACTION IS COMPLETE.
SO WE SPENT MANY YEARS PLAYING
WITH THE CHEMISTRY, DOING
INTERESTING THINGS AND
PUBLISHING WITH IT AND SO ON BUT
WE REALIZED PRETTY QUICKLY THAT
THIS WAS NOT THE IDEAL CHEMISTRY
FOR IMAGING APPLICATIONS.
FOR THE SIMPLE REASON IT'S A
RATHER SLUGGISH REACTION AND
GOES A LITTLE BIT TOO SLOW.
IT'S FINE FOR REACTIONS TO GO
SLOWLY IN THE ROUND BOTTOM FLASK
BECAUSE YOU CAN HEAT THEM UP AND
LET THEM GO FOR DAYS AND DAYS.
IN A MOUSE, YOU CAN'T HEAT IT
UP, OR YOU SHOULDN'T, ANYWAYS
AND FURTHERMORE YOU CAN'T JUST
WAIT FOR DAYS AND DAYS AND DAYS
BECAUSE THE MOUSE WILL CLEAR
THESE REAGENTS OUT OF ITS BODY
BECAUSE THE CHEMISTRY'S HAD A
CHANCE TO OCCUR IF THE REACTION
IS TOO SLOW.
SO IN THE EARLY 2000s WE WENT
BACK IT THE DRAWING BOARD AND
STARTED THINKING ABOUT OTHER
CHEMISTRIES WE COULD PERFORM
PRESERVING THE AZIDE AS AS A
PARTNER BECAUSE WE LOVED IT.
IT'S SMALL AND HAS GREAT
STABILITY IN BIOLOGICAL SYSTEMS.
LOTS OF HUMAN MEDICINES HAVE
AZIDES IN THEM SO WE KNOW A LOT
ABOUT THE METABOLISM AND THEY'RE
INERT IN THE BODY AND IT TURNS
OUT THEY HAVE ACTIVITIES WITH
OTHER GROUPS ONE BEING ALKINES.
NOW ALKINES WERE WELL KNOWN ALSO
SOME FOUNDATIONAL CHEMISTRY FROM
GERMANY IN THE '50s AND '60s AND
FORMED A TRIAZOL.
THIS WAS WELL KNOWN TO THE
CHEMICAL COMMUNITY BUT NOT IDEAL
FOR BIOORTHOGONAL APPLICATIONS
BECAUSE IT'S EVEN SLOWER THAN
THE STAUDINGER CHEMISTRY ON ITS
FACE.
BUT IN THE YEAR 2002 THESE TWO
SYNTHETIC ORGANIC CHEMISTS,
DR. MEIDAL AND SHARPLESS
INDEPENDENTLY AND SIMULTANEOUSLY
ZORCHD YOU CAN SPEED IT UP BY
ADDING A COPPER CATALYST AND
THIS AZIDE CYCLE EDITION BECAME
POPULARLY KNOWN AS CLICK
CHEMISTRY.
WE LEARNED FOR THE BIOLOGISTS IN
THE AUDIENCE YOU PROBABLY NOT
HEARD OF ANY OTHER REACTION BY
NAME OTHER THAN CLICK CHEMISTRY.
HOW MANY HEARD OF THIS?
AND HOW MANY HAVE DONE CLICK
CHEMISTRY?
YEAH.
THIS IS ONE OF THE FEW CHEMICAL
RAKES THAT EVEN A BIOLOGIST CAN
DO IN THE LABORATORY.
BUT THAT SAYS SOMETHING.
IT'S FOOL PROOF AND YOU CAN SPIT
ON THE REACTION, AND THAT'S AN
INSIDE CHEMISTRY JOKE.
WHEN THE PAPER CAME OUT WE GOT
EXCITED BECAUSE IT'S SUCH A FAST
REACTION.
IT DOESN'T TAKE DAYS BUT
MINUTES.
MAYBE AN HOUR AT MOST.
THE PROBLEM WAS THE COPPER
CATALYST.
THAT COPPER CATALYST IS TOXIC TO
CELLS.
SO WHILE CLICK CHEMISTRY IS
USEFUL TO JOIN THINGS TOGETHER
IN VERY COMPLEX EVEN BIOLOGICAL
SETTINGS, IT'S NOT USEFUL IF THE
OBJECTIVE IS TO STUDY SOMETHING
THAT IS ALIVE WHETHER IT'S CELLS
OR ORGANISMS OKAY, SO THAT
WASN'T GOING TO SOLVE THE
PROBLEM FOR US.
BUT IT TURNS OUT THERE ARE OTHER
WAYS OF ACCELERATING THE RATE OF
AN AZIDE ALKINE CYCLE EDITION
THAT DON'T REQUIRE THE CATALYST
AND THE STRATEGY WAS TO TAKE THE
TRIPLE BOND ALKINE AND CONFINE
IT IN AN EIGHT MEMBER DRAIN AND
YOU INDUCE RING STRAIN AND THE
CHEMISTS WILL KNOW WHEN THINGS
ARE CONSTRAINED THEY'RE
UNCOMFORTABLE.
MOLECULES WANT TO RELEASE IT AND
WHEN THIS REACTS WITH THE AZIDE
TO FORM THE TRIZOLE THAT DRIVES
THE KINETICS AND THE COPPER FREE
CLICK CHEMISTRY AS WE CALL IT IS
SOMETHING YOU CAN DO IN LIVING
SYSTEMS WITHOUT HAVING TO WORRY
ABOUT TEXASES.
-- TOXICITIES AND BADGE --
BARRY AND MYSELF AND OTHERS WERE
RECOGNIZED WITH A NOBEL PRIZE
FOR THIS IN 2022.
WE HAD A NEW BIOORTHOGONAL
CHEMISTRY AND THE OBJECTIVE WAS
TO GET THEM IN MOLECULES, CELLS,
ORGANISMS IN THE SUGAR AROUND
INTRODUCE THE PROBES WITH
FLUORESCENT DIES OR IMAGING
PROBES MORE BROADLY AND LET THEM
CLICK TOGETHER AND IMAGE THE
PRODUCTS.
IT TURNED OUT TO BE THE EASIEST
PART BECAUSE WE HAD THIS GREAT
PRECEDENT FROM THE PROFESSOR'S
EARLIER WORK.
WE HAD IT ON THE PRECURSOR.
WE FED THAT TO CELLS, THE CELLS
DID THE HARD WORK AND BUILT A
SIALIC ACID AND YOU CAN DO THIS
PRETTY MUCH ANY SUGAR.
YOU CAN PUT IT IN THE GLUCOSE.
THAT'S ANOTHER INTERESTING SUGAR
BIOLOGICALLY AND INTO GALNAC AND
FOLKS WHO STUDY THE MUSIN FAMILY
MAY HAVE AVEILED THEMSELVES OF
THIS TECHNOLOGY.
WHO HERE HAS FED CELLS GALNAZ.
ALL RIGHT, DR. HANOVER.
AND THE ORGANISM WE STARTED WITH
IS POPULAR FOR OPTICAL
MICROSCOPY IS ZEBRAFISH THEY'RE
TRANSLUCENT AND THEY'RE
VERTEBRATES ARE SIMILAR TO
HUMANS AND AS PEOPLE ARE AWARE,
THE PROGRAMS OF EMBRYO GENESIS
VERY MUCH MIRROR THE PROGRAMS OF
CANCER DEVELOPMENT AND
DISSEMINATION.
MANY OF THE SAME GENES WE
ASSOCIATE WITH MALIGNANCIES ARE
EXPRESSED AND THAT'S TRUE FOR
THE CELL SURFACE GLYCANS.
IT'S A GOOD PLACE TO START.
FOR THOSE WHO HAVE NOT SPENT
TIME LOOKING AT ZEBRAFISH
EMBRYOS IT ALL STARTS WITH A
SINGLE CELL YOU FERTILIZE IN A
DISH, IN VITRO FERTILIZATION AND
THE EMBRYO STARTS TO DEVELOP AND
AT THE END OF EIGHT DAYS YOU
HAVE A YOUNG ADULT FISH AND CAN
WATCH THE DEVELOPMENTAL PROGRAM
UNFOLD IN REAL TIME DURING THE
FIVE-DAY PERIOD.
SO SO WE STARTED DOING
EXPERIMENTS OF THIS TYPE WHERE
WE WOULD TAKE THESE EMBRYOS AND
WE WOULD BATHE THEM IN MEDIA
THAT HAD SUGARS WITH AZIDES IN
THEM.
WE WOULD LET THOSE SUGARS GET
TAKEN UP AND INTO THE GLYCANS
AND ADD DYE AND THIS HAPPENS TO
BE A TYPE AND DO IT AGAIN AND
AGAIN TO YOUR HEART'S DESIRE.
TAKE THE FISH AND DO A WASHING
STEP AND THEN BATHE THEM IN A
NEW BATCH OF SUGAR INTRODUCING A
NEW BATCH OF AZIDES AND THEN YOU
CAN CLICK ON A DIFFERENT COLOR.
THROUGH THIS EXPERIMENTATION YOU
CAN LOOK AT DIFFERENT SUGARS AND
TIME POINTS THROUGH THE COURSE
OF DEVELOPMENT.
SO IMAGES LIKE THESE BECAME LIKE
THE DOMINATING DATA SETS IN OUR
LAB AROUND THOSE YEAR.
THIS NOW TAKES ME UP TO 2004
THROUGH 2008.
UP HERE IS THE HEAD OF A
FIVE--DAY-OLD ZEBRAFISH THAT WAS
LABELLED WITH THREE DIFFERENT
SUGARS AT DIFFERENT TIME POINTS.
YOU CAN SEE THE COLORS ARE
DISTRIBUTED IN INTERESTING WAYS.
YOU CAN DISTINGUISH THE PATTERNS
OF GLYCOSYLATION IN THE UPPER
RESPIRATORY TRACT IN THE NOSE
WHICH IS DIFFERENT FROM THE
EPIDERMIS OR OTHER TISSUE.
YOU CAN BACK UP TO EARLY STAGES
OF EMBRYO GENESIS.
THE BALL OF CELLS IS A 24-HOUR
OLD EMBRYO AND A LOT OF CELL
DIVISION IS HAPPENING IN THE
EARLY STAGES.
CELLS ARE MAKING THEIR DISHES
ABOUT WHAT TISSUE FATE THEY WILL
ADOPT AND WE CAPTURED
INTERESTING PHENOMENA.
FOR EXAMPLE, EVERY TIME WE FOUND
A CELL GOING THROUGH A ROUND OF
MITOSIS WE DISCOVERED THE CELL
SURFACE SUGARS WERE
REDISTRIBUTING THEMSELVES AND
BELOW IS A BLOWUP OF ONE OF
THESE EVENTS OF MITOSIS.
TO SEE IF I CAN GET THIS TO
PLAY.
SO THIS IS A BLOWUP OF A MOTHER
CELL WHO HAS DUPLICATED HER
CHROMOSOMES AND DIVIDED TO TWO
DAUGHTER CELLS.
IT'S THE SAME FRAME IN EACH OF
THE PANELS BUT WE'RE LOOKING AT
DIFFERENT MOLECULES WITH
DIFFERENT DYES.
ON THE LEFT ARE CELLS IN WHICH
THE FLUORESCENT DYE IS ON THE
DNA AND YOU'RE LOOKING AT THE
CHROMOSOMES AND HERE THEY
DUPLICATED, CLASSIC MICROTUBIAL
DISCRIMINATION.
THAT'S WHAT IT LOOKS LIKE AND
JUMP OVER THE PANEL IN THIS
FRAME WITH WE HAVE A MEMBRANE
WITH A GENETIC FUSION YOU CAN
SEE THE MEMBRANE STARTS AS
BASICALLY CIRCLES THE MOTHER
CELL AND AS IT DIVIDES YOU GET
NEW MEMBRANE FORMING BETWEEN
ULTIMATELY TWO DAUGHTER CELLS.
THAT IS WHAT CELL DIVISION LOOKS
LIKE THROUGH THE LENS OF THE
MEMBRANE.
NOW FOR THE FIRST TIME WE GOT TO
SEE WHAT CELL DIVISION LOOKS
LIKE THROUGH THE LENS OF A CELL
SURFACE GLYCANS.
AND WHAT'S KIND OF SURPRISING
YOU SEE THEY LOOK LIKE A FUZZY
HALO.
PICTURE ALL THE TREES IN THE
GRASS AND LANDSCAPING ON THE
CELL SURFACE.
AS THE CELL GETS TO THE LATE
STAGES OF CELL DIVISION THE
GLYCANS ARE NO LONGER UNIFORMLY
DISTRIBUTED.
THEY START TO PILE UP AT THE
JUNCTION BETWEEN THE TWO
DAUGHTER CELLS AND TURNS OUT
THEY ARE CONTRIBUTING TO
SEGREGATION AT THIS STAGE OF
CELL DIVISION.
AND WE SPUN OUT A WHOLE PROJECT
SORT OF STUDYING THE FUNDAMENTAL
CONTRIBUTION OF THE GLYCAN TO
LATE STAGES OF CELL DIVISION.
IT'S LIKE YOU TURN ON YOUR
SPOTLIGHT AND YOU SHINE IT IN
PLACE AND SEE A NEW THING.
THAT WAS OUR EXPERIENCE USING
THESE BIOORTHOGONAL TOOLS.
BY THE WAY, THE LAST PANEL WAS
JUST AN OVERLAY OF THE THREE
PANELS WHERE THEY'RE JUST
COLORED.
SUFFICE IT SAY AT THIS POINT WE
DEVELOPED A TECHNOLOGY, A
STRATEGY FOR IMAGING SUGARS ON
THE SURFACE OF CELLS.
WE DEVELOPED SOME TOOLS WE COULD
USE IN ANIMAL MODELS AND WE SPUN
OUT A LOT OF THESE TECHNOLOGIES
INTO COMPANIES, SOME WERE
LICENSED BY COMPANIES INTERESTED
IN CLINICAL DIAGNOSTICS, A FEW
COMPANIES I CO-FOUNDED WITH SOME
STUDENTS FROM MY LAB.
BUT I THOUGHT IT WOULD BE FUN TO
FAST FORWARD AND TELL YOU A
RECENT STORY ALSO BASIC SCIENCE
DISCOVERY THAT WAS ENABLED BY
BIOORTHOGONAL CHEMISTRY AND IT
TAKES ME BACK TO THE GLYCOCALYX
OR THE CELL SURFACE.
EVERY TEXT YOU READ WILL TELL
YOU THERE'S TWO KINDS OF
GLYCOMOLECULES ON THE SURFACE,
GLYCOLIPIDS AND PROTEINS.
OKAY.
THAT HAS BEEN THE STANDARD STATE
OF THE ART OF KNOWLEDGE FOR MORE
ETHAN 100 YEARS NOW.
EVERY TEXTBOOK WILL TELL YOU
THIS.
IT TURNS OUT THAT'S NOT ALL.
A COUPLE YEARS AGO WE DISCOVERED
THAT THERE'S ANOTHER BIO
MOLECULE THAT ALSO GETS
GLYCOSYLATED AND ALSO PRESENTED
ON THE CELL SURFACE AND IT'S
RNA.
SO CHECK THIS OUT.
A FORMER POSTDOC IN MY LAB CAME
FROM A Ph.D. TRAINING IN HOWARD
CHANG'S LAB AT STANFORD WHO IS A
VERY FAMOUS RNA BIO CHEMIST NOW
HEAD OF RESEARCH FOR AMGEN AND
RECENTLY LEFT STANFORD TO AMGEN
AND CAME WITH DEEP EXPERTISE AND
ASKED ME HOW COME ALL THE OTHER
MOLECULE GLYCOSYLATED BUT NOT
RNA AND I SAID THAT'S THE WAY IT
IS AND HOW LIFE EVOLVED AND HE
SAID HAS ANYONE LOOKED?
AS FAR AS I KNOW NO ONE THOUGHT
TO LOOK AND THE CELL BIOLOGIST
WHY WOULD YOU LOOK AND WE KNOW
WHERE IT IS IN THE LUMEN OF THE
ER AND GOLGI COMPARTMENT AND
THEY TWO WORLDS DON'T TOUCH EACH
OTHER.
WHY WOULD YOU THINK RNA COULD BE
GLYCOSYLATED.
FORTUNATELY, RYAN THOUGHT IF NO
ONE'S LOOKED, LET ME BE THE ONE
AND ANSWER THE QUESTION ONCE AND
FOR ALL.
HE USED AZIDE SUGARS AND SAID IF
I GET IT IN MY RNA MAYBE IT'S
GLYCOSYLATED AND HE DID THAT AND
TOOK CELLS AND TOOK THE SUGARS
AND ACID AND DID RNA EXTRACTION
AND PURIFICATION STEPPED AND
CLICKED ON A BIO TIN PROBE USING
THE COPPER CLICK CHEMISTRY AND
ANALYZED THE CHEMISTRY USING
VARIOUS GELS AND BLOTTING
TECHNIQUES AND WAS SURPRISED TO
SEE THERE WAS A SPECIES THERE
THAT IS RNA CARRYING THE SUGAR.
HE RAN A SUCROSE GRADIENT TO
SEPARATE THEM OUT AND FOUND THE
SUGAR LABELLED RNA, IT'S
ACTUALLY MAPPING WITH THE SMALL
RNAs BUT IT WAS ON A GEL LIKE A
LARGE RNA.
IT HAS AN ANOMALOUS MIGRATION ON
THE GEL.
COULD IT BE GLYCOSYLATED RNA?
FOR THE NEXT TWO YEARS WE
BASICALLY SPENT EVERY DAY
THINKING HOW TO SHOOT DOWN THE
HYPOTHESIS BECAUSE IT SEEMED
IMPOSSIBLE BUT WE PROVED TO
OURSELVES AND ULTIMATELY THE
COMMUNITY THERE'S GLYCORNAs ON
THE SURFACE OF THE CELLS.
IN THE FIRST SET OF EXPERIMENTS
THE TYPE WE IDENTIFIED WERE ALL
SMALL STRUCTURED RNAs LIKE Y-RA
RNAs AND T RNAS AND THEY WERE
HIGHLY BRANCHED GLYCANS AND WE
PUBLISHED THIS IN '21 AND IT
TOOK FOUR ROUNDS OF REVIEWERS
AND SEVEN REVIEWERS TO GET IT
PUBLISHED IN CELL.
ONE THING WE REPORT IN THE PAPER
IS THE INTRIGUING OBSERVATION
THAT GLYCO RNAs ON THE CELL
SURFACE CAN ENGAGE A FAMILY OF
IMMUNE CHECKPOINT RECEPTORS.
IT'S EARLY DAYS BUT WE SUSPECT
IT MAY BE PART OF AN IMMUNE
MODULATORY OR TRAINING EFFECT
AND MAY PLAY A ROLE IN
REGULATION.
THAT'S AN OPEN QUESTION.
MOST IMPORTANTLY IS WE PUT WORK
INTO IDENTIFYING THE SPECIFIC
CHEMICAL LINKAGE THAT CONNECTS
THE GLYCOAND RNA.
ONE OF THE LINKAGES WE
IDENTIFIED WAS BASICALLY GLY
GLYCOSYLATION OF RTRNs AND THE
ACP3U MODIFIED BASE WHICH HAS
THE BIG CHEMICAL NAME AND IN A
NUTSHELL WE THE ACP3U RESIDUES
HIGHLY CONSERVED IN YOUR tRNA
AND VARIES FROM SPECIES TO
SPECIES BUT IN MAMMALS IT'S
ALWAYS IN THIS PARTICULAR LOOP.
HIRES THE CHEMICAL STRUCTURE.
IT'S THIS EXTRA AMINO ACID ABOUT
IT HANGING OFF OF IT AND THERE'S
A BIO SYNTHETIC PATHWAY WHEN
IT'S ALREADY IN THE TRNA SO YOU
GET THE MODIFICATION.
WITH A COLLABORATOR CALLED BEN
GARCIA A WORLD CLASS MASS
SPECTOMETERRIST THEY WERE ABLE
TO LINK IT AND HERE'S THE BASE,
HERE'S THE MODIFIED BASE AND
THERE'S AN AMINO GROUP AND FROM
THEN ON IT LOOKS LIKE A GLYCAN
YOU FIND ON YOUR FAVORITE
NEIGHBORHOOD PROTEIN.
SHOCKING.
AND RYAN HAS NOW DONE A LOT MORE
WORK ON THIS AND IS NOW A YOUNG
PROFESSOR AT HARVARD UNIVERSITY
AND HE IS BASICALLY BUILDING OUT
THE STORY OF GLYCORNA BIO
SYNTHESIS, TRAFFICKING, HOW THEY
DO THEY GET IN THE LUMEN AND
THAT'S WHY THEY'RE GETTING
GLYCOSYLATED?
IS A PRIMORDIAL FORM LATER
MIMICKED BY PROTEINS WITH THEIR
RESIDUES OR THE OTHER WAY
AROUND?
I MEAN, ACP3U IS HIGHLY
CONSERVED IN ALL DOMAINS OF
LIFE.
THIS MAY BE SOMETHING ANCIENT.
HIS LAB IS PICKING THIS APART
AND IF YOU DO A PUB MED SEARCH
OF GLYCORNA YOU'LL GET 40.
HE HAS A PIPELINE COMING OUT
NEXT MONTH WHICH WILL BLOW YOUR
MIND AND THE IMMUNE SYSTEM IS
PART OF THIS.
HOPEFULLY YOU'LL SEE THIS IS A
BASIC SCIENCE DISCOVERY THAT CAN
COME WHEN YOU HAVE A TOOL THAT
ALLOWS YOU TO OPEN A WINDOW TO A
NEW TYPE OF MOLECULE.
OKAY.
SO LET'S REVISE THE TEXTBOOKS.
HOPEFULLY IN THE NEXT GENERATION
OF TEXTBOOKS YOU WON'T JUST SEE
THE GLYCOPROTEINS AND
GLYCOLIPIDS BUT GLYCORNAs AND
ANCHORED BY RNA BINDING
PROTEINS.
SO IN THE LAST PART I'LL GIVE A
COUPLE VIGNETTES THAT ILLUSTRATE
HOW BIOORTHOGONAL AND CLICK
CHEMISTRIIES STARTED TRANSLATIN
INTO THE WORLD OF THERAPEUTICS.
YOU CAN GOOGLE WEBSITES AND A
NUMBER OF COMPANIES WILL
ADVERTISE THEMSELVES AS USING
BIOORTHOGONAL CHEMISTRY AS A
PLATFORM.
A VERY POPULAR APPLICATION IS IN
MAKING NEXT GENERATION ANTIBODY
DRUG CONJUGATES AND AN AREA MY
LAB HAS WORKED IN AND SOME
COMPANIES ARE USING THEM TO MAKE
VACCINES FOR PNEUMOCOCCUS AND
PEOPLE WHO MAKE ANY KIND OF
COMPLEX CONJUGATE PROBABLY ARE
AVAILING THEMSELVES OF THESE
CLICK CHEMISTRIES AND
BIOORTHOGONAL CHEMISTRIES.
THERE'S CONJUGATES BETWEEN A
TUMOR FIGHTING ANTIBODY AND
TOXIC PAY LOAD CONNECTED BY A
CHEMICAL LIQUOR.
THIS IS AN AREA I'VE BEEN
FOLLOWING SINCE IT CAUGHT FIRE
BACK IN THE 1990s AND FDA
APPROVALS IN THE 2000s.
RIGHT NOW THERE'S AT LEAST A
DOZEN ANTIBODY DRUG CONJUGATES
BY THE FDA AND CLINICAL TRIALS
IN CHINA ARE IN THE THOUSANDS
FOR NEXT GENERATION.
PEOPLE SPENT THE LAST 30 YEARS
SYSTEMICALLY SOLVING THE
PROBLEMS.
WE FIGURED OUT HOW TO MAKE
BETTER ANTIBODIES.
WE FIGURED OUT WHAT PAY LOADS
WORK BEST.
WE KIND CLEVER LINKERS STABLE IN
SERUM BUT FALL APART INSIDE
CANCER CELLS BUT THE AREA THAT
HASN'T BEEN DISRUPTED YET TO MY
MIND IS HOW YOU DO THE CHEMISTRY
BETWEEN THE PROTEINS WHICH COMES
FROM RECOMBINANT PRODUCTION AND
THE LINKER DRUG FROM CHEMICAL
SYNTHESIS.
AND SOME ARE DOING IT ON SISTINE
SIDE CHAINS WITHOUT MUCH CONTROL
OVER THE SITES AND STOICY STOIC
METRY.
THIS IS WHAT IT'S WELL
POSITIONED TO SOLVE.
IF YOU HAVE AN ANTIBODY WHICH
MIGHT TYPICALLY HAVE 70 OR 80
SURFACE EXPOSED LYSISES AND YOU
CAN'T HAVE 60 OR 70 BECAUSE
YOU'LL KILL THE MOLECULE.
IF THERE'S A WAY TO INTRODUCE A
NEW CHEMICAL GROUP,
BIOORTHOGONAL GROUP THAT NEITHER
INTERACTS OR INTERFERES WITH ANY
OTHER SIDE CHAIN YOU CAN DIRECT
THE CHEMISTRY WHERE YOU WANT IT.
YOU CAN PUT THE PAY LOAD HERE OR
HERE OR HERE OR HERE AS LONG AS
YOU HAVE A WAY TO INTRODUCE THE
CHEMICAL GROUP AT THE SITES AND
WITH THE RATIO OF INTEREST.
SO, WE PUT THOUGHT INTO HOW TO
LEVERAGE BIOORTHOGONAL CHEMISTRY
THIS WAY AND ULTIMATELY
DEVELOPED A TECHNIQUE WE CALL
THE ALDAHIDE TAG FOR MAKING SITE
SPECIFIC ANTIBODY DRUG
CONJUGATES STARTING WITH AN
ENZYME WHICH WILL OXIDIZE IT IN
A SHORT SEQUENCE ONLY FIVE
RESIDUES AND CONVERT IT TO WHAT
HAS AN ALDEHYDE AND CLONE THAT
INTO YOUR PROTEIN OF INTEREST
YOUR SITE OF INTEREST, THIS WILL
INTRODUCE AN ALDEHYDE AND YOU
CAN DO CHEMISTRY THAT DOESN'T
INTERFERE WITH THE REST OF THE
PROTEIN.
ON THE GROUND WHAT WE WOULD DO
WITH AN ANTIBODY IS CLONE THAT
MOTIF INTO THE GENE SEQUENCE OF
EITHER THE HEAVY CHAIN OR THE
LIGHT CHAIN, DEPENDING WHERE YOU
WANT IT.
EXPRESS THE PLASMIDS IN CELLS
WITH A PLASMID THAT BOOSTS THIS
ENZYME'S EXPRESSION AND THE
CELLS WILL SECRETE THE ANTIBODY
WITH ALDEHYDES READY TO GO AT
THE SITES YOU ENGINEER.
WE DEVELOPED A CHEMISTRY ALSO
BASED ON ANCIENT GERMAN
CHEMISTRY I WON'T GO INTO BUT
ALLOWS YOU TO ATTACH A LINKER
AND PAY LOAD DIRECTLY TO THE
SITES AND FOR THE CHEMISTS HERE
SOMEBODY ALWAYS ASKS WHAT'S THE
MECHANISM?
BASICALLY THIS REACTS AND YOU
GET AN INTRAMOLECULAR
SUBSTITUTION AND THE PEOPLE IN
THE AUDIENCE HARDCORE CHEMISTS
WILL RECOGNIZE THIS AND THAT
FORMS A CARBON LINKED PRODUCT
THAT'S ROCK STABLE.
WILL NOT FALL A PART EVEN IN THE
HUMAN BODY.
WE FOUND A COMPANY BY MYSELF AND
MY FORMER STUDENT AND EVENTUALLY
THAT COMPANY WAS ACQUIRED AND
NOW THIS IS ONE OF THE OFFERINGS
OF IT.
THEY PARTNERED WITH COMPANY WHO
WANT TO MAKE ADCs.
YOU E-MAIL THEM THE SEQUENCE OF
YOUR HEAVY CHAIN AND LIGHT CHAIN
AND FROM THERE THEY'LL MAKE THE
ADC AND SOME MOLECULES IN THE
CLINIC.
WHAT ABOUT CELLS?
WE CAN DO CHEMISTRY ON CELLS.
WE ALREADY KNOW THAT.
MAYBE WE CAN ADD SOMETHING TO
THE WORLD OF CELLULAR
THERAPEUTICS.
SO CAR-Ts.
EVERYONE'S EXCITE ABOUT THIS NEW
FORM OF IMMUNE THERAPY.
IT'S EXPENSIVE AND HARD TO SCALE
AND VERY BESPOKE.
THE TYPICAL PROTOCOL INVOLVES
FIRST REMOVING BLOOD FROM THE
CANCER PATIENT, TAKING THEIR T
CELLS EX VIVO AND INTRODUCE A
CHIMERIC RECEPTOR SO AFTER YOU
GROW THE CELLS AND REINTRODUCE
THEM IN THE PATIENT IN THE
PERFECT WORLD THEY HOLD TO THE
TUMOR AND KILL IT AND IT'S
REMARKABLE WHEN THEY WORK AND
FOR CERTAIN BLOOD CANCERS YOU
CAN GET A CURE WITH A CAR-T
THERAPY BUT LIKE I SAID, IT'S
EXPENSIVE AND DOESN'T SCALE AND
IS PATIENT BY PATIENT BY
PATIENT.
MOST PEOPLE DON'T HAVE ACCESS TO
THIS TECHNOLOGY.
EVEN PATIENTS WITH CANCERS THAT
COULD BENEFIT.
WELL, ONE OF MY FORMER STUDENTS
FORMED A STUDENT OF WHICH I
SERVE ON THE BOARD OF DIRECTORS
THAT IS USING BIOORTHOGONAL
CHEMISTRY TO CHEMICALLY ENGINEER
T CELLS TO HOME TO TUMORS RATHER
THAN GENETICALLY ENGINEER.
AS A CONSEQUENCE THEY CAN TAKE
OFF THE SHELF T CELLS THE
SO-CALLED GAMMA DELTA T CELLS
THAT ARE GENERALLY WELL
TOLERATED IN ALLOGENEIC FOLKS SO
THEY DON'T GET REJECTED.
SO TAY TAKE THE OFF THE SHELF T
CELLS AND DO BIOORTHOGONAL CHEM
TRY TO INTRODUCE THE ANTIBODY
FROM THE OUTSIDE AND IN EARLY
CLINICAL DATA THEY'RE EFFECTIVE
AT HOMING TO AND KILLING BLOOD
CANCERS IN PATIENTS.
HOPEFULLY IT WILL MAKE A CAR T
LIKE THERAPEUTIC CHEAPER AND
MORE AVAILABLE TO A BROADER
SPECTRUM OF PATIENTS.
AND THEN FINALLY I MENTIONED
EARLY ON WE HAD THIS DREAM OF
TAKING CHEMISTRY OUT OF THE
ROUND BOTTOM FLASK AND ONTO THE
CELL BUT CAN WE GO ALL THE WAY
TO THE CHEMISTRY IN THE HUMAN
BODY AND THE ANSWER IS YES.
THIS IS NOW HAPPENING IN THE
HANDS OF A COMPANY IN THE BAY
AREA.
SO THEY'RE USING A CHEMISTRY
CALLED TETRAZINE LIGATION AND
USES A HIGHLY STRAINED REAGENT
BUT IT'S A TRANS CYCLE OCTINE
AND THEY'LL REACT WITH REAGENTS
AND YOU CAN MANEUVER THE
CHEMISTRY SO AT THE SAME TIME
THESE TWO THINGS ARE CLICKING
TOGETHER THERE'S THE RELEASE OF
THIS SIDE CHAIN.
SO YOU CAN USE THE CHEMISTRY TO
NOT ONLY CLICK BUT RELEASE WHICH
MAKES IT NICE FOR A PRODRUG
APPLICATION.
TET -- THEY HAVE TAKEN DRUGS
THAT'S AN OLDER BUT GOODIE.
AND TAKE IT TO A PLACE WHERE IT
INACTIVATE THE DRUG AND THEY'LL
INTERJECT INTO THE INTRATUMORAL
ENVIRONMENT WITH THE TETRAZINE
ANCHORED TO IT AND THAT IS
INSERT IN THE HUMAN BODY AND
LATER WHEN YOU INFUSE THIS DRUG
WHENEVER THAT DRUG ENCOUNTER THE
TUMOR IT CAN REACT WITH THE
CLICK TO RELEASE AND SPIT OUT AT
HIGH LOSE LOCALLY IN THE TUMOR.
YOU CAN HAVE THIS DRUG COURSING
THROUGH THE BODY BUT IT'S NOT
TOXIC ONLY AS IT'S RELEASED IN
THE MICROTISSUE ENVIRONMENT AND
THEY'RE DOING A STUDY TO TREAT
SOFT TISSUE SARCOMA.
WHAT STARTED AS A SOLUTION TO A
SPECIFIC PROBLEM TO IMAGE CELL
SURFACE SUGARS TOOK OFF AND
FOUND ALL KIND OF INTERESTING
APPLICATIONS.
SOME IN MY LAB BUT MOST OUTSIDE
IN OTHER SETTINGS AND MOST
EXCITINGLY NOW IN WAYS THAT WILL
HOPEFULLY TRANSLATE TO
TREATMENTS FOR HUMAN DISEASES.
SO THERE'S A LOT OF NEW
APPLICATIONS AND I LOVE SEEING
HOW THE COMMUNITY HAS RUN WITH
THIS KIND OF SCIENCE.
WE CAN'T PREDICT THE FUTURE BUT
I THINK BIOORTHOGONAL AND CLICK
CHEMISTRY WILL BE USEFUL TO YOU
AND IF YOU SEE SOMETHING YOU
WANT TO TRY, SEND ME AN AGENT
AND WE'LL SEND YOU WHATEVER
REAGENTS WE HAVE, OF COURSE.
FOR THAT, THANK YOU VERY MUCH
AND I CAN'T THANK EVERYONE
BECAUSE IT'S ABOUT MANY OVER THE
DECADES BUT I'M INDEBTED TO MY
POSTDOCS FOR DOING THE WORK WITH
ME AND HAPPY TO ANSWER
QUESTIONS.
THANK YOU.
>> FOLKS, USE THE MICS.
>> HI.
SUPER COOL.
THE THING I'M THINKING ABOUT THE
ENTIRE I'M WATCHING IS THE EIGHT
MEMBER THING.
WHERE DOES THAT COME FROM?
DO YOU HAVE TO MAKE IT?
IS IT UNSTABLE AND EXPENSIVE?
THAT'S WHAT I WAS SWEATING
ABOUT?
>> IN THE EARLY DAYS THERE
REALLY WASN'T MUCH LITERATURE ON
CYCLE OCTINES AND THERE WERE A
COUPLE PAPERS AND PEOPLE PLAYED
WITH THEM BACK THEN.
WE DEVELOP SYNTHETIC ROUTES TO
TUNE THEIR REACTIVITIES OVER
MANY ORDERS OF MAGNITUDE AND
NOWADAYS YOU CAN BUY THEM FROM
SOURCES AND THAT WAS A MOM AND
POP SHOP OUT OF GEORGIA SELLING
THEM AND THEY GOT ACQUIRED BY
VECTOR LABS WHICH NOW SELLS THEM
BUT YOU CAN BUY FROM THERMAL
FISHER ALL THIS STUFF IS NOW
TOTALLY COMMERCIALIZED.
YOU MAY HAVE USED THEM AS
REAGENT A, B AND C.
>> ARE THEY STABLE?
>> YOU CAN MAKE THEM STABLE.
THE ONES WE USE ARE VERY STABLE.
WE STORE THEM ROOM TEMP ON THE
BENCH TOP.
THE MORE REACTIVE THEY ARE THE
LESS STABLE SO YOU CAN SOUP THEM
UP SO THEY'RE SUPER FAST BUT
THEN THEY DON'T HAVE A GOOD
SHELF LIFE.
THE ONES WE USE ARE FAST ENOUGH
AND FINE.
YEAH.
WE DON'T TREAT THEM IN A SPECIAL
WAY WITH KID GLOVES OR ANYTHING
AND AFTERWARDS BY THE WAY, COME
LOOK AT MY EARRINGS IF YOU CAN'T
TELL FROM THERE.
>> CAROLYN, BEAUTIFUL TALK.
THIS IS ABOUT NEW BIOORTHOGONAL
REACTION.
THERE'S SO MUCH CHEMICAL
REACTIVITY DATA PEOPLE ARE DOING
SYNTHESIS.
WE'RE USING A.I.
I'M CURIOUS IS THERE A WAY TO
GET THE REACTIVITY DATA AND USE
THE KNOWN BIOORTHOGONAL
REACTIONS TO PREDICT WHAT COULD
BE NEW REACTIONS.
DO YOU THINK THAT CAN BE DONE OR
IS BEING DONE
>> ABOUT QUESTION.
LOTS OF PEOPLE NOWADAYS MORE
HARDCORE CHEMISTRY LABS ARE
INTERESTED IN DEVELOPING NEXT
GENERATIONS OF BIOORTHOGONAL
CHEMISTRIES.
WHERE THEY COULD BE MOST USEFUL
IS ORTHOGONAL TO THE CURRENT SET
WITHOUT CROSS REACTIVITY.
MOST THE WORK I'VE SEEN HAS
TAKEN THE SAME THEME WE TOOK
WHICH IS DIAL THE CLOCK BACK TO
THE 1950s AND '60s WHERE
PHYSICAL AND ORGANIC CHEMISTRY
WAS THE THEME AND YOU COULD FIND
PEARLS FROM THAT LITERATURE AND
BRING IT UP AND USE IT THIS WAY.
THE FIELD IS SHIFTING NOW MORE
TOWARDS USING MODERN
COMPUTATIONAL METHODS BECAUSE WE
HAVE THE ABILITY NOW TO SIMULATE
TRANSITION STATES AND THIS IS
ALL ABOUT TRANSITION STATES.
HAVE YOU TWO YOU WANT TO BIND
TOGETHER THAN WHAT THEY COULD
ACHIEVE WITH A BIOLOGICAL
CHEMICAL.
THAT'S THE SELECTIVITY.
I KNOW SOME PEOPLE MODELLING
LIKE IF I TAKE THIS REAGENT
WHICH MAY FORM A LOW ENERGY
TRANSITION STATE.
I THINK THAT'S AN INTERESTING
AND POTENTIALLY EXCITING WAY TO
BREAK OPEN NEW INNOVATION IN
THIS SPACE.
>> THANK YOU.
>> I'LL SNEAK IN ONE QUESTION
FROM ONLINE IN THE FRONT ROW.
OVER HERE.
>> I CAN'T FIND YOU.
>> IT COMES FROM ONLINE THOUGH
SO YOU DON'T HAVE TO STARE
ANYWHERE WHERE WHEN YOU ANSWER.
AZIDES ARE KNOWN TO BE TOXIC TO
HUMANS SO HOW WOULD YOU STUDY
AZIDES IN HUMANS?
>> THIS IS MY FIRST GRANT
APPLICATION BACK IN 1999.
SO AZIDE AND THE SALT IS TOXIC.
IT'S USED AS A METABOLIC POISON.
WE PUT IT TO PREVENT BACTERIAL
GROWTH BUT WHERE THE N3 IS
ATTACHED TO AN ORGANIC MOLECULE
NOT A NEGATIVE CHARGE BUT BOND
TO A CARBON ARE HARMLESS,
TOTALLY FINE.
FOR EXAMPLE, THE DRUG AZT, THAT
WAS ONE OF THE FIRST GENERATION
AIDS DRUGS, THAT IS AN AZIDE.
PEOPLE TAKE GRAMS OR PEOPLE TOOK
GRAMS PER DAY AND
GLAXOSMITHKLINE DID THE STUDY
AND IT COMES OUT IN THE URINE.
IT'S VERY SAFE AS LONG AS
THEY'RE BIOORGANIC.
IT'S A TRANSPORT CHAIN.
>> EXCELLENT TALK.
THE REACTION OF A CLICK REACTION
HAS THE REACTION WHO INTRODUCED
IT?
>> WHO INVENTED IT?
>> [INDISCERNIBLE].
OF COURSE IT WAS A CLICK
REACTION AT ROOM TEMPERATURE.
AND SO YOU SET IT UP BEAUTIFUL.
FANTASTIC WITH THE CHEMISTRY BUT
THE ANTIBODY DRUG CONJUGATION WE
USE THE ENZYMES TO CONNECT THE
SUGARS BASED ON THE STRUCTURE
MUTATED SO IT CAN BE BASICALLY
CONNECTED VIA SUGARS, GLYCAN
SUGARS.
YOU DID NOT INCLUDE THAT S&P --.
>> OTHER PEOPLE HAVE.
YOU MAY ASK WHY DIDN'T WE PUT AN
AZIDE ON THE PROTEIN?
BECAUSE METHODS FOR INTRODUCING
AZIDES LIKE AN UNNATURAL AMINO
ACID EXIST.
PET
PETER SCHULTZ' LAB DID THAT AND
YOU HAVE TO DO THAT WITH
UNNATURAL AMINO ACIDS SO WE
THOUGHT THIS WAS AN EASIER WAY
TO DO IT WHERE IT'S NOT
UNNATURAL BUT REPURPOSED
POST-TRANSLATIONAL MODIFICATION
THAT CREATES A DIFFERENT KIND OF
CHEMICAL GROUP SO HONESTLY WE
THOUGHT WE COULD GET TO SITE
SPECIFIC ADCs ON SCALE FASTER
THIS WAY.
BUT OTHER PHARMACEUTICALS IF I
GO BACK TO THAT SLIDE WITH LOGOS
ON IT -- THIS COMPANY, THEY'RE
MAKING ANTIBODY DRUG CONJUGATES
WITH COPPER-FREE CLICK CHEMISTRY
AND PATENTED IT AND ANOTHER IS
MAKING SITE-SPECIFIC ADCs AND
THREES ARE DOING THAT BUT WE
HAVE A DIFFERENT CHEMISTRY.
>> AND THE ENZYME IS BEING USED.
THERE'S TRANSFERASE.
>> IN FACT SYNOFIX DOES IT WITH
THE ANTIBODY WITH THE ACID.
>> I'D LIKE TO SQUEEZE IN THREE
MORE QUESTIONS ONLINE.
ONE AND TWO AND THREE.
WE'RE AT THE HOUR NOW.
>> SO MAYBE I MISSED THIS BUT ON
ONE OF THE -- HI.
ON ONE OF THE LAST SLIDES WHEN
YOU'RE TALKING ABOUT THE
TETRAZINE LOCATION WHAT ABOUT
THE ENVIRONMENT CAUSE THE
BINDING AND RELEASE.
>> SORRY, I WENT THROUGH THAT SO
QUICKLY IN THE INTEREST OF TIME
BUT SINCE YOU ASKED, NOW I CAN
USE MORE TIME.
THE IDEA IS THIS.
SO, THIS IS A HYDROGEL POLYMER A
FORM OF HYALURONIC ACID.
WHICH IS JUVADERM AND THEY
MODIFY IT AND INJECT IT IN THE
TUMOR SO IT'S A PROBLEM IN THE
TUMOR SITTING THERE DOING
NOTHING.
WHEN IT ENCOUNTERS THE DRUG THEY
FIND EACH OTHER AND BOOM THE
REACTION HAPPENS SPONTANEOUSLY.
WHEN THAT OCCURS ALL THE
UNRAVELLING HAPPENS AND YOU
LIBERATE THIS MOLECULE WHICH IS
NOW TOXIC AND CONCENTRATED IN
THE MICROENVIRONMENT.
>> TO MORE AND WE'LL HAVE A
RECEPTION.
>> HELLO.
THIS IS A REALLY GENERAL
QUESTION, BUT TO THE POINT ABOUT
THE SPACE OF BIOLOGICALLY
OCCURRING CHEMICAL REACTIONS I'M
WONDERING WHAT IS KNOWN ABOUT
HOW QUICKLY BIOLOGICAL SYSTEMS
EVOLVE TO EXPLORE NEW --
>> IF WE WAIT LONG ENOUGH
REACTIONS EVOLVE IN NATURE.
GREAT QUESTION.
ONE THING I'VE LEARNED IS IT'S
UNWISE TO SAY NATURE DOESN'T DO
X.
BECAUSE WHAT WE KNOW ABOUT WHAT
NATURE DOES AND DOESN'T DO IS A
SCRATCH ON THE SURFACE OF
REALITY.
WHO KNOWS WHAT EXISTS THAT WE
DON'T KNOW ABOUT YET.
AND ALL THE MORE REASON TO STUDY
HUMAN BIOLOGY WITH CHEMICAL
TOOLS, I GUESS.
>> SO THIS IS ALSO VERY BROAD
QUESTION BUT WITH ALL THESE
BEAUTIFUL MOLECULES AND
EVERYTHING AND THESE REACTIONS,
ONCE THEY ACTUALLY OCCUR AND
REACT, WHAT'S HAPPENING
CLEARANCE WISE WITH THE BIO
AZIDES MAINLY CLEARED BUT ARE
THERE COMMON CLEARANCE PATHWAYS
FOR A LOT OF THESE OR HAS THAT
NOT BEEN EXPLORED?
>> I WISH I HAD MORE OF
SUBSTANCE TO SAY IN RESPONSE TO
THAT QUESTION.
THE TRUTH IS WE DON'T REALLY
KNOW HOW THE ADDUCTS WE MAKE ARE
GETTING METABOLIZED.
THEY GET CLEARED AND DISAPPEAR
AND PRESUMABLY BROKEN DOWN TO A
FRAGMENT THAT GETS CLEARED FROM
THE LIVER OR MAYBE KIDNEYS AND
SECRETED OUT IN THE URINE BUT
I -- EXCRETED THROUGH THE URINE.
IT THANKS FOR EXPOSING THE
IGNORANCE.
>> WE'RE GOING TO WRAP IT UP.
I INVITE YOU TO ATTEND A
RECEPTION ACROSS THE HALL IN THE
NIH LIBRARY AND PLEASE JOIN ME
IN THANKING DR. BERTOZZI FOR AN
AMAZING LECTURE.
>> THANK YOU.