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OUR SPEAKER TODAY IS DR. NATALIE
SHAW, DR. SHAW IS A LASKER
CLINICAL RESEARCH SCHOLAR AND
HEAD OF PEDIATRIC
NEUROENDOCRINOLOGY GROUP AT
NATIONAL INSTITUTE OF
ENVIRONMENTAL HEALTH SCIENCE.
DR. SHAW EARNED HER MEDICAL
DEGREE FROM SATE UNIVERSITY NEW
YORK MEDICAL SCHOOL AT BUFFALO
AND MASTERS IN MEDICAL SCIENCE
FROM HARVARD MEDICAL SCHOOL.
SHE COMPLETED HER PEDIATRIC
RESIDENCY AT JOE PEDIATRIC
HOSPITAL IN PITTSBURGH, AND
CLINICAL RESEARCH FELLOWSHIP IN
REPRODUCTIVE END KIN UNIT AT
MASSACHUSETTS GENERAL.
SHE SERVED ATTENDING
PEDIATRICIAN AT BOSTON
CHILDREN'S HOSPITAL UNTIL
RECRUITED TO THE NIH IN 2015 AS
A LASKER CLINICAL RESEARCH
SCHOLAR.
AS A INVESTIGATOR, IN THE
CLINICAL RESEARCH BRANCH OF
NIEHS DR. SHAW STUDIED
ENVIRONMENTAL GENETIC CONTROL
PUBERTAL DEVELOPMENT IF RESEARCH
ON EFFECT OF OBESITY ON PUBERTAL
INITIATION AND DEVELOPMENTAL
TRAJECTORY OF CYCLES THAT FOLLOW
MONARCHY TO ESTABLISHMENT OF
CYCLES IN YOUNG ADULTHOOD.
DR. SHAW CONDUCTS STUDIES IN
HEALTHY PEDIATRIC VOLUNTEERS
COMPLIMENTED IN PHENOTYPIC
STUDIES IN PATIENTS WITH RARE
FORMS OF HYPOGOD INISM AND
PATIENT DERIVED NEURAL STEM
CELLS CELLS.
SHE RECEIVED NUMEROUS
RECOMMENDATIONS TO INCLUDE
ENDOCRINE SOCIETY EARLY
INVESTIGATOR AWARD, AMERICAN
SOCIETY FOR CLINICAL
INVESTIGATION YON PHYSICIAN
SCIENTIST AWARD AND WOMEN IN
ENDOCRINOLOGY INVESTIGATOR
AWARD.
SHE IS BOARD CERTIFIED IN
PEDIATRICS AS MEMBER OF THE
ENDOR ENDOCRINE SOCIETY WOMEN IN
ENDOCRINOLOGY AND PEDIATRIC
ENDOCRINE SOCIETY.
HER TALK TODAY IS ENTITLED SMCHD
1 AND EPIGENETICS.
LESSONS LEARNED FROM PATIENTS
BORN WITHOUT A NOSE.
WELCOME OUR SPEAKER, DR. SHAW.
ALL RIGHT.
THANK YOU AGAIN FOR THE
INVITATION TO SPEAK TODAY.
AS DR. BURR CASH FLO I'M A PEDIATRIC
ENDOCRINOLOGIST AT NIEHS IN
NORTH CAROLINA AND TODAY I WILL
TELL YOU A RECENT EXCITING
GENETIC DISCOVERY THAT STARTED
OUT AS A SMALL SIDE PROJECT WHEN
I WAS AN ENDOCRINOLOGY
FELLOWSHIP BIBUT SINCE BLOSSOMED
TO INTERNATIONAL COLLABORATIVE
PROJECT CROSSING MULTIPLE
MEDICAL DISCIPLINES.
I HAVE NOTHING TO DISCLOSE.
HERE IS TODAY'S LEARNING
OBJECTIVE.
AS WITH MANY MEDICAL
DISCOVERIES, SORRY, I'M TRYING
TO MOVE -- THE PICTURES.
OKAY.
AS WITH MANY MEDICAL DISCOVERIES
THIS STARTED WITH AN INTRIGUING
PATIENT.
THE WOMAN PRESENTED TO THE
REPRODUCTIVE ENDOCRINE UNIT AT
MASS GENERAL HOSPITAL WITH A
CHIEF COMPLAINT OF AMENRRHEA OR
ABSENT MEDICAL CYCLES. SHE WAS
BORN WITH MULTIPLE DEFECTS
INCLUDING ABOUT NORMAL EYES AND
CONDITION CALLED ARYANIA OR
COMPLETE ABSENCE OF EXTERNAL
NOSE.
SHE ALSO REPORTED SHE HAD SOME
TYPE OF HYPOTHALAMIC PITUITARY
-- SO DO THESE TWO CONDITIONS
HAVE ANYTHING TO DO WITH
(INAUDIBLE) LET'S STEM BACK A
MOMENT AND REVIEW THE
GONADOTROPIN RELEASING HORMONE
AT BASE OF THE HYPOTHALAMUS.
GNRH SIMULATES PITUITARY
SECRETION WHICH THEN SIMULATE
GOD INIAL SECRETION OF
TESTOSTERONE AND EXTRA DIAL.
BECAUSE REPRODUCTION CONSUMES SO
MUCH ENERGY PARTICULARLY IN
WOMEN, THE ACCESS IS TIGHTLY
CONTROLLED BY THE HYPOTHALAMUS
INTEGRATING INTERNAL AND
EXTERNAL FACTORS.
FOR EXAMPLE, METABOLIC MUST
INDICATE ENERGY STORES FOR
REPRODUCTION AND ENVIRONMENTAL
QUEUES MUST INDICATE FAVORABLE
ENVIRONMENTAL CONDITIONS.
FOR REPRODUCTION.
NOW, THE REPRODUCTIVE ACCESS HAS
TWO VERY DISTINCT FEATURES.
FIRST IS GNRH MUST BE RELEASED
IN A PULSE FASHION AS SHOWN
HERE.
WE CAN'T MEASURE GNRH BECAUSE OF
SHORT HALF LIFE BUT AS YOU CAN
IT IS ALSO PULSE TILE AND IS A
VALIDATED MARKER OF GNRH
SECESSION THAT WE CAN MEASURE IN
THE PERIPHERAL BLOOD.
THE SECOND UNIQUE FEATURE IS
THAT GNRH NEURONS DON'T STARTS
OUT IN THE BRAIN.
THEY MIGRATE THERE FROM THE
PRIMITIVE NOSE.
SO WHEN WE WALK YOU THROUGH THIS
SLIDE, THIS IS A CARTOON SHOWING
THE -- WE OFFER THE ROOT OF THE
NASAL CAVITY WHICH IS SEPARATED
BY THE BRAIN -- BY THE STRUCTURE
CALLED THE CRIB FORM PLATE WHICH
IS ALSO SHOWN HERE.
VERY EARLY IN DEVELOPMENT THERE
IS AN ECTO DERMAL THICKENING
CALLED THE OLFACTORY PLATEAU
WHICH GAVE RISE TO OLFACTORY
NEURONS AND GNHR NEURONS AND
OLFACTORY NEURONS EXTEND AXONS
THROUGH THE TRIGGER FORM PLATE
TO SYNAPSE ANNUAL OLFACTORY BULB
AND THEY HITCH A RIDE ALONG THE
AXONS TO MIGRATE TO BRAIN AND
REACH FINAL DESTINATION IN THE
HYPOTHALAMUS.
AS THIS IS HAPPENING, THE CRIB
REFORM PLATE IS MORE OF A SWAMPY
MESENCHYME SO WHEN THE BONE --
YOU CAN SEE PERFORATIONS IN THE
CRIB FORM PLATE WHICH ARE RELICS
OF THE OLFACTORY NERVE PASSAGE.
SO GIVEN THIS EMBRYOLOGY YOU
MIGHT PREDICT A DEFECT IN
OLFACTORY STRUCTURE WOULD ALSO
LEAD TO GNRH DEFICIENCY AND YOU
WOULD BE CORRECT THERE IS A A
NAME FOR THAT, COMMON SYNDROME
DEFINED BY COMBINATION OF
OLFACTORY DEFECTS AND --
DEFECTS.
THESE PATIENTS HAVE ABSENT OR
HYPOPLASTIC OLFACTORY BULKS IN
MRI AND ABSENT SENSE OF SMELL
DIAGNOSED BY A SIMPLE -- SHOWN
HERE.
THE MALES ARE BORN WITH
MICROPENIS AND ORPHANISM AND
BOTH SEXES HAVE ABSENT PUBERTY
AND INFERTILITY.
IF YOU WERE TO DO A FREQUENT
BLOOD SAMPLING STUDY ON SUE
ADJUDICATION YOU WOULD SEE
EXTREMELY SLOW LH LEVELS HERE
SHOWN ON THE BOTTOM IN CONTRAST
TO THIS -- PATTERN IN PATIENTS
HAVE UNCOVERED MUTATIONS IN MORE
THAN 30 GENES THAT HAVE PROVIDED
CRITICAL INSIGHT INTO
REPRODUCTIVE PHYSIOLOGY IN
HEALTH AND DISEASE.
FOR EXAMPLE, SNPS IN MANY OF
THESE GENES HAVE SHOWN UP IN
GENOME WIDE ASSOCIATION STUDIES
FOR AGENT MONARCHY.
GETTING BACK TO OUR PATIENT SHE
SIMPLY THEN HAVE EXTREME FORM OF
KALLMANN SYNDROME.
LET ME TELL YOU MORE ABOUT THE
PRESENTATION.
SHE WAS BORN WITH ARYANIA O OR
SMALL EYES, CATARACTS AND A HOLE
IN THE IRIS, AT AGE 16 SHE HAD
NO BREAST DEVELOPMENT, NO
MENSTRUAL PERIODS AND HAD SMALL
OVARIES AND UTERUS ON THE
ULTRASOUND.
FREQUENT BLOOD SAMPLING
DEMONSTRATED HYPOGONADSISM WITH
UNDETECTABLE LH AND FSH LEVEL
BUS REMAINDER OF HER
HYPOTHALAMIC PITUITARY AXIS WAS
INTACT.
AT AGE 23 SHE HAD A FORMAL
OLFACTORY EVALUATION WHICH
REVEALED COMPLETE INOSINEIA AND
ABSENCE OF OLFACTORY BULBS ON
MRI.
THIS PATIENT WAS ALSO STUDIED IN
COLLABORATION WITH JANICE LEE AT
NIDCR.
AND SHE CAME TO THE NIH CLINICAL
CENTER TO UNDERGO SOMETHING
CALLED THE CONE BEAM CT WHICH
PROVIDES HIGH RESOLUTION IMAGES
OF THE HEART TISSUE STRUCTURES
OF THE FACE AND JAW.
THIS IS SHOWING ENDOCRINE
CONTROL PATIENT AND YOU CAN SEE
THE WELL AIR ATED SINUSES AND
SIGH ANY ORANGE ARROWS POINTING
TO PERFORATION -- PLATE.
HERE YOU CAN SEE -- FACTORY
NEVER GNRH NERVE PASSAGE.
GENERAL IT CAN TESTING WAS
NEGATIVE FOR ALL KNOWN COMMON
GENES.
SO AS I DUG INTO THE LITERATURE
BIT MORE TO TRY TO UNDERSTAND
WHAT THIS PATIENT MIGHT HAVE, I
REALIZED THAT SHE HAD A LITTLE
KNOWN SYNDROME CALLED BOSMA
ARYANIA MICROON THAT WILL EMEA
SYNDROME.
THIS WAS DELINEATED BAY
PHYSICIAN AT THE NIH WHAT USED
TO BE CALLED THE NATIONAL
INSTITUTE OF DENTAL RESEARCH.
AND HE IDENTIFIED THIS SYNDROME
AND CAME ACROSS TWO MALE
PATIENTS WITH A TRIAD OF ARYANIA
OCULAR DEFECT AND HYPOGONADSISM,
ARYANIA IS EXCEEDINGLY RARE WITH
FEWER THAN HUNDRED CASES OVER
THE PAST CENTURY.
THE ETIOLOGY WAS UNKNOWN.
BUT THE PRESENCE OF FAMILIAL
CASES AND THE ABSENCE OF ANY
UNIFYING ENVIRONMENTAL EXPOSURE,
SUGGESTED THAT THERE MAY BE A
GENETIC CAUSE.
SO RECOGNIZING THIS TO BE THE AN
EXCITING OPPORTUNITY FOR GENE
DISCOVERY AND DEFINE THE
REPRODUCTIVE PHENOTYPE FURTHER,
I PARTNERED WITH THIS PATIENT
WHO IS A VERY BRIGHT WOMAN AND
ACTUALLY VERY CURIOUS ABOUT HER
CONDITION ADDITIONAL CASES.
THE NEWSPAPER CLIPPING FROM MORE
THAN 20 YEARS AGO DESCRIBING A
LO DAHLMAN WITH THE SAME EXACT
SYNDROME.
HE HAD UNDERGONE REPRODUCTIVE
SURGERY AS WELL BUT THROUGH A
BIT ROLL GOOGLE DETECTIVE WORK,
I WAS ABLE TO LOCATE THIS MAN
WHO HAD SINCE PASSED AWAY BUT
WHO WAS A LAWYER IN ALBANY AND
WHO WAS ACTUALLY ONE OF THE
FIRST OF THE TWO PATIENTS
ORIGINALLY DESCRIBED BY DR.
JAMES BOSMA.
BY NETWORKING WITH COLLEAGUES IN
ENDOCRINOLOGY AND GENETICS AND
CONTACTING AUTHORS OF PREVIOUS
CASE REPORTS AND FINALLY BY
ENLISTING THE HELP OF A MOTHER
WHO RUNS AN ARYANIA FACEBOOK
PAGE I WAS ABLE TO AMASS A
COHORT OF PATIENTS OF ALMOST 45
CASES AND 51 FAMILY MEMBERS.
IN COLLABORATION WITH COLLEAGUES
AT MASS GENERAL WE NEXT
PERFORMED WHOLE EXOME OR WHOLE
GENOME SEQUENCING THE FIRST 21
PATIENTS AND WHEN YOU LOOK FOR
ULTRA RARE NON-SYNONYMOUS
VARIANTS WE FOUND 84% CASES HAD
MUTATION IN THE SAME GENE SMCHD
1.
ALL HETEROZYGOUS MISSENSE
MUTATIONS RESTRICTED TO AXONS
3-13 OF THIS LARGE 48 EXON
PROTEIN -- GENE.
WE THEN PERFORM SOMETHING CALLED
GENE BASED RARE VARIANT BURDEN
TESTING WHERE WE LOOK AT THE
FREQUENCY OF ALL RARE
NON-SYNONYMOUS VARIANTS AND
EVERY -- IN THE GENOME IN OUR
COHORT COMPARED WITH 60,000
CONTROLS IN WHAT USED TO BE
CALLED EXACT IS NOW REFERRED TO
AS NOMAD BUT AS YOU CAN SEE IN
THIS MANHATTAN PLOT THERE WAS
ONLY ONE GENE ON CHROMOSOME 18
HERE TO ACHIEVE GENOME WIDE
SIGNIFICANCE AND THAT AGAIN WAS
SMCHD 1.
SO WE HAD VERY STRONG
STATISTICAL EVIDENCE FOR ROLE OF
SMCHD 1 BUT ONE SLIGHT PROBLEM.
AND THAT WAS THAT MUTATIONS IN
SMCHD 1 MANY SOME CASES
IDENTICAL TO THOSE FOUND IN OUR
COHORT, HAD ALREADY BEEN SHOWN
TO CAUSE A COMPLETELY DIFFERENT
DISORDER, FASTIO SCAUP LOW
MUSCULAR DYSTROPHY TYPE 2.
A DEGENERATIVE MUSCLE DISORDER
WITH AVERAGE ONSET IN THE 20s.
AND IT CAUSES SLOWLY PROGRESSIVE
WEAKNESS THAT PREDOMINANTLY
AFFECTS THE FACE, SHOULDER
GIRDLE AND UPPER ARMS.
THIS IS THE PATIENT DEMON
INVESTIGATING SOMETHING CALLED
WING SCAPULA WHERE THE MUSCLES
THAT NORMALLY ANCHOR THE SCAPULA
TO THE RIB CAGE ARE WEAKENED
LEADING TO THIS WINGED
APPEARANCE.
SO IT IS NOT UNHEARD OF FOR
MUTATIONS IN ONE PART OF PROTEIN
IN ONE CONDITION AND ANOTHER
PART OF THE PROTEIN TO CAUSE A
DIFFERENT CONDITION BE BUT AS I
SHOW YOU FOR FSHD 2 AND ARYANIA
THAT IS NOT THE CASE.
THIS SLIDE SHOUGHS THE DOMAINS
OF SMCHD 1 PROTEIN, THE 48 EXONS
HERE AND THE DIFFERENT MUTATION
TYPES THAT HAVE BEEN IDENTIFIED
IN THE PATIENTS.
SO IN RED IS THE DELETION
NON-SENSE AND SPLICE SITE AND
BLACK IS MISSENSE.
THE SMCHD 1 MUTATION HAS A
UBIQUITIN LIKE DOMAIN AND FSC
TERMINAL A HINGE DOMAIN.
YOU CAN SEE IN FHSD 2 MUTATIONS
ARE EITHER LOSS OF FUNCTION OR
MISSENSE MUTATIONS.
LOSS OF FUNCTION MUTATION SPAN
THE ENTIRE GENE WHEREAS THE
MISSENSE MUTATIONS SEND TO BE
CLUSTERED IN THIS END TERMINAL
REGION.
THAT IS ALSO THE CASE FOR THE
ARYANIA MUTATIONS, THESE
MUTATIONS ARE ENTIRELY MISSENSE
AND THEY ARE RESTRICTED TO EXONS
3-13.
AT LEAST WE IDENTIFIED FIVE
MUTATIONS THAT ARE IDENTICAL IN
THESE TWO CONDITIONS.
SO YOU MIGHT BE ASKING YOURSELF
IS THIS ONE DISEASE OR TWO?
THAT IS DO PATIENTS WITH ARYANIA
GO ON TO DEVELOP MUSCULAR
DYSTROPHY?
THIS HAS BEEN A DIFFICULT
QUESTION TO ANSWER BECAUSE MANY
OF THE CASE REPORTS ARE IN
INCIDENCE FIRST MADE AND
PATIENTS MAYBE LOSS TO
FOLLOW-UP.
THE SECOND IS THE FACIAL
WEAKNESS THAT IS CHARACTERISTIC
OF FHSD TYPE 2 MULTIPLE
RECONSTRUCTIVE SURGERIES.
WHAT ABOUT PATIENTS WITH
MUSCULAR DYSTROPHY, SUS L
DEFECTS IN SMELL AND
REPRODUCTION?
LET ME TELL YOU MORE ABOUT SMCHD
1 BIOLOGY AND FSHD 2.
SO FSHD 1 STANDS STRUCTURAL
MAINTENANCE OF CHROMOSOME HINGE
DOMAIN CONTAINING PROTEIN ONE,
THE MAJOR FUNCTION IS SERVING AS
A EPIGENERAL IT CAN REPRESSER.
AS A REMINDER EPIGENETICS REFERS
TO CHANGES IN GENE EXPRESSION
WITHOUT CHANGES IN DNA SEQUENCE.
AND YOU CAN THINK OF THIS AS
GIVING OUR GENOME SOME AGILITY.
THAT IS THE ABILITY TO FINE TUNE
GENE EXPRESSION IN SPACE OVER
TIME AND IN RESPONSE TO
ENVIRONMENTAL QUEUES.
GENES ARE TURNED OFF BY ONE OF
THREE MAJOR MECHANISMS.
THE FIRST IS DNA METHYLATION,
SECOND IS BY POST TRANSLATIONAL
MODIFICATION OF HISTONE PROTEIN
TAILS AND THE THIRD IS THROUGH
RNA BASED MECHANISMS SUCH AS
LONG RNs.
SMCHD 1 REPRESSES GENES ON THE
INACTIVE X CHROMOSOME, IT ALSO
SILENCES A SUBSET OF AUTOSOMAL
GENES PARTICULARLY THOSE THAT
ARE IMPRINTED OR MONOLITHICALLY
EXPRESSED FOR OTHER REASONS AND
GENES THAT ARE HIDDEN WITHIN
REPETITIVE DNA SEQUENCES.
IT DOES THIS THROUGH ALL OF
THOSE MECHANISMS THAT I SPOKE
ABOUT BY STABILIZING DNA
METHYLATION REPRESSSIVE HISSTONE
MARKS AND NON-CODING RNEs, IT
IS THOUGHT TO CONTRIBUTE TO
HIGHER ORDER CHROMOSOMAL
STRUCTURE SUCH AS TAGS.
SO HOW DO THESE MUTATIONS LEAD
TO MUSCULAR DYSTROPHY?
IN FHDH 2 MUTATIONS ARE NULL OR
MISSENSE MUTATION MUTATIONS IN SMCHD 1.
WHAT HAPPENS IS THAT THE LOSS OF
REPRESSSIVE ACTIVITY HAS A MAJOR
EFFECTNA IS SPECIFIC LOCUST ON
CHROMOSOME 4 AND THAT IS CALLED
THE D 4Z 4 REPEAT ARRAY.
LOSS OF REPRESSSIVE ACTIVITY
LEADS TO ECTOPIC EXPRESSION OF
DOUBLE HOMEOBOX OR TRANSCRIPTS
FACTOR.
VERY EARLY IN DEVELOPMENT, DUX 4
IS TURNED ON AN PLAYS A ROLE IN
ZYGOTIC GENOME ACTIVATION.
THIS IS WHEN THE GENOME OF THE
ZYGOTE OR EMBRYO IS FIRST
ACTIVATED AND MATERNAL mRNA
AND PROTEINS FROM THE OO SITE
ARE CLEARED.
AT SOME POINT IN DEVELOPMENT
THIS REGION BECOMES METHYLATED
SMCHD 1 BINDS AND STABILIZES ITS
IN DUX 4 IS TURNED OFF.
BUT FOR REASONS THAT AREN'T
ENTIRELY CLEAR, IN FHSD 2
SOMETHING GOES WRONG IN MUSCLE
CELLS.
SO THERE'S LOSS OF METHYLATION,
LOSS OF SMCHD 1 REPRESSSIVE
ACTIVITY AND DUX 4 IS PRODUCED
LEADING TO MUSCLE CELL
APOPTOSIS.
SO BACK TO OUR QUESTIONS IS THIS
ONE DISEASE OR TWO?
WE HAVE STUDIED A COHORT OF FHSD
2 PATIENTS AND FOUND NO CRANIAL
FACIAL OLFACTORY OR REPRODUCTIVE
PHENOTYPES.
WE HAVE ALSO STUDIED A COHORT OF
ARYANNA PATIENTS AT THE CLINICAL
CENTER, IN COLLABORATION WITH
CLARKSTON DONOVAN'S GROUP AND
PERFORMED DEEP NEURAL MUSCULAR
PHENOTYPING STUDIES INCLUDING
MUSCLE US TRAY SOUND AND MUSCLE
MRI AND FOUND NO EVIDENCE OF
FHSD 2.
SO DOES THAT MEAN ARYANIA
PATIENTS DON'T MAKE ANY DUX
SCORE?
THAT IS -- THAT QUESTION HAS
BEEN DIFFICULT TO ANSWER.
THE REASON IS THAT DUX 4 EVEN IN
FSHD 2 MUSCLE IS PRESENT AT LOW
LEVELS AT BOTH THE mRNA AND
PROTEIN LEVEL.
BUT THERE'S TWO POTENTIAL WORK
AROUNDS.
THE FIRST IS THAT WE CAN TREAT
PATIENT FIBROBLASTS WITH TWO
DRUGS THAT RELIEVE EPIGENETIC
REPRESSION TO ASK WHETHER
REPRESSION OF DUX 4 IS DIFFERENT
IN PATIENTS WITH ARYANIA, IN
HEALTHY VOLUNTEERS AND IN
PATIENTS WITH FHSD 2, THAT WAS
THE FIRST -- AND HERE YOU CAN
SEE DUX 4 LEVELS IN HUMAN DERMAL
FIBROBLASTS IN EACH OF THE THREE
GROUPS.
YOU CAN SEE ARYANIA PATIENTS
MAKE SOME DUX 4 BUT IT TENDS TO
BE MUCH LESS THAN THE AMOUNT WE
SEE IN PATIENTS WITH FHSD 2.
WE ALSO OBTAINED MUSCLE BIOPSIES
AND DIFFERENTIATED MYOBLASTS
INTO MYOTUBES BECAUSE THIS IS
KNOWN TO INCREASE DUX 4
EXPRESSION AND HERE YOU CAN SEE
THIS SOMETHING DUX 4 IS HIGHLY
EXPRESSED IN MYOSITES FROM FHSD
2 AND LESS SO IN PATIENTS WITH
ARYANIA.
THIS TELLS US THAT THE CONTROL
OF DUX 4 EXPRESSION IS VERY
DIFFERENT IN THESE TWO
CONDITIONS.
AND PATIENTS WITH ARYANIA MAYBE
PROTECTED FROM DEVELOPING FHSD
2.
WHAT DO WE KNOW SO FAR?
THE FIRST IS THAT ARYANIA
APPEARS TO BE EXTREME FORM OF KA
WILLLMANN SYNDROME BUT NO
GENETIC OVERLAP.
WE SEE INSTEAD STATISTICAL
EVIDENCE MISSENSE MUTATIONS IN
SMCHD 1 ARE PRIMARY EARLY DRIVER
OF EARLY ARYANIA.
LOSS OF FUNCTION AND MISSENSE
MUTATIONS IN SMCHD 1 CAUSE FHSD
2 THE DUX 4 MUSCLE TO BE TOXIN
BUT ARYANIA IN 2 DESPITE
MUTATIONAL OVERLAP APPEARED TO
BE MUTUALLY EXCLUSIVE
CONDITIONS.
BEFORE WE JUMP HEAD FIRST ON TO
THE SMCHD 1 BAND WAGON, I NEED
TO SHARE SOME MORE INFORMATION
WITH YOU ABOUT THESE PATIENTS
THAT TELLS US THAT THE GENETIC
ARCHITECTURE OF THESE CONDITIONS
IS ACTUALLY MUCH MORE COMPLEX.
SO THE TRUTH IS THAT NOT
EVERYONE WITH A MISSENSE
MUTATION IN SMCHD 1 HAS ABNORMAL
PHENOTYPE AND THIS IS SOMETHING
CALLED INCOMPLETE PENETRANTS.
HERE YOU CAN SEE ONE MULTIPLEX
FAMILY WHERE THE MOTHER HAS A
MUTATION IN SMCHD 1 YET HAS NO
CRANIAL FACIAL OR REPRODUCTIVE
PHENOTYPE.
SHE PASSED THE VARIANT ON TO TWO
OF HER CHILDREN, THE DAUGHTER
HAS COMPLETE ARYANIA AND HYPOGOD
INISM AND HER SON HAS NASAL
HYPOPLASIA AND HYPOGONADSISM.
SO THIS ALSO DEMONSTRATES
SOMETHING CALLED VARIABLE
EXPRESSIVITY WHERE INDIVIDUALS
WHERE THE SAME MUTATION EXHIBIT
DIFFERENCES IN DISEASE SEVERITY.
AND YOU CAN SEE THAT WITH THIS
SECOND PEDIGREE AS WELL.
SO THIS FATHER WITH NASAL
HYPOPLASIA AND INOSINEIA PASSED
TO ONE DAUGHTER WITH COMPLETE
ARYANIA AND HYPOGOD INISM AND TO
ANOTHER DAUGHTER SHOWN HERE WHO
HAS NASAL HYPOPLASIA.
SO TO SUMMARIZE IN ARYANIA WE
SEE VARIABLE EXPRESSIVITY,
INCOMPLETE PENETRANTS AND
SOMETHING CALLED PEIO TROUGH PI,
INDIVIDUALS WITH THE SAME
MUTATION EXHIBIT COMPLETELY
DISTINCT DISEASES.
AND WHAT THIS TELLS US IS
MUTATIONS ARE NECESSARY BUT NOT
SUFFICIENT TO CAUSE ARHINIA OR
FSHD 2 AND POINTS TO A POTENTIAL
ROLE FOR GENETIC MODIFIERS
ENVIRONMENTAL EXPOSURES OR TO
RANDOMNESS DURING DEVELOPMENT.
SO HOW CAN WE IDENTIFY
MODIFIERS
SO WE WERE
TALKING HOW TO IDENTIFY THESE
GENETIC OR ENVIRONMENTAL
MODIFIERS.
S WHAT WE WERE HOPING TO DO IS A
FSHD VERSUS ARHINIA GWAS STUDY
BUT NOT SURE TO HAVE ENOUGH
PATIENTS TO HAVE STATISTICAL
POWER.
WE NEED TO OPEN A WET LAB.
THAT POSE AS SLIDE PROBLEM SINCE
I HAVEN'T USED A PIPETTE SINCE
MY COLLEGE DAYS BUT FORTUNATELY
MY SCIENTIFIC DIRECTOR DARRELL
ZELDON WAS SUPPORTIVE OF MY
PURSUING THIS WORK AND GAVE ME
THE RESOURCES AND PERSONNEL TO
OPEN A WET LAB SEVERAL YEARS
AGO.
IF WE ARE GOING TO MODEL THESE
MUTATION, THERE'S TWO CRITICAL
QUESTIONS TO ANSWER.
THE FIRST IS WHAT DO MISSENSE
MUTATIONS DO TO SMCHD 1
FUNCTION?
MISSENSE MUTATIONS ARE A BIT
TRICKIER THAN NULL MUTATIONS
THEY COULD MAKE PROTEIN LESS
ACTIVE MORE ACTIVE OR EVEN
GRANTED A NOVEL FUNCTION.
THE SECOND IS WHAT CELL TYPES
SHOULD WE STUDY?
SO WE CAN START WITH A HUMAN
EMBRYONIC STEM CELL, BECAUSE WE
KNOW THIS DEFECTIVE OCCURS EARLY
DURING EMBRYO GENESIS BUT THAT
IS UNLIKELY TO BE THE SPECIFIC
CELL TYPE AFFECTED OR WE WOULD
SEE ADDITIONAL DEFECTS IN OTHER
ORGAN SYSTEMS.
SO TO ANSWER THE FIRST QUESTION,
LET'S GO BACK FOR A MOMENT TO
BASIC BIOLOGY OF THE SMCHD 1
PROTEIN.
AS I MENTIONED, IT IS A LARGE
2005 AMINO ACID PROTEIN WITH
SEVERAL DOMAINS IN THE END
TERMINAL REGION AND SMCHD HINGE
AT THE C TERMINAL REGION.
IN THE CELL IT INTERESTS IN THIS
INTERESTING BARBELL LIKE
CONFIGURATION WITH DIMERIZATION
OF BOTH N AND C TERMINALS.
MY GROUP HAS BEEN VERY
INTERESTED IN THE END TERMINAL
REGION BECAUSE THAT'S WHERE THE
MUTATIONS LIE AND IN
COLLABORATION WITH LARS PETERSON
AT NIEHS WE CRYSTALLIZE A
PROTEIN CONSTRUCT ENCOMPASSING
AMINO ACIDS 24 TO 580 WHICH
DEMONSTRATES ATP DEPENDENT
DIMERIZATION, THIS IS THE ATP
MOLECULE SHOWN HERE.
THE DIMER STABILIZED BY THIS
SWATHING OF THIS STRAP IN PURPLE
AND UBIQUITIN LIKE DOMAIN.
SHOWN IN BEIGE.
IF WE LOOK AT THE MUTATIONS THE
ARHINIA MUTATIONS ARE IN PINK,
FSHD IN BLUE AND MUTATION SEEN
IN BOTH CONDITIONS ARE IN
ORANGE.
AND YOU CAN SEE THAT THE
MUTATIONS IN BOTH CONDITIONS ARE
CLUSTERED AROUND THE ATP ACE
DOMAIN OR AT THE DIMER
INTERFACE.
HOW DO THESE MUTATIONS AFFECT
PROTEIN FUNCTION?
WE LOOKED AT TWO DIFFERENT
FUNCTIONS ATP ACE ACTIVITY AND
DIMERIZATION.
SHOWN HERE ARE ALL OF THE
ARHINIA MUTANT CONSTRUCTS, WITH
THE EXCEPTION OF E 147A WHICH IS
OUR POSITIVE CONTROL WHICH NOW
ABOLISHES ATP ACE ACTIVITY.
AND THREE FSHD MUTATIONS.
AND YOU CAN SEE THAT THERE IS --
THERE ARE VERY MINOR EFFECTS ON
ATPASE ACTIVITY AND THERE
DOESN'T SEEM TO BE AN EFFECT IN
ONE DIRECTION AND ONE CONDITION
AND OPPOSITE EFFECT IN THE OTHER
CONDITION.
CONTRAST THAT WITH DIMERIZATION
WITH WHERE IN ARHINIA WE SEE NO
EFFECT ON DIMERIZATION, WHEREAS
ALL THREE FSHD 2 MUTATIONS LED
TO LOSS OF DIMERIZATION.
WHAT ABOUT LESSONS FROM SMCHD 1
MUTANT ANIMAL MODELS?
IN COLLABORATION WITH MIKO AND
ERICA DAVIS FORMERLY AT DUKE WE
SUPPRESSED SMCHD 1 IN ZEBRAFISH
AND SAW CHANGES IN FACIAL
CARTILAGE CAVITY AND SMALLER
EYES. HOWEVER A COMPETING GROUP
OVEREXPRESSED SMCHD 1 TO MODEL A
GAIN OF FUNCTION MECHANISM AND
ALSO EXERTS SMALLER EYES.
SMCHD 1 NULL MOUSE BUZZ REPORTED
IN 2005 BY LOUIS'S GROUP AND
INTERESTINGLY THESE MICE DO NOT
HAVE FSHD 2 OR ARHINIA, PERHAPS
BECAUSE THE MOUSE DOESN'T
EXPRESS THE HUMAN FORM OF DUX 4.
THE ONLY PHENOTYPE THAT WE HAVE
SEEN IS THAT THERE WAS INCREASE
EMBRYONICALLY LETHALITY IN
FEMALES LIKELY BECAUSE OF SMCHD
1 ROLE IN EXON ACTIVATION.
WE GENERATED AN SMCHD 1 KNOCK IN
MOUSE USING MUTATION WE SEE MOST
COMMONLY IN OUR COHORT, L 141F.
AND THESE MICE APPEAR TO
PHENOCOPY THE NULL MICE,
SUGGESTING A LOSS OF FUNCTION
MECHANISM IN ARHINIA.
THE STRONGEST EVIDENCE THESE
MUTATIONS MIGHT BE BEHAVING IN
THE A LOSS OF FUNCTION FASHION
CAME WHEN WE WENT BACK TO OUR
PATIENTS, AND USED A CLINICAL
TEST THAT USED -- IS USED TO
DIAGNOSE FSHD 2 AND THAT'S TO
LOOK AT THE METHYLATION LEVEL AT
THE DUX 4 PROMOTER AT THAT D 4Z
4 REGION.
IN FSHD 2 THOSE PATIENTS HAD
LESS THAN 25% METHYLATION.
AND WHAT WE SAW IS THAT THE
MAJORITY OF OUR PATIENTS WITH
SMCHD 1 MUTATION ALSO
DEMONSTRATED HYPOMETHYLATION,
UNAFFECTED FAMILY MEMBERS
WITHOUT THE MUTATION DID NOT.
SO IF WE GO BACK TO OUR
QUESTION, FIRST QUESTION, IT
SEEMS SAFE TO MODEL THESE
MUTATIONS AS A LOSS OF FUNCTION
MECHANISM.
BUT THAT LEAVES US WITH THE
SECOND QUESTION OF WHAT CELL
TYPE WE SHOULD BE STUDYING.
SO IF YOU RECALL THE CLINICAL
TRIAD OF THIS CONDITION, THAT IT
AFFECTS THE NOSE, EYES, AND
LEADS TO GNRH DEFICIENCY, THAT
SHOULD TAKE US BACK TO EARLY
EMBRYONIC DEVELOPMENT AROUND
WEEK 3 OR 4 AND HERE I'M SHOWING
THE DEVELOPING NEURAL PLATE
WHICH WILL -- IN PINK WHICH WILL
INVAGINATE AND FORM THE NEURAL
TUBE TO GIVE RISE TO THE BRAIN
AND SPINAL CHORD.
BUT THERE ARE THESE TWO CELLULAR
POPULATIONS RIGHT AT THE NEURAL
PLATE BORDER, THE NEURAL CREST
CELLS AND CRANIO CELLS.
NEURAL CREST CELLS MIGRATE
THROUGHOUT THE BODY GIVING RISE
TO DIVERSE ORGANS SUCH AS THE
HEART AND GANGLIA AND
MELANOCYTES.
HOWEVER THE CRANIAL SUBDIVISION
OF NEURAL CREST CELL GAVE RISE
TO FACIAL SKELETON, PERIOCULAR
MESENCHYME AND OLFACTORY NERVE
AND SHEATHING CELLS.
THE CRANIAL CELLS ARE PRIMITIVE
SENSORY ORGANS AND WE ALREADY
TALKED ABOUT HOW THE OLFACTORY
PLAQUE CODE GAVE RISE TO GNRH
NEURONS, OLFACTORY NEURONS AND
ALSO RELEASES GROWTH FACTORS
THAT INDUCE FORMATION OF THE
NASAL SKELETON.
SO JUST AS A REMINDER, THE GNRH
NEURONS HITCH ALONG THESE
OLFACTORY NERVE AXONS TO THE
BRAIN AND THESE ARE BLAH CODE
DERIVATIVES BUT THE SHEATHING
CELLS WHICH IS CRITICAL FOR THE
INTEGRITY OF OLFACTORY NERVES
ARE DERIVATIVES OF CRANIAL
NEURAL CREST CELLS SO WE NEED TO
CONSIDER BOTH CELLULAR
POPULATIONS.
AND IN FACT, WHEN WE GO TO OUR
ANIMAL MODELS BESEE EXPRESSION
IN BOTH OF THESE CELL TYPES SO
THIS IS XGAL STAINING OF EMBRYOS
AT E 9.5 AND YOU CAN SEE DEEP
STAINING AT THE NASAL PLA CODE,
THIS IS SHOWING XENOPUS EMBRYOS
STAINING ON THE EYES, THE NASAL
PLACODES AND FOUR MIGRATORY
STREAMS OF NEURAL CREST CELLS.
SO WE HAVE GOT OUR PATIENTS, AND
OUR CONTROLS AND SO WE OBTAIN
SKIN BIOPSIES AND CREATED INDUCE
PLURIPOTENT STEM CELLS FROM BOTH
PATIENTS AND CONTROLS.
AND WE ALSO STUDIED THE H 9
HUMAN EMBRYONIC STEM CELL LINE
BECAUSE IN THIS LINE WE WERE
ABLE TO KNOCK OUT SMCHD 1.
AND THESE -- THIS ES CELL MODEL
IS IMPORTANT BECAUSE THESE CELLS
ARE ISOGENIC, MEANING THEY HAVE
THE SAME GENETIC BACKGROUND.
SO IT TAKES AWAY SOME OF THE
VARIABILITY THAT WE WOULD SEE IN
OUR PATIENT CELL LINE SO WE USE
BOTH OF THESE FOR OUR APPROACH.
THIS IS JUST SHOWING A WESTERN
BLOT WITH SUCCESSFUL KNOCK OUT
OF SMCHD 1 IN H 9.
I DON'T HAVE TIME TO GO INTO ALL
THE DETAILS BUT I WILL TELL YOU
THAT WE STARTED BY DERIVING
NEURAL CREST CELLS FROM THESE
IPS CELLS AND THE H 9 LINE AND
SAW NO ABNORMAL PHENOTYPE.
THERE WAS NORMAL PROLIFERATION
AND NORMAL MIGRATION.
SO WE MOVED ON AND USED AND
ESTABLISHED CRANIAL PLA CODE
PROTOCOL THAT INVOLVES DUAL
INHIBITION OF TGF BETA AND BMP,
THIS TAKES 11 DAYS.
WHAT WE SAW RIGHT AWAY, IS
SIGNIFICANT CELL DEATH.
SO THIS IS CRYSTAL VIOLET
STAINING IN TWO OF OUR WILD TYPE
ES CELLS AND IN OUR KNOCK
OUTLINE.
AND YOU CAN SEE PURPLE MEANS
CELLS ARE ALIVE AND WHITE
INDICATES CELLS DEATH SO WE SAW
THIS AROUND DAY 3 TO DAY 4.
WHAT WAS UPREGULATED AT THIS
TIME?
NONE OTHER THAN THE DUX 4 MUZZLE
TOXIN WE TALKED ABOUT IN FHSD 2.
THIS IS SHOWING INCREASE
BEGINNING AT DAY 2 AT THE mRNA
LEVEL AND ROBUST EXPRESSION AT
THE PROTEIN LEVEL AT DAYS FOUR
AND FIVE WE ALSO PERFORMED A
TIME COURSE RNA SEQ EXPERIMENT
AND YOU CAN SEE IN THIS VOLCANO
PLOT THAT MANY GENES WERE
DOWN-REGULATED SHOWN IN GREEN IN
OUR KNOCK OUT CELLS AND MANY
WERE UPREGULATED SHOWN IN RED.
BUT ALL OF THESE GENES THAT ARE
LISTED HERE ARE KNOWN DUX 4
TARGETS.
AND I WILL JUST HIGHLIGHT ONE OF
THEM, TRIM 43 THAT WE WILL COME
BACK TO.
SO THE NEXT THING WE DID WAS TO
DEMONSTRATE THAT THE CELLULAR
PHENOTYPE WAS SPECIFICALLY
CAUSED BY LOSS OF SMCHD 1.
WE DID THAT BY KNOCKING IN OR
REINTRODUCING SMCHD 1 SHOWN HERE
ARE THE KNOCK OUTLINES WITH
INCREASE DUX 4 EXPRESSION NO
SMCHD 1 AND KNOCK IN LINE WHERE
THERE IS LOSS OF DUX 4 AND
RECOVERY OF SMCHD 1 AND HERE YOU
CAN SEE RECOVERY OF CELLULAR
VIABILITY IN THE KNOCK-IN LINE.
BEALSO CONFIRMED DUX 4
PATHOGENESIS, BY USING TWO
DIFFERENT DUX 4 INHIBITORS.
HERE WE USE SI RNA TO DIFFERENT
SI RNAs TO KNOCK DOWN DUX 4.
YOU CAN SEE WITH ONE OF THESE
SHOWN HERE AGAIN, RECOVERY OF
CELL VIABILITY.
WE ALSO USE AD PHARMACOLOGICAL
INHIBITOR CALLED IP 300W AND DUX
4 ACTIVATES TARGET GENES RECRUIT
RECRUITING THESE CO-ACTIVATORS
AND CRED BINDING PROTEIN.
WHEN YOU INHIBIT THESE PROTEINS
WITH THE DRUG, WE SAW LOSS OF
DUX 4 EXPRESSION, AND AGAIN
TREATMENT WITH INHUB TORR
RESCUES CELLS FROM CELL DEATH.
WHAT ABOUT THE PATIENT DERIVED
CELLS THAT I TOLD YOU ABOUT?
WELL, THESE CELLS ACTUALLY
BEHAVED QUITE SIMPLY TO THE
KNOCK OUT CELLS.
SO THIS IS SHOWING THE WILD TYPE
CLONES AND TWO KNOCK OUT CLONES
SHOWING CELL DEATH AT DAY 11 OF
PLACODE CELL DIFFERENTIATION.
HERE ARE IPS CELLS THAT WERE
DIFFERENTIATED INTO PL,ACODE
CELLS FROM HEALTHY PATIENT,
THREE WITH ARHINIA, AND TWO
PATIENTS WITH FSHD 2.
AND YOU CAN SEE THAT THIS -- THE
AMOUNT OF CELL DEATH CORRELATES
WITH EXPRESSION OF THAT DUX 4
TARGET I MENTIONED, TRIM 43 AS
SHOPE HERE.
AND I THINK WHAT IS INTERESTING
IS THAT THE SAME VARIABILITY IN
THESE CELLULAR PHENOTYPES THAT
THIS IS VERY REMINISCENT OF THE
VARIABILITY THAT WE SAW IN OUR
PATIENTS AS WELL AND THIS AGAIN
POINTS TO A ROLE FOR GENETIC OR
ENVIRONMENTAL MODIFIERS, AND
TELL US THAT THOSE MODIFIERS MAY
ACTUALLY BE CONTROLLING THE
AMOUNT OF DUX 4 EXPRESSION IN
PLACODE CELLS.
SO HOW ARE WE GOING TO FIND
THESE?
WELL, WE ARE PROPOSING TO
PERFORM A GENOME WIDE SI RNA
SCREEN.
SO WE CREATED A CONSTRUCT WHERE
DUX 4 BINDING TO ITS RESPONSIVE
ELEMENT DRIVES EXPRESSION OF
NANOLUCIFERASE.
AND AS A NEGATIVE CONTROL WE
HAVE JUST A PROMOTER LINKED TO
NANOLUCIFERASE.
AND WE TRANSFECTED OUR PLACODE
CELLS AT DAY THREE OF
DIFFERENTIATION, AND HERE YOU
CAN SEE KNOWN NANOLUCIFERASE,
STAYED NULL IN NEGATIVE CONTROL
BUT IN THE KNOCK OUT CELLS WHERE
WE EXPECTS TO SEE INCREASE DUX 4
PRODUCTION AND BINDING WE SEE AN
INCREASE IN NANOLUCIFERASE
ACTIVITY.
SO IN PERFORMING THIS SCREEN
WHAT WE ARE LOOKING FOR IS GENES
WHICH WHEN KNOCKED DOWN LEAD TO
DECREASE IN SIGNAL IN OUR KNOCK
OUT CELLS OR INCREASE IN SIGNAL
IN WILD TYPE CELLS TO DO THIS
FOR EVERY GENE IN THE GENOME WE
COLLABORATE WITH NCATS WITH HIGH
THROUGH PUT PLATE READER TO
DETECT LUMINESCENCE.
WHILE THIS IS COOKING WE HAVE
BEGUN TO THINK ABOUT POTENTIAL
ENVIRONMENTAL -- CAUSING
ARHINIA.
AND ONE SUCH CANDIDATE IS THE
HERPES VIRUS FAMILY.
THIS IS A RECENT PAPER BY FULL
ET ALL IN NATURE MICROBIOLOGY
DEMONSTRATING THAT TRIM 43 IS
UPREGULATED DURING HERPES VIRUS
INFECTION AS PART OF A GERM LINE
SPECIFIC TRANSCRIPTIONAL PROGRAM
MEDIATED BY NONE OTHER THAN DUX
4.
SO WE ASKED WHETHER HERPES VIRUS
INFECTION OF OUR PATIENT DERIVED
CELLS MIGHT SIMILARLY AUGMENT
DUX 4.
T THAT IS EXACTLY WHAT WE SAW.
ON THE LEFT IS SHOWING CRE --
PREINFECTION AND ON THE RIGHT
POST INFECTION WITH HSV 1, YOU
CAN SEE IN THE PRESENCE OF VIRAL
PROTEIN ICP 0 AND INDUCTION OF
DUX 4 NOT IN CONTROL IPS CELLS,
BUT IN CELLS FROM PATIENTS WITH
FSHD 2 AND ARHINIA.
HERE LOOKING AT TRIM 43 WE SEE
THE SAME PATTERN, SO NO INCREASE
PRE-INFECTION, AND A GREATER
INCREASE IN TRIM 43 AFTER HSV 1
INFECTION IN FSHD 2 AND PATIENTS
WITH ARHINIA CELLS.
LET'S PUT THIS ALL TOGETHER NOW.
WE STARTED WITH JUST ONE
PATIENT.
FORMED INTERNATIONAL CONSORTIUM
TO CREATE A LARGE COHORT OF
PATIENTS.
AND THROUGH NEXT GENERATION
SEQUENCING, WE WERE ABLE TO
DEMONSTRATE THAT MISSENSE
MUTATIONS IN SMCHD 1 ARE THE
PRIMARY DRIVER OF ARHINIA.
HOWEVER, THE HETEROGENEITY OF
PHENOTYPES THAT WE SAW IN OUR
PATIENTS, SUGGESTED THAT THERE
MAYBE ADDITIONAL GENETIC OR
ENVIRONMENTAL MODIFIERS.
WORK IN THE WET LAB DEMONSTRATED
THAT ARHINIA IS CAUSED BY DUX 4
MEDIATED PLACODE CELL DEATH AND
SUGGESTED A POTENTIAL ROLE FOR
HSV 1 IN PATHOGENESIS.
SO THERE ARE STILL SEVERAL OPEN
QUESTIONS THAT WE ARE TRYING TO
ADDRESS.
THE FIRST IS WHAT IS SPECIAL
ABOUT MUSCLE AND PLACODE CELLS
THAT ALLOWS FOR DUX 4
DEREPRESSION WITH LOSS OF SMCHD
1 ACTIVITY?
WHY IS IT LIMITED TO THESE CELL
TYPES?
AND WHY ARE THERE NO INDIVIDUALS
WITH BOTH DISORDERS?
DOES ONE PROTECT YOU FROM
GETTING THE OTHER DISORDER OR IS
IT THAT THE GENETIC CONDITIONS
ARE SO RARE THAT ONE INDIVIDUAL
IS UNLIKELY TO EVERY MEET
CRITERIA FOR BOTH CONDITIONS?
AND LASTLY, ARE THERE OTHER
ENVIRONMENTAL CONTRIBUTORS WE
SHOULD BE CONSIDERING?
I WANT TO THANK AND ACKNOWLEDGE
THE MANY COLLABORATORS WHO
HELPED THESE ARE ALL AUTHORS ON
OUR FIRST PAPER, IDENTIFYING
MUTATIONS IN SMCHD 1 AS CAUSE OF
ARHINIA.
AND THE MORE RECENT WORK
INVOLVING THE PHENOTYPING OF
PATIENTS AND THE PLACODE CELL
WORK PERFORMED BY THESE
INDIVIDUALS, I WANT TO THANK MY
COLLEAGUES AT THE NIH CLINICAL
CENTER AND AT NIEHS IN THE X-RAY
CRYSTALOGRAPHY FOR THE CLINICAL
RESEARCH UNIT AND
BIOINFORMATICS.
AND I WANT TO ESPECIALLY
HIGHLIGHT KAORU,NOUE WHO DID THE
PLA CODE AND STEM CELL WORK AND
COLLABORATORS AT MASS GENERAL.
THANK YOU FOR YOUR ATTENTION.
>> DR. SHAW, I WANT TO THANK YOU
FOR SHARING YOUR COMPLEX
INVESTIGATION INTO THIS UNUSUAL
PHENOTYPE AND INSIGHTS INTO THE
EPIGENETICS AS A FAST GAINING
PRESENTATION AND WE APPRECIATE
YOUR TIME.
I DO APOLOGIZE TO YOU AND THE
AUDIENCE FOR THE AUDIO BUT
THANKS TORR YOUR PATIENCE.
WE HAVE ONE QUESTION, LIKE TO
SHARE IT WITH YOU, SAYS DR.
SHAW, FANTASTIC PRESENTATION.
IF ARHINIA AND FSHD 2 ARE
MULTI-GENIC COMPLEX TRAITS,
WONDER IF STUDYING GENOME WIDE
PROFILES AND MODEL CELLS CAN BE
HELPFUL IN IDENTIFYING TADs
THAT ARE DIFFERENT FROM
FILTERING STRATEGY TO STUDY
EPISTATIC MACHINES.
>> THAT IS AN EXCELLENT POINT
AND I DON'T HAVE THE DATA TO
SHARE WITH YOU BUT WE HAVE
PERFORMED HIGH C STUDIES MANY
THE CRANIAL PLACODE CELLS AND WE
ARE ANALYZING THOSE DATA NOW BUT
THAT IS A GREAT SUGGESTION.
>> GREAT.
AND JUST ANY OTHER INSIGHTS INTO
-- YOU WERE LOOKING AT THE
ENVIRONMENTAL INFLUENCES AND
FOUND HERPES.
ARE THERE OTHER DIRECTIONS THAT
YOU ARE CURRENTLY LOOKING INTO
SPECIFICALLY RIGHT NOW?
>> YEAH.
WE DON'T -- WHEN I TALKED TO
THESE PATIENTS AND THEIR PARENTS
THERE'S REALLY NO -- THEY DON'T
MENTION ANY POTENTIAL EXPOSURES.
THEY HAVE BEEN TOLD DIFFERENT
THINGS THAT WERE CERTAINLY WRONG
LIKE MOTHER WAS TOO CLOSE TO THE
X-RAY -- COPY MACHINE AT HER
WORK OR THINGS LIKE THAT.
WHICH I JUST FEEL HORRIBLE WHEN
I HEAR -- BUT I THINK PROBABLY
WILL LOOK AT MORE VIRUS, BUT NO,
I DON'T HAVE ANY OTHER LEADS ON
POTENTIAL ENVIRONMENTAL
CONTRIBUTORS.
>> THANK YOU VERY MUCH.
LOOKS LIKE YOU HAVE ANSWERED ALL
THE QUESTIONS FROM THE AUDIENCE,
NOTHING ELSE IS COMING IN.
AGAIN, I WANT TO THANK YOU, VERY
MUCH FOR PRESENTING TODAY.
WE REALLY ENJOYED YOUR
PRESENTATION.
AND I WANT TO THANK -- WE HAVE
ONE MORE QUESTION JUST POPPED
IN.
IT IS A -- DR. SHAW, IS IT ALSO
POSSIBLE THESE TWO PHENOTYPES
HAVE NOT BEEN SEEN TOGETHER
BECAUSE OF SOME OTHER LINK THAT
LEAD TO LEE HALLTY IN UTERO?
-- LETHALITY IN UTERO?
>> I GUESS THAT'S POSSIBLE.
SO THAT WOULD -- YEAH, I MEAN,
SO YOU ARE SUGGESTING THAT IF
THERE WERE MORE SEVERE MUTATION,
THAT COULD CAUSE BOTH
CONDITIONS, IT WOULDN'T -- THE
CHILD WOULD NOT SURVIVE.
YEAH, I HAVEN'T THOUGHT OF THAT
BUT THAT ALSO COULD BE POSSIBLE.
THERE ARE NO -- IF YOU LOOK IN
NOMAD, THERE'S NO PATHOGEN I CAN
NULL MUTATION.
I SHOULDN'T SAY THAT, THAT CAUSE
ARHINIA, THEY ARE ALL MISSENSE.
BUT THERE ARE NO MUTATIONS THAT
CAUSE FSHD 2, THERE IS A SEVERE
FORM OF FSHD 2 THAT HAS A
NEONATAL ONSET THAT CAN CAUSE
MORE MUSCLE BUT ALSO
DEVELOPMENTAL DELAY BUT AGAIN,
THOSE PATIENTS HAVE NO CRANIAL
FACIAL ABNORMALITIES.
>> OKAY.
THANK YOU, VERY MUCH, DR. SHAW,
THANKS TO THE AUDIENCE.
I WISH EVERYONE A GREAT
AFTERNOON.
THANK YOU, VERY MUCH.
>> THANK YOU.