>>WELCOME TO THE NIH DNA
REPAIR INTEREST GROUP VIDEO
CONFERENCE.
I'M KEN KRAEMER, MY COHORT WILL
BOHR IS NOT AVAILABLE NOW BUT HE
WILL BE BACK NEXT MONTH.
WE DO HAVE OUR MONTHLY VIDEO
CONFERENCES NEXT MONTH AND JUNE
DR. TOM KONKLE FROM NIEHS WILL
BE SPEAKING ABOUT EXTRINSIC
PROOF READING OF DNA REPLICATION
ERRORS.
WE WILL NOT BE HAVING SEMINARS
IN JULY AND AUGUST.
AND WE WILL RESUME IN SEPTEMBER
AND DR. MARK WILL BE SPEAKING AT
THIS POINT.
THOSE OF YOU WHO KNOW ABOUT
THIS--IF YOU'RE NOT--IF YOU
WOULD LIKE TO KNOW ABOUT ALL OF
THESE SEMINARS YOU CAN GET ON
THE DNA REPAIR INTEREST GROUP
E-MAIL LIST AND JUST WRITE TO ME
AT KRAEMERK@NIHBNT GOFF.
SO OUR SPEAKER TODAY IS
DR. DONATA ORIOLI, FROM ITALY.
SHE WAS FROM ITALY AND DID GET
HER UNDERGRADUATE DEGREE AT
UNIVERSITY OF P A VIA WHERE SHE
IS NOW, BUT SHE WENT TO GERMANY
AND GOT HER Ph.D. AT THE
UNIVERSITY OF HIDELE BERG AND
THEN CAME BACK TO P A VIA AND
WORKED IN THE LABORATORY OF
DR. MARIA [INDISCERNIBLE] ON
GENETIC DNA REPAIR DEFICIENCY
DISEASES FOR MANY YEARS.
THAT WAS THE LABORATORY THAT WAS
IDENTIFYING ALL THOSE IN THE
COUNTRY OF ITALY AND THEY FOUND
PATIENTS WITH 0 DERMA AND FOR
THE FIRST TIME PATIENTS WITH
[INDISCERNIBLE] DYSTROPHY.
SO TODAY, DR. STEFANNI, ACTUALLY
GIVE A TALK IN THIS SERIES IN
1998 AND THAT IS ARCHIVED FOR
THOSE WHO WOULD LIKE TO SEE IT
ON THE VIDEOCAST.NIH.GOV
WEBSITE.
SO THEY WILL SPEAKING ABOUT THE
MULTIFACETED ROLE OF THE NER
PROTEINS AND THEIR IMPLICATIONS
IN HUMAN PATHOLOGY.
IF YOU HAVE ANY QUESTIONS PLEASE
PUT THEM IN THE CHAT BOX AND I
WILL READ THEM LATER.
THANK YOU VERY MUCH.
>> THANK YOU KEN, THANK YOU FOR
THE PRESENTATION, THA YOU BOTH
TO YOU TO YOU AND WILL FOR THE
INVITATIONS TO GIVE A TALK HERE.
BECAUSE THESE GIVES A GREAT
OPPORTUNITY TO PRESENT WHAT IS
OUR CONTRIBUTION IN THE FIELD OF
THE NOW NEURODEFECTIVE
DISORDERS.
SO, THE NUCLEOTIDE
EXCISION REPAIR, WE SAID WE KNOW
THE DNA REPAIR PATHWAY THAT
REMOVES THE BARRIER AND THIS
INCLUDES ALSO THE
[INDISCERNIBLE] USE OF LESIONS.
THERE ARE MANY PROTEINS THAT
CODE THE NER PROTEINS THAT ARE
IMPORTANT IN THIS MECHANISM
STARTING FROM THE DNA DAMAGE
RECOGNITION AND BASICALLY LIES
ON DIFFERENT MOTIVES DEPENDING
ON THE SUBPATHWAYS THAT ARE
USING GENOME REPAIR, RECOGNIZING
THE LESION THAT ARE DISTRIBUTED
ENTIRE GENOME WHAT IS
TRANSCRIPTIONAL COUPLED REPAIR.
AFTER THE DNA DAMAGE
RECOGNITION, THE 2 SUBPATHWAY
JOIN IN THE UNIQUE PATHWAY AND
THE NEXT, THE FOLLOWING STEP IS
THE FORMATION OF THE OPEN
COMPLEX AND THIS IS THANKS TO
THE HELIX ACTIVITY OF THE
COMPLEX THAT WE ARE GOING TO
SPEAK ABOUT TODAY.
AND AFTER THE INFORMATION OF THE
OPEN COMPLEX, SO THEN THERE IS
THE EXCISION OF THE DNA FRAGMENT
THAT CONTAINED LESIONS AND THE
FRAGMENTED GETS EXCISED,
ELIMINATED AND WITH THE ANSWER
WE HAVE THE DE NOVO SENTH SIS
WHERE THE POLYMERASE USES THE
UNDAMAGED STRAPPED AS A TEMPLATE
TO PERFORM THE SYNTHESIS.
SO AS I WAS SAYING, MANY GENES
WITH THE EXCISION REPAIR, MANY
PROTEINS, SO THE MUTATIONS IN
SEVERAL OF THE GENES THAT ARE
RESPONSIBLE FOR A
[INDISCERNIBLE] DISORDER.
OKAY, SO, THE AMOUNT IS OVER, WE
CAN DISTINGUISH THE DERMA
PIGMENTOSE UMKC, WHICH IS A
BONES, SORE SKIN TUMORS IN THE
AREA OF THE SKIN THAT ARE
EXPOSED TO THE SUN AND THEN THE
COCKAYNE SYNDROME AND
TRICHOTHIODYSTROPHY, BOTH
PATIENTS, THEY BOTH REPRESENT
SIGN OF PREMATURE AGING
INCLUDING SPECIAL IN THE CASE OF
COCKAYNE'S SYNDROME FOR
NEURODEGENERATION, THERE ARE
MULTISYSTEMIC DISORDERS, THEY
HAVE A WILD AMOUNT OF CLINICAL
FEATURES BUT IN THE PATIENTS AND
THE PATIENT WE CAN SAY THAT THEY
ALSO ARE CHARACTERIZED BY A WIDE
SPECTRUM OF SEVERITY.
BOAGHTD THE CS AND THE TTD, THE
CLINICAL HALLMARK THAT
CHARACTERIZE THIS PATIENT IS THE
FAYE NAIL [INDISCERNIBLE]
BECAUSE THEA PATIENTS ARE
CHARACTERIZED BY DEFECTS SO THEY
HAVE [INDISCERNIBLE], THEY
EASILY THEY HAVE REDUCED CONTENT
WITHED [INDISCERNIBLE] AND
DEVELOPMENT FOR THE ASPECT IS
WHEN THEY ARE ANALYZED UNDER THE
LIGHT OF THE POLARIZING SCOPE OF
THE HAIR AS SHOWN A SPECIFIC
PATTERN THAT IS CALLED THE THEME
PATTERN AND I WOULD SHE YOU A
PICTURE AFTERWARDS.
SO BUT CS AND TTD, THEY ARE BOTH
FACTOR IN THE EXCISION REPAIRS
SO THEY ACCUMULATE IN THE GENOME
AND THEN AND REPAIR THE LESIONS,
BUT THE DIFFERENT 3 FROM THESE
PATIENT DO NOT SHOW ANY CANCER
BONE IN THIS, IN THE AREA OF
THIS SCHEME THAT ARE SUPPOSED TO
BE THERE, SO THIS OPENS THE
QUESTION, HOW THEY ARE CAN GIVE
RISE TO SUCH DISEASE OF
HETEROGENERATED AITY, WHICH
PATIENTS PRESENT AN OPPOSITE
SKIN CANCER AND ALSO WITH SUCH
DIFFERENT APPEARANCE OF THE
FEATURES AND SO IN PARTICULAR
WITH THE GENE--THIS DISEASE
OBSERVED ALLOW US TO ANALYZE A
LARGER COHORT, SO IF WE LOOK AT
THE MUTATIONS, AT THE TOP,
FOUNDER IN THE PPD PATIENTS WE
CAN OBSERVE THAT THERE SEEMS TO
BE A DISEASE SPECIFIC MINORITY
THAT ARE DISTINCT AND THERE ARE
ACTIVITY AND THERE ARE BOTH OF
THEM DISTRIBUTED OVER KD--SALLY
TO THE PROTEIN.
THIS IS A SUBUNIT FOR THE
TRANSCRIPTION FACTOR TO AGE, IT
HAS AN ATPASE AND
[INDISCERNIBLE] ACTIVITY AND AS
I WAS SAYING THAT THE MUTATIONS
IN THIS GENE CAN BE RESPONSIBLE
FOR HPNTT, CONCERNING THE HBP
UNITS, THIS HAS ATPASE AND
TRANSLO CASE ACTIVITY AND
SIMILAR TO XPD, MUTATION IN THE
SUPPORTING GENE CAN BE
RESPONSIBLE BOTH FOR
EXPERIENCES.
WHEREAS THE MUTATIONS IN THE
GENE THAT ENCODES THE ATPASE,
ARE RESPONSIBLE FOR THE TTD
DIVISIONS, IN ADDITION, THE WAGE
ALSO SHOW A KINASE ACTIVITY
SUPERIOR TO THE COMPLEX.
SO AS I SAID IT IS A GENERAL
TRANSCRIPTION FACTOR THAT BINDS
TO THE GENE PROMOTERS, AND WE
THINK THAT THE ACTIVATE THAT CAN
PHOSPHORALATE, AND IT'S PART OF
THE COMPLEX AND WHEN IT IS
ASSEMBLED ON THE GENE, THROUGH
THE KINASE ACTIVITY, TF2 H
PHOSPHORALATE THE TERMINAL
DOMAIN OF CANNED WHAT.
SO IT ACTIVATES TRANSCRIPTION.
BUT AS I WAS SAYING BEFORE TF2 H
IS ALSO IMPORTANT KEY FACTOR IN
THE NUCLEOTIDE EXCISION REPAIR,
SO THANKS TO IN THE KAITION OFEA
IN THE REGION, THAT MUCH STRAND
IN THE REGION THAT CONTAINS THE
LESION IN THE WAY THAT OTHER
PROTEINS THEN CAN STABILIZE AND
CREATE AN OPEN COMPLEX.
SO WHAT IT HAS BEEN SHOWN IN THE
YOU HADITATIONS THAT THE FACT
THAT THAT ARE RESPONSIBLE FOR
BOTH EXPERIENCE TTD, RESULTING
IN THE ACTIVITY OF TF2 H WITH
THE CARE, SO EXPERIENCED TTD
PATIENTS REPAIR AND ACCUMULATE
IT IN THEIR GENOME.
THEY CAN EXPLAIN THE BONUS OF
SERVING THESE PATIENTS.
BUT IT DOESN'T EXPLAIN WHY TTD
PATIENTS MUTATED IN THE
[INDISCERNIBLE].
SO POSSIBLE EXPLANATION FOR THAT
CAN BE PROVIDED IF WE THINK
ABOUT THAT MUTATION SO THAT THAT
WAS RESPONSIBLE FOR TTT, MIGHT
ALSO IMPAIR THE TRANSCRIPTION
ACTIVITY OF TFIIH, SO THE MORE
TTD THAN THE XP MAY HAVE A FACT
IN THE TRANSCRIPTION OF TTD, AND
THERE ARE SEVERAL EVIDENCE THAT
MIGHT BE PROVIDED THAT THIS
MIGHT BE THE CASE AND THAT'S
WHAT WE'RE TRYING TO SHOW TODAY.
SO THE FIRST EVIDENCE, THE DRIVE
FROM WORK THAT HAS BEEN
PERFORMED--PART OF THOSE STUDIES
THAT HAVE BEEN PERFORMED IN OUR
LAB BY ANOTHER DOCTOR AND IN THE
LAB [INDISCERNIBLE] THEY BOTH
SHOWED THAT THE FIBRO BLAST FROM
TTD PATIENTS INDEPENDENTLYOT
GENE, MUTATED GENE, THEY ALL
SHOW A REDUCED [INDISCERNIBLE]
OF THE TF2 H, SO THIS CARRIES
MUTATION BUT ALSO ALL THE OTHER
SUBUNITS OF THE [INDISCERNIBLE].
THIS INDICATES THAT THESE
CONTAIN A REDUCED AMOUNT OF
MUTATED [INDISCERNIBLE]
TRANSCRIPTION FACTOR.
SO TRYING TO--WITH THE ATTEMPT
TO DEMONSTRATE THAT THE
TRANSCRIPTION DEFACT ARE REALLY
RESPONSIBLE FOR TTD, WE DECIDED
TO PERFORM A GENE EXPRESSION
ANALYSIS BY USING THE RNA SEQ
APPROACH AND WE SEQUENCE THE RNA
FIBRO BLAST FROM TTD PATIENT AND
TO EXPENSIVE LINKS, ALL
[INDISCERNIBLE] IN TTD, AND WE
DECIDED TO DO A 2 CLONE PAIR
WITH THE PATIENT EXPRESSION WITH
THE CORRESPONDING
[INDISCERNIBLE].
WE USE IT IN ALL PATIENTS, ALL
FEMALES, TO ORDER TO LIMIT THE
DIFFERENT [INDISCERNIBLE].
--IS HIGHER MUCH HIGHER IN THE
NUMBER OF GENES THAT ARE
REGULATED AND THE MORJORRITY OF
THE TTD REGULATED GENE ARE DOWN
REGULATED AS WE OBSERVE THE BLUE
BAR, BUT AS YOU CAN SEE, THERE
IS AN EQUAL NUMBER OF GENES THAT
ARE DOWN REGULATED OR
UPREGULATED.
SO THE SAME WE HAVE IT EVEN IF
THE GENE CELL IS DIFFERENT, WE
HAVE IT IF WE LOOK HERE AT
THIS--THE EXPRESSION OF
REGULATION AFTER UV RADIATION
WITH THESE FIBERS.
SO WE CAN THINK AND WE CAN SAY
THAT BY LOOKING AT THESE AT THE
NUMBER OF REGULATED GENE THAT
TTD MORE THAN THE XP5 ROUGH ATOM
BLASTIN ARE AFFECTED BY WIDE
TRANSCRIPTIONAL IMPAIRMENTS.
SO THEN WHEN WE TRY TO LOOK AT
ALL THE GENES THAT WE FOUND THAT
ARE REGULATED, THOSE IN THE
CONDITION OR UV RADIATION AND WE
DID THE ANALYSIS IN A LARGER
COHORT OF PATIENTS, ALL MUTATED
IN XPD, FOR EITHER TTD OR XP AND
AS A CONTROL WE USE THE
CORRESPONDING [INDISCERNIBLE]
AND THE RESULT WAS THAT WE
OBTAIN A REDUCED NUMBER OF GENE,
A HALF OF THEM, GENE THAT
RESPONDS TO THE UV RADIATION
BECAUSE WE CAN OBSERVE IN THE
CONTROL FIBRO BLAST IN THE
CURRENT THAT THE GENE BECOMES
EITHER UPREGULATED OR DOWN
REGULATED AFTER UV REGULATION.
THESE REGULATION NORMALLY IS
LOST IN TTD AND IN SOME OF--FOR
SOME OF THE GENE TRANSCRIPTION
REGULATION, ALSO IS CLEAR IN THE
XP PATIENTS THAT IN OTHER CASES
SEEMS TO ONLY AFFECT THE TTD AND
NOT THE [INDISCERNIBLE].
SO WE DECIDED TO GO FURTHER AND
TO UNDERSTAND WHETHER THE
TRANSCRIPTION REGULATION CAN
REALLY HAVE AN IMPACT ON THE
CLINICAL FEATURE OF THE PATIENTS
THAT WE EXPECT TO HAVE AN
ALTERATION OF THE LEVEL OF THE
PROTEIN COMPETENT, BUT TO OUR
SURPRISE WHEN WE CHECKED THE
AMOUNT OF PROTEIN OF THE
DISREGULATED GENES, EVEN THOUGH
TOO SHOW A VERY HIGH
TRANSCRIPTION REGULATION, WE
COULD NOT [INDISCERNIBLE] ANY
DIFFERENT, DIFFERENCE BETWEEN
THE TTD, THE XP OR THE CONTROL.
INDICATING THAT MOST OF THE
TRANSCRIPTION REGULATIONS, THE
SIMILAR LEVEL EITHER BY PROTEINS
THROUGH THE PROTEIN STABILITY.
NEVERTHELESS, WE WERE ABLE TO
IDENTIFY SOME TRANSCRIPTION
REGULATION THAT THE SPECIFICALLY
A FACTOR TO TTD PATIENTS, THE
TTD CELLS.
IN THE FIRST GENE WE IDENTIFIED
WAS SOMETIMES GOING IN 2013 IT'S
A COLLAGEN 6 A 1 GENE THAT IS
ALSO WITH THE COLLAGEN 6, WE
OBSERVE THAT FIBRO BLASTS WHEN
FRESHLY CULTURED AT CONFLUENCE
OF THE FIBRO BLAST REACH THE
CONFLEW ENSEL, THE TTD FIBRO
BLAST ARE EXTREMELY REUSE DOOD
AMOUNT OF TYPE 6 COMPARED TO THE
CONTROL BUT THE DEFACTOR IS NOT
OBSERVED IN THE CULTURE IN
DENSITY SO WE INVESTIGATED THE
MECHANISM THAT WAS RESPONSIBLE
FOR THIS TRANSCRIPTION
REGULATION AND WE WERE ABLE TO
OBSERVE THAT IN NORMAL FIBRO
BLASTOT PROMOTER OF THE COLLAGEN
6 IS PRESENT FOR THE COALESCE
BP1, WHICH MAINTAINS THE
PRESENCE OF THE PROTEIN,
MAINTAINS LOW RATE OF
TRANSCRIPTION.
WHEN THE FIBRO BLAST REACH THE
CELL, AND THE TF2 H
PHOSPHORALATE, AND THE PROTEIN
IS RELEASED FROM THE PROMOTER
AND NOW THE PROMOTER CAN START
TO TRANSCRIBE AT A HIGH RATE.
IN TTD, NOT EXPRESS,
SOPHISTICATED THE TTD SPECIFIC
MUTATION IN TF2 H, WE OBSERVE
THAT THE CELL DENSITY, SREBP1 IS
PRESENT SO THE AGENT CANNOT GET
TO THE PROTEIN SO THE GENE IS
NOT UPREGULATED.
BESIDE COLLAGEN 6, WE ALSO FOUND
THIS QUESTION, IN TTD WITH THE 1
GENE, THIS IS 1 OF THE GENE OF
THE FEW GENES THAT WE SHOW A
TRANSCRIPTION UPREGULATION IN
TDRKS, IT IS SPECIFIC FOR TDRKS,
IT IS ALSO THE FIBRO BLAST AND
THE DMP1 AND THE COLLIN ACE THAT
HAS THE ABILITY TO DEGRADE THE
SPECIFIC COLLAGENS IN
PARTICULAR, IN THIS CASE, THE
COLLAGEN AND FURTHER WHEN WE
LOOK AT NORMAL CULTURE,
THE,AMOUNT OF THE COLLAGEN TYPE
1, WE CAN SEE THAT THE TTD, BUT
NOT XP FIBRO BLAST CONTAIN THE
REDUCED AMOUNT OF THIS TYPE OF
COLLAGEN COMPARED TO THE
CONTROL.
WHEREAS RGTD TYPES OF COLLAGENS
ARE PRESENT IN THE TTD FIEB
PROBLAST.
THIS IS ALSO CLEAR WHEN WE LOOK
AT THE 3 B CULTURES IN WHICH THE
FIBRO BLAST ARE EMBEDDED IN THE
METRICS THAT CONTAIN THE
COLLAGEN TYPE 1.
THE COLLAGEN IS FISCHERIBLE BY
THE BLUE AND GREEN COLOR, AND
CLEARLY IN THE OTHER THAT
SURROUNDS THE TTD FIBRO BLAST,
BUT THE CONTROL, WE LOSE THE
BLUE STAINING INDICATING THAT
THE THERE IS A LOSS OF COLLAGEN
TYPE 1.
SO, WE ALSO LOOKED AT THE DERMAL
LAYER OF THE SKIN BIOPSIES OF
FORMER TTD PATIENTS AND WE CAN
CLEARLY OBSERVE THAT THERE IS A
REDUCED CONTENT OF COLLAGEN TYPE
1, BUT NOT AT A REDUCED COUNT OF
OTHER TYPE OF COLLAGENS AND THE
REDUCTION, WE OBSERVED THE
REDUCTION COMPARED POETIC THE
AMOUNT OF COLLAGEN TYPE THAT WE
OBSERVE IN KNOWN TTD
INDIVIDUALSEE USE DIFFERENT AGES
BECAUSE WE HAVE TO REMEMBER THAT
THE TTD IS A PROTEIN DISORDER,
SO IT'S AFFECTED BY A PREMATURE
AGENT.
SO THIS INDICATES THAT THE TTD
FIBRO BLAST IN THE AREA THAT
SURROUNDS THE FIEB PROBLAST NEXT
IN THE EXTRA CELLULAR REGION
THAT IS THE PRESENCE OF A LOOSE
COLLAGEN AMOUNT SO IT IS A
REDUCED COLLEGIA GENERATED
AMOUNT THEY ALL CONTROL FIBRO
BLAST, USING THOSE THAT MIGRATE
MIGRATE AT THE LOW RATE AND WE
CULTURE THEM MAINTAINING
EITHER--SORRY WE CULTURE THEM
WITH A NORMAL FIBRO BLAST OR
FIBRO BLAST MAINTAINING THE XP
AND THE SECOND FIBRO BLAST IN
THE SAME MEDIUM.
WE COULD OBSERVE THAT 1
EXPERIENCE WHEN THEY ARE CO
CULTURE WITH THE FORMAL FIBRO
BLAST.
SO THIS INDICATES THAT THE TTD
SECRETE AND PRODUCE THE FACTORS
THAT CAN INFLUENCE THE MIGRATION
RATE OF THE FIBRO BLAST, SO
THOSE SAY THAT THIS SECRETE
FACTOR IS INDEED AN NP 1 AND WE
USE AN INHIBITOR OF DMT, AND
ENZYMES THAT THE WE CAN ALSO
SLOW DOWN THE FIBRO BLAST
SURROUNDED BY TTP.
SO BECAUSE THE MUTATION IN THE
COLLAGEN TYPE 6 GENES THAT ARE
IN THE COLLAGEN TYPE 1 GENES ARE
RESPONSIBLE FOR HUMAN GENETIC
DISORDERS THAT WE DECIDED TO
COMPARE THE PHENOTYPE OF THE
COLLAGEN 6 PATIENTS, THEY ARE
AFFECTED BY THE CONGENIT ONAL
MUSCLE, OR THE MUSCLE DYSTROPHY,
SO WE ARE NOW WONDERING AND ASK
WHETHER THE TRANSCRIPTION
REGULATION, COLLAGEN 6 THAT WE
OBSERVE ONLY SPECIFIC CELL
CULTURE CONDITION MAY AFFECT FOR
THE JOIN CHAPTERS THAT BEEN
REPORTED THAT MEAN SOME TO TCB
PATIENTS AND WHETHER THE
OVEREXPRESSION OF AN NTP 1 COULD
EXPLAIN THE PROGRESSIVE BONE
ACTIVATIONS THAT HAVE BEEN
DESCRIBED IN TTD.
A THIRD GENE THAT WE FOUND
TRANSCRIPTIONOT REGULATOR SYSTEM
PPGIS AND THIS IS COMPARED TO
THE CONTROL FIBRO PLAOF THE AND
A REDUCED PROTEIN AMOUNT IN THE
TTD PATIENTS, MOVING MUTATION
BUT NOT IN THE EAST MUTATION.
SO THIS ENZYME BELONGS TO
PATHWAY THAT'S AGE 2 AS A
PRECURSOR AND THEN USES THE
LAMINA A2 AS A SUBSET THE PROOF
THE O GLANDIN I2, SO THOSE ARE
THE ACTIVE THAT ARE IMPORTANT
FOR FUNCTION THIS HAS BEEN SHOWN
TO HAVE IMPORTANT AS AN
IMPORTANT REGULATOR OF THE
CARDIOVASCULAR SYSTEM, IT IS AN
INHIBITOR OF THE PLACE OF
OBLIGATIONS, AND IT ALSO
IMPORTANT IT IS IMPORTANT
ENDOGENOUS VASE O DILATOR.
PLGA2 HAS SHOWN IT CAN BE IN THE
IMMUNE SYSTEM, THE INNATE AND
INDUSTRIOUSIVE IMMUNE SYSTEM AS
WELL AS A FACTOR ON THE OLIGIO
CITES DIFFERENTIATION AND IT HAS
BEEN FOUND OVER A SPLICE IN THE
STROMA THAT SURROUNDS A SPECIFIC
TYPE OF TUMOR CELL, MAINLY AT
THE LEVEL OF THE LUNGS.
SO CONSIDERING THAT TTD, THE
FREQUENTLY PRESENT A WEAK IMMUNE
SYSTEM AND THEY ALSO CANNOT
ACCUMULATE THE SUBCUTANEOUS
FACT.
WE ARE NOW WONDERING WHETHER THE
PGIS TRANSCRIPTION REGULATION
WILL SOMEHOW ACCOUNT FOR THIS
FEATURE IN TD, AND THEN
EVENTUALLY IT WILL SOMEHOW
CONTRIBUTE TO THE LACK OF CANCER
FORMATION IN T, TD.
SO IN TRYING TO UNDERSTAND IT
AND PARTIALLY ADDRESS THIS
QUESTION, WE EXTENDED THE
ANALYSIS TO ANOTHER GROUP OF
PATIENTS, THESE ARE TTD PATIENTS
BECAUSE THEY PRESENT THE TIGER
TAIL OF THE PATH, BUT THEY ARE
NOT MUTATED IN THE COMPLEX, THIS
REPRESENT IN A NORMAL NUCLEOTIDE
EXCISION REPAIR ACTIVITY AND THE
CELLS RESPOND NORMALLY TO THE UV
RADIATION.
SO, WE OBSERVED THE AMOUNT OF
THIS PATIENT THAT IS A WIDE
HETEROGENERATEDDITY THAT CAN BE
MUTATE INDEED SEVERAL DIFFERENT
GENES AND WE COULD OBSERVE THAT
BOTH THE PROTEIN LEVEL AND THE
TRANSCRIPT LEVEL INDEPENDENTLY
ON THE GENE, THE MUTATED GENE,
THE CELLS OF THE PATIENT CONTAIN
REDUCED AMOUNT OF THE PLGIS, AND
THIS IS CANNED WHAT BY
RECRUITMENT OF BOTH THE TF2 H
AND THE OLIGMERACE 2 ON THE GENE
PROMOTER.
BOTH IN THE PATIENTS, MUTATING
ACTIVATE AND ALSO IN THE
PATIENTS THAT ARE NOT MUTATING.
SO THEY HAVE A NORMAL--DNA
REPAIR ACTIVITY.
SO INDICATING THAT THIS IS THE
FIRST TIME AND THATY WE FOUND A
TRANSCRIPTION D-FACTOR THAT
ACCOUNTS AND IS PRESERVED, THE
REGULATION IN ALL TTD PATIENTS.
SO, IT'S WHAT WE CALL A GOLD
STANDARD MARKER FOR THIS
DISORDER AND WE HOPE THAT IT
MIGHT CONTRIBUTE TO THE CLINICAL
DIAGNOSIS.
CONCERNING THE PATIENT, THE TTD
PATIENTS PRESENT NORMAL DNA
REPAIR ACTIVITY.
IMPORTANT STUDIES HAVE BEEN
PERFORMED AND IN SUPPORT OF THE
FACT THAT TTD CAN REALLY BE
ASSOCIATED WITH TRANSCRIPTIONAL
IMPAIRMENT THIS CAME FROM A
STUDY THAT WE HAVE PERFORMED IN
COLLABORATION WITH KEN KRAEMER,
WE BOTH FOUND PATIENTS THAT WERE
MUTATING WITH THE GENE, WE FOUND
2 CASES OTHERS BOTH CONTAINING
THE PATIENT TO SHOW THE
DIFFICULT TYPE OF
[INDISCERNIBLE] AND THE 2
PATIENTS WERE HOMOZYGOUS FOR
THIS MUTATION IN THE SAME GENE
WHICH IS THE GTF2 E2 GENE WHICH
IS ENCODES THE BETA UNITS OF THE
TRANSCRIPTION FACTDOR 3.
SO THE TRANSCRIPTION FACTOR 3
MADE THE SUBUNITS AND IS ALSO
LIKE TF2 H AND IMPORTANT AND
ESSENTIAL ELEMENT IN THE
FORMATION OF THE INITIATION
COMPLEX, THE INTERACTION BETWEEN
TF2 H AND EXTREMELY IMPORTANT
ALSO TO ACTIVATE THE KINASE
ACTIVITY OF TF2 H AND THEN THE
SUBSEQUENT PHOSPHORYLATION AND
ACTIVATION OF [INDISCERNIBLE].
SO TOGETHER WE CAN, WE ARE ABLE
TO DEMONSTRATE THAT THE
MUTATIONS, BOTH MUTATIONS
AFFECTING, AFFECTERRING THE
STABILITY OF THE TF2 BETA UNIT
BUT ALSO THE TF2 IN OUR SUBUNIT,
INDICATING THAT SIMILARLY TO THE
TF2 H AND MUTATED THE CELLS,
ALSO IN THIS CASE, THERE IS A
REDUCED AMOUNT OF THE BASAL
TRANSCRIPTION FACTOR.
SO THESE WERE SURPRISING SOME
SUPPORT AND SUGGESTING THAT TTD
CLINICAL FEATURE ARE REALLY
RELATED TO THE TRANSCRIPTIONAL
DEFECT.
BUT THEN IMMEDIATELY AFTER THAT,
IN OUR LAB, AND IN COLLABORATION
WITH [INDISCERNIBLE] IN THE
[INDISCERNIBLE] LAB, IDENTIFY
ANOTHER FEW CASES OF THE
PATIENTS THAT ARE NOW PROFICIENT
SO THEY DON'T HAVE
[INDISCERNIBLE].
THEY PRESENT THE TIGER TAIL PAIR
OF ABNORMALITY AND THESE FEW
CASES OF PATIENTS ARE MUTATED IN
GENES SO THAT ENCODE FOR A GROUP
OF PROTEIN THAT ARE CALLED THE
TRNA PIECE, THE SPECIFIC TRNA
SYNTH TAIS INHIBITORS HAVE BEEN
FOUND MUTATED AND THESE ARE
ENZYMES SO THAT ARE EASIER TO
UPLOAD THE PROGRAM CORRECT AMINO
ACIDS ON THE CORRESPONDING TRNA,
SO WHEN THE TRNA IS CHARGED WITH
THE IMMUNO, IT'S USED TO--IN THE
PROTEIN SYNTHESIS PROCESS BY THE
RIBOSOMES FOR THE PROTEIN
SYNTHESIS.
SO WHAT WE OBSERVE IS THAT THE
OTHER YOU HADITATIONS THAT WE
HAVE IDENTIFIED IN THESE
PATIENTS ALSO RESULT IN REDUCED
PROTEIN SYNTHESIS.
SO THESE CELLS CONTAIN REDUCED
AMOUNT OF THE [INDISCERNIBLE]
TRA SYNTH TAIS INHIBITOR, BUT
BECAUSE OF THAT WE HAD TO
RECONSIDER AND REVISE OUR IDEA
OF CHILD DYSTROPHY AND NOW WE
ARE USING A BROADER DEFINITION
SAYING THAT TTD IS NOT ONLY
TRANSCRIPTIONAL BUT THE GENE
EXPRESSION DISORDER.
AND THE SECOND PART OF MY TALK,
I WOULD LIKE TO FOCUS ON THE
THIRD, THE DISORDER ASSOCIATED
WITH DNA REPAIR FACTOR WHICH IS
THE COCKAYNE SYNDROME, IT'S THE
MAJORITY OF PATIENTS ARE CSA OR
CSG GENE AND THESE PLAY AN
IMPORTANT ROLE IN TRANSCRIPTION
REPAIR, SO THEY ARE SHOWN TO
REMOVE IT, TO RECOGNIZE THE
LESIONS THAT INTERFERE WITH THE
TRANSCRIPTIONAL PROCESS.
IF WE LOOK HERE AT THE PICTURES
OF THE CS PATIENTS, OF THE SAME
PATIENTS OVER THE EAR, WE CAN
CLEARLY OBSERVE THE APPEARANCE
OF THE GENERAL FEATURES THAT
AFFECT THE PATIENTS, SO THIS IS
AN ITALIAN PATIENT, SO THAT'S
CARRYING A MUTATION IN THE CSA
GENE, SHE DIED AROUND THE AGE
EVER 18 YEARS, WITH THE PROBLEMS
THAT MANY DIFFERENT ORGANS.
SO WOO CAN--BASED ON THAT BY
LOOKING AT THE PROGRESSION OF
THE DISORDER WE CAN CLAIRLY SAY
THAT CSA AND IN A SIMILAR WAY
CSB PLAY AN IMPORTANT ROLE IN
PRESERVING THE FEATURE AND THE
ORGAN FUNCTIONALITY AND THEY
COME TO THE APPEARANCE OF THE
DEGENERATIVE FEATURES.
BUT HOWEVER, WE WERE VERY
SURPRISED THAT WHEN IN 2009,
ANOTHER NOW IN COLLABORATION
WITH THE [INDISCERNIBLE].
THEY WERE CHARACTERIZING A
PATIENT [INDISCERNIBLE] PATIENT
THAT WAS CLASSIFIED AS A UV
SENSITIVE, AFFECTED BY THE UV
SYNDROME, BECAUSE THE ONLY ONLY
SHOWED SEBSATIVITY AND AND THEN
THE PATIENTS SHOW A NORMAL
PHYSICAL AND MENTAL DEVELOPMENT.
BUT WHEN THEY PERFORM, WHEN IN
THE LAB THEY PERFORM THE
ANALYSIS TO IDENTIFY WHICH GENE
WAS RESPONSIBLE FOR THESE NEW
SENSITIVITY, SHE FOUND OUT THAT
LIMITATION WAS IN THE CSA GENE,
IN PARTICULAR IT WAS NOVEL
MUTATIONS THAT WAS FOUND IN
HOMOSIGNIFY SWROGGITY IN THE
PATIENTS AND IT'S LOCATED IN THE
C-TERMINAL OF THE PROTEIN.
SHE LOOKED AT THE AND THE
SENSITIVITY TO A RADIATION
HOWEVER, THE CELLS THAT ARE
DAMAGE INDICATING THE SPECIFIC
MUTATION SOMEHOW CAN AFFECT THE
TRANSCRIPTION CAP LANREPAIR BUT
IT DOESN'T AFFECT OTHER PATHWAY
IN THAT IN WHICH THE CSA IS
INVOLVED.
SO, WE CAN ALSO SAY THAT BASED
ON THAT, THAT THE DEFECTIVE CNA
PROBABLY MAINLY RESULTS IN THE
ACTIVATION BUT WE HAVE TO
CONSIDER THE POSSIBILITY THAT IT
WAS MOTOR LOGICAL AND PHYSICAL
DETERIORATIONS AND PREMATURE
AGING FEATURES THAT WE SEE IN
THE CS PATIENTS, THEY PROBABLY
REFLECT ADDITIONAL ROLES THAT
THE CSA PLAYS OUTSIDE
TRANSCRIPTION REPAIR, SO WITH
THE IDEA TO ADDRESS, WELL, FIRST
I HAVE TO SAY THAT THERE IS THE
FACT THAT THE CS PROBING LATER
ROLE OUTSIDE TRANSCRIPTION HAS
BEEN DEMONSTRATED AND TESTED BY
THE WORK OF MANY DIFFERENT
GROUPS, IN FACT, THE CS PROTEIN
ARE INVOLVED IN TRANSCRIPTION
BOTH DNA POLYMERASE AND RNA
POLYMERASE 1, THEY HAVE BEEN
SHOWN TO PLAY AN IMPORTANT ROLE
IN THE FUNCTIONALITY BECAUSE OF
THE CSA PATIENT, ACCUMULATING
THE POLARIZED MITOCHONDRIA O
CHONDRIAL AND MITOCHONDRIAL FRAG
M-TEBURKEULOSEISATIONS, CSB
IMPACTS SO WITH A KEY PLAYER IN
THE BASIC EXCISION REPAIR, AND
MORE SIMPLY IT HAS BEEN
DEMONSTRATED THAT CSB AND ALSO
PLAY A ROLE IN THE MITOTIC
PROCESS, SO IT IS NOT SURPRISING
TO THINK THAT THEY PLAY A ROLE
OUTSIDE TRANSCRIPTION REPAIR,
BUT PROBABLY WHAT WE CAN LEARN
FROM THE SPECIFICALLY SENSITIVE
PATIENT IS THAT CSA PROBABLY
INTERACTS WITH DIFFERENT
PLATFORMS AND THE INVITATION
WITH THE TERMINAL REGION WITH
THE FACT OF CSA, WE ARE NORMALLY
INTERACTING IN TRANSCRIPTION CAP
LANREPAIR BUT DOESN'T AFFECT THE
INTERACTION OF CSA WITH OTHER
PARTNERS, THAT ARE RELEVANT FOR
THE OUTER FUNCTION OF CSA.
>> SO WITH THE IDEA TO IDENTIFY
WHICH ARE OTHER PROTEINS
INTERACTING PROTEINS WITH THE
CSA FOR C SA FUNCTIONALITY, WE
DEVELOP THE PANEL OF ISOGENIC
CELLS, WE--BY USING FIRST WE USE
THE FOUND EFFECTIVE IN CSA, AND
ORGANIZE THE CELLS AND WE
INTEGRATED A SINGLE POSSIBILITY
OF THE CHROMOSOME 9, THAT
CONTAINS THAT INCLUDING THE WILD
TYPE FORM OF CSA.
WE ALSO USE THE SAME APPROACH TO
EXPRESS THE MUTATED SPECIFICALLY
MUTATED OF CSA, BUT TODAY I WILL
ONLY FOCUS ON THE WILD TYPE CSA.
SO WE FIRST VERIFY THIS QUESTION
OF THE COMBINATION OF THE TOPIC
GENE.
THE TRANSGENE WE VERIFIED THAT
PROPOSED DISCLOSURE TEEN WAS
LOCALIZING IN THE CASE, AND WE
SHOULD CHECK THE FUNCTIONALITY
OF THIS PROTEIN, BECAUSE THE C
SA DEFACTOR CELLS IN THE WILD
TYPE FORM OF CSA, COULD RECOVER
BOTH OF THE UV SENSITIVITY AND
ALSO THE SENSITIVITY TOWARDS THE
STEPS.
SO, THE PROTEIN IS FUNCTIONAL
AND THANKS TO THE FACT THAT THE
PROTEIN HAS DUAL TARGET ON THE
C-TERMINAL REGION.
WE COULD PERFORM
AUTHENTIFICATIONS TO IDENTIFY
AND ISOLATE CSA INTERACTION AND
THEN BY MASS SPEC TROMETRY WE
IDENTIFY WHICH PROTEINS INTERACT
WITH THE CSA, WE END UP WITH A
CONSIDERABLE NUMBER OF CSA
INTERACTIVES, ALL THE PROTEINS
THAT HAVE KNOWN TO ISHT ACT WITH
THE CSA TRANSCRIPTION REPAIR
HAVE BEEN IDENTIFIED BUT ALSO
SEVERAL OF THE NOVEL PROTEINS
AND AMOUNT THEM THE FIBRO
CHELATASE, SO THAT WAS AN
INTERESTING PROTEIN CLASS
BECAUSE IT IS FIRST AS A MIGHT
ON CHONDRIAL PROTEIN AND IT IS
KNOWN, IT'S AN ENZYME THAT IS
IMPORTANT TO UPLOAD THE
[INDISCERNIBLE] INTO THE 2 TEEN
9 SO IT CATALYZES AND MEDIATES
THE BIOSIPGHT SIS OF THE LAST
[INDISCERNIBLE].
THE MUTATION IN FACT ARE FOUND
IN PATIENTS AFFECTED BY THE
[INDISCERNIBLE].
THE PATIENT SHOW SEVERE AND PAIN
EMPLOY PHOTO TOXIC REACTIONS SO
THEY ARE PHOTO SENSITIVE, BOTH
AT THE LEVEL OF THE SKIN BUT
ALSO THE LEVEL OF THE LIVER.
AND THIS IS PROBABLY BECAUSE IT
IS DUE TO THE FACT THAT THE
PATIENTS ACCUMULATE
[INDISCERNIBLE] IN THEIR BLOOD.
SO WE DECIDE TO INVESTIGATE THE
INTERACTIONS BETWEEN FECH AND
THE DNA, OOH EXPOO FOUND IT'S
NOT SO MUCH INSIDE THE
MITOCHONDRIA, BUT MORE OCCURRING
OUTSIDE THE MITOCHONDRIA, BY
FORMING THE LIGATION ASSAY,
THERE IS INTERACTION BETWEEN THE
2 PROTEINS INSIDE AND IN
PARTICULARLY IN THE AREA THAT
THE [INDISCERNIBLE].
SO WE WENT FURTHER AND
WE--VERIFIED THAT FACTOR WAS IN
FACT PRESENT INSIDE THE
[INDISCERNIBLE].
AND THEN WE WENT BACK TO OUR
MASS [INDISCERNIBLE] ANALYSIS,
TO SEE WHETHER OTHER PROTEINS
INTERACT WITH CSA THAT WERE ALSO
FOUND INSIDE [INDISCERNIBLE],
AND WE FOUND SOME SPECIFIC
RIBOSOMAL PROTEIN THAT BELONGED
TO THE SMALL SUBUNIT OF THE
RIBOSOME [INDISCERNIBLE], SO WE
STARTED TO CHECK THE INFECTION
BETWEEN FECH AND THESE RIBOSOMAL
UNITS BUT ALSO THE POSSIBLE
INTERACTION BETWEEN FECH AND DNA
POLYMERASE 1 AND BY PERFORMING A
2 STEP [INDISCERNIBLE] IN THE
OUTER--BY USING THE CHROMATINS
AND [AUDIO CUTS OUT ] WE WANT TO
SHOW THAT ON THE CHROMATIN OF
NORMAL FIBRO BLAST, THERE IS THE
PRESENCE OF THE PROTEIN COMPLEX,
WHICH CONTAINS BOTH THE
[INDISCERNIBLE] CSA, CSB AND THE
SPECIFIC RIBOSOMA PROTEIN.
BY PERFORMING OF THESE
PRECIPITATION EXPERIMENTS WE
WERE ABLE TO OBSERVE THAT THE
COMPLEX IS BOUND TO THE
CHROMATIN, BUT WHEN CSA IS
MISSING IT ALSO BINDS DUE TO
CHROMATIN BUT THE AMOUNT OF THE
COMPLEX IS REDUCED AS WE CAN
OBSERVE WHEN WE CONVERT THE WIDE
BAR THAT ARE THE FACTOR WITH C
SA COMPARED TO THE BLACK BAR OF
THE WILD TYPE CSA PROTEIN.
BEFORE WE DO THE UV LIGHT WE
OBSERVE THAT NORMAL FIBRO BLAST
THERE IS A PARTIAL DESEGREGATION
OF THE CONFLICTS AS WE CAN
OBSERVE BY REDUCTION IN ALL
THE--IN ALL THE ELEMENT OF THE
COMPLEX, WHEREAS IN 1 CSA IS
DEFECTIVE, THERE IS NO
ALTERATION IN THE AMOUNT
OF--THERE IS NO DESEGREGATION,
SO THE FACT THAT THE CSA IS
IMPORTANT FOR THE FORMATION OF
THE PROTEIN OF THIS PROTEIN
COMPLEX ON THE KRONISHEMMA TIN,
IT HAS ALSO BEEN DEMONSTRATED BY
PERFORMING THE PRECIPITATION,
WHERE WE CAN OBSERVE THE
EQUIVALENCY OF THE DNA
POLYMERASE 1 ON THE RIBOSOMAL
DNA, AND CLEARLY WE CAN OBSERVE
THAT 1 CSA IS EXPRESSED, THE
BLACK BARS, THERE IS A HIGHER
[INDISCERNIBLE] OF DNA
POLYMERASE, SO IN NORMAL CELLS
IF WE SILENCE CSA, WE ALSO AGAIN
FIND A REDUCTION IN THESE
OTHERS.
BUT THE SAME RESULT WE OBTAIN
WHEN WE SILENCE THE FACT DURING
THE CANNED WHAT INDICATING THAT
MULTIPLE CSA, BUT ALSO THE
[INDISCERNIBLE] ARE IMPORTANT
FOR THE FORMATION OF THE RNA
POLYMERASE COMPLEX ON THE
RIBOSOMAL DNA.
HOWEVER, WHEN WE CONTINUE THE
EXPERIMENT TO VERIFY WHAT IF
THERE WAS A REDUCTION IN THE
SYNTHESIS OF THE ROBO STUDIES OF
MULTIPLE ENDOCRINAL RNA, WE WERE
SURPRISED TO OBSERVE THAT WHEN
WE SILENCE CSA, IN FACT, WE
OBLIGATIONS STAIN A REDUCTION IN
THE RNA SIPGHT SIS, BUT WHEN WE
SILENCE THAT, WE OBSERVE AN
ACCUMULATION AND INCREASE AMOUNT
OF RIBOSOMAL KD--SALLY RNA.
SO THE SIM THING IF WE LOOK IN
PATIENT CELLS SO THESE ARE
PRIMARY FIBRO BLAST FROM EITHER
NORMAL INDIVIDUAL OR PATIENTS
WITH CSA AND CLEAR WE WE HAVE A
REDUCTION IN THE RNA SYNTHESIS
AND THESE WAS EXPECTED BECAUSE
IT WAS PREVIOUSLY DEMONSTRATED
BY A GROUP, EVEN THEY ALL SHOW
THAT THE CS CELLS CONTAINA A
REDUCED RIBOSOMAL TRANSCRIPTION.
BUT AGAIN, WHEN WE LOOK AT THE
MUTATIONS OF THOSE IN THE GENE,
WE CAN OBSERVE AN INCREASE
AMOUNT OF RIBOSOMAL
[INDISCERNIBLE].
SO THIS IS IN COMONT WITH WHAT
WE OBSERVE FOR THE RNA
POLYMERASE 1 ON THE PROMOTER SO
TRY TO UNDERSTAND THAT THE
REASON OF THIS FREQUENCY, WE
WE--WE--YOU FORM A REALTIME PC R
ANALYSIS, IN PATIENTS FIBRO
BLAST, AND THAT WE COULD CLEARLY
OBSERVE THAT WHEN WE TRACK THE
EXPRESSION OF THE 47 AS A
RECURSOR OF THE RIBOSOMAL RNA,
WE ARE REDUCED SYNTHESIS, BOTH
EFFECTIVE AND CSA EFFECTIVE
CELLS.
BUT HOWEVER, WHEN WE CHECK THE
AMOUNT OF THE 18 S RIBOSOMAL
RNA, WE CAN OBSERVE IN FACT
PATIENT NOT CSA AND ACCUMULATION
OF THE THE SPECIFIC
[INDISCERNIBLE].
A SIMILAR RESULT HAVE BEEN
ACHIEVED BY PERFORMING THE
NORTHERN BLOT ANALYSIS IN THE
LOCALIZED CELLS IN WHICH WE
SILENCE EITHER THE FACT GENE OR
CSA.
THE NORTHERN BLOT HAS BEEN DONE
IN COLLABORATION WITH
[INDISCERNIBLE], AND WE CAN
OBSERVE THAT THERE IS REDUCED
AMOUNT OF THE 47 [INDISCERNIBLE]
RNA RECURSOR BUT WITH WE COMPARE
AND WE ANALYZE THE AMOUNT OF THE
18 S COMPARED TO THE AMOUNT OF
THE 47 S, WE CAN AGAIN OBSERVE
THE TENDENCY IN CELLS THAT ARE
DEFECTIVE, TO ACCUMULATION THE
18 S [INDISCERNIBLE].
SO, WE CAN SAY THAT BOTH THE CSA
IN FACT PLAY A ROLE IN
REGULATING THAT DNA POLYMERASE 1
AND IN THE CASE OF CSA, IT'S
ALSO PLACING AN IMPORTANT ROLE
IN THE RESPONSE TO UV
IRRADIATION, HOWEVER, THEY ARE
BOTH FOR THE COMPLEX CS AND THE
TTD PATIENTS SO WE SEE DIFFERENT
RIBOSOMAL BIOGENESIS
[INDISCERNIBLE].
SO CONSIDERING AND KNOWING THAT
THE CS, AND EPP STRESS AND THE
RIBOSOME BIOGENESIS ALERATIONS
ARE EARLY EPIIMENETTIC EVENT
THAT ARE CHARACTERIZED BY EITHER
A GENERATIONAL MATURE AGING
FEATURES, WE ARE NOW DEALING
WITH THESE--THESE PREROGATIVE
QUESTIONS WHETHER THEY ARE A
FUNCTION OF CS PROTEIN INSIDE
THE NUCLEOULAR.
SO I WILL CONCLUDE MY TALK AND I
WOULD WILL WOULD LIKE TO THANK
ALL THE PEOPLE THAT CONTRIBUTED
TO THE WORK BUT BEFORE DOING
THAT, I WOULD LIKE TO THANKS TO
GIVE A SPECIAL THANKS TO
[INDISCERNIBLE] BECAUSE SHE IS
THE PERSON WHO STARTED THE GROUP
WORKING ON DEFECTIVE DISORDERS
HERE IN P A VIA, AND SHEES ALSO
THE 1 WHO STARTED TO CREATE THE
IMPORTANT NETWORK OF
COLLABBUATION WITH THE
CLINICIANS AND SCIENTISTS AND
ALSO THE PATIENT FAMILIES.
I WOULD LIKE TO ACKNOWLEDGE A
NUMBER OF [INDISCERNIBLE].
SHE MADE A GREAT WORK HERE WITH
THE TTD PATIENTS.
BOTH OF THOSE THAT HAVE YOU
HADITATION IN TF2 H AND MORE
RECENTLY THOSE THAT ARE NOT
MUTATED IN TF2 H AND
[INDISCERNIBLE] THEY BOTH WORK
ON THE CS, AND THE RIEB OR
STUDIES OF MULTIPLE ENDOCRINE
ALTERATIONS IN CS.
MANUELA WAS A Ph.D. STUDENT IN
THE LAB AND HE DID HER FIRST
POST DOCKER WITH ME, WORKING ON
THE [INDISCERNIBLE] CSA
INTERACTION AND THEN AFTER A
PERIOD ABROAD, SHE WENT TO
BASUL, NOW SHE'S BEEN THERE, AND
SHE'S BACK NOW AND SHE HAS A
PERMANENT POSITION HERE WITH US
AT [INDISCERNIBLE].
JULIA HAS FINISHED HER Ph.D.
AND NOW SHE'S STARTED--SHE'S
STARTING THE POST DOC TO TRYING
TO CHARACTERIZE THE RIBOSOME
BIODPEN SIS ALTERATION IN CS,
AND APP PATIENTS.
ERICA IS A NEW Ph.D. STUDENT
THAT JUST JOINED US AND SHE'S
WORKING ON THE XP PATIENTS.
TIZIA NA I HAVE TO GREATLY
ACKNOWLEDGE HER FOR ALL THE WORK
SHE HAS ALWAYS DOEN WITH THE
PATIENTS.
THOSE ARE TO DEFINE THE--WHICH
THE CELLULAR DEFECT OF THE
PATIENT THAT ARE ALSO, SHE'S
VERY MUCH INVOLVED IN THE
IDENTIFICATION OF POSSIBLE NOVEL
CAUSATIVE GENES.
WE HAVE TO PERFORM THE MOLECULAR
ANALYSIS TO IEE, AUDIENCE
DENTIFY THE MUTATIONS IN THE
PATIENTS.
AND I ALWAYS HAVE TO THANK THE
PAST MEMBER IN THE LAB, LAVENIA,
WITH HER CONTRIBUTION IN THE
[INDISCERNIBLE] WORKING AND
ANITA LOMBARDI FOR THE WORK ON
THE PGIS AND DEBORAH AND MARTINA
WHO GAVE STRONG CONTRIBUTION TO
THE LAB EVEN IF I HAD NO TIME
TODAY TO SPEAK ABOUT THEIR WORK.
OUR COLLABORATORS ARE IGM,
[INDISCERNIBLE] AND
[INDISCERNIBLE] AND A SPECIAL
THANKS TO ALL OUR SEVERAL
COLLABORATORS AND MANY OF THEM
HAVE ALREADY BEEN MENTIONED BUT
I WILL JUST REMEMBER IN ADDITION
TO THE PREVIOUS PLAN,
[INDISCERNIBLE] THEY HAVE BEEN A
GREAT SUPPORTER IN DEFINING THE
AGE TO ALTER THE MECHANISM THAT
OUR RESPONSIBILITY FOR THE JEERN
EXPRESSION REGULATION, AND THEN
[INDISCERNIBLE] FROM THE UK,
BECAUSE THEY ARE GREAT SUPPORT
AND ALSO FOR THE--BECAUSE THEY
SHARED, THAT WE'VE LIFE--THEY
FREQUENTLY SHARE DATA WITH US.
AND I WANT TO THANK
[INDISCERNIBLE] BECAUSE HE HAS
PERFORMED EXOME SEQUENCING IN
SEVERAL OF OUR STILL UNSOLVED
CASES.
OF COURSE BIG THANKS TO ALL THE
COLLABORATORS, CLINICIANS,
PATIENTS AND I WOULD LIKE ALSO
TO MENTION THE ITALIAN
ASSOCIATION OF XP PATIENT THAT
WAS BORN LESS THAN 3 YEARS AGO
BUT IT WORKS QUITE NICELY AND IN
THE LAST FIELD, THEY WERE ALSO
ABLE TO SUPPORT AS IF FROM THE
ECONOMICAL POINT OF VIEW AND ALL
THE OTHER URGENCY THAT SUPPORT
[INDISCERNIBLE].
AND THANK YOU FOR ATTENDING.
>> OKAY, THANK YOU SO MUCH FOR
THIS WONDERFUL TALK.
>> THANK YOU.
>> THERE ARE A COUPLE QUESTIONS
HERE 1 WAS FROM JOHN DEAD WHAT,
FANTASTIC TALK, THE ABNORMALITY
IN PROOF THE GLANDIN I2 THAT YOU
SHOWED IN TTD, IS PROBABLY
CLINICALLY IMPORTANT.
DO THE PATIENTS WITH MUTATIONS
IN THE LESS COMMON TTD GENES
SPECIFICALLY TTD A, TRNA SYNTH
TAIS INHIBITORS, XPV, R113 A,
GTFEE2, DO THESE PATES ALSO HAVE
AN ABNORMALITY IN THE GOLD
STANDARD OF PROOF THE GLANDIN
I2?
>> GOOD QUESTION.
YES, WE CHECK THEM AND ALSO
THOSE [INDISCERNIBLE] IN THE
TRNA SYNTH TAIS INHIBITOR THEY
HAVE A DEFECT IN THE TGIS.
ALSO THOSE MUTATED IN THE TF3 OR
TBN 1 SO THEY ALL SHOW THE
DEFAULT, ALL 3 OF THOSE THAT WE
HAVE ANALYZED, SO THERE IS A
REDUCED AMOUNT.
SO FOR THAT REASON, I REALLY
HOPE THAT IT MIGHT BE RELEVANT
FOR THE CLINICAL FUTURES.
>> OKAY.
LET'S SEE WHAT'S HAPPENING HERE.
OH, HERE WE GO.
IT SAYS FROM ALAN, I ASSUME
THAT'SAL AN LAMAN, THANKS FOR
THE VERY NICE TALK.
TWO QUESTIONS.
ONE, DO NONPHOTO SENTHATIVE
TTDs SHOW THE SAME COLLAGEN
DEFECTS AND MIGRATION CHANGES.
WELL, WE WILL DO 1 AT A TIME.
>> YEAH, THEY DON'T.
THEY DON'T HAVE EITHER--WE
DIDN'T CHECK THE MIGRATION
FEATURE, I HAVE TO SAY BECAUSE
WE CHECKED THE TRANSCRIPTION
REGULATION AND IT'S NOT
SURPRISING TO IN
[INDISCERNIBLE].
EVEN MORE IMPORTANTLY, THE MMP1
OVEREXPRESSION SEEMS TO BE
SPECIFIC FOR THE FIBRO BLAST, WE
CHECK THE KERATINOCYTES OF THE
TTG PATIENTS IN XPD AND WE
COULDN'T FIND THE TRANSCRIPTION
REGULATION THERE.
SO IT SEEMS TO BE RELATED TO
FIBERS.
I CANNOT SAY WHICH ARE THE CELL
TYPES BUT THAT THE LEVEL OF THE
SKIN, IT AFFECTS THE FIBRO
BLASTS IN THE DERMAL LAYER WHICH
ARE THE CELLS THAT MAINLY
PRODUCE THE EXTRA CELLULAR
MATRIX, THEN.
>> OKAY.
>> THAT'S WONDERFUL.
OKAY, THEN, THE SECOND QUESTION
IS, I PROBABLY ASKED YOU THIS
BEFORE, THIS IS FROM ALAN, BUT
WHERE DOES CSB FIT INTO THE
FERIA KEEL LOW TAIS INHIBITOR
RP1 STORY?
>> WELL, WE FIND THE CSB IN THE
COMPLEXES SO THE PROTEIN IS
THERE, WHAT WE STILL DON'T KNOW
IS WHAT HAPPENS WHEN CSB IS
DEFECTIVE BECAUSE WE DID CHECK
IN CELLS THAT ARE KNOWN TO BE
DEFECTIVE FOR CSB, EVEN WITH
THIS TRUNCATED FORM OF CSB IS
LESS VULNERABILITY AS FAST, WHEN
WE ISOLATE THE COMPLEX BY USING
FACH AND THE CSB 1 WE FIND IT
THERE SO I CANNOT SAY WHETHER IN
THE ABSENCE OF CSB, THE
CONSEQUENCE WILL BE THE SAME AS
IF WE DON'T HAVE CSA, THIS I
DON'T KNOW.
>> OKAY, BUT WE ARE WORKING ON
THIS, WE ARE TRYING TO
IMMORTALIZE THIS CELL LINE THAT
IS DEFECTIVE FOR CSB, SO THEN
MAYBE WE WILL HAVE THE ANNOUNCER
AT A CERTAIN POINT.
>> GREAT.
WELL, THANK YOU SO MUCH.
JUST A FEW MINUTES LEFT, I MIGHT
AS WELL ASK A QUESTION, YEARS
AGO, WE'RE LOOKING AT PHOTO
PRODUCT REPAIR, DIFFERENTIAL
PHOTO PRODUCT REPAIR AND TTK
VERSUS XP AS A POSSIBLE
EXPLNATION FOR THE DIFFERENCE
BETWEEN THE FACT THAT THE XP
PATIENTS GOT A HIHIGH FREQUENCY
OF SKIN CANCER AND THE TTD
PATIENTS DIDN'T AFTER UV
RADIATION AND [INDISCERNIBLE]
ALSO FOWBD THAT THE TTD PATIENTS
DO REPAIR 6 [INDISCERNIBLE]
PHOTO PRODUCTS BUT NOT SO MUCH
THE CYCLOBUTANE DIMERS BUT THE
XP DOESN'T REPAIR ANY OF THOSE,
DO YOU HAVE ANY INFORMATION
ABOUT THAT ASPECT OF THE
DIFFERENCE?
>> WELL, NO, I KNOW YOUR STUDIES
AND I AM CONFIDENT THEY ARE
CORRECT BUT STILL, I DON'T
BELIEVE--I MEAN, I DON'T THINK
THIS MIGHT CONTRIBUTE MAYBE TO
DELAY THE APPEARANCE OF THE
TUMORS IN TTD, BUT THE FACT THAT
WE NEVER FIND TUMORS, WE HAVE
PATIENTS THAT CAN LIVE LONGER
WITH A [INDISCERNIBLE] CHILD
DYSTROPHY AND SO FAR THEY NORMAL
EXPOSE TO THE SUN MUCH MORE THAN
NORMALLY, NORMALLY, THEY DON'T
BEHAVE LIKE THE XP PATIENTS SO
EVENTUALLY THEY WOULD EXPLAIN A
DELAY IN THE APPEARANCE OF THE
TUMOR.
LIKE IF WE THINK OF THE VARIANT
FORM OF THE XP PATIENT, THEY
HAVE A NORMAL LAYER DEFECT.
THE NORMAL ACTIVITY BUT THEY
STILL, THERE IS--THERE ARE
PROBLEMS DOING AGAIN THE
APPLICATION AND THEY SHOW THE
APPEARANCE OF TUMOR RELATED TIME
DURING THEIR AGE.
SO I DON'T THINK--I MEAN IT
MIGHT CONTRIBUTE BUT IT'S NOT
SUFFICIENT TO REALLY EXPLAIN WHY
TTD DID NOT CANNED WHAT SCHEME,
THAT'S MY CONTRIBUTION, AND I
HONESTLY BELIEVE THAT
TRANSCRIPTIONAL IMPAIRMENT MIGHT
BE IMPORTANT.
YOU HAVE A LOW AMOUNT OF DEPOSIT
TRANSCRIPTION FACTOR SO PROBABLY
THOSE CELLS CANNOT REALLY
SUSTAIN THE HIGH RATE OF
TRANSCRIPTION THAT IS ACQUIRED
DURING THE PROLIFERATION OF
CANCER CELLS.
SO.
>> YEAH, THAT'S GOOD.
THANK YOU AND--COUPLE MINUTES
MORE, 1 MORE QUESTION, YOU
MEKSED BONE ABNORMALITIES IN
TTD, AND ACTUALLY JOHN AND I
RECENTLY SUBMITTED A MANUSCRIPT
FOR REVIEW, DESCRIBING THE
UNUSUAL SITUATION IN THE BONES
OF THE HIPS OF THE PATIENTS WITH
TTD, THAT DEVELOP PREMATURE
DEGENERATION OF THEIR HIPS THAT
LOOK LIKE ASEPTIC NECROSIS SO
THESE CHILDREN HAVE BEEN ABLE TO
WALK AND THEN SUDDENLY OVER A
RELATIVELY SHORT PERIOD OF TIME
DEVELOP PAIN IN THE HIP, FIRST 1
HIP AND THEN THE OTHER THEN THEY
BECOME UNABLE TO WALK AND IT
TURNS OUT THE HIPS THEMSELVES
ARE DEGENERATING AND THEY TRY TO
DO SURGICAL PROCEDURES ON THEM
BECAUSE THE PATIENTS ARE SO
WEAK, TO BEGIN WITH, THEY HAVE
HAD FOR RECOVERY VERY DIFFICULT
RECOVERY EXCEPT FOR THE CASE OF
1 PATIENT HAD PHYSICAL THERAPY
BEFORE THEY DID THAT.
BUT THAT'S AN UNUSUAL SITUATION
OCCURRING IN THESE LIGHT WEIGHT
CHILDREN WITH THAT.
NORMALLY THE ASEPTIC NECROSIS OF
BONE OCCURS AT A MUCH, MUCH
OLDER AGE WITH PEOPLE WHO MAY
HAVE INCREASED WEIGHT OR HAVE A
IMMUNO LOGIC PROBLEMS.
SO, THE NNT ABNORMALITIES MIGHT
PLAY A ROLE IN THAT BUT
UNFORTUNATELY WE HAVEN'T BEEN
ABLE TO GET ANY TISSUE TO
MEASURE THAT BUT IT IS POSSIBLE
THAT THEY DO HAVE CERTAINLY BONE
ABNORMALITIES AND THEY ALSO HAVE
AN UNUSUAL RADIO LOGIC SITUATION
IN THAT THERE'S INCREASED BONE
DENSITY IN THE SPINE, THE
CENTRAL SPINE OF THE PATIENTS
WITH TTD, AND DECREASED BONE
DENSITY, THEY CALL THAT IN THE
PERIPHERAL PORTION OF THE LEGS
AND THE ARM AND THE LOWER AND
THE HANDS IN THERE.
THEY CALL THIS CENTRAL
OSTEOSCLEROSIS AND PERIPHERAL
OSTERPENIA, AND WHICH IS UNUSUAL
FEATURES IN THE BONES OF
CHILDREN WITH TTD.
IN ACCOUNT FACAS WE HAD A REPORT
OF A RADIOLOGIST WHO MADE THE
DIAGNOSIS OF TTD JUST ON THE
BASIS OF THESE ABNORMALITIES IN
THE X-RAY.
SO THERE IS A LOT GOING ON.
>> , I WOULD LIKE TO KNOW
WHETHER IT IS NOVEL EXPRESSION
OF [INDISCERNIBLE] AT THE LEVEL
OF THE BONE.
IF YOU MANAGE TO HAVE SOME
TISSUE WE CAN CHECK.
>> YES, YES.
WELL THE DIFFICULTY IS GETTING
THE TISSUE.
>> RIGHT.
>> OKAY.
WELL, IF THERE'S NO MORE
QUESTIONS, WELL, THANK YOU.
>> THANK YOU.
>> THANK YOU SO MUCH FOR THIS
WONDERFUL TALK, YES.
>> IT'S VERY NICE, THANK YOU.
>> OKAY.