"The use of patient's fibroblasts in the evaluation of different therapeutic approaches
including enzyme replacement therapy “
Ann Saada (Reisch) PhD
Metabolic and Enzyme LaboratoryDepartment of Genetics and Metabolic Diseases
Hadassah Medical Center & Hebrew University Medical School
Jerusalem
1956 1966 1976 1986 1996 2006 20160
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4500
Year
num
ber o
f pub
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ons (
PubM
ed)
Human genome project WES(WGS)
SNP arrays/automated sequencing
mtDNA sequence
mtDNA Mutation
1988Luft’sDisease
1962
nDNA Mutation
1995
Respiratory chain biochemistry
Prevalence ~1:5000 ~1:8000 ~1:10000 ~1:4300
Mitochondrial disease are increasingly common inborn disorders
mtDNA encodedMRC subunits
mtDNA encodedt-RNA r-RNA
mtDNA
deletion/duplication
SECONDARY DEFECTScofactor bio synthesis/transport
lipid biosynthesismitochondrial fission/fusion
detoxificationapoptotic factors
agingneurodegenerative disease
environment
mtDNA-nDNA communicationreplication fork/nucleotide metabolism
nDNA encodedtranslation factors/ribosomal proteins/
t-RNA modifying enzymes
nDNA encoded
assembly factors
nDNA encoded
MRC subunits
mtDNA Depletion
mtDNA multiple deletions
SINGLE DEFECTCI,CII,CIII,CIV,CV
COMBINED DEFECTCI+CIII+CIV+CV
Mitochondrial diseases are caused by many factors
MGM2012
mtDNA encodedMRC subunits
mtDNA encodedt-RNA r-RNA
mtDNA
deletion/duplication
SECONDARY DEFECTScofactor bio synthesis/transport
lipid biosynthesismitochondrial fission/fusion
detoxificationapoptotic factors
agingneurodegenerative disease
environment
mtDNA-nDNA communicationreplication fork/nucleotide metabolism
nDNA encodedtranslation factors/ribosomal proteins/
t-RNA modifying enzymes
nDNA encoded
assembly factors
nDNA encoded
MRC subunits
mtDNA Depletion
mtDNA multiple deletions
SINGLE DEFECTCI,CII,CIII,CIV,CV
COMBINED DEFECTCI+CIII+CIV+CV
Treatment and Clinical trials are limited
Kanabus M, Heales SJ, Rahman S.Br J Pharmacol. 2014 171:1798-817
patient derived cells
microorganisms
animal models
MOUSE
C.elegans
D.melongaster
+ manipulation- phenotype
- maintenance
+mammal+ manipulation+/-phenotype- maintenance
- cost
+ manipulation+ eukaryote
- phenotype - maintenance
E.coli
S.cerevisiae
+ manipulation- prokaryote
- phenotype
MODEL systems For testing treatments
iPSCs
+ tissue specificity- generation/growth
- normal controls- ethics- cost
cybrids
+ mtDNA+/- tissue specificity- generation/growth - nuclear background
LYMHOCYTES/ LYMHOBLASTS
+ Accessible- phenotype
- manipulation
FIBROBLASTS(SKIN)
+ accessible
-limited passages (- )phenotype
+ growth, maintenance
MYOBLASTS
+ phenotype/muscle- sampling /timing
- growth- normal controls
STEMCELLS
+ tissue specificity-sampling
- generation/growth - -genetic instability
- normal controls- ethics- cost
IJCBC 2014
MITOCHONDRIAL DISEASE models
ROS? oxidative stress
ΔΨ(disruption of mitochondrial
membrane potential)
ATP depletionenergy deficit
Many parameters/consequences=what to measure?
Ca2+
(disrupted calcium
homeostasis)
Oxidative damage(lipids, proteins, nucleic acids)
DEFECTIVE OXPHOS
SECONDARY DEFECT
Vicious?
Circle
CELLULAR DYSFUNCTION
AUTOPHAGY/MITOPHAGYCELL DEATH/APOPTOSIS
mtDNA nDNA mutations
Patient’s FIBROBLASTS
Many “therapeutic approaches “
IMPROVED OXPHOS
IMPROVED CELL FUNCTION
Protein replacement therapy?? Mitochondrial therapy???Small molecules?
Patient’s FIBROBLASTS
Heteroplasmic shifting ?
Gene therapy ?Detoxification ?
PGC1α
INCREASE MITOCHONDRIAL
BIOGENESIS
Many “molecules and pathways“
ANTIOXIDANTS
COFACTORS, VITAMINS
IMPROVED OXPHOS
IMPROVED CELL FUNCTIONCa2+
Channelblockers
AUTOPHAGY/MITOPHAGYmodulators
APOPTOSIS inhibitors
PPARγactivators
AMPKactivators
Chemical chaperonesNucleotides etc.
Small molecules?
Patient’s FIBROBLASTS
PGC1α
INCREASE MITOCHONDRIAL
BIOGENESIS ANTIOXIDANTS
COFACTORS, VITAMINS
IMPROVED OXPHOS
IMPROVED CELL FUNCTIONCa2+
Channelblockers
AUTOPHAGY/MITOPHAGYmodulators
APOPTOSIS inhibitors
PPARγactivators
AMPKactivators
Chemical chaperonesNucleotides etc.
ROS? oxidative stress
ΔΨ(disruption of mitochondrial
membrane potential)
ATP depletionenergy deficit
Ca2+
(disrupted calcium
homeostasis)
Oxidative damage(lipids, proteins, nucleic acids) CELLULAR DYSFUNCTION
AUTOPHAGY/MITOPHAGYCELL DEATH/APOPTOSIS
DEFECTIVE OXPHOS
RiboflavinNiacin
ThiaminLipoate
AscorbateCoenzymeQ
Vitamin-EAICAR
bezafibrateResveratrol
genistein
Sodium phenylbutyrateUridine
OltiprazECGCNAC
Devorah SoifermanLiza DouievAnna Golubitzky
Small molecules?
Many “molecules and pathways“& Limited amount of cells/passages
PGC1α
INCREASE MITOCHONDRIAL
BIOGENESIS ANTIOXIDANTS
COFACTORS, VITAMINS
IMPROVED OXPHOS
IMPROVED CELL FUNCTIONCa2+
Channelblockers
AUTOPHAGY/MITOPHAGYmodulators
APOPTOSIS inhibitors
PPARγactivators
AMPKactivators
Chemical chaperonesNucleotides etc.
ROS? oxidative stress
ΔΨ(disruption of mitochondrial
membrane potential)
ATP depletionenergy deficit
Ca2+
(disrupted calcium
homeostasis)
Oxidative damage(lipids, proteins, nucleic acids) CELLULAR DYSFUNCTION
AUTOPHAGY/MITOPHAGYCELL DEATH/APOPTOSIS
DEFECTIVE OXPHOS
RiboflavinNiacin
ThiaminLipoate
AscorbateCoenzymeQ
Vitamin-EAICAR
bezafibrateResveratrol
genistein
Sodium phenylbutyrateUridine
OltiprazECGCNAC
Devorah SoifermanLiza DouievAnna Golubitzky
Small molecules?
Many “molecules and pathways“& Limited amount of cells/passages
x
0 20 40 60 80 100 120 1400.1
0.2
0.3
0.4
0.5
0.6
0.7
MB A620 Cont-GLU
Cont-GAL
NDUFS2-GLU
NDUFS2-GAL
time (hrs)
E
Methylene blue
5000 cells 72 hours
Patient’s fibroblasts that grow normally in high glucose medium show defective growth in glucose free medium (GAL)
Controls (n=5)
CI (NDUFS2)
CI (FOXRED1)
CI (LHON)
CIV (COX 6B1)
Trans. (EFTs)
Trans. (GFM1) Trans. (MRPS22)
Sec. (DNM1L)
0
0.1
0.2
0.3
0.4
0.5
0.6
GLUGAL
E
GROWTH
Most Patient’s fibroblasts show defective growth in GAL medium
nDNACI subunits
mtDNACI subunit
nDNACIV subunits
nDNAtranslation
nDNAsecondary
MB A620
E
Controls (n=5)
CI (NDUFS2)
CI (FOXRED1)
CI (LHON)
CIV (COX 6B1)
Trans. (EFTs)
Trans. (GFM1) Trans. (MRPS22)
Sec. (DNM1L)
0
500
1000
1500
2000
2500
3000
3500
GLUGAL
ROS
GROWTH
Controls (n=5)
CI (NDUFS2)
CI (FOXRED1)
CI (LHON)
CIV (COX 6B1)
Trans. (EFTs)
Trans. (GFM1) Trans. (MRPS22)
Sec. (DNM1L)
0
0.1
0.2
0.3
0.4
0.5
0.6
GLUGAL
MB A620
DCF RFU:MB A620
Most Patient’s fibroblasts show defective growth in GAL mediumMany show increased ROS and decreased ATP production
E
Controls (n=5)
CI (NDUFS2)
CI (FOXRED1)
CI (LHON)
CIV (COX 6B1)
Trans. (EFTs)
Trans. (GFM1) Trans. (MRPS22)
Sec. (DNM1L)
0
500
1000
1500
2000
2500
3000
3500
GLUGAL
ROS
GROWTH
Controls (
n=5)
CI
(NDUFS2
)
CI (FO
XRED1)
CI
(LH
ON)
CIV
(COX 6B1)
Trans.
(E
FTs)
Trans. (
GFM1)
Trans. (
MRPS22)
Sec.
(D
NM1L)0
40000
80000
120000 ATP-content (GAL)
Controls (n=5)
CI (NDUFS2)
CI (FOXRED1)
CI (LHON)
CIV (COX 6B1)
Trans. (EFTs)
Trans. (GFM1) Trans. (MRPS22)
Sec. (DNM1L)
0
0.1
0.2
0.3
0.4
0.5
0.6
GLUGAL
MB A620
DCF RFU:MB A620
ATP RLU:MB A620
Most Patient’s fibroblasts show defective growth in GAL mediumMany show increased ROS and decreased ATP production
E
Screening 10 compounds on 6 complex I deficient cellsAICAR has a positive effect on some cells
control NDUFS20
0.1
0.2
0.3
0.4
0.5
MB A620GLU GAL GAL+AICAR
control NDUFS20
500
1000
1500
2000
2500
3000
DCF RFU:MB A620
GLU GAL GAL+AICAR
control NDUFS20
100000
200000
300000
400000
500000
600000
ATP RLU:MB A620
GLU GAL GAL+AICAR
ROS
PLOSone2011
GROWTH
ATP-content (GAL)
Small moleculesNuclear encoded
CI subunits
E
AICAR enhances mito-biogenesis via AMPK
control NDUFS20
500
1000
1500
2000
2500
3000
DCF RFU:MB A620
GLU GAL GAL+AICAR
control NDUFS20
100000
200000
300000
400000
500000
600000
ATP RLU:MB A620
GLU GAL GAL+AICAR
ROS
ATP-content (GAL)
Small moleculesNuclear encoded
CI subunits
Mitochondrial contentNDUFS2 NDUFS2+AICAR
PLOSone2011
Phospho-AMPK
GROWTH
ATP-content (GAL)
Average Cell Growth
0.000.200.400.600.801.001.201.401.601.80
Ave
fold
GAL
no
addi
tive
Ave ROS Production/Cell
0.000.200.400.600.801.001.201.401.601.80
Ave
fold
GAL
no
addi
tive
Ave ATP Content/Cell
0.000.200.400.600.801.001.201.40
Ave
fold
GAL
no
addi
tive
EJHG 2016
MB A620
DCF RFU:MB A620
ATP RLU:MB A620
ROS
Small moleculesNuclear encoded
COX subunit COX6B1
Ascorbate, AICAR and resveratrol are beneficial to complex IV (COX6B1) deficient cells
ATP-content (GAL)
Ave ROS Production/Cell
0.000.200.400.600.801.001.201.401.601.80
Ave
fold
GAL
no
addi
tive
Ave ATP Content/Cell
0.000.200.400.600.801.001.201.40
Ave
fold
GAL
no
addi
tive
EJHG 2016
DCF RFU:MB A620
ATP RLU:MB A620
ROS
Small moleculesNuclear encoded
COX subunit COX6B1
Ascorbate, decrease ROS, increases ATP, mito-content and oxygen consumption
Ave Mitochondrial Content
0.000.200.400.600.801.001.201.401.60
GAL
AICAR 0.5mM
Ascorb
ate 10u
M
Bezafib
rate
0.1mM
NAC 2mM
Oltipraz
20uM
RSV 25uM
Ave
fold
GAL
no
addi
tive
Mitochondrial content(mitotracker green)
No a
dditi
ve
No a
dditi
ve
+ AI
CAR
0.5m
M
+ As
corb
ate
10uM
+ Re
sver
a-tr
ol
0.02
5mM
Control COX6B1
0.0000
0.0050
0.0100
0.0150
0.0200
0.0250 Maximal OCR
pMO
2/ce
ll/m
in
Oxygen consumption
ATP-content (GAL)
Ave ROS Production/Cell
0.000.200.400.600.801.001.201.401.601.80
Ave
fold
GAL
no
addi
tive
Ave ATP Content/Cell
0.000.200.400.600.801.001.201.40
Ave
fold
GAL
no
addi
tive
EJHG 2016
DCF RFU:MB A620
ATP RLU:MB A620
ROS
Small moleculesNuclear encoded
COX subunit COX6B1
BUT !! N-acetyl cysteine (NAC) reduces ROSNOT ONLYbut also ATP and mitochondrial content
Ave Mitochondrial Content
0.000.200.400.600.801.001.201.401.60
GAL
AICAR 0.5mM
Ascorb
ate 10u
M
Bezafib
rate
0.1mM
NAC 2mM
Oltipraz
20uM
RSV 25uM
Ave
fold
GAL
no
addi
tive
Mitochondrial content(mitotracker green)
Mito-translation disorders show individual results
Biochim2013
Small moleculesMitochondrial translation
Bezafibrate and idebenone are beneficial for Mitochondrial fission in DNM1L mutated cells
AJMG 2016 and in preparation
Small moleculesFission defect
DNM1L FOXRED1 C6ORF66 NDUFS4 NDUFS2 COX6B1 TRMU MRPS22 GFM1 EFTs
+/+/- ns + +/- +/+ +/-/+ ns +/+/ns +/ns +/ns BEZA
ns + +/+ - +/+/+ + +/ns +/ns +/ns ns AICAR
ns + ns +/+ ns +/ns ns ns ns OLTI
-/+/+ + + ns +/- +/-/- +/+ ns ns +/+ SBP
-/+ +/-/+ -/+ +/- +/+/- +/+ +/+ ns +/- +/+ RSV
-/-/+ +/- +/+ +/ns/- +/- +/- NAC
+/+/- +/+ +/ns +/ns ns +/+ Asc
+ -/+ +/+/- +/- GENI
+/- +/- GSE
The effect of ANTIOXIDANTS is variable Decreased ROS combined with decreased, unaffected, increased ATP
Small moleculesSummary
“beneficial” = green “mixed response”=gray“detrimental”= red ns=not significant
DNM1L FOXRED1 C6ORF66 NDUFS4 NDUFS2 COX6B1 TRMU MRPS22 GFM1 EFTs
+/+/ns ns + +/- +/+ +/-/+ ns +/+/ns +/ns +/ns BEZA
ns + +/+ - +/+/+ + +/ns +/ns +/ns ns AICAR
ns + ns +/+ ns +/ns ns ns ns OLTI
-/+/+ + + ns +/- +/-/- +/+ ns ns +/+ SBP
-/+ +/-/+ -/+ +/- +/+/- +/+ +/+ ns +/- +/+ RSV
-/-/+ +/+ +/+ +/ns/- +/- +/- NAC
+/+/- +/+ +/ns +/ns ns +/+ Asc
+ -/+ +/+/- +/- GENI
+/- +/- GSE
“beneficial” = green “mixed response”=gray“detrimental”= red ns=not significant
The effect of MITOBIOGENESIS inducers is variable Is increased biogenesis beneficial or does it “exacerbate stress” ?
Small moleculesSummary
DNM1L FOXRED1 C6ORF66 NDUFS4 NDUFS2 COX6B1 TRMU MRPS22 GFM1 EFTs
+/+/ns ns + +/- +/+ +/-/+ ns +/+/ns +/ns +/ns BEZA
ns + +/+ - +/+/+ +/+/+ +/ns +/ns +/ns ns AICAR
ns + ns +/+ ns +/ns ns ns ns OLTI
-/+/+ + + ns +/- +/+ ns ns +/+ SBP
-/+ +/-/+ -/+ +/- +/+/- +/+ +/+ ns +/- +/+ RSV
-/-/+ +/+ +/+ +/ns/- +/- +/- NAC
+/+/- +/+ +/ns +/ns ns +/+ Asc
+ -/+ +/+/- +/- GENI
+/- +/- GSE
“beneficial” = green “mixed response”=gray“detrimental”= red ns=not significant
The response is individual
Unpublished, EJHG 2015, Biochim2013, PLOSone2011
Small moleculesSummary
DNM1L FOXRED1 C6ORF66 NDUFS4 NDUFS2 COX6B1 TRMU MRPS22 GFM1 EFTs
+/+/ns ns + +/- +/+ +/-/+ ns +/+/ns +/ns +/ns BEZA
ns + +/+ - +/+/+ +/+/+ +/ns +/ns +/ns ns AICAR
ns + ns +/+ ns +/ns ns ns ns OLTI
-/+/+ + + ns +/- +/+ ns ns +/+ SBP
-/+ +/-/+ -/+ +/- +/+/- +/+ +/+ ns +/- +/+ RSV
-/-/+ +/+ +/+ +/ns/- +/- +/- NAC
+/+/- +/+ +/ns +/ns ns +/+ Asc
+ -/+ +/+/- +/- GENI
+/- +/- GSE
“beneficial” = green “mixed response”=gray“detrimental”= red ns=not significant
The response is individual Could results obtained in fibroblasts be relevant for personalized treatment?
Unpublished, EJHG 2015, Biochim2013, PLOSone2011
Small moleculesConclusion?
?
Some other “therapeutic approaches “
IMPROVED OXPHOS
IMPROVED CELL FUNCTION
Protein replacement therapy??
Patient’s FIBROBLASTS
Prof. Haya Lorberboum-GalskiMatan RapoportDana Marcus
Small molecules
Protein replacement therapies are currently used in some diseasesBut there are still obstacles
Protein replacement
COMMON OBSTACLES: delivery by IV infusion
mis-targeting of recombinant enzymeDifficulty reaching target tissue
Immune reactions
Enzyme replacement therapy (ERT)In lysosomal storage diseases (LSD’s):
Gaucher diseasePompe (GSDII) disease
Fabry diseaseMucopolysaccharidoses I,II, IV
Protein replacement
A recombinant mitochondrial protein should: Penetrate not only cellular membrane,
BUT also both the OUTER and INNER mitochondrial membranes AND integrate into a multi-subunit complex
COMMON OBSTACLES: delivery by IV infusion
mis-targeting of recombinant enzymeDifficulty reaching target tissue
Immune reactions
Protein replacement in mitochondrial diseasesis even more complex
Protein replacement
A recombinant mitochondrial protein should: Penetrate not only cellular membrane,
BUT also both the OUTER and INNER mitochondrial membranes AND integrate into a multi-subunit complex
Protein replacement in mitochondrial diseasesis even more complex
PROTEIN TRANSDUCTION DOMAINS PTD’s
Amino acid domains serving asDelivery vectors
Protein replacement
A recombinant mitochondrial protein should: Penetrate not only cellular membrane,
BUT also both the OUTER and INNER mitochondrial membranes AND integrate into a multi-subunit complex
Protein replacement in mitochondrial diseasesis even more complex
MITOCHONDRIAL protein+ mitochondrial target sequence
MTS
Trans Activator of Transcription TAT
11 amino acids from HIV-1(Arginine rich, positively charged)
TAT+MTS+MITOPROTEINFUSIONPROTEIN produced in vitro
Protein replacement
A recombinant mitochondrial protein should: Penetrate not only cellular membrane,
BUT also both the OUTER and INNER mitochondrial membranes AND integrate into a multi-subunit complex
Protein replacement in mitochondrial diseasesis even more complex
Trans Activator of Transcription TAT
11 amino acids from HIV-1(Arginine rich, positively charged)
MITOCHONDRIAL protein+ mitochondrial target sequence
MTS
TAT+MTS+MITOPROTEINFUSIONPROTEIN produced in vitro
Protein replacement-LAD
Lipoamide dehydrogenase (PHDc-E3 aKDH-E3 BCKA-E3)The pure TAT-MTS-LAD recombinant protein is active
TAT+MTS+LADFUSIONPROTEIN
Molec Ther 2008
PDHcE3
Protein replacement-LAD
TAT+MTS+LADFUSIONPROTEIN
(native protein)(control )
Molec Ther 2008
Lipoamide dehydrogenase (PHDc-E3 aKDH-E3 BCKA-E3)The pure TAT-MTS-LAD recombinant protein is active
The TAT-MTS-LAD recombinant protein enters mitochondria is processed and
improves activity in LAD deficient Patient’s fibroblasts
Patient (G229C/Y35X )Fibroblasts
TAT+MTS+LADFUSIONPROTEIN
-processed TAT-LAD and native mutant LAD-TAT-LAD
LAD activity
Protein replacement-LAD
Molec Ther 2008
The TAT-MTS-LAD recombinant protein enters mitochondria co-localized with PDHc
and improves PDHc activity in LAD deficient Patient’s fibroblasts
Molec Ther 2008
TAT+MTS+LADFUSIONPROTEIN
Patient (G229C/Y35X )Fibroblasts
anti-PDHc1α FITC- TAT-LAD Overlay
PDHc activity
Protein replacement-LAD
In theory, intermittent treatment couldSuffice in mild cases/during crisis
BBRC 2000
Muscle/LiverIntermittent“Normal life”
Liver dysfunctionInfantile, fatalLiver failureencephalopathy
ATP-production
Protein replacement-LAD
Protein replacement-CI
TAT+MTS+C6ORF66 (NDUFAF4)FUSIONPROTEIN
The TAT-MTS-NDUFAF4 recombinant protein enters mitochondria and improves CI activity
and mitochondrial functions in Patient’s fibroblasts
GROWTH
ATP-content (GAL)
ROS
FITC- TAT-NDUFAF4
1h 3h
Mol Med 2013
Enzymatic activity
TAT+MTS+LADFUSIONPROTEIN
Next step-proof of concept in animal models
Rapoport et al J Mol Med 2011,
Protein replacement-next step
Another “therapeutic approach “
IMPROVED OXPHOS
Mitochondrial therapy???
Patient’s FIBROBLASTS
Protein replacement-next step
Eyal Kesner
NATURE1982
Mitochondrial therapy?
IMPROVED OXPHOS
Mitochondrial therapy???
Patient’s FIBROBLASTS
Eyal Kesner
NATURE1982
Sci Rep 2016,
Isolated mitochondria from HeLa cells quicklyenter recipient cells
HeLa-dsRed2
Recipient cells
Sci Rep 2016,
Mitochondrial therapy???
NATURE1982
Isolated mitochondria from HeLa cells quicklyenter fibroblasts, maintaining identity & improves growth
GROWTH(GAL)
Enzymatic activity
Mitochondrial therapy???
Sci Rep 2016,
Isolated mitochondria from HeLa cells quicklyenter fibroblasts, maintaining identity & improves growth
Mechanism of mitochondrial import: Intact mitochondrial membrane
MacropinocytosisProteoglycan involvement
Mitochondrial therapy???
Sci Rep 2016,
Biochemical Diagnostics
Corinne BelaicheRosi Shwartz
Sarah weissmanPolina Ilin
Elena fatale
Molecular diagnostics Prof.Orly Elpeleg
Dr.Avraham ShaagProf.Vardiella Meiner
Collaborators
Prof. Chaya Lorberboum-Galski
Dr. Matan RapaportEyal Kesner
Dana Markus
Experimental Devorah Soiferman
Lisa DouievDr Chaya Miller
Anna Golubitzky (alumni)Dr Phyllis Dan (alumni)
Dr. Maskit Bar-Meir (alumni)
Clinical collaboratorsProf.Orly Elpeleg
Dr.Simon EdvardsonDr. Ruth Sheffer
Prof.Ronen SpiegelDr Itai Beger
Protein replacement therapy?? Mitochondrial therapy???Small molecules?
Thank you …………….
COFACTORS, VITAMINS
RiboflavinNiacin
ThiaminLipoate
COFACTORS, VITAMINS
One fibroblast one parameter-riboflavin beneficial
AscorbateCoenzymeQ
Vitamin-E
ANTIOXIDANTS
Bar-Meir J Ped 2001
Small molecules
Mitochondrion 2009
the response is individual depending on the defect
+gentamicin +gentamicin +gentamicin
nDNAtranslation Individual mito translation disorders are sensitive to
translation targeted antibiotics
AICARAminoImidazole Carboxamide Ribonucleotide
Adapted from: Scarpulla RC. 2011 BBA :1813
Resveratrol
GenisteinBezafibrate
Same pathway DIFFERENT EFFECTS Mitochondrial BIOGENESIS
Nicotinamide(Ribo nucleoside)
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