next generation sequencing technologies (n gs) for whole...
TRANSCRIPT
Next generation sequencing technologies (NGS) forwhole exome/genome sequencing (WES/WGS):
impact on daily clinical practice, in dementia and beyond
9ο Πανελλήνιο Διεπιστημονικό Συνέδριο Νόσου AlzheimerΘεσσαλονίκη 14-17/5/2015
Ζαγανάς ΙωάννηςΝευρολόγος,Επίκουρος ΚαθηγητήςΝευρολογίας
Βογιατζή Εμμανουέλλα,Βιολόγος-Γενετίστρια
Λατσούδη Λένα,Βιολόγος-Γενετίστρια
Introduction: Clinical whole exome sequencing as a tool to end the“diagnostic Odyssey”
Ζαγανάς ΙωάννηςΝευρολόγοςΕπίκουρος Καθηγητής Νευρολογίας,Τμήμα Ιατρικής Πανεπιστημίου Κρήτης /Πανεπιστημιακό Γενικό Νοσοκομείο Ηρακλείου
9ο Πανελλήνιο Διεπιστημονικό Συνέδριο Νόσου AlzheimerΘεσσαλονίκη 14-17/5/2015
Η δική μας εμπειρία
Πρόγραμμα«Γενετικές αναλύσεις νέας γενιάς σε ασθενείς
με νευρολογικές και άλλες παθήσεις»
18-year old woman with limb-girdlemuscular dystrophy
• Her sister similarly affected, but not herparents
• Central-type muscle weakness, severelydepressed tendon reflexes, calf hypertrophy,scapular winging, waddling gait
• Subject to multiple tests, including musclebiopsy, with no diagnostic results obtained
Sarcolemma and proteins involved in muscular dystrophies (Mercuri and Muntoni, 2013)
ISPD
Diagnosedby us(UOC)
Sarcolemma and proteins involved in muscular dystrophies (Mercuri and Muntoni, 2013)
ISPD
Diagnosedby us(UOC)
2-year old girl with recurrent lacticacidosis, epilepsy
• Hypotonia• Seizures• Mental retardation• Microcephaly
62-year old man with progressivelyworsening muscle weakness (A)
• About 15 years ago, weakness and muscularatrophy in the periphery of his extremities wasnoted (progressively worsening since then)
• Deep tendon reflexes abolished in all 4extremities
• loss of sensory function in a stocking-glovedistribution (pain, temperature, vibration)
• FH: Brother died at a young age from an, furtherun-specified, neurological disorder that led toquadriplegia
62-year old man with progressivelyworsening muscle weakness (B)
• Electrophysiology: sensorimotor demyelinating-typepolyneuropathy (later on with a patchy distribution)
• LP: moderately increased protein• Testing for PMP22 duplication: negative• Patient subjected to several tests (at various
centers) with no diagnostic results• Treated with immunotherapy with no improvement
The application of whole exome sequencing in dementia and beyond:the clinician’s viewpoint
Ζαγανάς ΙωάννηςΝευρολόγοςΕπίκουρος Καθηγητής Νευρολογίας,Τμήμα Ιατρικής Πανεπιστημίου Κρήτης /Πανεπιστημιακό Γενικό Νοσοκομείο Ηρακλείου
9ο Πανελλήνιο Διεπιστημονικό Συνέδριο Νόσου AlzheimerΘεσσαλονίκη 14-17/5/2015
Γενετικοί παράγοντες συνδεόμενοι με βεβαιότητα με τη νόσο Alzheimer
Χρωμό-σωμα
Αριθμόςμεταλλάξεων(οικογένειες)
Ηλικίαέναρξης
(έτη)
% περιπτώσεωνπρώϊμηςέναρξης
%συνολικών
περιπτώσεων
APP 21 33 (90) 45-65 <1 <0.1
PS1 14 185 (405) 30-60 50 1-2
PS2 1 13 (22) 40-80 <1 <0.1
Source: AD and FTD Mutation Database (Nov 2014)
Γονίδια σχετιζόμενα με FTLD ( AD and FTD Mutation Database, Nov 2014)
Μεταλλαγμένο Γονίδιο Φαινότυπος Locus Μεταλλάξεις(οικογένειες)
Expansion of a non-codinghexanucleotide repeat (C90RF72) FTLD/ALS 9p21 1 (336)
Progranulin(PGRN) FTLD 17q21-q22 69 (231)
Microtubule-associated proteintau (MAPT)
FTLD (FTDP-17) 17q21-q22 44 (134)
TDP43 protein (TADPR) ALS/FTLD 1p36 34 (131)
Fused in sarcoma (FUS) ALS/FTLD 16p11.2 23 (49)
Valosin-containing protein gene(VCP) IBMPFD 9p21-p12 18 (48)
Charged multivesicular bodyprotein 2B (CHMP 2B) FTLD 3p13-3p12 4 (5)
18-year old woman with progressivemyoclonic epilepsy and dementia
– At the age of 14 y.o. visual phenomena– Since 2-3 years myoclonus and possibly absence
seizures– Gradual decline in cognitive functions– FH: negative for neurologic disease– MRI: Normal– Initial response to levetiracetam, later evolution to
generalized seizures despite addition of valproate
PME Type Gene Locus Protein
Unverricht- Lundborgdisease
EMP1 (CSTB) 21q22 Cystatin (proteaseinhibitor)
Lafora disease EMP2AEMP2B/NHLRC1
6q246p22
LaforinMalin
Neuronal ceroidlipofuscinosis (CLN1-8, CLN10)
PPT1 (CLN-1)……
1p32 Palmitoyl-proteinThioesterase- 1
MERRF syndrome A8344G Mitochondrial DNA Lys tRNA
Sialidosis Type Ι NEU1 6p21.33 Neuraminidase(glycoproteinmetabolism)
DRPLA ATN1 12p13.31 Atrophin 1 (nuclearprotein)
Progressive myoclonic epilepsy and dementia
Τύπος PME Γονίδιο Locus Πρωτείνη
Unverricht- Lundborgdisease
EMP 1 (CSTB) 21q22 Cystatin (proteaseinhibitor)
Lafora disease EMP2AEMP2B/NHLRC1
6q246p22
LaforinMalin
Neuronal ceroidlipofuscinosis (CLN1-8, CLN10)
PPT1 (CLN-1)……
1p32 Palmitoyl-proteinThioesterase- 1
MERRF syndrome A8344G Mitochondrial DNA Lys tRNA
Sialidosis Type Ι NEU 1- 6p21.33 Neuraminidase(glycoproteinmetabolism)
DRPLA ATN 1 12p13.31 Atrophin 1 (nuclearprotein)
Τύπος PME Γονίδιο Locus Πρωτείνη
Unverricht- Lundborgdisease
EMP 1 (CSTB) 21q22 Cystatin (proteaseinhibitor)
Lafora disease EMP2AEMP2B/NHLRC1
6q246p22
LaforinMalin
Neuronal ceroidlipofuscinosis (CLN1-8, CLN10)
PPT1 (CLN-1)……
1p32 Palmitoyl-proteinThioesterase- 1
MERRF syndrome A8344G Mitochondrial DNA Lys tRNA
Sialidosis Type Ι NEU 1- 6p21.33 Neuraminidase(glycoproteinmetabolism)
DRPLA ATN 1 12p13.31 Atrophin 1 (nuclearprotein)
Τύπος PME Γονίδιο Locus Πρωτείνη
Unverricht- Lundborgdisease
EMP 1 (CSTB) 21q22 Cystatin (proteaseinhibitor)
Lafora disease EMP2AEMP2B/NHLRC1
6q246p22
LaforinMalin
Neuronal ceroidlipofuscinosis (CLN1-8, CLN10)
PPT1 (CLN-1)……
1p32 Palmitoyl-proteinThioesterase- 1
MERRF syndrome A8344G Mitochondrial DNA Lys tRNA
Sialidosis Type Ι NEU 1- 6p21.33 Neuraminidase(glycoproteinmetabolism)
DRPLA ATN 1 12p13.31 Atrophin 1 (nuclearprotein)
65 year old man with muscular weakness,fibrillations and behavioral disorder
– Since 2010, according to his wife, language problems(“poor speech”), inability with rational associations,change in behavior, recklessness, poor grooming, flataffect, inability to perform his work, loss of interests
– On 2011 weakness of the left upper extremity, graduallyworsening and extending centripetally and then to theother extremities and bulbar muscles, fibrillations,atrophies (on examination increased tendon reflexes)
– FH: Negative for neurological disorders
Γονίδια σχετιζόμενα με FTLD ( AD and FTD Mutation Database, Nov 2014)
Μεταλλαγμένο Γονίδιο Φαινότυπος Locus Μεταλλάξεις(οικογένειες)
Expansion of a non-codinghexanucleotide repeat (C90RF72) FTLD/ALS 9p21 1 (336)
Progranulin(PGRN) FTLD 17q21-q22 69 (231)
Microtubule-associated proteintau (MAPT)
FTLD (FTDP-17) 17q21-q22 44 (134)
TDP43 protein (TADPR) ALS/FTLD 1p36 34 (131)
Fused in sarcoma (FUS) ALS/FTLD 16p11.2 23 (49)
Valosin-containing protein gene(VCP) IBMPFD 9p21-p12 18 (48)
Charged multivesicular bodyprotein 2B (CHMP 2B) FTLD 3p13-3p12 4 (5)
• Whole exome sequencing
Pure FTD
MAPT
Progranulin
The middlegroundC9ORF72
FUS
VCP
TDP- 43
Pure ALS
SOD 1
65 year old man with muscular weakness,fibrillations and behavioral disorder
• Whole exome sequencing
Pure FTD
MAPT
Progranulin
The middlegroundC9ORF72
FUS
VCP
TDP- 43
Pure ALS
SOD 1
65 year old man with muscular weakness,fibrillations and behavioral disorder
• Whole exome sequencing
Pure FTD
MAPT
Progranulin
The middlegroundC9ORF72
FUS
VCP
TDP- 43
Pure ALS
SOD 1
65 year old man with muscular weakness,fibrillations and behavioral disorder
• Whole exome sequencing
Pure FTD
MAPT
Progranulin
The middlegroundC9ORF72
FUS
VCP
TDP- 43
Pure ALS
SOD 1
65 year old man with muscular weakness,fibrillations and behavioral disorder
- Clinical application of Whole Exome / Genome Sequencing for rare oryet undiagnosed diseases has dramatically increased our diagnosticsuccess rates
- A final diagnosis, even without changing their therapeuticmanagement, gives significant information and comfort to the patientand his family by terminating a long stressful diagnostic process(often involving several different physicians)
- Financially beneficial, as a time-consuming process (diagnosticOdyssey), involving several tests (MRIs, biopsies, blood tests etc) isavoided for a relative minimal cost
Conclusions
"To learn how to treat a disease, one must learn howto recognize it. The diagnosis is the best trump in thescheme of treatment.”
“Για να μάθει κανείς πως να θεραπεύσει μία νόσο,πρέπει πρώτα να μάθεις πως να την αναγνωρίζεις. Ηδιάγνωση είναι το καλύτερο ατού στο σχήμα τηςθεραπείας.»
Jean-Martin Charcot (1825 – 1893)
Ευχαριστίες (Πρόγραμμα: «Γενετικές αναλύσεις νέας γενιάςσε ασθενείς με νευρολογικές και άλλες παθήσεις»)
• Εμμανουέλλα Βογιατζή• Λένα Λατσούδη• Νευρολογική Κλινική ΠΑΓΝΗ (Γ. Αμοιρίδης, Α. Πλαϊτάκης)• Κλινικοί που μας εμπιστεύτηκαν τα δείγματα των ασθενών
τους (Α. Ευαγγελίου, Β. Μαστοροδήμος, Δ. Κοτζαμάνη, Μ.Τζαγκουρνισάκης, Μ. Μαυρίδης, Π. Βοργιά, Δ. Ζαφειρίου, Α.Μόντη, Γ. Μπριασούλης, κτλ.)