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Transcript of Next generation sequencing technologies (N GS) for whole ... · PDF file Next generation...

  • Next generation sequencing technologies (NGS) for whole exome/genome sequencing (WES/WGS):

    impact on daily clinical practice, in dementia and beyond

    9ο Πανελλήνιο Διεπιστημονικό Συνέδριο Νόσου Alzheimer Θεσσαλονίκη 14-17/5/2015

    Ζαγανάς Ιωάννης Νευρολόγος, Επίκουρος Καθηγητής Νευρολογίας

    Βογιατζή Εμμανουέλλα, Βιολόγος-Γενετίστρια

    Λατσούδη Λένα, Βιολόγος-Γενετίστρια

  • Introduction: Clinical whole exome sequencing as a tool to end the “diagnostic Odyssey”

    Ζαγανάς Ιωάννης Νευρολόγος Επίκουρος Καθηγητής Νευρολογίας, Τμήμα Ιατρικής Πανεπιστημίου Κρήτης / Πανεπιστημιακό Γενικό Νοσοκομείο Ηρακλείου

    9ο Πανελλήνιο Διεπιστημονικό Συνέδριο Νόσου Alzheimer Θεσσαλονίκη 14-17/5/2015

  • Lu et al, N Engl J Med, 2014

  • Η δική μας εμπειρία

    Πρόγραμμα «Γενετικές αναλύσεις νέας γενιάς σε ασθενείς

    με νευρολογικές και άλλες παθήσεις»

  • 18-year old woman with limb-girdle muscular dystrophy

    • Her sister similarly affected, but not her parents

    • Central-type muscle weakness, severely depressed tendon reflexes, calf hypertrophy, scapular winging, waddling gait

    • Subject to multiple tests, including muscle biopsy, with no diagnostic results obtained

  • Sarcolemma and proteins involved in muscular dystrophies (Mercuri and Muntoni, 2013)

  • ISPD: isoprenoid synthase domain containing

  • Sarcolemma and proteins involved in muscular dystrophies (Mercuri and Muntoni, 2013)

    ISPD

    Diagnosed by us (UOC)

  • Sarcolemma and proteins involved in muscular dystrophies (Mercuri and Muntoni, 2013)

    ISPD

    Diagnosed by us (UOC)

  • 2-year old girl with recurrent lactic acidosis, epilepsy

    • Hypotonia • Seizures • Mental retardation • Microcephaly

  • Diagnosed by us (UOC)

    2 y.o. f., lactic acidosis, epilepsy

  • 2 y.o. f., lactic acidosis, epilepsy

    13 y.o. f., myalgias, ↑ CPK

    6 y.o. f., myopathy

    Diagnosed by us (UOC)

  • 62-year old man with progressively worsening muscle weakness (A)

    • About 15 years ago, weakness and muscular atrophy in the periphery of his extremities was noted (progressively worsening since then)

    • Deep tendon reflexes abolished in all 4 extremities

    • loss of sensory function in a stocking-glove distribution (pain, temperature, vibration)

    • FH: Brother died at a young age from an, further un-specified, neurological disorder that led to quadriplegia

  • 62-year old man with progressively worsening muscle weakness (B)

    • Electrophysiology: sensorimotor demyelinating-type polyneuropathy (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

  • Rossor et al, Nat Rev Neurol,

    2013

  • Rossor et al, Nat Rev Neurol, 2013

    Diagnosed by us (UOC)

  • Jean-Martin Charcot (1825 – 1893)

    Pierre Marie (1853-1940)

    Howard Henry Tooth (1856–1925)

  • The application of whole exome sequencing in dementia and beyond: the clinician’s viewpoint

    Ζαγανάς Ιωάννης Νευρολόγος Επίκουρος Καθηγητής Νευρολογίας, Τμήμα Ιατρικής Πανεπιστημίου Κρήτης / Πανεπιστημιακό Γενικό Νοσοκομείο Ηρακλείου

    9ο Πανελλήνιο Διεπιστημονικό Συνέδριο Νόσου Alzheimer Θεσσαλονίκη 14-17/5/2015

  • Γενετικοί παράγοντες συνδεόμενοι με βεβαιότητα με τη νόσο Alzheimer

    Χρωμό- σωμα

    Αριθμός μεταλλάξεων (οικογένειες)

    Ηλικία έναρξης

    (έτη)

    % περιπτώσεων πρώϊμης έναρξης

    % συνολικών

    περιπτώσεων

    APP 21 33 (90) 45-65

  • Γονίδια σχετιζόμενα με FTLD ( AD and FTD Mutation Database, Nov 2014)

    Μεταλλαγμένο Γονίδιο Φαινότυπος Locus Μεταλλάξεις(οικογένειες)

    Expansion of a non-coding hexanucleotide repeat (C90RF72) FTLD/ALS 9p21 1 (336)

    Progranulin (PGRN) FTLD 17q21-q22 69 (231)

    Microtubule-associated protein tau (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 body protein 2B (CHMP 2B) FTLD 3p13-3p12 4 (5)

  • Genetic testing for familial dementia with additional neurological features (Loy et al, 2014)

  • 18-year old woman with progressive myoclonic 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- Lundborg disease

    EMP1 (CSTB) 21q22 Cystatin (protease inhibitor)

    Lafora disease EMP2A EMP2B/NHLRC1

    6q24 6p22

    Laforin Malin

    Neuronal ceroid lipofuscinosis (CLN1- 8, CLN10)

    PPT1 (CLN-1) ……

    1p32 Palmitoyl-protein Thioesterase- 1

    MERRF syndrome A8344G Mitochondrial DNA Lys tRNA

    Sialidosis Type Ι NEU1 6p21.33 Neuraminidase (glycoprotein metabolism)

    DRPLA ATN1 12p13.31 Atrophin 1 (nuclear protein)

    Progressive myoclonic epilepsy and dementia

  • Τύπος PME Γονίδιο Locus Πρωτείνη

    Unverricht- Lundborg disease

    EMP 1 (CSTB) 21q22 Cystatin (protease inhibitor)

    Lafora disease EMP2A EMP2B/NHLRC1

    6q24 6p22

    Laforin Malin

    Neuronal ceroid lipofuscinosis (CLN1- 8, CLN10)

    PPT1 (CLN-1) ……

    1p32 Palmitoyl-protein Thioesterase- 1

    MERRF syndrome A8344G Mitochondrial DNA Lys tRNA

    Sialidosis Type Ι NEU 1- 6p21.33 Neuraminidase (glycoprotein metabolism)

    DRPLA ATN 1 12p13.31 Atrophin 1 (nuclear protein)

  • ΕΠ: Μ. Τζαρδή

  • Τύπος PME Γονίδιο Locus Πρωτείνη

    Unverricht- Lundborg disease

    EMP 1 (CSTB) 21q22 Cystatin (protease inhibitor)

    Lafora disease EMP2A EMP2B/NHLRC1

    6q24 6p22

    Laforin Malin

    Neuronal ceroid lipofuscinosis (CLN1- 8, CLN10)

    PPT1 (CLN-1) ……

    1p32 Palmitoyl-protein Thioesterase- 1

    MERRF syndrome A8344G Mitochondrial DNA Lys tRNA

    Sialidosis Type Ι NEU 1- 6p21.33 Neuraminidase (glycoprotein metabolism)

    DRPLA ATN 1 12p13.31 Atrophin 1 (nuclear protein)

  • Τύπος PME Γονίδιο Locus Πρωτείνη

    Unverricht- Lundborg disease

    EMP 1 (CSTB) 21q22 Cystatin (protease inhibitor)

    Lafora disease EMP2A EMP2B/NHLRC1

    6q24 6p22

    Laforin Malin

    Neuronal ceroid lipofuscinosis (CLN1- 8, CLN10)

    PPT1 (CLN-1) ……

    1p32 Palmitoyl-protein Thioesterase- 1

    MERRF syndrome A8344G Mitochondrial DNA Lys tRNA

    Sialidosis Type Ι NEU 1- 6p21.33 Neuraminidase (glycoprotein metabolism)

    DRPLA ATN 1 12p13.31 Atrophin 1 (nuclear protein)

  • 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, flat affect, inability to perform his work, loss of interests

    – On 2011 weakness of the left upper extremity, gradually worsening and extending centripetally and then to the other extremities and bulbar muscles, fibrillations, atrophies (on examination increased tendon reflexes)

    – FH: Negative for neurological disorders

  • 65 year old man with muscular weakness, fibrillations and behavioral disorder

  • Turner et al, 2013

  • Γονίδια σχετιζόμενα με FTLD ( AD and FTD Mutation Database, Nov 2014)

    Μεταλλαγμένο Γονίδιο Φαινότυπος Locus Μεταλλάξεις(οικογένειες)

    Expansion of a non-coding hexanucleotide repeat (C90RF72) FTLD/ALS 9p21 1 (336)

    Progranulin (PGRN) FTLD 17q21-q22 69 (231)

    Microtubule-associated protein tau (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) IBM