neurodevelopmental disorders focus: rare diseases neurodevelopmental disorders focus: rare diseases

Download Neurodevelopmental Disorders Focus: Rare Diseases Neurodevelopmental Disorders Focus: Rare Diseases

Post on 21-Jun-2020

0 views

Category:

Documents

0 download

Embed Size (px)

TRANSCRIPT

  • Neurodevelopmental Disorders

    Focus: Rare Diseases

    Margot Mayer-Proschel, John Foxe, Jonathan Mink,

    Mark Noble, Alex Paciorkowski, Chris Proschel

  • Alex Paciorkowski

    Chris Proschel

    Jonathan Mink

    Margot

    Mayer-Proschel

    Mark Noble

    FOXG1

    Disorders

    MECP2

    disorders

    Developmental

    Brain

    Disorders

    Fragile-

    X

    Vanishing White Matter

    16p11.2

    Ataxia Telangiectasia

    MLD Gaucher

    Krabbe Batten Disease

    Cystinosis

    NPC

    Rett

    Syndrome

    • Drug discovery through identification of common

    denominators • Broad impact

    John Foxe

  •  Congenital diseases

     Variable disease onset

     Secondary insults

    modulate disease

    Misfolded proteins

    ER stress

    Mitochondrial defects

    Inflammation

    Oxidative stress

    A Common Denominator

    Lysosomal dysfunction Drugs

  •  Congenital diseases

     Variable disease onset

     Secondary insults

    modulate disease

     Genetic predisposition

     Variable disease onset

     Secondary insults

    contribute to disease

    Misfolded proteins

    ER stress

    Mitochondrial defects

    Inflammation

    Oxidative stress

    A Broader Impact

    Lysosomal dysfunction Drugs

  • Cellular

    endpoints

    • Proliferation

    • Differentiation

    • Survival

    • Neurite outgrowth

    Drug screen

    • FDA approved drug screen

    • Confirm candidate “hits”

    • Verification in relevant human

    iPSC cells

    Structural Analysis

    • Identify BBB permeable drug • Test in animal model

    • Test for off target effects

    Clinical application

    • Identification of clinical trial

    cohorts

    • Analysis of carriers

    • Relevant “recovery” readouts

    Genetic

    • Disease phenotype

    • Clinical history

    • Identification of new mutations

    • Cohort access

    General Concept

  • Oligodendrocyte

    Progenitors

    • Proliferation

    • Differentiation

    • Lysosomal

    Function

    Drug Screen

    • Identify candidates

    • Confirm cellular rescue

    • Effect on lysosome

    • Test outcome in iPSCs

    • Identify harmful drugs.

    Structural Analysis

    • Identified BBB permeable drugs

    • Test in animal model (Twi)

    • No reported CNS toxicity

    Clinical application

    • Identification of clinical trial

    cohorts

    • Clinical history of patients

    receiving the “bad drug”

    • Analysis of carriers?

    Krabbe Disease

    • Lysosomal storage disease

    • Known mutation in GALC

    • Heterozygosity associated

    with mild phenotype

    A Specific Example

  • P10 P40P25P20P15 P35P30

    symptom onset

    twitching weight loss /

    muscle wasting death

    twi/twi P5P0

    Brain Psy detected

    Postnatal age (days)

    Krabbe Disease:

    • Severe, progressive loss of myelin & neurodegeneration • Enzymatic deficiencies (mutations) in galactocerebrosidase (GALC) • Accumulation of the toxic lipid Psychosine (Psy) • Twitcher mouse: a pathologically/genetically authentic murine model

    W T

    tw i

    DAPI fluoromyelin

    P10 P40P25P20P15 P35P30

    symptom onset

    twitching weight loss /

    muscle wasting death

    twi/twi P5P0

    Brain Psy detected

    Postnatal age (days)

    Krabbe Disease:

    • Severe, progressive loss of myelin & neurodegeneration • Enzymatic deficiencies (mutations) in galactocerebrosidase (GALC) • Accumulation of the toxic lipid Psychosine (Psy) • Twitcher mouse: a pathologically/genetically authentic murine model

    W T

    tw i

    DAPI fluoromyelin

    *

    WT twi

    P40 OLs

    **

    P40 OPCs

    WT twi

    *

    WT twi

    P15 OPCs

    *

    WT twi

    P40 OLs

    **

    P40 OPCs

    WT twi

  • Oligodendrocyte

    Progenitors

    • Proliferation

    • Differentiation

    (Lysosomal

    Function)

    Drug Screen

    • Identify candidates

    • Confirm cellular rescue

    • Effect on lysosome

    • Test outcome in iPSCs

    • Identify harmful drugs.

    Structural Analysis/In vivo

    • Identified BBB permeable drugs

    • Test in animal model (Twi)

    • No reported CNS toxicity

    Clinical application

    • Identification of clinical trial

    cohorts

    • Clinical history of patients

    receiving the “bad drug”

    • Analysis of carriers?

    Krabbe Disease

    • Lysosomal storage disease

    • Known mutation in GALC

    • Heterozygosity associated

    with mild phenotype

    A Specific Example….

  • …and also disrupts endocytosis, lipid metabolism, and the

    activity of resident enzymes.

    PND 17

    WT twi

    OPC pH

    *

    Isolated progenitor cells recapitulate in vivo defects

    Proliferation Lysosomal pH

  • Oligodendrocyte

    Progenitors

    • Proliferation

    • Differentiation

    (Lysosomal

    Function)

    Drug Screen

    • Identify candidates

    • Confirm cellular rescue

    • Effect on lysosome

    • Test outcome in iPSCs

    • Identify harmful drugs.

    Structural Analysis/In vivo

    • Identified BBB permeable drugs

    • Test in animal model (Twi)

    • No reported CNS toxicity

    Clinical application

    • Identification of clinical trial

    cohorts

    • Clinical history of patients

    receiving the “bad drug”

    • Analysis of carriers?

    Krabbe Disease

    • Lysosomal storage disease

    • Known mutation in GALC

    • Heterozygosity associated

    with mild phenotype

    A Specific Example….

  • Focus on correcting important cellular behaviors, not on specific proteins or genes

    Unbiased screening

    Psy 1052

    Drugs

    Cell division

    Validation of

    ‘hits’

    Restoration

    of lysosomal

    function?

    calcein-AM (live) PI (dead)

    + vehicle (DMSO) + psychosine 0h 24h 48h 72h

    342 632 1630 3192

    Elapsed time:

    Cell count:

    + psy

    1 1 0 4

    ++++

    1 3 0 3

    1 3 0 5

    1 1 0 7

    c c c

    b

    +++++++

    8 0 8

    1 0 0 2

    7 0 4

    6 1 1

    9 0 7

    9 0 6

    ++++++ 2 0 8

    2 1 1

    1 0 5

    2 1 0

    2 1 1

    c c c c

    c c c

    a a b

    a ba

    +

    6 1 0

    + veh

    + Psy

    Reduce Psy toxicity

    2400 drugs @ 3 concentrations

    : + Psy

    Enhance Psy toxicity?

    psy

    N K

    H c lo

    f IG

    F -1

    +++ +

    **

    a b a

    CathD

    activity

    b a a

    + psy

    N K

    H

    +

    IG F -1

    +

    c lo

    f

    +

    Endocytosis Lipid accumulation

    c lo

    f N

    K H

    IG F -1

    + + + + psy

    b

    psy

    9 1 0

    +

    9 0 6

    +

    5 0 5

    +

    1 0 4

    +

    N T -3

    +

    IG F -1

    +

    N K

    H

    +

    c lo

    f

    ++

    ** ** * * * *

    a a a

    Lysosomal pH

    Top hits rescue cellular defect associated with lysosomal

    dysfunction in vitro

  • Oligodendrocyte

    Progenitors

    • Proliferation

    • Differentiation

    (Lysosomal

    Function)

    Drug Screen

    • Identify candidates

    • Confirm cellular rescue

    • Effect on lysosome

    • Test outcome in iPSCs

    • Identify harmful drugs.

    Structural Analysis/In vivo

    • Identified BBB permeable drugs

    • Test in animal model (Twi)

    • No reported CNS toxicity

    Clinical application

    • Identification of clinical trial

    cohorts

    • Clinical history of patients

    receiving the “bad drug”

    • Analysis of carriers?

    Krabbe Disease

    • Lysosomal storage disease

    • Known mutation in GALC

    • Heterozygosity associated

    with mild phenotype

    A Specific Example….

  • Top hits rescue cellular defect associated with

    lysosomal dysfunction in vivo

    (A) Stance time (B) Break time (C) Propel time

    c

    † †

    a

    c

    WT twi WT twi WT twi

    Weight gain

    *!

    BMT

    AAV

    v e h

    N K

    H

    twi

    Survival

    *

    N K

    H

    v e

    h

    WT

    v e

    h

    twi

    W T

    v e

    h T

    w i

    v e

    h T

    w i

    N K

    H

    DAPI fluoromyelin

    *

    N K

    H

    v e

    h

    WT

    v e

    h

    twi

    (A) PND40 My

Recommended

View more >