The ALS Association Research Where are the gaps and Opportunities

Presentation Overview

• Brief overview of ALS

• Opportunities and Challenges for Therapy Development

Aging

Genes Environment

? APOE

SOD1

GenderRis

k

Low

Hig

h ? retrovirus

? pesticides

smoking

high glut diet

Ris

k

Age

Youth

Ris

k

Gene-Environment Interactions and Aging

5-10% familial

G37R G93C G93CG37R

G85R

H48Q H48Q

H46R

Zn++

Cu++A4V

I113T

Zn++

> 100

different

mutations

known

Mutant- Copper/Zinc SOD1 mediated Toxicity (include genetics summary slide)

The ALS Association Research Department

Model Systems for ALS drug discovery

Ilieva et al, J. Cell Biol., 2009 6

Jeff Lichtman

Involvement of astrocytes, microglia and muscle

Familial ALS

C9orf72

SOD1

FUS TDP43

0

5

10

15

20

25

30

19931994

19951996

19971998

19992000

20012002

20032004

20052006

20072008

20092010

20112012

2013

SOD1

senataxinVAPBalsin

dynactin

TDP43NTE

ANG

OPTND-AAVCPAtxn2

FUSPON1-3KIFAP3ELP3CHGBUNC13A

C9orf72TAF15ubqIn2p62sigR-1EphA4

hnRNPA2B1 hnRNPA1

Pfn1

Each gene defines pathways and treatment targets.

The rate of gene discovery in ALS is increasing:>25 ALS genes

# o

f G

en

es

Research Program Strategies

• Establishment of a translational program TREAT ALS™

• Fund academic-industry partnerships

• Provide infra-structure for multi-center clinical trials

• Consortium initiatives and Global research

TARGET

IDENTIFICATION

EARLY

DISCOVERY

VALIDATED

TARGETS

THERAPEUTIC

DEVELOPMENT

PRE-CLINICAL

STUDIES

CLINICAL

TRIALS

TREAT ALS PROGRAM

GLOBAL BIOMEDICAL RESEARCH

CLINICAL

MANAGEMENT

TOOLS

MODEL

SYSTEMSBIOMARKERS

Laboratory bedside

SOD1 antisense trial-UCSD/Isis Pharmaceuticals

Industry and Biotech focused on ALS

• Biogen Idec Synapse

• Bristol Myers Squibb Avanir

• Pfizer Amorfix

• Genzyme/Sanofi Aventis ALS Biopharma

• Genentech/Roche Santhera

• Cytokinetics UBC

• Neuraltus Biofocus

• Neuralstem Osainix

Public-Private Partnerships

Fishburn 2012

NGO’sThe ALSAssociation

5’’

Endogenous RNase H mediated degradation of the mRNA in a DNA/RNA duplex

G37R G93C G93CG37R

G85R

H48Q H48Q

H46R

Zn++

Cu++A4V

I113T

Zn++

ALSA-initiated Study 2003

Antisense Approach to Lower SOD1

Cleveland, Miller, Smith

Collaboration with ISIS

• Intrathecal drug administration – Lumbar Puncture bolus injection

• ASOs have long half-lives (several months) in CNS tissue, with even longer duration of action, so enables infrequent dosing

• For more frequent dosing, there are implantable devices that can also be used

Intrathecal ASO Drug Delivery to the CNS

IHC against drug in monkey spinal cord following intrathecal delivery of ASOneedle

15 Courtesy of Frank Bennett, ISIS

Repeat Primed PCR

Expanded

Southern:

Up to 1500 repeats

C9orf72 gene

Normal

(GGGGCC)n

C9orf72 gene

Modified from DeJesus-Hernandez Neuron (2011)

Intrathecal injection of AAV9 and AAVrh10 leads to transgene

expression in the entire spinal cord and brainstem

Intraventricular injection of AAV9 and AAVrh10 leads to

widespread transduction in the forebrain including the motor cortex

TREAT™ ALS/NEALS Clinical Network

…the ALS pipeline has many treatments in development for ALS

• Phase II

– Tirasemtiv (CK-2017357)– Immunosuppression (4) – Gilenya– Mexiletine– Nuedexta– Ursodiol– NurOwn™ – Neuralstem– NP001– GSK1223249– High fat nutrition– Exercise– Tamoxifen/Creatine– Diaphragm pacing– GM604

• FALS SOD1– ISIS333611– Arimoclomol

• FALS/SALS – C90rf72

www.clinicaltrials.gov

www.alsconsortium.org

ALS diagnostic: Ratio of pNFH to complement C3 Levels

A cut-off value was obtained that was 90% accurate for ALS in 106 subjects in a retrospective study.

J Neurochem 117: 528-537 (2011)

Cervical Cord Magnetic Resonance Spectroscopy in ALS

Michael Benatar

Charlie Stagg

Charlie Stagg

EAAT2 Rat PET Imaging: WT vs. ALS Model

Frontal Cortex (FrCTx)

Caudate/Putamen (CP)

Lumbar spine

coronal

Cerebellum (CE)

Motor nuclei (MNuc)

FrCTx

Thalamus (TH)

Fr & Cing

CTx

TH

Decreasing spine

signal In ALS

rat model

0.25SUV

5.0SUV

Sprague-Dawley rat

0.50 mCi of tracer

15 min post injection

sagittal

Identification of dysregulated microRNA signature in blood monocytes and spinal cord from ALS subjects

Blood monocytes Spinal cord

J Clin Invest. 2012

Spinal cordBlood monocytes

Generation of Induced Pluripotent Stem

(iPS) Cell Lines From Human Skin Cells

Human Skin Biopsy

yes

Dr. Shinya Yamanaka’s

Technology (2012 Nobel Prize)

Stem CellsSkin Cells

Motor Neurons

Use of Patient-Specific iPS Cells to Investigate Pathogenic Mechanisms of GGGGCC Repeats in C9ORF72

-- GGGGCC repeat expansion is the most common genetic mutation in ALS

-- We have generated multiple iPS cell lines from 3 patients with GGGGCC repeats

-- We found RNA foci in iPSC-derived cortical neurons and motor neurons

-- We are investigating how to reduce repeat toxicity in human motor neurons

Control Patient

Fen-Biao Gao

Automated Microscopy: A Powerful Tool for Unraveling Cause and

Effect Relationships and Doing Clinical Trials in a Dish

Steve Finkbeiner

102-103 improved sensitivity Phenotypic screens Z’ > 0.8

Provided by Gendron T. and Petrucelli L.Unpublished observation

Detection of RAN products in iPS cell culture supernatants

RAN products may be released into supernatants and could therefore be used a alternative biomarker readout

The ALS Association Opportunities• Centralize and co-ordinate consortium

efforts

• Pursue the most promising targets and therapeutic approaches and partner with industry to bring to the clinic

• Innovative academic-industry pre-competitive partnerships-especially biomarkers

• Exploit technologies both for people living with the disease now and future therapies

• Large Data initiatives-Project MINE

• Phase II clinical trials

Milton-Safenowitz Post-Doctoral Fellowships