John E. Landers, Ph.D. Professor of Neurology University ... · Background of ALS • A progressive, fatal, disease caused by the degeneration of motor neurons • As motor neurons

John E. Landers, Ph.D.

Professor of Neurology

University of Massachusetts Medical School

Background of ALS

• A progressive, fatal, disease caused by the

degeneration of motor neurons

• As motor neurons die, muscles are unable

to function and gradually become weaker

• Eventually all voluntary movement is lost

and muscles become paralyzed

• Typically develops between ages 40-70

• ~50% affected die within 3 years/~75%

within 5 years

• ~5,000 people are newly diagnosis in the

US every year

• ~30,000 total affected people in the US

• Family History of ALS

• 10% of all ALS cases

• Typically due to a single mutation

in a single gene

• Researchers have identified ~2/3 of

the genes contributing to Familial

ALS

• Stronger Genetic / Weaker

Environmental Influence

• No Family History of ALS

• 90% of ALS cases

• Multiple genes may be contributing

to your risk of ALS + Environmental

Factors

• Few Risk Genes Contributing to

ALS have been Identified

• Weaker Genetic / Stronger

Environmental Influence

ALS

Genetics Opens Several Avenues of

Research for Therapeutic Treatments

Therapeutic Treatments

Genetics

Pathways/PathogenesisGene Therapy Model Organisms

Improvements in Any of These Areas Will Facilitate

The Development of Therapeutic Treatments

• All Affected Family Members are due

to the Same Genetic Mutation

• Segregation of DNA regions to

Multiple Affected Family Members to

Narrow Genomic Region

• Typically due to a single “causative”

mutation in a single gene

• Familial ALS Genes: SOD1, FUS,

TARDBP, UBQLN2

• Identification of common SNP

alleles that are more frequent in

ALS vs. control population

• Does not necessary identifying the

contributing factor but rather an

associated marker

• Usually identify variants with small

impact of disease risk (odds ratios

< 2)

• Sporadic ALS Genes: UNC13A,

SARM1, C21orf2

Overview of Positional Cloning

2-20 MbUp to >100

Genes

Pediatr Nephrol (2007) 22:2023–2029http://portal.ccg.uni-koeln.de/ccg/index.php?id=49

Gene Mapping

GeneIdentification

MutationScreening

Require Multiple Affected

Family Members

Linkage

Analysis

Human Genome Project

Novel Approaches Needed to Identify Causative

Genes for Difficult Families

• For some disorders, families are too small for traditional positional cloning

• Late-onset disease- difficult to obtain multiple generations

• Complete penetrance of the disease is not always observed

• Results in Numerous genomic regions which are too large and expensive to sequence by Sanger sequencing

Exome sequencing has resulted in a massive boost

in gene identification for Mendelian diseases

Exome Sequencing = Technique to Deep Sequence

only the ~2% of the genome coding for protein

Perform Exome Sequencing on Affected Family Members

Identify Variants in Each Sample

Filter Variants Based on Several Criteria

Identify Causative Mutation

Wu et al. Nature 488: 499–501. (2012).

Variant identification:

► Exome sequence two affected members from two ALS families

Variant Filtering:► Observed in both family members

► Alters amino acid

►Within a linkage peak

► Absent/Rare in SNP databases

Family 1

282,792

SNPs

C71G

Family 2

382,751

SNPs

2 SNPs 3 SNPs

M114T

PFN1

Combined LOD Score of Families #1- 2 = 4.51

(Chance of observing: 30,000:1)

Wu et al. Nature 488: 499–501. (2012).

• Conversion of monomeric (G)-actin to

filamentous (F)-actin

• Poly-L-Proline Binding Domain

• Reported to bind >50 proteins

• Regulation of Actin Polymerization

(VASP, Formin)

• VCP

• SMN

• Huntingtin

W. Witke. Trends in Cell Biol. 14: 461-469 (2004).

S=Soluble

I=Insoluble

Primary Motor Neurons

N2A Cells

Wu et al. Nature 488: 499–501. (2012).

Primary Motor Neurons

Wu et al. Nature 488: 499–501. (2012).

F-actin = Filamentous Actin

G-actin = Monomeric ActinWu et al. Nature 488: 499–501. (2012).

Problem: How to Identify Genes Associated with ALS

Without Multiple Family Members?

No additional affected members are available

for ~90% of FALS DNA collected

• Exome sequence of individual cases will not

identify causative genes:

• ~80 Novel non-synonymous variants

• ~800 Rare (<0.5%) non-synonymous variants

Linkage Studies

GWAS

Nature 461, 747-753 (2009)

Linkage Studies

GWAS

Rare Variant Association Analysis

For Every Gene, Compare the Frequency of All Rare Deleterious Variantsin a Cohort of Cases and Controls

Nature 461, 747-753 (2009)

ALS Patient

DNA Sequences

Control

DNA Sequences

Sequence #

1.

2.

3.

4.

5.

1.

2.

3.

4.

5.

Variant

ALS Patient

DNA Sequences

Control

DNA Sequences

Sequence #

1.

2.

3.

4.

5.

1.

2.

3.

4.

5.

Gene #1 Gene #2

ALS Patient

DNA Sequences

Control

DNA Sequences

Sequence #

1.

2.

3.

4.

5.

1.

2.

3.

4.

5.

Gene #1 Gene #2

ALS Patient

DNA Sequences

Control

DNA Sequences

Sequence #

1.

2.

3.

4.

5.

1.

2.

3.

4.

5.

Gene #1 Gene #2

ALS Patient

DNA Sequences

Control

DNA Sequences

Sequence #

1.

2.

3.

4.

5.

1.

2.

3.

4.

5.

Gene #1 Gene #2

ALS Patient

DNA Sequences

Control

DNA Sequences

Sequence #

1.

2.

3.

4.

5.

1.

2.

3.

4.

5.

Gene #1 Gene #2

ALS Patient

DNA Sequences

Control

DNA Sequences

Sequence #

1.

2.

3.

4.

5.

1.

2.

3.

4.

5.

Gene #1 Gene #2

ALS Patient

DNA Sequences

Control

DNA Sequences

Sequence #

1.

2.

3.

4.

5.

1.

2.

3.

4.

5.

Gene #1 Gene #2

Variants in ALS: 3

Variants in Controls: 4

No Differences Observed in

ALS versus controls

ALS Patient

DNA Sequences

Control

DNA Sequences

Sequence #

1.

2.

3.

4.

5.

1.

2.

3.

4.

5.

Gene #1 Gene #2

Variants in ALS: 3 3

Variants in Controls: 4 1

Higher Rate of Variants

in ALS versus controls

Rare Variant Association (RVA) Analysis

• Advantages

• Does not require related family members for analysis

• Can identify risk factors of greater effect than through GWAS

• Multiple software packages available to allow for

weighting/filtering variants by:

• Allele frequency

• Functional annotations

• Both positive and negative effects on phenotypes

• Software packages: CAST, CMC, VT, SKAT

Rare Variant Association (RVA) Analysis

• Disadvantages

• Although, associated genes can be identified, it does not necessarily identified those mutations that are pathogenic

• May require large sample sizes to identify associated genes based on genetic contribution

• Samples must be exome or whole genome sequenced

• Many different parameters are established by user• Allele frequency threshold to define as “rare variant”• Various packages/methods of statistical testing• Many different methods of defining a variant as “deleterious”• Application of multiple parameters increases the level of multiple test

correction to be applied

PolyPhen-2 FATHMM CADD

LRT MetaLR MetaSVM

phyloP PROVEAN MutationAssessor

SIFT SiPhy MutationTaster

Exome-wide Rare Variant Analysis To Identify

Novel Familial ALS Associated Gene

• Familial ALS Exomes:Australia 68

Belgium 13

Canada 39

Germany 202

Ireland 18

Italy 151

Netherlands 47

Turkey 131

UK 216

USA 448

Total 1376 Exomes (1,022 pass QC)

(~90% devoid of known mutations)

• Control Exomes: 13,883 Exomes

(7,315 pass QC)

Optimization of Parameters for Rare

Variant Analysis Using Known ALS Genes

1) Analysis performed on 10 Known ALS

Genes using 308 different combinations of

parameters:- 7 Allele Frequencies

- 44 Functional Filters

2) For each analysis, plotted how many

genes could be detected at different P-

value thresholds

3) Determined the optimal set of parameter

by measuring the area under the curve

4) No one test is best for all genes

Cases: n=1,022

Controls: n=7,315

Known Genes Tested: SOD1, FUS,

TARDBP, VCP, VAPB, TUBA4A,

TBK1, OPTN, PFN1, UBQLN2

FATHMM

MAF < 0.001

Kenna et al. Nature Gen. 48: 1037-1042

Rare Variant Analysis Using

Optimized Parameter Settings

1) Analysis detected 7 of 10 Known

ALS Genes with P values <5 x 10-4

2) One gene, NEK1, detected with a

significant P value

OR=8.2; P= 1.7 x 10-6)

OPTN, VAPB, PFN1 displayed less

significant P-values (ALS genes lower in

frequency in the population)

Cases: n=1,022

Controls: n=7,315Kenna et al. Nature Gen. 48: 1037-1042

Signal From FALS Rare Variant Analysis

is Primarily Driven by NEK1 LOF Variants

ALS Cases (n=1,022): 12 LOF Variants (1.17%)

Controls (n=7,315): 14 LOF (0.19%)

P = 7.3 x 10-7 Odds Ratio: 8.9

p.I10T(1)

p.Y30C(1)

c.217+1C>T*(1)

p.R91X(1)

p.R91Q(1)

p.N93D(1)

p.V98I(3)

p.I105T(1)

p.A115S(1)

p.Q132R*(1)

p.R161X(1)

p.R161X(1)

p.D185G(1)

p.L193F(1)

p.E204G(1)

p.V223M(1)

p.Y229C(12)

p.N250S(1)

p.R261H(17)

p.R261H(52)

c.878+1C>G*(1)

p.I308M(1)

p.A313T(1)

p.K337N(1)

p.A341T(7)

p.A341T(36)

p.T344A(1)

p.R356K(1)

p.D379E(4)

p.Q380E*(1)

p.L384S(1)

c.1153+3T>G*(1)

p.Q393X(1)

p.W409X(1)

p.W409X(1)

p.R440Q(1)

p.E462V(1)

p.A463V(105)

p.A463V(790)

p.G478A(1)

c.1446+1C>A*(1)

c.877− 1C >A *(1)

p.K511Q(1)

p.A512V(1)

p.Q525K(1)

p.R540Q(1)

p.M545T(1)

p.R550X(1)

c.1683+2A>G*(1)

p.F597I(1)

p.N598S(2)

p.N598S(2)

p.E610K(1)

p.A626T(165)

p.A626T(1119)

p.R630H(1)

p.V646I(1)

p.K648E(1)

p.K648E(2)

p.K654M(1)

p.S681F(1)

p.P682L(1)

p.Q696X(1)

p.V704I(2)

p.V713M(3)

p.V713M(19)

p.R742C(6)

p.N745K(10)

p.N745K(77)

p.E752G(216)

p.E752G(1520)

p.H769R(2)

p.H775Y(2)

p.D784E(1)

p.G791R(1)

p.G792D(1)

p.G792D(1)

p.G825V(1)

p.G825V(1)

p.P835L(2)

p.A846T(1)

p.K870N(1)

c.2 6 12 − 2 T >C *(1)

c.2 45 8− 2 T >C *(1)

p.Q911E(1)

p.K939E(1)

p.C945Y(1)

p.Q962R(1)

p.L986F(1)

p.P993A(1)

p.Q1031R(1)

p.S1036X(3)

p.S1036X(5)

p.T1065A(5)

p.M1075L(1)

p.T1096S(1)

p.T1096S(1)

p.D1099H(1)

p.E1142Q(1)

p.R1146K(1)

p.D1180A(1)

p.F1211S(1)

p.H1265Y(1)

p.I1270S(1)

p.D1283G(1)

p.N1284D(2)

FALS (n=1,022)

Controls (n=7,315)

Domain key

PKD

Coiled−coil

a

Variants observed in

ALS

Variants observed

in Controls


ALS Cases (n=2,303): 23 LOF Variants

Controls (n=1,059): 0 LOF Variants

P = 1.5 x 10-4

p.K3E(1)

p.I10T(1)

p.V64A(1)

p.N93D(1)

p.E158fs(1)

p.D185G(1)

p.F220fs(1)

p.S224F(1)

p.Y229C(1)

p.R232H(1)

p.P243A(1)

p.R261C(1)

p.R261H(39)

p.R261H(6)

c.877− 1C>A*(1)

p.H330fs(1)

p.A341T(7)

p.A341T(9)

p.Q343X(1)

p.R356K(1)

p.D379E(4)

p.K422E(1)

p.Y443C(1)

p.E444D(1)

p.A463V(277)

p.A463V(124)

p.R484H(2)

p.R540X(1)

p.P569R(1)

p.N598S(1)

p.E624K(1)

p.A626T(362)

p.A626T(152)

p.V646I(1)

p.R655fs(1)

p.E662fs(2)

p.V675I(1)

p.P682L(3)

p.P682L(1)

p.V704I(3)

p.V704I(2)

p.V713M(1)

p.R742C(2)

p.R742C(2)

p.N745K(36)

p.N745K(14)

p.E752G(458)

p.E752G(222)

p.S761fs(1)

p.G792D(1)

p.K841E(1)

p.Y871fs(1)

p.S902G(1)

p.L920X(1)

p.S1036X(10)

p.T1065A(3)

p.L1091fs(1)

p.E1129G(1)

p.N1189K(1)

p.N1284D(2)

SALS (n=2,303)

Controls (n=1,059)

Domain key

PKD

Coiled-coil

bVariants observed in

ALS

Variants observed in Controls

Increased LOF Variants in NEK1 Protein

Replicates in Sporadic ALS

Analysis of ALS within an Isolated

Community in the Netherlands

Population: <25,000

Genome Sequencing Was

Performed on Four ALS Patients

• Patients in isolated

communities may be the

result of the homozygosity

of a genetic mutation

• The region of the mutation

can be identified by

identifying regions of

homozygosity of affected

patients

• Similar approach used to

identify FUS and OPTN


Homozygosity Mapping Identifies NEK1

As a Candidate Risk Factor for ALS

Identified 4 Rare Non-synonymous Variants Within Regions of Homozygosity

R261H Variant within NEK1 Remained and Replicated as a Risk Factor for ALS


ALS

Cases

#

Controls

%

Cases

%

ControlsOdds Ratio P

7,194 11,732 1.62 0.70 2.41 1.2 x 10-7

NEK1 Displays Several Functional Properties

• NEK1 is a primary regulatory of the formation of non-motile primary

cilium

• Cilia are microtubule-based sensory organelles present on the surface

of non-proliferating cells, including neurons.

• ~90% of Ciliopathies display neurobehavioural or neurodevelopmental

deficits

• Homozygotes for NEK1 LOF cause Short Rib Polydactyly Syndrome

Type II

• Characterized by constricted thoracic cage, short ribs, shortened tubular

bones

• Fibroblast from patients display defect of cilia formation and

cytoskeleton microtubule architecture

• Disruption of NEK family members disrupts neuronal morphology,

neurite outgrowth, microtubule stability and microtubule dynamics

• Regulates DNA damage repair through interactions with C21ORF2

(recently identified as a GWAS hit)

Improvements Towards the Identification of Novel Familial

ALS Genes Using Rare Variant Association Analysis

• Compared the Number of Variants in Every Gene Within:

• 1,138 Index Familial ALS (FALS) Exomes (one sample per family)

• Increased from 1,022 FALS Exomes

• 19,494 Control Exomes

• Increased from 7,315 control exomes

• Restricted to Variants:

• Loss of Function (LOF) Mutations (nonsense or splice altering)

• Last 2 ALS genes (NEK1, TBK1) displayed LOF mutations

• Minor Allele Frequency < 0.1%

• Exome-Wide Analysis

• 11,472 genes passed all QC parameters

Loss of Function (LOF) Rare Variant Analysis

Identifies KIF5A as an ALS Gene

1,138 Index Familial ALS (FALS) Cases, 19,494 Controls

Nonsense/Splice Altering Variants

• 6/1,138 FALS Cases vs 3/19,494 Controls

• Odds Ratio = 32.1; P = 5.6 x 10-7

• Segregation observed in two siblings for one variant and a

one sibling from a second variant

• Three heavy chain isoforms (KIFA5A, KIF5B,

KIF5C) are all expressed in neurons

• Hetero/homodimerize through Stalk Domain

• Axonal Transport many cargos by binding to

distinct adaptor proteins

• KIF5 transports RNA and RNA binding proteins

– Includes ALS proteins FUS and hnRNPA1

• Transports Neurofilaments and Mitochondria

– Abnormal NFs, Mitochondrial dysfunction and Impaired

Axonal Transport are all commonly observed in ALS

• Mutations in KIF5A identified in hereditary spastic paraplegia

(SPG10) and Charcot-Marie-Tooth, type 2 disease

Kinesin Family Member 5A (KIF5A)

SPG10/CMT2p.Y63C, p.D73N, p.R162W, p.M198T,

p.S202N, p.S203C, p.R204Q, p.R204W, p.V231L, p.D232N, p.G235E,

p.E251K, p.K253N, p.K256del, p.N256S, p.K257N, p.S258L, p.L259Q, p.Y276C, p.P278L, p.R280H, p.R280C, p.R280L, p.R323W, p.A361V, p.E755K

ALSp.Pro986Leu**, c.2993-3C>T, p.Arg1007Gly, p.Arg1007Lys,c.3020+1G>A, c.3020+2T>A,c.3020+3A>G, p.Asp996fs,

p.Asn999fs, p.Asn997fs,c.2993-1G>A, p.Asn999del

Motor DomainMicrotubule Binding, Kinesin Motor

(9-327)

Stalk

Heavy Chain Dimerization

(331-906)

Tail

Cargo Binding

(907-1032)

https://www.youtube.com/watch?v=y-uuk4Pr2i8

CAGGAAATGCCACAGATATCAATGACAATAGGTA

ConsensusSplice Sequence:

G N A T D I N D N R

T A G A A A G

Normal Splice mRNA:

Normal Protein:

Mutant Splice mRNA:

Mutant Protein:

Normal Splice

RNA

B.

C.

A.

MDNGNATDINDNRSDLPCGYEAEDQAKLFPLHQETAAS MDNGVTCRVAMRLRTRPSFSLSTKRQQPANLPHPRLHTCTFSF

26 27 28

26 27 28 26 28

Transla: onStop

AG ... Exon 27 AG GT...GC ACT A A G

Mutant Splice

Transla: onStop

100

200

(+) RT (-) RT

Co

ntr

ol

#1

Co

ntr

ol

#2

AL

S

Wa

ter

Co

ntr

ol

#1

Co

ntr

ol

#2

AL

S

Wa

ter

Co

ntr

ol

#1

Co

ntr

ol

#2

AL

S

Wa

ter

Co

ntr

ol

#1

Co

ntr

ol

#2

AL

S

Wa

ter

(+) RT (-) RTD.

Variants Identified in FALS are Predicted

Bioinformatically to Create a Truncated Protein

28 Coding Exons

29 Total Exons

1032 Amino Acids

a.a. 998-1007

• Deletes 34 C-terminal a.a. and adds novel 39 a.a. sequence

• Confirmed by RT-PCR using Patient-Derived RNA

• The C-terminal region functions to bind cargo

Analysis of Small Insertions/Deletions (indels)

Reveals Additional LOF Variants in FALS

Position Variant Exon cDNA Description Predicted Exon

Skipping

Control Variants

57,963,470 A>G 11 c.1117+4A>G 3' Splice Junction P

57,966,423 C>T 15 c.1630C>T p.Arg544* -

57,976,884 G>C 28 c.3021G>C 5' Splice Junction N

FALS Variants 57,975,729 GA>A 26 c.2987delA p.Asp996fs -

57,976,382 C>T 27 c.2993-3C>T 5' Splice Junction Y

57,976,385 GA>G 27 c.2996delA p.Asn999fs -

57,976,411 A>G 27 c.3019A>G p.Arg1007Gly Y

57,976,412 G>A 27 c.3020G>A p.Arg1007Lys Y

57,976,413 G>A 27 c.3020+1G>A 3' Splice Junction Y

57,976,414 T>A 27 c.3020+2T>A 3' Splice Junction Y

57,976,415 A>G 27 c.3020+3A>G 3' Splice Junction Y

• 2/1,138 FALS Cases vs 0/19,494 Controls (indels)

• Combined results: 8/1,138 FALS Cases vs 3/19,494 Controls

• Combined Odds Ratio = 41.2; P = 3.8 x 10-9

GWAS Identifies an Association of the

KIF5A Region and Sporadic ALS Risk

• C9orf72, TBK1, UNC13a, C21orf2, TNIP1 Previously Identified

• KIF5A: Pro986Leu (Odds Ratio = 1.38; P = 6.43 x 10-10)

Cohort Size: 20,806 Cases 59,804 Controls

10,031,630 genotyped and imputed variants tested for

association

Replication Studies Confirm Pro986Leu

as a Risk Factor for ALS

Replication samples contributed in part by:

1) Project Mine

2) New York Genome Center

3) CReATe Consortium

4) AnswerALS

5) Genomic Translation for ALS Care Consortium

Odds Ratio = 1.38; P = 2.31 x 10-12

KIF5A Loss of Function Mutations Display Higher

Prevalence in Familial vs. Sporadic ALS

• Additional 9,046 ALS cases (predominantly sporadic) and 1,955

controls were screened for LOF variants

• 2 LOF variants (0.022%) Identified in sporadic ALS cases

–c.2989delA p.Asn997fs

–c.2993-1G>A 5' Splice Junction

• No LOF variants were identified in controls

• Rate is much lower (~30 fold) than the observed rate in FALS

cohort (0.703%)

• Suggests LOF variants display a high penetrance

ALS Associated Mutations in KIF5A

are Distinct from SPG10/CMT2 Mutations







(9-327)

Stalk


(331-906)

Tail

Cargo Binding

(907-1032)

Missense

Mutations LOF

Mutations

Advantages of Whole Genome Sequencing

Over Exome Sequencing

• Exome sequencing does not cover all genes

• ENCODE Project has revealed that substantially more of the “functional genome” lies within non-coding sequences

• e.g. miRNAs, lncRNAs, promoter regions

• Exome sequencing is not suited to identify copy number variations/ indels that may contribute to human disease

• Whole genome sequencing is considerably more amenable to phasing of variants than exome sequencing

• Distinguishing cis variants and compound heterozygosity for recessive models and eQTL analysis

• Imputation of missing variants

To Comprehensively Analyze WGS,

Tens of Thousands of Cases/Controls are Required

Project MinE: An International Consortium for ALS

Whole Genome Sequence

• Goal: To sequence 15,000 ALS and 7,500

control genomes

• Consists of 17 countries:

• Australia, Belgium, Brazil, Canada, France, Germany,

Ireland, Israel, Italy, Netherlands, Portugal, Spain,

Sweden, Switzerland, Turkey, UK, USA

• Project MinE USA Directors (Oct. 2014)

• Jonathan Glass (Emory U.) / John Landers

• Approximately 35% completed

• Funding for 8,338 samples

Genetics Opens Several Avenues of Research for



Genetics

Pathways/PathogenesisGene Therapy Model Organisms

Improvements in Any of These Areas Will Facilitate

The Development of Therapeutic Treatments

Paralysis Beginning at ~140 days

Weight Loss

Decreased Grip Strength

Motor Neuron Loss

5X Overexpression of

No Phenotype in WT Overexpression

Transgenic Mice Expressing Mutant PFN1

Represent a Mouse Model for ALS

Additional Efforts to Develop

Mouse Models for ALS

• Several Mouse Models are Currently in Development

In Collaboration with The Jackson Laboratory

• Mouse Models are Based on Genes Identified by Our

Laboratory

• Profilin-1 (PFN1)

• Tubulin, alpha 4A (TUBA4A)

• NEK1

• KIF5A

• For Each Gene, Inducible Transgenic and CRISPR

Models are in Development for Multiple Mutations

Numerous Pathways are Impaired in the Motor

Neurons of ALS Patients

Ferraiuolo…Shaw, Nature Reviews Neurology (2011) 7:616

Genetics Pinpoints the Primary Causes and

Pathways for Human Disease

Eykens & Robberecht, Advances in Genomics and Genetics(2015) 5:327

• RNA Processing

• FUS

• TDP-43

• hnRNPA1

• Protein Degradation

• UBQLN2

• VCP

• OPTN

• Cytoskeletal Dynamics

• PFN1

• TUBA4A

• KIF5A

Cytoskeletal Dynamics, RNA Processing and Protein

Degradation May Represent Overlapping Pathways in ALS

Cytoskeletal

Dynamics

RNA

Processing

Protein

Degradation

Motor

Neuron

Death

Future Directions

• Continued Efforts in ALS Genetics

• Collaborative Efforts in Whole Genome Sequencing with

numerous groups

• Development of Mouse Models in Collaboration with Jackson

Laboratory

• Profilin-1 (PFN1)

• Tubulin, alpha 4A (TUBA4A)

• NEK1

• KIF5A

• Development of Therapeutics for ALS Using High Throughput

Screening of Small Compounds and Biological Compounds

• Obtaining Compound Libraries from Several Sources

• Developing Novel Methods to Screen Knockdown

Expression of All Genes for Therapeutics

• Using Mouse Models to Validate Therapeutics for ALS

Acknowledgements

UMass Medical School

Kevin Kenna

Brendan Kenna

Aoife Kenna

Pamela Keagle

Janice Dominov

Robert Brown

National Institutes of Health

Bryan Traynor

Aude Nicolas

Alan Renton

Faraz Faghri

Ruth Chia

University of Milan

Nicola Ticozzi

Vincenzo Silani

University of Turin

Adriano Chiò

Emory University

Jonathan Glass

Umea University

Peter Andersen

University Medical Centre

Utrecht

Jan Veldink

Leonard van den Berg

King’s College London

Ammar Al-Chalabi

Chris Shaw

Jackson Laboratory

Cat Lutz

Project Mine

Guy Rouleau

Nazli Basak

Karen Morrison

Kevin Talbot

Markus Weber

Martin Turner

Michael A. Van Es

Orla Hardiman

Pamela J. Shaw

Philip Van Damme

Russell L. McLaughlin

Vincenzo Silani

Wim Robberecht

University of Helsinki

Pentti Tienari

Merck Research Labatories

David Stone

Answer ALS

Jeff Rothstein

Steve Finkbeiner

CReATe Consortium

Michael Benatar

J. Paul Taylor

NYGC

Hemali Phatnani

Genomic Translation for ALS Care

Matt Harms

David Goldstein

SLAGEN

ITALSGEN

Funding

ALS Association (Lucie Bruijn)

NIH / NINDS

Packard Center for ALS Research

• Three heavy chain isoforms (KIFA5A, KIF5B,

KIF5C) are all expressed in neurons

• Hetero/homodimerize through Stalk Domain

• Axonal Transport many cargos by binding to

distinct adaptor proteins

• KIF5 transports RNA and RNA binding proteins

– Includes ALS proteins FUS and hnRNPA1

• Transports Neurofilaments and Mitochondria

– Abnormal NFs, Mitochondrial dysfunction and Impaired

Axonal Transport are all commonly observed in ALS

• Mutations in KIF5A identified in hereditary spastic paraplegia

(SPG10) and Charcot-Marie-Tooth, type 2 disease

Kinesin Family Member 5A (KIF5A)







(9-327)

Stalk


(331-906)

Tail

Cargo Binding

(907-1032)

Documents

John E. Landers, Ph.D. Professor of Neurology University ... · Background of ALS • A progressive, fatal, disease caused by the degeneration of motor neurons • As motor neurons