Reduced expression of the Kinesin-Associated
Protein 3 (KIFAP3) gene increases survival
in sporadic amyotrophic lateral sclerosis
John E. Landersa,1, Judith Melkib,2, Vincent Meiningerc, Jonathan D. Glassd, Leonard H. van den Berge,
Michael A. van Ese, Peter C. Sappa,f, Paul W. J. van Vughte, Diane M. McKenna-Yaseka, Hylke M. Blauwe, Ting-Jan Choa,
Meraida Polakd, Lijia Shia, Anne-Marie Willsa, Wendy J. Brooma, Nicola Ticozzia,g, Vincenzo Silanig, Aslihan Ozoguzh,
Ildefonso Rodriguez-Leyvaa,i, Jan H. Veldinke, Adrian J. Ivinsonj, Christiaan G. J. Sarise, Betsy A. Hoslera,
Alayna Barnes-Nessaa, Nicole Couturea, John H. J. Wokkee, Thomas J. Kwiatkowski, Jr.a, Roel A. Ophoffk,l,
Simon Croninm, Orla Hardimanm, Frank P. Diekstran, P. Nigel Leighn, Christopher E. Shawn, Claire L. Simpsonn,
Valerie K. Hansenn, John F. Powelln, Philippe Corciao, Franc ¸ois Salachasc, Simon Heathp, Pilar Galanq, Franck Georgesb,
H. Robert Horvitzf, Mark Lathropp, Shaun Purcellr, Ammar Al-Chalabin,3, and Robert H. Brown, Jr.a,1,3
aCecil B. Day Neuromuscular Research Laboratory, Massachusetts General Hospital-East, Building 114, Navy Yard, Charlestown, MA 02129;bLaboratoire de
Neurogenetique Moleculaire, Institut National de la Sante ´ et de la Recherche Me ´dicale U-798, Universite d’Evry et Paris 11, 2 rue Gaston Cre ´mieux, CP5724,
91057 Evry France;cFe ´de ´ration des maladies du syste `me nerveux, Assistance Publique—Ho ˆpitaux de Paris, Ho ˆpital de la Salpe ˆtrie `re, 75651 Paris, France;
dDepartment of Neurology, Emory University, Atlanta, GA 30322; Departments ofeNeurology andkMedical Genetics, Rudolf Magnus Institute of
Neuroscience, University Medical Center Utrecht, 3584 CX, Utrecht, The Netherlands;fHoward Hughes Medical Institute, Department of Biology,
Massachusetts Institute of Technology, Cambridge, MA 02139;gDepartment of Neurology and Laboratory of Neuroscience, ‘Dino Ferrari‘ Center, University
of Milan Medical School—Istituto Di Ricovero e Cura a Carattere Scientifico Istituto Auxologico Italiano, 20149 Milan, Italy;hDepartment of Molecular
Biology and Genetics, Neurodegeneration Research Laboratory, Bogazici University, Istanbul, Turkey;iFaculty of Medicine, Universidad Autonoma de San
Luis Potosi, San Luis Potosi, Mexico S.L.P., CP 78210;jHarvard NeuroDiscovery Center, Harvard Medical School, Boston, MA 02115;lNeuropsychiatric Institute,
University of California, Los Angeles;mDepartment of Neurology, Beaumont Hospital, Dublin 9, Ireland;nMedical Research Council Centre for
Neurodegeneration Research, Department of Clinical Neuroscience, PO43, Institute of Psychiatry, King’s College London, London SE5 8AF, United Kingdom;
oService de Neurologie, Centre Hospitalier Universitaire Bretonneau, 37044 Tours, France;pCentre National de Ge ´notypage, Institut Ge ´nomique,
Commissariat a ` l’E´nergie Atomique, 91057 Evry, France;qUnite ´ de Recherche en Epide ´miologie Nutritionnelle, l’UFR Sante Me ´decine et Biologie Humaine,
74 rue Marcel Cachin, 93017 Bobigny, France; andrCenter for Human Genetics Research, Massachusetts General Hospital, Richard B. Simches Research
Building, CPZN-6254, 185 Cambridge Street, Boston, MA 02114
Edited by David E. Housman, Massachusetts Institute of Technology, Cambridge, MA, and approved April 1, 2009 (received for review December 22, 2008)
Amyotrophic lateral sclerosis is a degenerative disorder of motor
neurons that typically develops in the 6th decade and is uniformly
fatal, usually within 5 years. To identify genetic variants associated
with susceptibility and phenotypes in sporadic ALS, we performed
a genome-wide SNP analysis in sporadic ALS cases and controls. A
total of 288,357 SNPs were screened in a set of 1,821 sporadic ALS
susceptibility were further screened in an independent sample set
gene (encoding a kinesin-associated protein) yielded a genome-
wide significant result (P ? 1.84 ? 10?8) that withstood Bonferroni
correction for association with survival. Homozygosity for the
favorable allele (CC) conferred a 14.0 months survival advantage.
Sequence, genotypic and functional analyses revealed that there is
linkage disequilibrium between rs1541160 and SNP rs522444
within the KIFAP3 promoter and that the favorable alleles of
rs1541160 and rs522444 correlate with reduced KIFAP3 expression.
No SNPs were associated with risk of sporadic ALS, site of onset, or
age of onset. We have identified a variant within the KIFAP3 gene
that is associated with decreased KIFAP3 expression and increased
survival in sporadic ALS. These findings support the view that
genetic factors modify phenotypes in this disease and that cellular
motor proteins are determinants of motor neuron viability.
genome-wide association study ? single nucleotide polymorphism
ops in the 6th decade and is uniformly fatal, usually within 5 years
20% of these are caused by mutations in the gene encoding
copper/zinc superoxide dismutase 1 (SOD1) (4); mutations in the
TARDBP gene (5, 6) account for ?5% of cases. Rare familial cases
arise from mutations in genes encoding the vesicle-associated
membrane associated protein B (7), alsin (a RAB5-guanine nu-
myotrophic lateral sclerosis (ALS) is an age-dependent, de-
generative disorder of motor neurons (1) that typically devel-
cleotide exchange factor) (8, 9), senataxin (10) or dynactin (11).
Recently, we reported that ?5% of familial ALS cases are due to
and RNA, as does TARDBP. The cause of sporadic ALS is thought
to be multifactorial, with environmental, infectious and genetic
etiologies. Reported associations with variants in diverse genes
(14–25) have proven difficult to replicate. Advances in the tech-
nology for large-scale genotyping of single nucleotide polymor-
phisms (SNPs) have facilitated unbiased, genome-wide association
studies. Examples include the identification of IL2RA and IL7RA
variants as risk factors for multiple sclerosis (26–28) and the recent
reports of 6 new gene regions associated with type 2 diabetes
(29–35). To test the hypothesis that commonly occurring genetic
Author contributions: J.E.L., J.M., V.M., J.D.G., L.H.v.d.B., M.A.v.E., P.C.S., P.W.J.v.V.,
D.M.M.-Y., H.M.B., T.-J.C., M.P., L.S., A.-M.W., W.J.B., N.T., V.S., A.O., I.R.-L., J.H.V., A.J.I.,
S.H., P.G., F.G., H.R.H., M.L., S.P., A.A.-C., and R.H.B. designed research; J.E.L., M.A.v.E.,
P.C.S., P.W.J.v.V., D.M.M.-Y., H.M.B., T.-J.C., M.P., L.S., A.-M.W., W.J.B., N.T., V.S., A.O.,
I.R.-L., J.H.V., C.G.J.S., B.A.H., A.B.-N., N.C., T.J.K., R.A.O., S.C., O.H., F.P.D., C.L.S., V.K.H.,
J.D.G., L.H.v.d.B., W.J.B., I.R.-L., A.J.I., P.N.L., C.E.S., J.F.P., S.P., A.A.-C., and R.H.B. contrib-
uted new reagents/analytic tools; J.E.L., J.M., V.M., L.H.v.d.B., M.A.v.E., P.C.S., P.W.J.v.V.,
D.M.M.-Y., H.M.B., T.-J.C., M.P., L.S., A.-M.W., W.J.B., N.T., V.S., A.O., I.R.-L., J.H.V., A.J.I.,
C.G.J.S., B.A.H., A.B.-N., N.C., J.H.J.W., T.J.K., R.A.O., S.C., O.H., F.P.D., P.N.L., C.E.S., C.L.S.,
V.K.H., J.F.P., P.C., F.S., S.H., P.G., F.G., H.R.H., M.L., S.P., A.A.-C., and R.H.B. analyzed data;
and J.E.L., S.P., A.A.-C., and R.H.B. wrote the paper.
The authors declare no conflict of interest.
This article is a PNAS Direct Submission.
Freely available online through the PNAS open access option.
1To whom correspondence should be addressed at the present address: Department of
Neurology, University of Massachusetts Medical School, Worcester, MA 01605. E-mail:
2Present address: Department of Human Genetics, Hadassah University Hospital, P.O. Box
12000, 91120 Jerusalem, Israel.
3A.A.-C. and R.H.B. contributed equally to this work.
This article contains supporting information online at www.pnas.org/cgi/content/full/
June 2, 2009 ?
vol. 106 ?
variants modify susceptibility, survival, site of onset or age of onset
of 1,821 sporadic ALS cases (SALS) and 2,258 controls.
Genotypes were obtained from 3 sources in the U.S. (Boston,
Atlanta, National Institute of Neurological Disorders and
Stroke) and 3 in Europe (London, France, Netherlands), using
Illumina BeadArrays. Survival information was not available for
samples from the NINDS and France. No SNPs generated
significant P values for association with susceptibility, site of
onset, and age of onset of disease after Bonferroni correction
(288,357 SNPs ? 4 phenotypes) (Fig. 1 C–E, Table 1, see also
Table S1). In a further attempt to reveal any SNPs that were
associated with susceptibility of sporadic ALS, we elected to
genotype those SNPs that yielded a P ? 5.0 ? 10?4(153 in total)
in a confirmatory (Stage 2) panel consisting of 538 ALS cases
of the samples. Successful genotypes were obtained for 139
(90.8%) of the SNPs; none of the variants yielded a significant
P value after Bonferroni multiple test correction (Table S1).
Although our study failed to confirm recent reports that sus-
ceptibility to sporadic ALS may be mediated by variants in the
inositol-triphosphate receptor (ITPR2) (21), DPP6 (22, 23) or a
novel, brain-expressed gene (FLJ10986) (24), these discrepan-
cies may reflect differences in methodology or case populations
In contrast to susceptibility, site of onset, and age of onset, SNPs
rs1541160 and rs855913 generated significant P values after Bon-
ferroni correction (288,357 SNPs ? 4 phenotypes) for association
with disease survival, using linear regression (Fig. 1A and Table 1).
SNPs (including imputed SNP alleles) yielded a cluster of positive
values; 4 of the imputed SNPs were significant after Bonferroni
correction (Fig. 1B). SNP rs1541160 maps within intron 8 of the
KIFAP3 gene (encoding a kinesin-associated protein) on chromo-
some 1. For SNP rs855913, the nominal and Bonferroni-corrected
of the ZNF746 gene. This gene was not further characterized for 3
reasons. First, a sensitivity analysis of this SNP revealed that it does
not replicate within the individual Boston population (P ? 0.264).
Second, in our sensitivity analyses, had we analyzed the U.S. as the
Stage 1 population, we would not have identified this variant due
is in contrast with SNP rs1541160 that emerges as significant in our
study, whether considering the aggregate of all cases or each
individual population. Finally, for the ZNF746 gene variant in
question, the homozygotes for the minor allele are rare (0.7%) so
that it is difficult to ascertain the reliability of the results (despite
having ?1,821 ALS cases in our screening study).
The genotype frequencies of rs1541160 are 9.9% (CC), 39.7%
(CT) and 50.4% (TT); the minor allele frequency is 29.7% (Table
cases P ? 0.527, all P ? 0.970). Haplotypes defined by 3 SNPs,
rs2750014, rs4656729 and rs12123693, but excluding rs1541160,
yielded association with survival comparable to that of rs1541160
(P ? 1.35 ? 10?9), indicating that genotyping artifacts specific to
rs1541160 are not generating the association. Further tests con-
regression analysis of survival. SNPs rs1541160 (circled) and rs855913 were significant after Bonferroni correction. (B) A closer view of the rs1541160 region is
shown. Dark points represent SNPs typed in the study, and light points represent SNPs whose genotypes were imputed. (Lower) Imputation certainty for each
imputed SNP, defined as the average maximum posterior genotype call probability. The chromosomal region spans 5 Mb on either side of SNP rs1541160.
Positions are in National Center for Biotechnology Information build-35 coordinates.
Plot of ?log10(P) for survival, age of onset, site of onset and susceptibility of sporadic ALS. Analysis for survival, age of onset, site of onset and
Landers et al.PNAS ?
June 2, 2009 ?
vol. 106 ?
no. 22 ?
SNPs distributed across the locus defined by KIFAP3 and 5 neigh-
boring genes (SCYL3, C1orf156, Clorf112, and Selectins E and L)
revealed disequilibrium that spanned ?155 kb from marker
rs2750014 to rs1216443 but was centered on rs1541160 within the
gene KIFAP3 (Fig. S1).
Our approach to identify variants associated with increased
survival was based on a joint analysis of 4 DNA sets. This approach
is more powerful than a 2-staged method in which a set of SNPs
within an initial population below a cutoff P value is verified within
a secondary confirmation population (36). However, because sev-
eral genome-wide association studies (GWAS) have used 2-stage
analyses (21, 22, 24, 30, 32, 33), we have investigated how such an
approach would influence our results. We performed a sensitivity
the study and computed the P value associated with both the
remaining populations (i.e., simulating a Stage 1 study) and the
removed population (i.e., simulating a Stage 2 study). In each case,
this sensitivity testing (Table 2) revealed that rs1541160 remained
in the top 5 survival-associated SNPs. The largest increase in the P
value (to 5.50 ? 10?6) was observed after removal of the Boston
the P values for rs1541160 from each individual site (the simulated
Stage 2 study) also yielded significant P values, with the exception
of samples (n ? 90). Interestingly, the median survival increased at
a minimum of 20.51% (Netherlands) and a maximum of 78.54%
(London) in individuals harboring a CC genotype for rs1541160 as
compared with a TT genotype (Table 2). We also performed the
sensitivity analysis by grouping the population into U.S. (Atlanta
and Boston) and Europe (London and Netherlands). By this
approach, both the U.S. and Europe yielded a high ranking for
rs1541160 if used as a Stage 1 population (20th and 31st, respec-
tively). Furthermore, both the U.S. and Europe yielded significant
individual P values (5.55 ? 10?5and 7.70 ? 10?5, respectively)
(Table 2). These results also confirm that the observed association
is not due to population stratification, which would not be expected
to yield a significant P value for each individual population.
The absolute median survival for the CC, CT, and TT genotypes
were 3.96, 2.84 and 2.67, respectively. The absolute mean survival
for the CC, CT, and TT genotypes were 4.60, 3.40, and 3.07,
analysis with SNP rs1541160 alone. With genotypic-based survival
curve analysis using the Peto-Prentice generalized Wilcoxon text
and deceased ALS cases (Fig. 2A), the P value for the rs1541160
SNP is 3.87 ? 10?6(n ? 1,014). A censored analysis considering all
(n ? 1,321).
Sequence analysis of the KIFAP3 coding region and exon/intron
boundaries of 8 individuals homozygous for the CC and 4 for the
TT rs1541160 genotype (12 individuals) did not reveal variants in
strong LD with rs1541160, suggesting that the KIFAP3-mediated
increase in survival is not due to an alteration in its protein
sequence. To determine whether the expression of KIFAP3 is
modified by the genotype of rs1541160, we performed real-time
PCR on lymphoblastoid cell lines harboring either a CC (n ? 38)
or TT (n ? 40) genotype for rs1541160. KIFAP3 expression in the
CC genotypes was 31.9% less than that in the TT genotypes (Fig.
3A) (P ? 0.0084, Wilcoxon 2-sample test). A comparison of
Table 1. SNPs yielding best P-values in four tested categories
AllelesRaw Corrected Case Cont.Limb Bulbar
Site of Onset
Age of Onset
Table 2. Sensitivity analysis of rs1541160 within four populations. Median Survival is represented in years
population Stage 1 P
rank Stage 2 P
Stage 2 median
Stage 2 median
(CC vs. TT)
1.84 ? 10?8
www.pnas.org?cgi?doi?10.1073?pnas.0812937106Landers et al.
occipital lobe brain samples homozygous for either the C (n ? 9)
or T (n ? 17) alleles again revealed a decrease in expression of
KIFAP3 (41.1%) in the CC as compared with TT samples (Fig.
3B) (P ? 0.025, Wilcoxon 2-sample test). A second real-time
PCR probe for KIFAP3 confirmed the findings for both lym-
phoblast and brain samples. Western blotting of the brain
samples confirmed a decrease in KIFAP3 protein (69.8%) in CC
samples compared with TT samples (Fig. S2) (P ? 0.014,
Wilcoxon 2-sample test).
Given that rs1541160 is located in the eighth intron of
KIFAP3, we decided to examine SNPs within the promoter
region which may influence gene regulation to determine
whether any were in linkage disequilibrium with rs1541160.
Sequencing of ?1.6 kb of this promoter within 8 individuals
homozygous for the CC and 4 for the TT rs1541160 genotype
revealed a variant (C/G, previously identified as rs522444)
located at ?25 bp relative to the transcription start site (Fig.
3C). This variant is in complete linkage disequilibrium with
rs1541160 for all 24 chromosomes (r2? 1.00). Genotypes
derived from the HapMap project for rs1541160 and rs522444
further documented that these 2 SNPs are in complete LD
(r2? 1.00) in Caucasian and SubSaharan African populations
(evident through analysis of 116 and 118 chromosomes, re-
spectively). We further confirmed this high level of LD by
genotyping an additional 1,017 individuals (r2? 0.99).
Analysis of the KIFAP3 promoter using transcription element
search system (TESS) (37) revealed that the KIFAP3 gene lacks a
TATA box and that rs522444 lies within a putative Sp1 binding site
(log-likelihood score ? 12) (Fig. 3C). Moreover, the C allele of
rs522444, which is in linkage disequilibrium with the lower express-
ing C allele of rs1541160, creates the putative Sp1 binding site,
the promoter region in other primates demonstrates that the G
allele is evolutionarily conserved suggesting this is the ancestral
allele and that the KIFAP3 gene is not normally regulated by a Sp1
binding site. Because Sp1 family members binding to cognate Sp1
binding sites can both repress and activate gene expression (38), we
sought to define the influence of variants of rs522444 on KIFAP3
promoter activity by subcloning the promoter region and 5? UTR
of KIFAP3 upstream of the firefly luciferase gene. The constructs
(Fig. 3D), differing only at the variant position, were transfected
into the neuroblastoma cell line SKN-AS and the resulting lucif-
erase activity was measured. This revealed that the promoter
(Fig. 3E). This result is analogous to the decreased expression of
lymphoblast tissues. We conclude that rs522444 C variant alone
suffices to attenuate expression of the KIFAP3 gene.
In a set of 4,079 DNA samples from sporadic ALS cases and
distributed across the genome and identified a SNP (rs1541160)
within the KIFAP3 gene that is associated with reduced KIFAP3
expression and longer survival. Other SNPs in the region of
curve plots percentage survival versus duration for each of the 3 genotypes
observed for rs1541160. To better visualize the differences between the geno-
the CT (green) and TT (blue) genotypes. (A) uncensored data; (B) censored data.
In B, small vertical marks superimposed on the survival curve show censored
points at which individuals were lost to further assessment.
Influence of alleles of rs1541160 in KIFAP3 on survival in sporadic ALS.
brain tissue harboring (B) either a CC or TT genotype for rs1541160. Relative
expression of KIFAP3 was determined by real-time PCR. As shown, individuals
harboring the CC genotype display decreased expression compared with
individuals with the TT genotype. Error bars represent the 95% C.I. (C) The
sequence of the KIFAP3 promoter region is shown. The arrow indicates the
transcriptional start site. SNP rs522444 is indicated at the ?25 position. The
box represents the location of the putative Sp1 binding site. (D) The KIFAP3
promoter region and 5? UTR (633 bp) were amplified from individuals har-
boring either the CC or TT genotype and subcloned upstream of the firefly
luciferase gene. The schematic (not drawn to scale) represents the resultant
constructs, which differ only at a single base pair located at rs522444. (E) The
resultant constructs were transfected into SKN-AS cells and relative luciferase
activity was measured. The error bars represent the 95% C.I. A promoterless
vector yielded ?1% relative activity. The construct containing the G allele
Association of rs1541160/rs522444 with expression of KIFAP3. (A and
Landers et al. PNAS ?
June 2, 2009 ?
vol. 106 ?
no. 22 ?
rs1541160 trended toward association (Fig. 1B); the 4th and 10th
highest SNPs in the survival analysis were also within the KIFAP3
gene (Table S1). The failure to detect other significant SNP
genotypes is subject to multiple interpretations. Perhaps most
importantly, this suggests that there is not a single, readily detect-
able genetic variant that exerts a preponderant influence on either
the risk of developing sporadic ALS or ALS phenotypes other than
survival. In the present study, the absence of strongly associated
SNPs other than rs1541160 may reflect other factors including
inherent heterogeneity in the populations studied, locus and allelic
heterogeneity, the inability of our present study design to detect
study to detect genes of small effect. (For susceptibility studies,
assuming conservatively a genome-wide significance level of 5.0 ?
10?8and a minor allele frequency of 0.3, for genotypic relative
risks of 1.3 and 1.5 the corresponding powers are 53% and
100%). (Table S4).
That homozygosity for SNP rs1541160 confers survival variation
of ?14 months is of clinical importance in a disorder with a mean
survival of only 3–5 years. Why attenuation of KIFAP3 expression
should slow progression in ALS is unclear. With the kinesin motor
proteins KIF3A and KIF3B, KIFAP3 forms a trimeric motor
complex, KIF3, that mediates binding between the motor proteins
and their cargoes, serving multiple functions such as chromosomal
cytokinesis and anterograde transport (39, 40). Presumably, re-
duced levels of KIFAP3 modulate survival by favorably affecting
both the stoichiometry of KIFAP3 and the KIF3 complex and one
is beneficial. Heightened expression of KIFAP3 is reportedly an
early marker of disease in transgenic mutant SOD1 mice (41),
suggesting that levels of KIFAP3 reflect adverse events within
motor neurons. In human sporadic ALS, expression of 2 other
kinesin-related proteins (KIF3Ab and KIF1Bb) is reportedly re-
duced (42). A recent report documents that KIFAP3 (also de-
scribed as KAP3) binds mutant SOD1 protein, slowing axonal
KIFAP3/KAP3 colocalizes with mutant SOD1 in human motor
neurons at autopsy (43). It is conceivable that diminished expres-
sion of KIFAP3/KAP3 has a beneficial impact on the SOD1-motor
protein interaction in ALS. More generally, mutations in motor
proteins are implicated in multiple motor neuron degenerative
disorders in both humans (11, 44) and mice (45).
It is encouraging that investigations employing complex genetics
may provide fresh insight into sporadic ALS (21–24, 46). Few
genetic factors that modify ALS survival are reported (19, 20, 47);
none were identified in the previous ALS genome analyses (21–24,
46). The identification of KIFAP3 as a determinant of progression
rate of sporadic ALS is therefore promising; insights into this
pathway may provide new targets for therapies to slow this devas-
or modifying its interactions with 1 or more protein binding partners.
Materials and Methods
Genotypes were obtained from 3 sources in the U.S. (917 ALS, 912 controls)
and 3 in Europe (904 ALS, 1,346 controls) (Table S5). To maximize the power
of this study, we combined these into a single set of cases and controls (36).
Duration information was obtained from 4 of the sites. A set of 307,776 SNPs
common to all sites was used for this analysis. Multiple quality control mea-
sures were applied to the set of DNAs and SNPs. 10,360 SNPs were eliminated
because they were not in Hardy–Weinberg equilibrium (P ? 10?6in controls)
or demonstrated call rates ?0.95 or minor allele frequencies ?0.001. An
additional 9,059 SNPs were eliminated by tests for divergence of cases and
control call rates and for nonrandom missing genotype data (to determine
whether genotypes are missing with respect to the true genotype as defined
rates were ?95%; if genotypes revealed duplicate samples, relatedness (pro-
portion of genome IBD ?0.2), or excess homozygosity or heterozygosity
the genotypically-assessed gender. The sample set was additionally subjected
to stratification analysis; based on the distribution of pairwise genome-wide
identity-by-state distances, we applied complete linkage hierarchical cluster
analysis and classical multidimensional scaling. As a result, 72 outlier samples,
defined as 3 standard deviations from the group mean, were eliminated,
leaving the cases and controls as in Table S5. For the final analysis, 288,357
SNPs were evaluated (Table S6). After applying quality control metrics to the
full set of DNAs and SNPs (SI Methods), 288,357 SNPs remained that were
Genotypic and phenotypic data are available through the dbGaP database at
the National Center for Biotechnology Information web site (www.ncbi.nlm-
.nih.gov/UH). Genotypes were used for the analysis of 4 SALS phenotypes:
susceptibility, site of onset, age of onset and survival of disease. Multiple
cohorts (see SI Methods).
ACKNOWLEDGMENTS. We dedicate this article to the memories of Frieda R.
Abraham, Donna Roberts, and Jennifer Estess. We thank Daniel Mirel (Broad
Institute) for all of his assistance, all of the clinicians who assisted in recruiting
support and use of the Hewlett-Packard computing facilities. The Boston-based
investigations were supported by the ALS Therapy Alliance, Project ALS, the
ALS Research Foundation (R.H.B., J.E.L., D.M.M.-Y., I.R.-L, T.J.K.), the ALS Family
Charitable Foundation, the American Academy of Neurology Foundation/ALS
Association (A.-M.W.), the National Institute of Neurological Disorders and
Stroke (R.H.B., T.J.K.), the Harvard Business School Class of 2007, the Yamner
family, Avichai Kremer, Guy Yamen, Nate Boaz, and Professor Robert S. Kaplan.
London-based investigations were supported by the Motor Neurone Disease
Council (A.A.-C.), the Wellcome Trust (P.N.L. and C.E.S.), and Psychiatry Research
Sante ´ et de la Recherche Me ´dicale, GIS-Institut des Maladies Rares and the
Ministere de l’Enseignement Superieur et de la Recherche. V.S. was partially
suported by a donation of the Peviani Family. We thank DNA banks maintained
by the Coriell Institute (with support from the National Institute of Neurological
at the National Institute of Neurological Disorders and Stroke Human Genetics
tigations in The Netherlands were supported by The Netherlands Organisation
for Scientific Research, the ‘‘Prinses Beatrix Fonds,’’ the Kersten Foundation, the
Adessium Foundation, J. R. van Dijk and VSB Fonds (L.H.v.d.B.). This work was
supported by Howard Hughes Medical Institute (P.S. and H.R.H.), the Packard
Center for ALS Research (J.D.G. and M.P.), The Muscular Dystrophy Association
(U.S.) (S.C. and O.H.), and National Center for Research Resources Grant U54
is Howard Hughes Medical Institute Investigator.
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