Neurobiology of Aging 33 (2012) 210.e9 –210.

e10
www.elsevier.com/locate/neuaging

Mutation analysis of the optineurin gene in familial amyotrophic

lateral sclerosis
Jennifer A. Solskia, Kelly L. Williamsa,b, Shu Yanga, Garth A. Nicholsona,b,c, Ian P. Blaira,b,*
a
Northcott Neuroscience Laboratory, ANZAC Research Institute, Sydney, 2139, Australia
b
Sydney Medical School, the University of Sydney, Sydney, 2006, Australia
c
Molecular Medicine Laboratory, Concord Hospital, Concord, 2139, Australia
Received 9 August, 2011; received in revised form 7 September 2011; accepted 16 September 2011

Abstract

Mutations in the optineurin gene (OPTN) have been reported in rare familial and sporadic amyotrophic lateral sclerosis (ALS) cases. It
is yet to be established whether mutations segregate with dominantly inherited familial ALS. We therefore performed mutation analysis in
a cohort of 96 autosomal dominant ALS families. A novel heterozygous nonsynonymous variant (c.218C ⬎ T, S73L) was identified in one
patient; however, analysis in the extended pedigree demonstrated that this variant was inherited from an unaffected parent. The variant was
absent in 480 control individuals. The affected serine residue is highly conserved and its substitution is predicted to alter phosphorylation.
Despite this, our evidence indicates that this variant is unlikely to play a pathogenic role in the disease. Cell and animal models will be
required to functionally support the pathogenic role of OPTN mutations.
© 2012 Elsevier Inc. All rights reserved.

Keywords: Amyotrophic lateral sclerosis; Optineurin; Gene; Mutation

1. Introduction from consanguineous marriages, and a heterozygous mis-

sense mutation. Other OPTN variants have since been
Amyotrophic lateral sclerosis (ALS) is a neurodegenera-
identified in both familial and sporadic ALS Patients (van
tive disorder caused by the loss of motor neurons in the
Blitterswijk et al., 2011). It is yet to be established
brain and spinal cord leading to paralysis of voluntary
whether mutations segregate with dominantly inherited
muscles. ALS is usually rapidly progressive, frequently
familial ALS. Therefore we performed mutation analysis
leading to death within 2 to 5 years of onset. Around 10%
of OPTN in an Australian autosomal dominant familial
of ALS cases are familial, most with autosomal dominant
ALS cohort.
inheritance. Genetic analyses of ALS families have identi-
fied several genes, including SOD1, TARDBP, and FUS, but
these account for only approximately 20 –25% of familial
cases (Ticozzi et al., 2011). 2. Results and discussion
Mutations in the optineurin gene (OPTN) are known to
Methods are available as online Supplementary material.
cause primary open-angle glaucoma (POAG) (Rezaie et
To examine OPTN for mutations, all 13 coding exons of
al., 2002). More recently, OPTN mutations were identi-
OPTN were sequenced in an affected individual from 96
fied in Japanese ALS families (Maruyama et al., 2010).
autosomal dominant ALS families. A novel nonsynony-
These included a homozygous deletion, a homozygous
mous variant in exon 5 (NM_001008211, c.218C ⬎ T,
nonsense mutation where both patients were offspring
S73L) was identified in one patient (Suppl. Fig. 1, individ-
ual III:1). This variant was absent from public SNP data-
* Corresponding author. Tel: ⫹61 2 9767 9118. bases including dbSNP 132 and the 1000 Human Genomes
E-mail address: iblair@med.usyd.edu.au (I.P. Blair). Project. Using a custom TaqMan genotyping assay, this

0197-4580/$ – see front matter © 2012 Elsevier Inc. All rights reserved.
doi:10.1016/j.neurobiolaging.2011.09.023
210.e10 J.A. Solski et al. / Neurobiology of Aging 33 (2012) 210.e9 –210.e10
variant was shown to be absent in a further 480 ethnically
matched control individuals.
Analysis of protein conservation demonstrated that the
serine residue at Position 73 is highly conserved (Suppl.
Fig. 1). Bioinformatic analysis of protein phosphorylation
predicts that the variant destroys a protein kinase CKI phos-
phorylation site at residue threonine 76.
No DNA was available from the patient’s affected
mother. However, sequencing of the patient’s unaffected
father (Suppl. Fig. 1, individual II:1) revealed that he carries
the S73L variant. Because ALS in this family shows an
autosomal dominant inheritance pattern with no evidence of
consanguinity (Suppl. Fig. 1), this variant is unlikely to be
pathogenic.
To date, OPTN mutations have largely been restricted to
ALS patients of Japanese and Italian descent. Our results
indicate that OPTN mutations are not a common cause of
familial ALS in Australia.
It is typically difficult to establish segregation over mul-
tiple generations in ALS families because the disease has a
late onset, is rapidly progressive, and is fatal. When a new
ALS family is established for research, the affected parents
are often deceased and ascertainment of affected DNA sam-
ples may take decades. To date, reported variants have been
shown to be absent in study control cohorts; however, it is
yet to be established whether mutations segregate with dom-
inantly inherited familial ALS. Cell and animal models will
be required to functionally support the pathogenic role of
OPTN mutations.
Acknowledgments
This work was supported by the NHMRC of Australia
(1004670, 511941, 570957), Bill Gole fellowship (to SY)
and Peter Stearne grant from the MND Research Institute of
Australia.
Appendix. Supplementary data
Supplementary data associated with this article can be
found, in the online version, at doi:10.1016/j.neurobiolaging.
2011.09.023.
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