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Ann Neurol. 2016 Nov;80(5):741-753. doi: 10.1002/ana.24782. Epub 2016 Oct 19.

Disruption of a miR-29 binding site leading to COL4A1 upregulation causes pontine autosomal dominant microangiopathy with leukoencephalopathy.

Author information

1
Inserm U1161, Genetics and Physiopathology of Cerebrovascular Diseases, Paris, France.
2
Inserm U1161, Paris7 Diderot University, Sorbonne Paris Cité, Paris, France.
3
AP-HP Lariboisière Neurology Department, CERVCO Reference Center for Rare Vascular Diseases of the Eye and Brain, Paris, France.
4
Neurology department, CHRU of Lille, Lille 2 University, Inserm U1171, Lille, France.
5
AP-HP Lariboisière Molecular Genetics Department, CERVCO Reference Center for Rare Vascular Diseases of the Eye and Brain, Paris, France.
6
Neurology Department, Saint Philibert Hospital, Lomme, France.
7
Neurology Department, Gonesse Hospital, Gonesse, France.
8
Neurology Department, Henri Mondor Hospital, Creteil, France.
9
Neurology Department, Valenciennes Hospital, Valenciennes, France.
10
Neurology Department, Coulommiers Hospital, Coulommiers, France.
11
Institute of Neuropathology, University Medical Center, Hamburg, Germany.
12
Department of Neurology, University Hospital Schleswig-Holstein, Kiel, Germany.
13
Neurology Department, Agen Hospital, Agen, France.
14
CMAbio3, Applied Microscopy and Biology Center, University of Caen, Caen, France.
15
Institute of Cardiovascular and Medical Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United Kingdom.
16
Pathology Department, CHRU of Caen, University of Caen Normandy, Inserm, U1075.

Abstract

OBJECTIVE:

Cerebral small vessel disease (cSVD) is a heterogeneous group of disorders. Screening of known cSVD genes identifies the causative mutation in <15% of familial cSVD cases. We sought to identify novel causes of cSVD.

METHODS:

We used linkage analysis and exome sequencing to identify the causal mutation in a French cSVD family. The identified candidate gene was then screened in 202 cSVD unrelated probands, including 1 proband from the first reported pontine autosomal dominant microangiopathy with leukoencephalopathy (PADMAL) family. Sanger sequencing was used to confirm variants in all mutated probands and analyze their segregation in probands' relatives. Mutation consequences were assessed with luciferase reporter assays and real-time quantitative polymerase chain reaction (RT-qPCR).

RESULTS:

A candidate heterozygous variant located in a predicted miR-29 microRNA binding site, within the 3' untranslated region of COL4A1, was identified in the large French cSVD family. Five additional unrelated probands, including the PADMAL proband, harbored heterozygous variants in this microRNA binding site. Variants cosegregated with the affected phenotype, and cumulative logarithm of odds score reached 6.03, establishing linkage to this locus. A highly significant difference was observed when comparing the number of variants within this binding site in cases and controls (p = 1.77 × 10E-12). RT-qPCR analyses of patients' primary fibroblasts and luciferase reporter assays strongly favor an upregulation of COL4A1 mediated by disruption of miR-29 binding to its target site. Magnetic resonance imaging features were characterized by the presence of multiple pontine infarcts in all symptomatic mutation carriers.

INTERPRETATION:

Mutations upregulating COL4A1 expression lead to PADMAL, a severe early onset ischemic cSVD, distinct from the various phenotypes associated with COL4A1 missense glycine mutations. Ann Neurol 2016;80:741-753.

PMID:
27666438
DOI:
10.1002/ana.24782
[Indexed for MEDLINE]

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