The recent advances in sequencing technology
make it a relatively simple prospect to sequence the entire exome of any given
individual with an inherited disorder. What is significantly less
straightforward is how to filter this huge amount of data to find the causative
variant. One approach is to use linkage analysis in large families affected
with the disorder to hone the search for the disease variant. We describe a unique pedigree (Aus-12) that presents with dementia and
motor deficits most consistent with frontotemporal dementia-motor neuron
disease (FTD-MND), but with additional corticobasal pathology. DNA from affected Aus-12 individuals was screened and found to be
negative for mutations in all known dementia and MND genes (APP, PSEN1,
PSEN2, MAPT, GRN, VCP, CHMP2B,
SOD1, TARDBP, FUS, TAF15, C9ORF72,
OPTN). We performed genome-wide linkage analysis on 13 pedigree members,
five of whom were classed as affected. Linkage analysis provided highly suggestive evidence (maximum multipoint
LOD score of 2.9) of a locus on chromosome 16p12.1-16q12.2. Haplotype
construction of chromosome 16 revealed a shared haplotype in all affecteds
flanked by D16S3103 and D16S489 and spanning a maximum of 37.9 Mb. A smaller
suggestive disease haplotype, flanked by D16S753 and D16S3112 and spanning 24.4
Mb, is defined by recombination in an elderly unaffected individual. The Aus-12
critical region overlaps with a separate locus
on 16q12.1-q12.2 reported in two independent MND families, indicating that this
region may harbour a major locus for neurodegeneration. We performed whole-exome sequencing of five Aus-12 individuals (four
affected and one elderly unaffected). From 16,384 non-dbSNP variants identified
initially we have identified four variants present in the maximal critical
region that segregate with disease. Our study highlights the effectiveness of
combining both traditional and next-generation genetic techniques for
positional cloning of Mendelian disorders.