Objective
Much of the genetic basis for Alzheimer disease (AD) is unexplained. We sought to identify novel AD loci using a unique family-based approach that can detect robust associations with infrequent variants (minor allele frequency <0.10).
Methods
We conducted a genome-wide association study in the Framingham Heart Study (FHS) (discovery) and NIA-LOAD (replication) family-based cohorts using an approach that accounts for family structure and calculates a risk score for AD as the outcome. Links between the most promising gene candidate and AD pathogenesis were explored in silico as well as experimentally in cell-based models and in human brain.
Results
Genome-wide significant association was identified with a PLXNA4 SNP (rs277470) located in a region encoding the semaphorin-3A (SEMA3A) binding domain (meta-analysis p value [meta-P]=4.1×10−8). A test for association with the entire region was also significant (meta-P=3.2×10−4). Transfection of SH-SY5Y cells or primary rat neurons with full-length PLXNA4 (TS1) increased tau phosphorylation when stimulated by SEMA3A. The opposite effect was observed when transfected with shorter isoforms (TS2 and TS3). However, transfection of any isoform into HEK293 cells stably expressing APP did not result in differential effects on APP processing or Aβ production. Late-stage AD cases (n=9) compared to controls (n=5) had 1.9-fold increased expression of TS1 in cortical brain tissue (P=1.6×10−4). Expression of TS1 was significantly correlated with the Clinical Dementia Rating score (ρ=0.75, P=2.2×10−4), plaque density (ρ=0.56, P=0.01) and Braak stage (ρ=0.54, P=0.02).
Interpretation
Our results indicate that PLXNA4 has a role in AD pathogenesis through isoform-specific effects on tau phosphorylation.