Functional validation of EIF2AK4 (GCN2) missense variants associated with pulmonary arterial hypertension.
Giulia EmanuelliJiaYi ZhuWei LiNicholas W MorrellStefan John MarciniakPublished in: Human molecular genetics (2024)
Pulmonary arterial hypertension (PAH) is a disorder with a large genetic component. Biallelic mutations of EIF2AK4, which encodes the kinase GCN2, are causal in two ultra-rare subtypes of PAH, pulmonary veno-occlusive disease and pulmonary capillary haemangiomatosis. EIF2AK4 variants of unknown significance have also been identified in patients with classical PAH, though their relationship to disease remains unclear. To provide patients with diagnostic information and enable family testing, the functional consequences of such rare variants must be determined, but existing computational methods are imperfect. We applied a suite of bioinformatic and experimental approaches to sixteen EIF2AK4 variants that had been identified in patients. By experimentally testing the functional integrity of the integrated stress response (ISR) downstream of GCN2, we determined that existing computational tools have insufficient sensitivity to reliably predict impaired kinase function. We determined experimentally that several EIF2AK4 variants identified in patients with classical PAH had preserved function and are therefore likely to be non-pathogenic. The dysfunctional variants of GCN2 that we identified could be subclassified into three groups: misfolded, kinase-dead, and hypomorphic. Intriguingly, members of the hypomorphic group were amenable to paradoxical activation by a type-1½ GCN2 kinase inhibitor. This experiment approach may aid in the clinical stratification of EIF2AK4 variants and potentially identify hypomorophic alleles receptive to pharmacological activation.
Keyphrases
- pulmonary arterial hypertension
- copy number
- pulmonary hypertension
- pulmonary artery
- end stage renal disease
- chronic kidney disease
- genome wide
- intellectual disability
- tyrosine kinase
- coronary artery
- dna methylation
- extracorporeal membrane oxygenation
- high resolution
- health information
- peritoneal dialysis
- social media
- patient reported outcomes
- mass spectrometry