Data from: Pathogenic effect of TP73 gene variants in people with amyotrophic lateral sclerosis

Objective: To identify novel disease associated loci for amyotrophic
lateral sclerosis (ALS), we utilized sequencing data and performed in
vitro and in vivo experiments to demonstrate pathogenicity of mutations
identified in TP73. Methods: We analyzed exome sequences of 87 sporadic
ALS patients and 324 controls, with confirmatory sequencing in independent
ALS cohorts of >2,800 patients.  For the top hit, TP73, a
regulator of apoptosis, differentiation, and a binding partner as well as
homolog of the tumor suppressor gene TP53, we assayed mutation effects
using in vitro and in vivo experiments.  C2C12 myoblast
differentiation assays, characterization of myotube appearance, and
immunoprecipitation of p53-p73 complexes were perform in vitro. 
In vivo, we used CRISPR/Cas9 targeting of zebrafish tp73 to assay motor
neuron number and axon morphology. Results: Four heterozygous rare,
nonsynonymous mutations in TP73 were identified in our sporadic ALS
cohort. In independent ALS cohorts, we identified an additional 19 rare,
deleterious variants in TP73.   Patient TP73 mutations
caused abnormal differentiation and increased apoptosis in the myoblast
differentiation assay, with abnormal myotube appearance.
 Immunoprecipitation of mutant ΔN-p73 demonstrated that patient
mutations hinder ΔN-p73’s ability to bind p53.  CRISPR/Cas9
knockout of tp73 in zebrafish led to impaired motor neuron development and
abnormal axonal morphology, concordant with ALS pathology. 
Conclusion: Together, these results strongly suggest that variants in TP73
correlate with risk for ALS and indicate a novel role for apoptosis in ALS
disease pathology.