Table 3.

Summary of variants of each patient and the corresponding amino acid change, American College of Medical Genetics and Genomics (ACMG) criteria met and Combined Annotation Dependent Deletion (CADD) for each variant.

Patient/s	Variant	Amino Acid Change	ACMG classification	ACMG criteria met	*CADD Score
1	de novo c.879C>G; p.I293M	Isoleucine is a nonpolar, non-charged, branched chain amino acid, while methionine is a nonpolar, non-charged, sulfur-containing amino acid with no branched chain characteristics. This substitution is a non-conservative change in a well-conserved residue. In silico analysis, including SNAP2, predicts that this variant likely impacts the secondary structure of its respective protein.	Pathogenic	PM2, PP2, PP5, PS2	25.4
2	de novo c.2998G>A; p.Glu1000Lys	Glutamate is a negatively charged amino acid with high tendency for interactions, while lysine is a positively charged and basic amino acid. This substitution is a non-conservative change in a well-conserved residue. In silico analysis predicts that tliis variant likely impacts the secondary structure of its respective protein.	Likely pathogenic	PM2, PP3, PP2, PS2	32
3	c.1017+5G>A paternally inherited, father has hemiplegia migraine	Splicing mutation in the ATP1A2 gene. Contributing to the pathogenicity of the c. 1017+5G>A splicing variant is the fact that the patient’s father has the same variant and has hemiplegic migraine. When examining the splicing change of the intron. the effect of +5G>A is expected to impact the splice efficiency and thus reduce the cellular level of the correctly spliced mRNA.	VUS	PM2, PP5, PP3	22.5
4	de novo status unknown c.2426T>G; p.Leu809Arg	Leucine is a nonpolar, non-charged, branched chain amino acid, while arginine is a positively charged and basic amino acid. This substitution is a non-conservative change in a well-conserved residue. In silico analysis predicts that this variant likely impacts the secondary structure of its respective protein.	VUS	PM2, PP3, PP2	25.3
5, 6, 7	de novo c.2438T>A; Met813Lys	Methionine is a nonpolar, non-charged, sulfur containing amino acid, while lysine is a positively charged and basic amino acid. This substitution is a non-conservative change in a well-conserved residue. In silico analysis predicts that this variant likely impacts the secondary structure of its respective protein.	Pathogenic	PM2, PP5, PP3, PP2, PS2	27

^*CADD score >20 indicates that these variants are among the 1% most delirious substitutions in humans