The c.533G>T variant in the glucokinase gene, GCK, causes an amino acid change of glycine to valine at codon 178 (p.(Gly178Val)) of NM_000162.5. GCK is defined by the ClinGen MDEP as a gene that has a low rate of benign missense variation and has pathogenic missense variants as a common mechanism of disease (PP2). This variant is predicted to be deleterious by computational evidence, with a REVEL score of 0.987, which is greater than the MDEP VCEP threshold of 0.70 (PP3). This variant is absent in gnomAD v4.1 (PM2_Supporting). This variant was identified in three unrelated probands with hyperglycemia; however, PS4_Moderate cannot be applied because this number is below the ClinGen MDEP threshold (PMID: 36257325, internal lab contributors). At least one of these individuals did have a clinical history highly specific for GCK-hyperglycemia (FBG 5.5-8 mmol/L and HbA1c 5.6 - 7.6% and OGTT with minimal increment <3mmol/L) (PP4_Moderate; Internal lab contributor). This variant segregated with hyperglycemia with two informative meioses in a single family; however, this does not meet the thresholds for PP1 set by the ClinGen MDEP (Internal lab contributor). Another missense variant at the same residue, c.533G>A p.Gly178Glu, has been interpreted as pathogenic by the ClinGen MDEP, and p.Gly178Val has a greater Grantham distance (PM5). In summary, c.533G>T meets the criteria to be classified as likely pathogenic for monogenic diabetes. ACMG/AMP criteria applied, as specified by the ClinGen MDEP (specification version 3.1.0, approved 10/10/2025): PP2, PP3, PM2_Supporting, PM5, PP4_Moderate.