PM2 (supporting): This variant is absent from gnomAD v2.1 and present in gnomAD v4.1 at extremely low frequency (AF=0.000062%, 10/1,610,852 alleles, 0 homozygotes), well below the PM2 threshold of less than 0.1%.¹ BS2 (supporting): This variant is observed in 10 heterozygous individuals in gnomAD v4.1. TSC is a dominant disorder with near-complete penetrance and early childhood onset; observation in a population database of presumably healthy adults is inconsistent with a fully penetrant pathogenic variant.² BP4 (supporting): BayesDel score 0.161 is in the benign range. REVEL score 0.496 is borderline (just below 0.5 pathogenic threshold). SpliceAI max delta 0.02 predicts no splicing impact. Multiple computational lines of evidence suggest no deleterious effect.³ PVS1 not applicable: Variant is a missense substitution (p.Pro341Leu), not a null variant eligible for PVS1 assessment.⁴ PM1 not met: This variant does not lie in a statistically significant mutational hotspot per cancerhotspots.org. PM5 not met: No same-residue pathogenic missense comparator variants were identified.⁵ PS3 not met: No functional studies have been reported for this variant. OncoKB reports Unknown Oncogenic Effect.⁶ PP3 not met: REVEL 0.496 is borderline; BayesDel 0.161 is benign; SpliceAI max delta 0.02 shows no splice impact. In silico tools do not converge on a damaging prediction.⁷ PP5 not met: ClinVar review status is 1-star (criteria provided, single submitter), not 3-star expert panel. Classification is Uncertain significance with mixed submitters.⁸ BP6 not met: ClinVar review status is 1-star, not 3-star expert panel. A single submitter classifies as Benign, insufficient for BP6.⁹ No publications reviewed contained variant-specific evidence for NM_000368.4:c.1022C>T. All PMIDs are guideline/review papers that do not mention this variant.¹⁰