Case# Monnier 2:3, male, French

CasePresentingHPOs HP:0002047

CasePreviousTesting N/A

CaseMethod1 sequenced all cDNA

CaseGenotypingMethod single strand conformation analysis and sequencing

Variant NM_000069.2(CACNA1S):c.3257G>A (p.Arg1086His)

ClinVar 17626

gnomAD 0.00002978 link<br /> gnomAD did not include the G>A mutation, only C>T but it was the same amino acid change

VariantEvidence N/A, all of these case studies claim ICVT verifies the phenotype but that doesn't seem like enough to pass for variant evidence since it is tested in patient muscle tissue.

CaseAddInfo Family not counted, too few segregations for the R1086H variant.

NonPatientCell Mouse cell line, EFO:0002887 "Myotubes of the homozygous CaVα1S-null (dysgenic) mouse cell line GLT were cultured on 35-mm culture dishes as described previously (32)."

GeneAlteration Mouse GLT line myotubes were transfected with GFP-tagged α1S cDNA constructs at the onset of myoblast fusion using FuGENE 6 transfection reagent according to the manufacturer's protocols (Roche Diagnostics).

FunctionalEvidence Fig 3: faster voltage-gated activation than WT. Fig 6: increased release of calcium ions in the presence of caffeine vs WT.

FAVariants NM_000069.2(CACNA1S):c.4060A>T (p.Thr1354Ser), ClinVar ID: 161208

Family#PresentingHPOs HP:0002047

CountryOrigin Italy

Ethnicity NonHispanic

Race White

Family#PreviousTesting no

Family#Method1 PCR

Affected 7

Unaffected 7

Segregations 5

PublishedLOD l.76

eLOD 1.5

ProbandLabel Pirone 2:1, male

Variant NM_000069.2(CACNA1S):c.4060A>T (p.Thr1354Ser)

ClinVar 161208 link

Case# Monnier 3:12, died age 30, male, French

CasePresentingHPOs HP:0001649, HP:0006682, HP:0012416, HP:0001695, HP:0001945

CasePreviousTesting N/A

CaseMethod1 PCR

CaseGenotypingMethod PCR and sequencing gel for CACNL1A3.PCR1 amplimer

Variant NM_000069.2(CACNA1S):c.3257G>A (p.Arg1086His)

ClinVar 17626

gnomAD 0.00002978 link

VariantEvidence N/A

Monnier VE2GR

Family#PresentingHPOs HP:0002047

CountryOrigin France

Ethnicity NonHispanic/Latino

Race White

Family#PreviousTesting Reverse transcription–PCR

Family#Method1 PCR

Affected 11

Unaffected 6

Segregations 9

PublishedLOD 4.38 eLOD 2.7

ProbandLabel Monnier 3:12

Variant NM_000069.2(CACNA1S):c.3257G>A (p.Arg1086His)

ClinVar 17626

gnomAD 0.00002978 link

NonPatientCell Mouse cell line, EFO:0002887 "Myotubes generated from the homozygous dysgenic (mdg/mdg) mouse cell line GLT were cultured as described previously (44)."

GeneAlteration GFP tagged point mutation transfected into mouse cells

FunctionalEvidence Variant increased channel sensitivity to caffeine/voltage (fig. 2)

FAVariants NM_000069.2(CACNA1S):c.3257G>A (p.Arg1086His), ClinVar ID: 17626

NM_000445.4(PLEC):c.3719G>A (p.Arg1240His), ClinVar ID: 448061