rs148044781

Positions:

• chr15-42401791-T-C

Variant summary

Our verdict is Likely pathogenic. Variant got 7 ACMG points: 7P and 0B. PM1 PP3_Strong PP5

The NM_000070.3(CAPN3):​c.1505T>C​(p.Ile502Thr) variant causes a missense change involving the alteration of a conserved nucleotide. The variant allele was found at a frequency of 0.000424 in 1,613,672 control chromosomes in the GnomAD database, with no homozygous occurrence. In-silico tool predicts a pathogenic outcome for this variant. 12/21 in silico tools predict a damaging outcome for this variant. Variant has been reported in ClinVar as Conflicting classifications of pathogenicity (no stars).

• Frequency:
  • Genomes: 𝑓 0.00024 (0 hom., cov: 32)
  • Exomes 𝑓: 0.00044 (0 hom.)
• Consequence: CAPN3 NM_000070.3 missense
• Clinical Significance: Conflicting classifications of pathogenicity criteria provided, conflicting classifications P:10 U:3
• Conservation: PhyloP100: 7.96

Genes affected

CAPN3 (HGNC:1480): (calpain 3) Calpain, a heterodimer consisting of a large and a small subunit, is a major intracellular protease, although its function has not been well established. This gene encodes a muscle-specific member of the calpain large subunit family that specifically binds to titin. Mutations in this gene are associated with limb-girdle muscular dystrophies type 2A. Alternate promoters and alternative splicing result in multiple transcript variants encoding different isoforms and some variants are ubiquitously expressed. [provided by RefSeq, Jul 2008]

ENSG00000258461 (HGNC:):

ACMG classification

Verdict is Likely_pathogenic. Variant got 7 ACMG points.

• PM1: In a region_of_interest Domain III (size 168) in uniprot entity CAN3_HUMAN there are 132 pathogenic changes around while only 0 benign (100%) in NM_000070.3
• PP3: MetaRNN computational evidence supports a deleterious effect, 0.952
• PP5: Variant 15-42401791-T-C is Pathogenic according to our data. Variant chr15-42401791-T-C is described in ClinVar as [Conflicting_classifications_of_pathogenicity]. Clinvar id is 282512.We mark this variant Likely_pathogenic, oryginal submissions are: {Uncertain_significance=3, Pathogenic=2, Likely_pathogenic=6}. Variant chr15-42401791-T-C is described in Lovd as [Pathogenic]. Variant chr15-42401791-T-C is described in Lovd as [Likely_pathogenic].

Transcripts

RefSeq

Gene	Transcript	HGVSc	HGVSp	Effect	#exon/exons	MANE	Protein	UniProt
CAPN3	NM_000070.3	c.1505T>C	p.Ile502Thr	missense_variant	11/24	ENST00000397163.8	NP_000061.1
CAPN3	NM_024344.2	c.1505T>C	p.Ile502Thr	missense_variant	11/23		NP_077320.1
CAPN3	NM_173087.2	c.1361T>C	p.Ile454Thr	missense_variant	10/21		NP_775110.1

Ensembl

Gene	Transcript	HGVSc	HGVSp	Effect	#exon/exons	TSL	MANE	Protein	Appris	UniProt
CAPN3	ENST00000397163.8	c.1505T>C	p.Ile502Thr	missense_variant	11/24	1	NM_000070.3	ENSP00000380349.3
ENSG00000258461	ENST00000495723.1	n.*1301T>C		non_coding_transcript_exon_variant	15/26	2		ENSP00000492063.1
ENSG00000258461	ENST00000495723.1	n.*1301T>C		3_prime_UTR_variant	15/26	2		ENSP00000492063.1

Frequencies

GnomAD3 genomes AF: 0.000237 AC: 36 AN: 152174 Hom.: 0 Cov.: 32

Gnomad AFR AF: 0.0000724

Gnomad AMI AF: 0.00

Gnomad AMR AF: 0.0000654

Gnomad ASJ AF: 0.00

Gnomad EAS AF: 0.00

Gnomad SAS AF: 0.00

Gnomad FIN AF: 0.00

Gnomad MID AF: 0.00

Gnomad NFE AF: 0.000456

Gnomad OTH AF: 0.000478

GnomAD3 exomes AF: 0.000148 AC: 37 AN: 250192 Hom.: 0 AF XY: 0.000148 AC XY: 20 AN XY: 135284

Gnomad AFR exome AF: 0.00

Gnomad AMR exome AF: 0.000203

Gnomad ASJ exome AF: 0.00

Gnomad EAS exome AF: 0.00

Gnomad SAS exome AF: 0.00

Gnomad FIN exome AF: 0.00

Gnomad NFE exome AF: 0.000256

Gnomad OTH exome AF: 0.000163

GnomAD4 exome AF: 0.000443 AC: 648 AN: 1461498 Hom.: 0 Cov.: 33 AF XY: 0.000435 AC XY: 316 AN XY: 727010

Gnomad4 AFR exome AF: 0.0000597

Gnomad4 AMR exome AF: 0.000157

Gnomad4 ASJ exome AF: 0.00

Gnomad4 EAS exome AF: 0.00

Gnomad4 SAS exome AF: 0.00

Gnomad4 FIN exome AF: 0.00

Gnomad4 NFE exome AF: 0.000551

Gnomad4 OTH exome AF: 0.000431

GnomAD4 genome AF: 0.000237 AC: 36 AN: 152174 Hom.: 0 Cov.: 32 AF XY: 0.000121 AC XY: 9 AN XY: 74346

Gnomad4 AFR AF: 0.0000724

Gnomad4 AMR AF: 0.0000654

Gnomad4 ASJ AF: 0.00

Gnomad4 EAS AF: 0.00

Gnomad4 SAS AF: 0.00

Gnomad4 FIN AF: 0.00

Gnomad4 NFE AF: 0.000456

Gnomad4 OTH AF: 0.000478

Alfa AF: 0.000294 Hom.: 0

Bravo AF: 0.000310

TwinsUK AF: 0.000809 AC: 3

ALSPAC AF: 0.000519 AC: 2

ESP6500AA AF: 0.00 AC: 0

ESP6500EA AF: 0.000582 AC: 5

ExAC AF: 0.000157 AC: 19

Asia WGS AF: 0.000289 AC: 1 AN: 3478

EpiCase AF: 0.000491

EpiControl AF: 0.000416

ClinVar

Significance: Conflicting classifications of pathogenicity

Submissions summary: Pathogenic:10 Uncertain:3

Revision: criteria provided, conflicting classifications

Submissions by phenotype

not provided Pathogenic:4 Uncertain:1

Likely pathogenic, criteria provided, single submitter	clinical testing	Athena Diagnostics	Aug 27, 2018	The best available variant frequency is uninformative because it is below the disease allele frequency. Found in at least one symptomatic patient. Predicted to have a damaging effect on the protein. Occurs in three or more cases with a recessive pathogenic variant in the same gene. -
Uncertain significance, flagged submission	clinical testing	Eurofins Ntd Llc (ga)	May 17, 2018	- -
Likely pathogenic, criteria provided, single submitter	clinical testing	Revvity Omics, Revvity	Aug 09, 2023	- -
Likely pathogenic, criteria provided, single submitter	clinical testing	GeneDx	Sep 27, 2024	In silico analysis supports that this missense variant has a deleterious effect on protein structure/function; This variant is associated with the following publications: (PMID: 22926650, 26810512, 10330340, 30564623, 30919934, 32994280, 31555977, 35734998, 18337726, 17562833, 39258114, 38494715) -
Likely pathogenic, criteria provided, single submitter	clinical testing	CeGaT Center for Human Genetics Tuebingen	Aug 01, 2023	CAPN3: PM1, PM5, PM2:Supporting, PP3 -

Autosomal recessive limb-girdle muscular dystrophy type 2A Pathogenic:3 Uncertain:1

Likely pathogenic, no assertion criteria provided	clinical testing	Natera, Inc.	May 22, 2020	- -
Pathogenic, criteria provided, single submitter	clinical testing	Labcorp Genetics (formerly Invitae), Labcorp	Jan 22, 2024	This sequence change replaces isoleucine, which is neutral and non-polar, with threonine, which is neutral and polar, at codon 502 of the CAPN3 protein (p.Ile502Thr). This variant is present in population databases (rs148044781, gnomAD 0.02%). This missense change has been observed in individuals with clinical features of autosomal recessive CAPN3-related conditions (PMID: 10330340, 26810512, 30919934; Invitae). ClinVar contains an entry for this variant (Variation ID: 282512). Advanced modeling of protein sequence and biophysical properties (such as structural, functional, and spatial information, amino acid conservation, physicochemical variation, residue mobility, and thermodynamic stability) performed at Invitae indicates that this missense variant is expected to disrupt CAPN3 protein function with a positive predictive value of 80%. This variant disrupts the p.Ile502 amino acid residue in CAPN3. Other variant(s) that disrupt this residue have been observed in individuals with CAPN3-related conditions (PMID: 18337726), which suggests that this may be a clinically significant amino acid residue. For these reasons, this variant has been classified as Pathogenic. -
Uncertain significance, flagged submission	clinical testing	Counsyl	Mar 20, 2018	- -
Pathogenic, criteria provided, single submitter	clinical testing	Victorian Clinical Genetics Services, Murdoch Childrens Research Institute	Sep 23, 2024	Based on the classification scheme VCGS_Germline_v1.3.4, this variant is classified as Pathogenic. Following criteria are met: 0102 - Loss of function is a known mechanism of disease in this gene and is associated with recessive limb-girdle muscular dystrophy 1 (MIM#253600). Dominant negative mechanism is a suggested mechanism for dominant limb-girdle muscular dystrophy 4 (MIM#618129) associated with milder phenotypes and later age of onset (PMID: 27259757, 28881388, 32342993). (I) 0108 - This gene is associated with both recessive and dominant disease. It is predominantly reported as a recessive condition, with the dominant condition reported with only a few missense variants and one in-frame deletion (PMID: 27259757, 28881388, 31066050). (I) 0200 - Variant is predicted to result in a missense amino acid change from isoleucine to threonine. (I) 0251 - This variant is heterozygous. (I) 0304 - Variant is present in gnomAD (v2) <0.01 (41 heterozygotes, 0 homozygotes). (SP) 0309 - An alternative amino acid change at the same position has been observed in gnomAD (v2) (13 heterozygotes, 0 homozygotes). (I) 0501 - Missense variant consistently predicted to be damaging by multiple in silico tools or highly conserved with a major amino acid change. (SP) 0600 - Variant is located in the annotated calpain large subunit, domain III (DECIPHER). (I) 0710 - Other missense variants comparable to the one identified in this case have inconclusive previous evidence for pathogenicity. ClinVar contains three VUS entries for p.(Ile502Val), and p.(Ile502Ser) has also been identified in the literature in an individual with limb-girdle muscular dystrophy 2A (PMID: 18337726). (I) 0801 - This variant has strong previous evidence of pathogenicity in unrelated individuals. This variant has been observed in compound heterozygous individuals (PMID: 30919934), and in heterozygotes without a second hit (PMID: 35734998, 22926650), with limb-girdle muscular dystrophy. It has also been reported as pathogenic and likely pathogenic in ClinVar, along with some older entries with VUS classifications. (SP) 1205 - This variant has been shown to be maternally inherited by trio analysis). (I) Legend: (SP) - Supporting pathogenic, (I) - Information, (SB) - Supporting benign -

Muscular dystrophy, limb-girdle, autosomal dominant 4 Pathogenic:1

Likely pathogenic, criteria provided, single submitter

clinical testing

Baylor Genetics

Mar 14, 2024

- -

Autosomal recessive limb-girdle muscular dystrophy Pathogenic:1

Likely pathogenic, criteria provided, single submitter

clinical testing

Women's Health and Genetics/Laboratory Corporation of America, LabCorp

Dec 13, 2023

Variant summary: CAPN3 c.1505T>C (p.Ile502Thr) results in a non-conservative amino acid change located in the Peptidase C2, calpain, large subunit, domain III (IPR022682) of the encoded protein sequence. Five of five in-silico tools predict a damaging effect of the variant on protein function. The variant allele was found at a frequency of 0.00015 in 250192 control chromosomes (gnomAD). This frequency is not significantly higher than estimated for a pathogenic variant in CAPN3 causing Limb-Girdle Muscular Dystrophy, Autosomal Recessive (0.00015 vs 0.0032), allowing no conclusion about variant significance. c.1505T>C has been reported in the literature in individuals affected with Limb-Girdle Muscular Dystrophy, Autosomal Recessive (example, Lahoria_2016, Ten Dam_2019, Barp_2020, Nallamilli_2018, Richard_1999, Becker_2022). These data indicate that the variant is likely to be associated with disease. To our knowledge, no experimental evidence demonstrating an impact on protein function has been reported. The following publications have been ascertained in the context of this evaluation (PMID: 31555977, 26810512, 30564623, 10330340, 30919934, 35135626). Eight clinical diagnostic laboratories have submitted clinical-significance assessments for this variant to ClinVar after 2014. Multiple laboratories reported the variant with conflicting assessments (Pathogenic/Likely pathogenic, n=5; VUS, n=3). Based on the evidence outlined above, the variant was classified as likely pathogenic. -

CAPN3-related disorder Pathogenic:1

Likely pathogenic, no assertion criteria provided

clinical testing

PreventionGenetics, part of Exact Sciences

May 31, 2024

The CAPN3 c.1505T>C variant is predicted to result in the amino acid substitution p.Ile502Thr. This variant was reported along with a second known pathogenic CAPN3 variant in multiple individuals with a limb-girdle muscular dystrophy (LGMD) phenotype (van der Kooi AJ et al 2007. PubMed ID: 17562833, E-Table 1; Nallamilli BRR et al 2018. PubMed ID: 30564623; Ten Dam L et al 2019. PubMed ID: 30919934, Supplementary Table 1). It was also reported in an individual with a confirmed calpainopathy based on tissue pathology and Western blot analysis. No second CAPN3 variant was identified in that patient, who was was described as more mildly affected (Luo SS et al 2012. PubMed ID: 22926650). The c.1505T>C (p.Ile502Thr) variant was also reported along with a second rare CAPN3 variant in a patient with rhabdomyolysis (Lahoria R et al 2015. PubMed ID: 26810512). An alternate substitution at the same amino acid was also reported in a patient with suspected LGMD (Duno M et al 2008. PubMed ID: 18337726). CAPN3 expression was reported to be severely reduced in patients with either the p.Ile502Thr or p.Ile502Ser substitutions and a second CAPN3 variant (van der Kooi AJ et al 2007. PubMed ID: 17562833; Duno M et al 2008. PubMed ID: 18337726). This variant is reported in 0.026% of alleles in individuals of European (Non-Finnish) descent in gnomAD (http://gnomad.broadinstitute.org/variant/15-42693989-T-C). This variant is entered in ClinVar with conflicting interpretations of uncertain and likely pathogenic. Based on the collective evidence, we interpret this variant as likely pathogenic. -

Inborn genetic diseases Uncertain:1

Uncertain significance, criteria provided, single submitter

clinical testing

Ambry Genetics

Sep 16, 2014

The c.1505T>C (p.I502T) alteration is located in exon 11 (coding exon 11) of the CAPN3 gene. This alteration results from a T to C substitution at nucleotide position 1505, causing the isoleucine (I) at amino acid position 502 to be replaced by a threonine (T). This nucleotide position is conserved in available vertebrate species.This amino acid position is conserved in available vertebrate species. Based on insufficient or conflicting evidence, the clinical significance of this alteration remains unclear. -

Computational scores

Source: dbNSFP v4.3

Name	Calibrated prediction	Score	Prediction
AlphaMissense	Pathogenic	0.61
BayesDel_addAF	Pathogenic	0.28	D
BayesDel_noAF	Pathogenic	0.46
CADD	Pathogenic	27
DANN	Uncertain	1.0
DEOGEN2	Uncertain	0.71	D;.;.;D
Eigen	Pathogenic	0.86
Eigen_PC	Pathogenic	0.73
FATHMM_MKL	Pathogenic	0.98	D
LIST_S2	Pathogenic	0.99	D;D;D;D
M_CAP	Pathogenic	0.53	D
MetaRNN	Pathogenic	0.95	D;D;D;D
MetaSVM	Pathogenic	1.1	D
MutationAssessor	Pathogenic	3.7	.;H;.;H
PrimateAI	Pathogenic	0.83	D
PROVEAN	Pathogenic	-4.9	.;D;D;D
REVEL	Pathogenic	0.96
Sift	Uncertain	0.0010	.;D;D;D
Sift4G	Uncertain	0.0020	D;D;D;D
Polyphen		1.0	.;D;D;D
Vest4		1.0
MVP		0.99
MPC		0.70
ClinPred		0.95	D
GERP RS		4.9
Varity_R		0.80
gMVP		0.89

Splicing

Name	Calibrated prediction	Score	Prediction
SpliceAI score (max)		0.0		Details are displayed if max score is > 0.2

Find out detailed SpliceAI scores and Pangolin per-transcript scores at spliceailookup.broadinstitute.org

Publications

LitVar

Other links and lift over

dbSNP: rs148044781; hg19: chr15-42693989;