Submissions for variant NM_000527.5(LDLR):c.1586+5G>A

dbSNP: rs781362878

Total submissions: 14

Submitter	RCV	SCV	Clinical significance	Condition	Last evaluated	Review status	Method	Comment
LDLR-LOVD, British Heart Foundation	RCV000237174	SCV000295512	likely pathogenic	Hypercholesterolemia, familial, 1	2016-03-25	criteria provided, single submitter	literature only
Centre de Génétique Moléculaire et Chromosomique, Unité de génétique de l'Obésité et des Dyslipidémies, APHP, GH Hôpitaux Universitaires Pitié-Salpêtrière / Charles-Foix	RCV000237174	SCV000503370	uncertain significance	Hypercholesterolemia, familial, 1	2016-12-16	criteria provided, single submitter	clinical testing	subjects mutated among 2600 FH index cases screened = 2
U4M - Lille University & CHRU Lille, Université de Lille - CHRU de Lille	RCV000237174	SCV000583852	likely pathogenic	Hypercholesterolemia, familial, 1	2017-03-30	criteria provided, single submitter	clinical testing
Fundacion Hipercolesterolemia Familiar	RCV000237174	SCV000607609	uncertain significance	Hypercholesterolemia, familial, 1	2016-03-01	criteria provided, single submitter	research
Women's Health and Genetics/Laboratory Corporation of America, LabCorp	RCV000587894	SCV000697204	uncertain significance	not provided	2017-04-25	criteria provided, single submitter	clinical testing	Variant summary: The LDLR c.1586+5G>A variant involves the alteration of a conserved intronic nucleotide that is 5 nucleotides away from the exon-intron junction. 4/5 splice prediction tools predict a significant impact on normal splicing and ESE finder predicts that the variant may cause an SRp55 ESE site to be introduced at the locus. One functional study has shown that the variant causes two aberrant splicing transcripts in several heterozygous patient cell lines and that the variant segregates with disease within two families, although several family members are discordant. This study also showed that LDL receptor was not properly trafficked to the cell surface in transfected COS7 cells (Jensen_CG_1999). However, a second functional study also detected aberrant splicing, but assessed LDL receptor protein function in heterozygous patient-derived lymphoblast cell lines and found that ~75% or greater of LDLR was at the cell surface and LDL was internalized similarly to controls, suggesting the variant transcripts do not have a significant biological impact (Holla_MGM_2009). Additionally, several studies have cited the variant in hypercholesterolemia patients, though cosegregation data is lacking in most cases. Based on the literature data, the variant may represent a mild mutation and may have a potentiated effect in compound heterozygosity with a truly pathogenic mutation, but have a less severe impact in heterozygous patients. This variant was found in the large control database ExAC at a frequency of 0.000034 (4/117662 control chromosomes), which does not exceed the estimated maximal expected allele frequency of a pathogenic LDLR variant (0.0010005). Multiple clinical diagnostic laboratories/reputable databases have classified this variant with conflicting interpretations, including likely pathogenic and uncertain significance. Taken together, this variant is classified as VUS until more conclusive family segregation data and/or control data can be assessed.
Invitae	RCV001044315	SCV001208106	pathogenic	Familial hypercholesterolemia	2023-10-28	criteria provided, single submitter	clinical testing	This sequence change falls in intron 10 of the LDLR gene. It does not directly change the encoded amino acid sequence of the LDLR protein. RNA analysis indicates that this variant induces altered splicing and likely results in a shortened protein product. This variant is present in population databases (rs781362878, gnomAD 0.02%). This variant has been observed in individuals with hypercholesterolemia (PMID: 7635461, 10668928, 17539906, 19208450, 19446849, 25463123). This variant is also known as c.1592+5G>A. ClinVar contains an entry for this variant (Variation ID: 251909). Variants that disrupt the consensus splice site are a relatively common cause of aberrant splicing (PMID: 17576681, 9536098). Studies have shown that this variant results in skipping of 10, but is expected to preserve the integrity of the reading-frame (PMID: 10668928, 19208450). This variant disrupts the c.1586+5 nucleotide in the LDLR gene. Other variant(s) that disrupt this nucleotide have been determined to be pathogenic (PMID: 17964958; Invitae). This suggests that this nucleotide is clinically significant, and that variants that disrupt this position are likely to be disease-causing. For these reasons, this variant has been classified as Pathogenic.
Brunham Lab, Centre for Heart and Lung Innovation, University of British Columbia	RCV000237174	SCV001432658	uncertain significance	Hypercholesterolemia, familial, 1	2019-06-05	criteria provided, single submitter	research
GeneDx	RCV000587894	SCV001789378	likely pathogenic	not provided	2023-03-27	criteria provided, single submitter	clinical testing	Not observed at significant frequency in large population cohorts (gnomAD); While mRNA studies confirm the presence of one or more alternatively-spliced transcripts, functional studies at the protein level do not agree on the effect of this variant on receptor function (Jensen et al., 1999; Holla et al., 2009); Also known as c.1592+5 G>A; This variant is associated with the following publications: (PMID: 27578104, 27821657, 25463123, 19208450, 16250003, 17539906, 19446849, 31447099, 32793292, 17964958, 30710474, 7635461, 32977124, 32660911, 33740630, 34037665, 33418990, 34456049, 35913489, 31947532, 10668928)
MGZ Medical Genetics Center	RCV000237174	SCV002581306	likely pathogenic	Hypercholesterolemia, familial, 1	2022-02-17	criteria provided, single submitter	clinical testing
Ambry Genetics	RCV002401938	SCV002705210	likely pathogenic	Cardiovascular phenotype	2018-01-15	criteria provided, single submitter	clinical testing	The c.1586+5G>A intronic variant results from a G to A substitution 5 nucleotides after coding exon 10 in the LDLR gene. This alteration has been detected in individuals with confirmed familial hypercholesterolemia (FH) and in FH cohorts (Fouchier SW et al. Hum. Mutat., 2005 Dec;26:550-6; Taylor A et al. Clin. Genet., 2007 Jun;71:561-8; Guardamagna O et al. J. Pediatr., 2009 Aug;155:199-204.e2; Mollaki V et al. Atherosclerosis, 2014 Dec;237:798-804). Segregation with hypercholesterolemia has been observed with this alteration (reported as c.1592+5G>A); of note, in at least one family some heterozygotes had normal or near normal cholesterol levels at the time of testing (Jensen HK et al. Clin. Genet., 1999 Nov;56:378-88). RNA splicing studies have revealed exon-skipping and use of a cryptic donor site in mutant transcripts, while functional studies demonstrated some reduction in LDLR activity and conflicting membrane-trafficking results (Jensen HK et al. Clin. Genet., 1999 Nov;56:378-88; Holla ØL et al. Mol. Genet. Metab., 2009 Apr;96:245-52). This nucleotide position is highly conserved in available vertebrate species. Using the BDGP and ESEfinder splice site prediction tools, this alteration is predicted to abolish the native splice donor site; however, direct evidence is unavailable. Based on the majority of available evidence to date, this variant is likely to be pathogenic.
Illumina Laboratory Services, Illumina	RCV000237174	SCV002762690	likely pathogenic	Hypercholesterolemia, familial, 1	2022-04-29	criteria provided, single submitter	clinical testing	The LDLR c.1586+5G>A variant, also known as c.1592+5G>A, occurs in a splice region and results in the substitution of a guanine at nucleotide position c.1586+5 with an adenine. Across a selection of the available literature, the c.1586+5G>A variant has been identified in at least seven individuals with clinically diagnosed familial hypercholesterolemia and was shown to segregate with the disorder (Ekstrom et al. 1995; Jensen et al. 1999; Fouchier et al. 2005; Mollaki et al. 2016; Bertolini et al. 2020; Meshkov et al. 2020; Lamiquiz-Moneo et al. 2021; Leren et al. 2021; Sturm et al. 2021). Another variant at the same nucleotide position, has been reported as c.1592+5G>C in a heterozygous state in three individuals with familial hypercholesterolemia (Yang et al. 2007; Defesche et al. 2017). This variant is reported in the Genome Aggregation Database in six alleles at a frequency of 0.000196 in the South Asian population (version 2.1.1). Analysis of mRNA from patient cells with sequence analysis found that the c.1586+5G>A variant results in alternate RNA splicing mechanisms that cause the activation of a cryptic splice site within intron 10 or skipping of exon 10 and causes defects in receptor transport in vitro (Jensen et al. 1999). Based on the available evidence, the c.1586+5G>A variant is classified as likely pathogenic for familial hypercholesterolemia.
Color Diagnostics, LLC DBA Color Health	RCV001044315	SCV004358535	pathogenic	Familial hypercholesterolemia	2023-10-16	criteria provided, single submitter	clinical testing	This variant causes a G to A nucleotide substitution at the +5 position of intron 10 of the LDLR gene. Splice site prediction tools predict that this variant may have a significant impact on RNA splicing. Molecular studies of the impact of this variant on RNA splicing demonstrated two aberrant mRNAs due to either in-frame skipping of exon 10 or the activation of a cryptic splice site in intron 10 inserting 66 intronic base pairs (PMID: 10668928). Additionally, this variant was shown to cause aberrant LDL receptor trafficking in transfected COS7 cells (PMID: 10668928). This variant has been reported in over 10 individuals affected with familial hypercholesterolemia (PMID: 7635461, 10668928, 16250003, 17539906, 19446849, 25463123, 31947532, 32660911, 32977124, 33418990, 34037665, 36960729). It has been shown that this variant segregates with disease in multiple affected individuals in one family (PMID: 10668928). This variant has been identified in 8/250846 chromosomes in the general population by the Genome Aggregation Database (gnomAD). Loss of LDLR function is a known mechanism of disease (clinicalgenome.org). A different variant occurring at the same position, c.1586+5G>C, is known to be pathogenic (ClinVar variation ID: 440652), indicating that c.G nucleotide at this position is important for normal RNA splicing. Based on the available evidence, this variant is classified as Pathogenic.
Laboratorium voor Moleculaire Diagnostiek Experimentele Vasculaire Geneeskunde, Academisch Medisch Centrum	RCV000237174	SCV000606461	pathogenic	Hypercholesterolemia, familial, 1		no assertion criteria provided	research
Stanford Center for Inherited Cardiovascular Disease, Stanford University	RCV000587894	SCV000925125	pathogenic	not provided	2017-12-11	no assertion criteria provided	provider interpretation	c.1586+5G>A in intron 10 of the LDLR gene (NM_00527; chr19-11224443-G-A) SCICD Classification: pathogenic variant based on strong case data, strong segregation data and strong functional data. We do feel it is suitable for assessing risk in healthy relatives ("predictive genetic testing"). Gene-level evidence: LDLR: LDL receptors are located on the surface of the liver and play an important role in LDL recycling. Pathogenic variants in LDLR account for 80% of cases of familial hypercholesterolemia. Both missense and truncating/frameshift variants can be pathogenic. Case data (not including our patient): at least 25 cases, plus other family members. · ClinVar: § LDLR-LOVD, British Heart Foundation - likely pathogenic - 2 out of 2600 patients with FH have this variant § Centre de Génétique Moléculaire et Chromosomique, Unité de génétique de l'Obésité et des Dyslipidémies,APHP, GH Hôpitaux Universitaires Pitié-Salpêtrière / Charles-Foix - VUS § U4M - Lille University & CHRU Lille,Université Lille 2 - CHRU de Lille - likely pathogenic § Fundacion Hipercolesterolemia Familiar - VUS § Never seen at Invitae § Color Genomics has this variant in their "John Doe" report: they indicate that many algorithms predict a significant impact on the intron 10 donor · Laboratorium voor Moleculaire Diagnostiek Experimentele Vasculaire Geneeskunde,Academisch Medisch Centrum - pathogenic · Cases in the literature: · Yang et al 2007: 87 Taiwanese individuals with FH from 30 unrelated families: seen in 2 families. Authors report that this variant may result in the skipping of exon 10. This variant segregated with disease in family members; however, the number of family members with which it segregates is unknown. · Ekström et al 1994: this variant (noted as c.1596+5G>A in this paper) was detected in 1 out of 7 children with FH. · Fouchier et al 2005: 1 out of 325 patients in the Dutch cohort · Taylor et al 2007: Found this variant in one out of 400 patients with FH. This particular patient had possible FH. · Guardamagna et al 2009: This variant was seen in one patient of the following cohort: 264 children from 201 families, 148 affected parents and 100 unaffected siblings. · Mollaki et al 2014: Variant detected in 1 out of 561 patients from 262 families · Jensen et al 1999: This variant is known as c.1596+5G>A in this paper. This variant was present in a 36yo Danish man presenting with tendon xanthomas, severe CAD, CABG at 24yo and very high LDL cholesterol. This man had another variant as well - p.T383P. This variant segregated with disease in his family members (pedigree A, FH-DK 8). Another family had only this variant and it segregated with disease. From the first family, this variant segregated with 8 affected individuals. From the second family, this variant segregated in 4 individuals. Segregation data: See above, Jensen et al 1999 Functional data: Jenner et 1999: sequence analysis of cDNA indicates that either i) a cryptic splice site is activated or ii) exon 10 is skipped: i) the donor splice site at the +5 position is activated to create a cryptic splice site which results in an in-frame insertion of 22 amino acids; ii) in-frame deletion of 75 amino acids in the EGF precursor homology domain of the LDL receptor protein. The authors predicted about 50% of these variants were spliced correctly, and 50% spliced incorrectly. To measure the amount of LDL receptor protein on the surface of the cells, fluorescence was used. Fluorescence of both of these mutant cell lines was markedly reduced in both of these cell lines compared to wildtype. Splice data (splice variants only): Buratti et al 2007: "...point mutations leading to cryptic 5' splice site activation were most common in the first intron nucleotide, followed by the fifth nucleotide. Substititons at position +5 were exclusively G>A transitions... The frequency of point mutations at position +5 was significantly higher than observed in[HGMD] suggesting that alterations of this position are particularly prone to aberrant splicing, possibly due to a requirement for sequential interactions with U1 and U6 snRNAs." Functional data above by Jensen and colleagues corroborates predictions that this variant impacts splicing. Conservation data: The G at position c.1586+5 is conserved among species. Population data: Highest MAF in South Asian population: 0.0195%. The variant was reported online in 8 of 122,672 individuals in the Genome Aggregation Consortium Dataset (gnomAD; http://gnomad.broadinstitute.org/), which currently includes variant calls on >140,000 unrelated individuals of African, Asian, European, Ashkenazi, Latino descent. Specifically, the variant was observed in 6 of 15,376 individuals of South Asian descent (MAF=0.0195%) and 2 of 55,674 individuals of European descent. The phenotype of those individuals is not publicly available. The dataset is comprised of multiple cohorts, some of which were recruited from the general population, others were enriched for common cardiovascular disease.