Dermal fibroblasts in Hutchinson-Gilford progeria syndrome with the lamin A G608G mutation have dysmorphic nuclei and are hypersensitive to heat stress

Paradisi, Mauro; McClintock, Dayle; Boguslavsky, Revekka L.; Pedicelli, Christina; Worman, Howard J.; Djabali, Karima

doi:10.1186/1471-2121-6-27

Cited by 62 publications

(38 citation statements)

References 47 publications

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“…The nuclear abnormalities observed in the p.(Arg331Gln) fibroblasts are in line with the nuclear irregularities observed in fibroblasts of other established pathogenic mutations in LMNA, thereby supporting its pathogenicity. 22,[36][37][38] Like the abnormal nuclear structures in the fibroblasts observed using immunofluorescence, ultrastructural investigation on diseased cells in cardiac tissue with EM also support pathogenicity. The convoluted shapes of the nuclei, blebs, discontinuous layer of heterochromatin, and possible enlarged nuclear pores are features commonly seen in other LMNA mutations.…”

Section: Hoorntje Et Al Mild Phenotype In Lmna P(arg331gln) Carriersmentioning

confidence: 99%

Lamin A/C -Related Cardiac Disease

Hoorntje¹,

Bollen²,

Barge-Schaapveld³

et al. 2017

Circ Cardiovasc Genet

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T he LMNA gene encodes for the intermediate filament proteins lamin A and C. LMNA mutations are associated with a wide spectrum of phenotypes ranging from progeroid syndromes, muscular disease, and lipodystrophy to isolated cardiac disease (dilated cardiomyopathy [DCM] and conduction disorders) and phenotypes consisting of combinations of these different features.1 Although their precise role is unknown, LMNA proteins are believed to play an important role in the structural integrity of the cell nucleus and in gene regulation. 2LMNA is one of the genes most frequently involved in genotyped DCM.3 Sinus node dysfunction, atrioventricular conduction disorders, and supraventricular and ventricular arrhythmias often precede or accompany DCM.4 LMNArelated cardiac disease is associated with a high incidence of major cardiac events like sudden cardiac death, appropriate implantable cardioverter-defibrillator (ICD) therapy, or endstage heart failure. DCM patients with an LMNA mutation are, in general, believed to have a poor prognosis compared with non-LMNA mutation DCM patients. 5,6 Currently, with all the new DNA-sequencing technologies implemented in routine patient care, increasing numbers Background-Interpretation of missense variants can be especially difficult when the variant is also found in control populations. This is what we encountered for the LMNA c.992G>A (p.(Arg331Gln)) variant. Therefore, to evaluate the effect of this variant, we combined an evaluation of clinical data with functional experiments and morphological studies. Methods and Results-Clinical data of 23 probands and 35 family members carrying this variant were retrospectively collected. A time-to-event analysis was performed to compare the course of the disease with carriers of other LMNA mutations. Myocardial biopsies were studied with electron microscopy and by measuring force development of the sarcomeres. Morphology of the nuclear envelope was assessed with immunofluorescence on cultured fibroblasts. The phenotype in probands and family members was characterized by atrioventricular conduction disturbances (61% and 44%, respectively), supraventricular arrhythmias (69% and 52%, respectively), and dilated cardiomyopathy (74% and 14%, respectively). LMNA p.(Arg331Gln) carriers had a significantly better outcome regarding the composite end point (malignant ventricular arrhythmias, end-stage heart failure, or death) compared with carriers of other pathogenic LMNA mutations. A shared haplotype of 1 Mb around LMNA suggested a common founder. The combined logarithm of the odds score was 3.46. Force development in membranepermeabilized cardiomyocytes was reduced because of decreased myofibril density. Structural nuclear LMNA-associated envelope abnormalities, that is, blebs, were confirmed by electron microscopy and immunofluorescence microscopy. Conclusions-Clinical, morphological, functional, haplotype, and segregation data all indicate that LMNA p. (Arg331Gln) is a pathogenic founder mutation with a phenotype reminiscent of other LMNA mutations b...

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Section: Hoorntje Et Al Mild Phenotype In Lmna P(arg331gln) Carriersmentioning

confidence: 99%

Lamin A/C -Related Cardiac Disease

Hoorntje¹,

Bollen²,

Barge-Schaapveld³

et al. 2017

Circ Cardiovasc Genet

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“…Primary cultures of dermal fibroblasts were processed for indirect immunofluorescence as described in ref. 17. Rabbit Abs directed against A-type and B-type lamins were kindly provided by N. Chaudhary (Ridgeway Biosystems, Inc., Cleveland) (33,34).…”

Section: Methodsmentioning

confidence: 99%

“…We obtained three primary cultures of dermal fibroblasts derived from HGPS patients carrying the LMNA mutation G608G: HGADFN001, HGADFN003, and HGAFN127 from the Progeria Research Foundation (Peabody, MA) and PT001 recently identified in ref. 17. We used in parallel three previously established control dermal fibroblast cultures.…”

Section: Methodsmentioning

confidence: 99%

“…Because the endoproteolytic cleavage site is lost in the truncated lamin A (progerin), it was predicted to be permanently prenylated (3). Direct and indirect analyses have recently confirmed that progerin retains the farnesyl group (12)(13)(14)(15).Previous studies of A-type lamin distribution in primary dermal fibroblasts from HGPS patients showed nuclear abnormalities in size and shape in a subpopulation of cells in culture (3,16,17). Farnesylated progerin appears to be responsible for the nuclear deformations because administration of farnesyltransferase inhibitors to the HGPS fibroblast cultures normalized the nuclear shape for the majority of the cells in vitro (12-15).…”

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confidence: 95%

“…A rabbit polyclonal Ab was raised against a short peptide overlapping the 50-aa deletion region in the mutant lamin A G608G (progerin; see Fig. 6, which is published online as supporting information on the PNAS web site) (17). The immune sera, denoted anti-LMNA G608G, were tested on primary dermal fibroblast cultures at early and late population doublings (PPDs; 20-50) by using four different HGPS fibroblast cultures (see Materials and Methods).…”

Section: Mutant Lamin a G608g Accumulates Within The Nucleus In A Celmentioning

confidence: 99%

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Hutchinson–Gilford progeria mutant lamin A primarily targets human vascular cells as detected by an anti-Lamin A G608G antibody

McClintock

Gordon

Djabali

2006

Proc. Natl. Acad. Sci. U.S.A.

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183

192

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Hutchinson-Gilford progeria syndrome (HGPS; Online Mendelian Inheritance in Man accession no. 176670) is a rare disorder that is characterized by segmental premature aging and death between 7 and 20 years of age from severe premature atherosclerosis. Mutations in the LMNA gene are responsible for this syndrome. Approximately 80% of HGPS cases are caused by a G608 (GGC3 GGT) mutation within exon 11 of LMNA, which elicits a deletion of 50 aa near the C terminus of prelamin A. In this article, we present evidence that the mutant lamin A (progerin) accumulates in the nucleus in a cellular age-dependent manner. In human HGPS fibroblast cultures, we observed, concomitantly to nuclear progerin accumulation, severe nuclear envelope deformations and invaginations preventable by farnesyltransferase inhibition. Nuclear alterations affect cell-cycle progression and cell migration and elicit premature senescence. Strikingly, skin biopsy sections from a subject with HGPS showed that the truncated lamin A accumulates primarily in the nuclei of vascular cells. This finding suggests that accumulation of progerin is directly involved in vascular disease in progeria.aging ͉ atherosclerosis H utchinson-Gilford progeria syndrome (HGPS) is a rare, fatal genetic disorder that is characterized by accelerated aging in children. The LMNA gene encoding the A-type lamins A and C is the causative gene of HGPS (1-3). Approximately 80% of HGPS cases carry the heterozygous silent point mutation G608G within exon 11 of LMNA (3). This mutation creates an abnormal splice donor site, which produces a truncated protein (progerin) lacking residues 607-656 of prelamin A but retaining the C-terminal CAAX box, a target for prenylation (1-3).The nuclear lamina is a scaffold, which provides structural and mechanical stability for the nuclear envelope (NE); it consists primarily of type V intermediate filament proteins (A-and B-type lamins) and many inner-nuclear membrane proteins (4-7). Lamins interact with heterochromatin and transcriptional regulators, suggesting their important role in the maintenance of chromatin organization and gene expression (8).At the NE periphery, lamin precursors undergo a series of posttranslational modifications. B-type lamins are permanently isoprenylated, whereas prelamin A loses its modification after incorporation into the lamina by lamin A-specific processing steps involving Zmpste24 endoprotease (9-11). Because the endoproteolytic cleavage site is lost in the truncated lamin A (progerin), it was predicted to be permanently prenylated (3). Direct and indirect analyses have recently confirmed that progerin retains the farnesyl group (12)(13)(14)(15).Previous studies of A-type lamin distribution in primary dermal fibroblasts from HGPS patients showed nuclear abnormalities in size and shape in a subpopulation of cells in culture (3,16,17). Farnesylated progerin appears to be responsible for the nuclear deformations because administration of farnesyltransferase inhibitors to the HGPS fibroblast cultures normalized the nucle...

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An LMNA synonymous variant associated with severe dilated cardiomyopathy: Case report

Gao¹,

Mumme‐Monheit²,

Chen³

et al. 2021

American J of Med Genetics Pt A

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Dilated cardiomyopathy (DCM) is one of the most common cardiac phenotypes caused by mutations of lamin A/C (LMNA) gene in humans. In our study, a cohort of 57 patients who underwent heart transplant for dilated cardiomyopathy was screened for variants in LMNA. We identified a synonymous variant c.936G>A in the last nucleotide of exon 5 of LMNA in a DCM family. Clinically, the LMNA variant carriers presented with severe familial DCM, conduction disease, and high creatine‐kinase level. The LMNA c.936G>A variant is novel and has not been reported in current genetic variant databases. Sanger sequencing results showed the presence of LMNA c.936G>A variant in the genomic DNA but not in the cDNA derived from one family member's heart tissue. Real‐time quantitative polymerase chain reaction showed significantly lower LMNA mRNA levels in the patient's heart compared to the controls, suggesting that the c.936G>A LMNA variant resulted in reduced mRNA and possibly lower protein expression of LMNA. These findings expand the understanding on the association between synonymous variant of LMNA and the molecular pathogenesis in DCM patients.

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Dermal fibroblasts in Hutchinson-Gilford progeria syndrome with the lamin A G608G mutation have dysmorphic nuclei and are hypersensitive to heat stress

Cited by 62 publications

References 47 publications

Lamin A/C -Related Cardiac Disease

Lamin A/C -Related Cardiac Disease

Hutchinson–Gilford progeria mutant lamin A primarily targets human vascular cells as detected by an anti-Lamin A G608G antibody

An LMNA synonymous variant associated with severe dilated cardiomyopathy: Case report

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