Loss-of-function variants in KLF4 underlie autosomal dominant palmoplantar keratoderma

Malovitski, Kiril; Sarig, Ofer; Assaf, Sari; Mohamad, Janan; Malki, Liron; Bergson, Shir; Peled, A.; Eskin‐Schwartz, Marina; Gat, Andrea; Pavlovsky, Mor; Sprecher, Eli

doi:10.1016/j.gim.2022.01.009

Cited by 4 publications

(3 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Exome sequencing was conducted at the Tel Aviv Sourasky Genomic Center using the IDT xGen capture reagent (Integrated DNA Technologies, Coralville, IA) and Illumina NovaSeq 6000 platform (Illumina, San Diego, CA). Sequencing and analytical methodologies have been previously described elsewhere (Malovitski et al, 2022). Validation of the variants and co‐segregation analysis was performed by direct sequencing.…”

Section: Methodsmentioning

confidence: 99%

“…Genomic DNA was PCR‐amplified using oligonucleotide primer pairs spanning the entire coding sequence as well as intron–exon boundaries of CLDN1 (Table S2). PCR cycling conditions have been previously described (Malovitski et al, 2022).…”

Section: Methodsmentioning

confidence: 99%

“…For cell immunofluorescence studies, HaCaT cells were grown on glass coverslips in 12 wells plates (Malovitski et al, 2022) and stained with the use of a mouse monoclonal anti V5 epitope tag antibody (dilution 1:500; Invitrogen, Carlsbad, CA, R960-25) as primary antibody and Rhodamine RED anti-mouse (Life Technologies/Invitrogen) as secondary antibody. Image analysis was done with an LSM 510 confocal microscope (Zeiss, Jena, Germany).…”

Section: Immunofluorescence Analysismentioning

confidence: 99%

See 2 more Smart Citations

Autosomal recessive congenital ichthyosis caused by a pathogenic missense variant in CLDN1

Mohamad

Samuelov

Assaf

et al. 2022

American J of Med Genetics Pt A

Self Cite

View full text Add to dashboard Cite

Autosomal recessive congenital ichthyosis (ARCI) refers to a large and genetically heterogenous group of non‐syndromic disorders of cornification featuring diffuse scaling. Ichthyosis, leukocyte vacuoles, alopecia, and sclerosing cholangitis (ILVASC) syndrome is a rare autosomal recessive syndromic form of ichthyosis. The disease usually results from premature termination codon‐causing pathogenic variants in CLDN1 encoding CLAUDIN‐1 (CLDN1). We used whole exome sequencing (WES), Sanger sequencing, 3D protein modeling, Western blotting, and immunofluorescence confocal microscopy to delineate the genetic basis of ichthyosis in two siblings with ichthyosis but no other ectodermal abnormalities. One of the two siblings underwent liver transplantation in early childhood due to biliary atresia. Both patients were found to carry a homozygous missense pathogenic variant, c.242G>A (p.Arg81His), in CLDN1. The variant resulted in decreased CLDN1 expression in patient skin. 3D protein modeling predicted that p.Arg81His induces deleterious conformational changes. Accordingly, HaCaT cells transfected with a construct expressing the mutant CLDN1 cDNA featured decreased levels and mislocation of CLDN1 as compared with cells expressing the wildtype cDNA. In conclusion, we describe the first pathogenic missense variant in CLDN1 shown to result in ARCI.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Immunofluorescence Analysismentioning

confidence: 99%

See 1 more Smart Citation

Autosomal recessive congenital ichthyosis caused by a pathogenic missense variant in CLDN1

Mohamad

Samuelov

Assaf

et al. 2022

American J of Med Genetics Pt A

Self Cite

View full text Add to dashboard Cite

show abstract

Proteolytic and Antiproteolytic Activity in the Skin: Gluing the Pieces Together

Peled,

Sprecher

2024

Journal of Investigative Dermatology

View full text Add to dashboard Cite

Defective cathepsin Z affects EGFR expression and causes autosomal dominant palmoplantar keratoderma

Malovitski

Sarig²,

Feller

et al. 2023

British Journal of Dermatology

View full text Add to dashboard Cite

Background Abnormal function of the epidermal growth factor receptor (EGFR) has been recently shown to underlie various disorders of cornification. Objectives Here we aimed at delineating the genetic basis of a novel dominant form of palmoplantar keratoderma (PPK). Methods We used whole exome and direct sequencing, RT-qPCR, protein modelling, confocal immunofluorescence microscopy, immunoblotting, three-dimensional skin equivalents and an enzyme activity assays. Results Whole exome sequencing revealed heterozygous variants (c.274T > C and c.305C > T) in CTSZ encoding cathepsin Z in four individuals with focal PPK, who belong to three unrelated families. Bioinformatics and protein modeling predicted the variants to be pathogenic. Previous studies suggested that EGFR expression may be subject to cathepsin regulation. Immunofluorescence staining revealed reduced cathepsin Z expression in the upper epidermal layers and concomitant increased epidermal EGFR expression in patients harboring CTSZ variants. Accordingly, human keratinocytes transfected with constructs expressing PPK-causing variants in CTSZ displayed reduced cathepsin Z enzymatic activity as well as increased EGFR expression. In line with the role played by EGFR in the regulation of keratinocyte proliferation, human keratinocytes transfected with the PPK-causing variants, showed significantly increased proliferation which was abolished upon exposure to erlotinib, an EGFR inhibitor. Similarly, downregulation of CTSZ resulted in increased EGFR expression and increased proliferation in human keratinocytes, suggestive of a loss-of-function effect of the pathogenic variants. Finally, 3-dimensional organotypic skin equivalents grown from CTSZ-downregulated cells showed increased epidermal thickness and EGFR expression as seen in patient skin; here too, erlotinib was found to rescue the abnormal phenotype. Conclusions Taken collectively, these observations attribute to cathepsin Z a hitherto unrecognized function in epidermal differentiation.

show abstract

Loss-of-function variants in KLF4 underlie autosomal dominant palmoplantar keratoderma

Cited by 4 publications

References 40 publications

Autosomal recessive congenital ichthyosis caused by a pathogenic missense variant in CLDN1

Autosomal recessive congenital ichthyosis caused by a pathogenic missense variant in CLDN1

Proteolytic and Antiproteolytic Activity in the Skin: Gluing the Pieces Together

Defective cathepsin Z affects EGFR expression and causes autosomal dominant palmoplantar keratoderma

Contact Info

Product

Resources

About