Characterization of immortalized human epidermolysis bullosa simplex (KRT5) cell lines: trimethylamine N-oxide protects the keratin cytoskeleton against disruptive stress condition.

Abstract

BACKGROUND:

Epidermolysis bullosa simplex (EBS) is an autosomal inherited mechano-bullous disease, characterized by intraepidermal blistering and skin fragility caused by mutations in the keratin (KRT) 5 or 14 genes. Despite a vast knowledge about the intermediate filament pathology in this disease, the progress in therapy has been slow. Animal models and well-characterized continuous cell culture models of EBS are needed prior to clinical testing.

OBJECTIVES:

Our aim was to generate immortalized cell lines as an in vitro model for the study of EBS and test a chemical chaperone, trimethylamine N-oxide (TMAO), as a putative novel therapy.

METHODS:

We generated four immortalized cell lines, two each from an EBS patient with a KRT5-mutation (V186L) and a healthy control, using human papillomavirus 16 (HPV16) E6E7 as transducer. Cell lines were established in serum-free and serum-containing medium and assessed for growth characteristics, keratin expression profiles, ability to differentiate in organotypic cultures, and response to heat stress with and without the presence of TMAO.

RESULTS:

All cell lines have been expanded >160 population doublings and their cellular characteristics are similar. However, the formation of cytoplasmic keratin filament aggregates in response to heat-shock treatment differed between EBS and normal cell lines. Notably, serum-free established EBS-cell line was most vulnerable to heat shock but both cell lines exhibited significant reduction in the number of keratin aggregates containing cells by TMAO.

CONCLUSION:

The immortalized cell lines represent a suitable model for studying novel therapies for EBS. TMAO is a promising new agent for future development as a novel EBS therapy.

• Authors: Jean Christopher Chamcheu, Elizabeth Pavez Lorié, Baki Akgul, Elin Bannbers, Marie Virtanen, Luke Gammon, Aristidis Moustakas, Harshad Navsaria, Anders Vahlquist, Hans Törmä
• Journal: Journal of dermatological science (J Dermatol Sci) Vol. 53 Issue 3 Pg. 198-206 (Mar 2009) ISSN: 0923-1811 [Print] Netherlands
• PMID: 19157792 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• KRT5 protein, human
• Keratin-5
• Methylamines
• Oxidants
• Keratins
• trimethyloxamine

Topics

• Apoptosis (drug effects)
• Biopsy
• Case-Control Studies
• Cell Differentiation (drug effects)
• Cell Line
• Cytoskeleton (drug effects, metabolism)
• Epidermolysis Bullosa Simplex (genetics, metabolism, pathology)
• Hot Temperature (adverse effects)
• Humans
• In Vitro Techniques
• Keratin-5 (genetics, metabolism)
• Keratinocytes (metabolism, pathology)
• Keratins (metabolism)
• Methylamines (pharmacology)
• Models, Biological
• Mutation (genetics)
• Oxidants (pharmacology)
• Skin (metabolism, pathology)