# EPIDERMOLYSIS BULLOSA:  MECHANISMS OF PATHOGENESIS

> **NIH NIH R01** · UNIV OF NORTH CAROLINA CHAPEL HILL · 1989 · $110,097

## Abstract

Recent understanding of factors regulating collagenase production
facilitates new approaches to the study of recessive dystrophic
epidermolysis bullosa (RDEB), in which collagenase production is elevated,
and other forms of epidermolysis bullosa (EB).  1) Following leads
suggested by this new information, we wish to determine whether RDER
fibroblasts have a defect in regulation of production of collagenase such
that they appear constitutively stimulated and therefore not only secrete
high levels of collagenase but fail to respond to stimulation.  In order to
evaluate this hypothesis, we will determine whether RDEB fibroblasts show
reduced sensitivity compared to normal fibroblasts on exposure to
interleukin-1 and other stimulators of collagenase production, including
such agents produced by keratinocyte cultures.  We will examine culture
supernatants for excessive production of factors capable of stimulating
production of collagenase.  Since extracellular matrix, especially
collagen, can increase collagenase production, we will assess collagenase
modulation in normal and affected cells plated with or without collagen or
fibronectin and on collagen gels.  We have recently documented production
of collagenase by cultures of human keratinocytes exposed to the phorbol
ester, TPA.  We will compare production of collagenase in keratinocytes
from controls and patients with various types of EB.  We will search for
physiologic stimuli for the production of collagenase by keratinocytes and
will characterize the collagenase.  We will examine supernatants from EB
keratinocytes in an assay designed to detect activity dissociating
keratinocytes, as occurs in vivo.  2) Since abnormal cytoskeletal
regulation may underly elevated collagenase in RDEB fibroblasts, we will
use antibodies to vinculin, actin, talin, spectrin, and desmoplakin  to
examine the cytoskeleton of normal and affected cells by
immunofluorescence, immunoblotting, and immunoprecipitation.  We have
recently shown alterations in distribution of vinculin in keratinocytes
after an increase in Ca+ in the medium from 0.1 to 1.0 mM and will
similarly examine the keratinocyte cytoskeleton under these two conditions
in cells from patients with other types of EB.  3) Finally, we will explore
collagenase production by an in vitro model of skin made from a
fibroblast-populated.  Collagen lattice covered by added cultured
keratinocytes.  We will attempt to produce basement membrane antigens and
lamina densa in vitro with this model.  These studies may help elucidate
the abnormal regulation of collagenase in EB.

## Key facts

- **NIH application ID:** 3157630
- **Project number:** 5R01AR036497-03
- **Recipient organization:** UNIV OF NORTH CAROLINA CHAPEL HILL
- **SAM.gov UEI:** D3LHU66KBLD5
- **Principal Investigator:** EDWARD J O'KEEFE
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1989
- **Award amount:** $110,097
- **Award type:** 5
- **Project period:** 1987-01-01 → 1990-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3157630

## Citation

> US National Institutes of Health, RePORTER application 3157630, EPIDERMOLYSIS BULLOSA:  MECHANISMS OF PATHOGENESIS (5R01AR036497-03). Retrieved via MORVS 2026-10-24 from https://api.morvs.ai/grant/nih/3157630. Licensed CC0.

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