Exonic mutations associated with atopic dermatitis disrupt lympho-epithelial Kazal-type related inhibitor action and enhance its degradation.
Cytokines
/ metabolism
Dermatitis, Atopic
/ genetics
Epidermis
/ metabolism
Exons
HaCaT Cells
Haplotypes
Humans
Inflammation
/ metabolism
Kallikreins
/ chemistry
Keratinocytes
/ metabolism
Mutation
Polymorphism, Single Nucleotide
Protein Domains
/ genetics
Proteolysis
Serine Peptidase Inhibitor Kazal-Type 5
/ genetics
Signal Transduction
/ genetics
Trypsin
/ metabolism
KLK5 protease
LEKTI activity
atopic dermatitis
exonic mutation
mesotrypsin degradation
Journal
Allergy
ISSN: 1398-9995
Titre abrégé: Allergy
Pays: Denmark
ID NLM: 7804028
Informations de publication
Date de publication:
02 2020
02 2020
Historique:
received:
02
04
2019
revised:
09
07
2019
accepted:
24
07
2019
pubmed:
14
8
2019
medline:
20
1
2021
entrez:
14
8
2019
Statut:
ppublish
Résumé
Skin desquamation is facilitated by serine proteases KLK5 and KLK7, which are tightly regulated by lympho-epithelial Kazal-type related inhibitor (LEKTI). LEKTI itself is controlled through degraded by mesotrypsin. Here, we sought to determine whether LEKTI exonic mutations associated with atopic dermatitis (AD) affect the protease inhibitory activity of LEKTI or its susceptibility to mesotrypsin degradation. The inhibitory activities of the LEKTI domain 4 (D4) and D6 WT and AD-associated mutants on the enzyme activities of KLK5 and KLK7 were compared using fluorogenic substrates. A keratinocyte cell culture system using HaCat cells was established to assess the role of D6 WT and D386N on triggering inflammation via the induction of thymic stromal lymphopoietin (TSLP). A degradation assay was used to assess the susceptibility of D4 and D6 mutants to mesotrypsin degradation. Enzymatic assays revealed that the D6 D386N mutation affected the inhibitory activity of LEKTI on KLK5 but not KLK7. Other exonic mutations on D6 (N368S, V395M, and E420K) and D4 (R267Q) did not alter LEKTI inhibition. The D386N mutation disrupted the role of D6 in suppressing TSLP induction by KLK5 in HaCat cells. Although WT D4 is more susceptible to mesotrypsin degradation than WT D6, the D4 R267Q mutant was more resistant to mesotrypsin degradation, whereas the D6 E420K mutant showed enhanced mesotrypsin-mediated degradation. Exonic mutations in D6, which previously have been associated with AD, may cause a disruption of inhibitory activity on KLK5 or enhance the degradation by mesotrypsin.
Sections du résumé
BACKGROUND
Skin desquamation is facilitated by serine proteases KLK5 and KLK7, which are tightly regulated by lympho-epithelial Kazal-type related inhibitor (LEKTI). LEKTI itself is controlled through degraded by mesotrypsin. Here, we sought to determine whether LEKTI exonic mutations associated with atopic dermatitis (AD) affect the protease inhibitory activity of LEKTI or its susceptibility to mesotrypsin degradation.
METHODS
The inhibitory activities of the LEKTI domain 4 (D4) and D6 WT and AD-associated mutants on the enzyme activities of KLK5 and KLK7 were compared using fluorogenic substrates. A keratinocyte cell culture system using HaCat cells was established to assess the role of D6 WT and D386N on triggering inflammation via the induction of thymic stromal lymphopoietin (TSLP). A degradation assay was used to assess the susceptibility of D4 and D6 mutants to mesotrypsin degradation.
RESULTS
Enzymatic assays revealed that the D6 D386N mutation affected the inhibitory activity of LEKTI on KLK5 but not KLK7. Other exonic mutations on D6 (N368S, V395M, and E420K) and D4 (R267Q) did not alter LEKTI inhibition. The D386N mutation disrupted the role of D6 in suppressing TSLP induction by KLK5 in HaCat cells. Although WT D4 is more susceptible to mesotrypsin degradation than WT D6, the D4 R267Q mutant was more resistant to mesotrypsin degradation, whereas the D6 E420K mutant showed enhanced mesotrypsin-mediated degradation.
CONCLUSION
Exonic mutations in D6, which previously have been associated with AD, may cause a disruption of inhibitory activity on KLK5 or enhance the degradation by mesotrypsin.
Substances chimiques
Cytokines
0
SPINK5 protein, human
0
Serine Peptidase Inhibitor Kazal-Type 5
0
TSLP protein, human
0
KLK5 protein, human
EC 3.4.21.-
KLK7 protein, human
EC 3.4.21.-
Kallikreins
EC 3.4.21.-
PRSS3 protein, human
EC 3.4.21.4
Trypsin
EC 3.4.21.4
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
403-411Subventions
Organisme : Ministry of Education Singapore
ID : MOE2011-T2-2-002
Pays : International
Organisme : Ministry of Education Singapore
ID : R-154-000-531-112
Pays : International
Informations de copyright
© 2019 EAACI and John Wiley and Sons A/S. Published by John Wiley and Sons Ltd.
Références
Milstone LM. Epidermal desquamation. J Dermatol Sci. 2004;36:131-140.
Egelrud T, Hofer P-Å, Lundström A. Proteolytic degradation of desmosomes in plantar stratum corneum leads to cell dissciation in vitro. Acta dermato-venereologica. 1988;68:93.
Lundwall Å, Brattsand M. Kallikrein-related peptidases. Cell Mol Life Sci. 2008;65:2019-2038.
Serge JA. Epidermal barrier formation and recovery in skin disorders. J Clin Invest. 2006;116:1150-1158.
Deraison C, Bonnart C, Lopez F, et al. LEKTI fragments specifically inhibits KLK5, KLK7, and KLK14 and control desquamation through a pH-dependent interaction. Mol Biol Cell. 2007;18:3607-3619.
Miyai M, Matsumoto Y, Yamanishi H, Yamamoto-Tanaka M, Tsuboi R, Hibino T. Keratinocyte-specific mesotrypsin contributes to the desquamation process via kallikrein activation and LEKTI degradation. J Invest Dermatol. 2014;134:1665-1674.
Bitoun E, Chavanas S, Irvine AD, et al. Netherton syndrome: disease expression and spectrum of SPINK5 mutations in 21 families. J Invest Dermatol. 2001;118:352-361.
Oiso N, Kawada A. LEKTI: netherton syndrome and atopic dermatitis. In: Oiso N, ed. Current Genetics in Dermatology. London, UK:InTechOpen; 2013:67-72.
Smith F, Irvine AD, Terron-Kwiatkowski A, et al. Loss-of-function mutations in the gene encoding filaggrin cause ichthyosis vulgaris. Nat Genet. 2006;38(3):337-342.
Palmer C, Ismail T, Lee SP, et al. Filaggrin null mutations are associated with increased asthma severity in children and yound adults. J Allergy Clin Immunol. 2007;120:64-68.
Kasparek P, Ileninova Z, Zbodakova O, et al. KLK5 and KLK7 ablation fully rescues lethality of netherton syndrome-like phenotype. PLoS Genet. 2017;13:e1006566.
Markova NG, Mrekov LN, Chipev CC, Gan SQ, Idler WW, Steinert PM. Profilaggrin is a major epidermal calcium-binding protein. Mol Cell Biol. 1993;13:613-625.
Cork MJ, Robinson DA, Vasilopoulos Y, et al. New perspectives on epidermal barrier dysfunction in atopic dermatitis: gene-environment interactions. J Allergy Clin Immunol. 2006;118:3-21.
Morar N, Cookson W, Harper JI, Moffatt MF. Filaggrin mutations in children with severe atopic dermatitis. J Invest Dermatol. 2007;127:1667-1672.
Matsubara Y, Matsumoto T, Koseki J, Kaneko A, Aiba S, Yamasaki K. Inhibition of human kallikrein 5 protease by triterpenoids from natural sources. Molecules. 2017;22:1829.
Briot A, Deraison C, Lacroix M, et al. Kallikrein 5 induces atopic dermatitis-like lesions through PAR2-mediated thymic stromal lymphopoietin expression in Netherton syndrome. J Exp Med. 2009;206:1135-1147.
Soumelis V, Reche PA, Kanzler H, et al. Human epithelial cells trigger dendritic cell-mediated allergic inflammation by producing TSLP. Nat Immunol. 2002;3:673-680.
Yoo J, Omori M, Gyarmati D, et al. Spontaneous atopic dermatitis in mice expressing an inducible thymic stromal lymphopoietin transgene specifically in the skin. J Exp Med. 2005;202:541-549.
Chavanas S, Bodemer C, Rochat A, et al. Mutations in SPINK5, encoding a serine protease inhibitor, cause Netherton syndrome. Nat Genet. 2000;25:141-142.
Zhao LP, Di Z, Zhang L, et al. Association of SPINK5 gene polymorphisms with atopic dermatitis in Northeast China. J Eur Acad Dermatol Venereol. 2012;26:572-577.
Kato A, Fukai K, Oiso N, Hosomi N, Murakami T, Ishii M. Association of SPINK5 gene polymorphisms with atopic dermatitis in the Japanese population. Br J Dermatol. 2003;148:665-669.
Walley AJ, Chavanas S, Moffatt MF, et al. Gene polymorphism in Netherton and common atopic disease. Nat Genet. 2001;29:175-178.
Namkung J-H, Lee J-E, Kim E, et al. Hint for association of single nucleotide polymorphisms and haplotype in SPINK5 gene with atopic dermatitis in Koreans. Exp Dermatol. 2010;19:1048-1053.
Egelrud T, Brattsand M, Kreutzmann P, et al. hk5 and hk7, two serine proteinases abundant in human skin, are inhibited by LEKTI domain 6. Br J Dermatol. 2005;153:1200-1203.
Fortugno P, Furio L, Teson M, et al. The 420K LEKTI variant alters LEKTI proteolytic activation and results in protease deregulation: implications for atopic dermatitis. Hum Mol Genet. 2012;21(19):4187-4200.
Lan C-C, Tu H-P, Wu C-S, et al. Distinct SPINK5 and IL-31 polymorphisms are associated with atopic eczema and non-atopic hand dermatitis in Taiwanese nursing population. Exp Dermatol. 2011;20:975-979.
Schechter I, Berger A. On the size of the active site in protease. I. Papain. 1967. Biochem Biophys Res Commun. 2012;425:497-502.
Hovnanian A. Netherton syndrome: skin inflammation and allergy by loss of protease inhibition. Cell Tissue Res. 2013;351:289-300.
Furio L, Pampalakis G, Michael IP, Nagy A, Sotiropoulou G, Hovnanian A. KLK5 inactivation reverses cutaneous hallmarks of Netherton syndrome. PLoS Genet. 2015;11:e1005389.
Roelandt T, Thys B, Heughebaert C, et al. LEKTI-1 in sickness and in health. Int J Cosmet Sci. 2009;31:247-254.
Bitoun E, Micheloni A, Lamant L, et al. LEKTI proteolytic processing in human primary keratinocytes, tissue distribution and defective expression in Netherton syndrome. Hum Mol Genet. 2003;12:2417-2430.
Wilson S, Thé L, Batia L, et al. The epithelial cell-derived atopic dermatitis cytokine TSLP activates neurons to induce itch. Cell. 2013;155:285-295.
Nyaruhucha CN, Kito M, Fukuoka SI. Identification and expression of the cDNA-encoding human mesotrypsin(ogen), an isoform of trypsin with inhibitor resistance. J Biol Chem. 1997;272:10573-10578.
Fölster-Holst R, Stoll M, Koch WA, Hampe J, Christophers E, Schreiber S. Lack of association of SPINK5 polymorphisms with nonsyndromic atopic dermatitis in the population of Northern Germany. Br J Dermatol. 2005;152:1365-1367.
Jongepier H, Koppelman GH, Nolte IM, et al. Polymorphisms in SPINK5 are not associated with asthma in a Dutch population. J Allergy Clin Immunol. 2005;115:486-492.
Ramesh K, Lama D, Tan KW, et al. Homologous lympho-epithelial Kazal-type related inhibitor domains delay blood coagulation by inhibiting factor X and XI with differential specificity. Structure. 2018;26:1-9.