Elevated Human Tissue Kallikrein Levels in the Stratum Corneum and Serum of Peeling Skin Syndrome-Type B Patients Suggests an Over-desquamation of Corneocytes
2006; Elsevier BV; Volume: 126; Issue: 10 Linguagem: Inglês
10.1038/sj.jid.5700379
ISSN1523-1747
AutoresNahoko Komatsu, Yasushi Suga, Kiyofumi Saijoh, Chang Amber Liu, Saba Khan, Yuki Mizuno, Shigaku Ikeda, Hua-Kang Wu, Arumugam Jayakumar, Gary L. Clayman, Fumiaki Shirasaki, Kazuhiko Takehara, Eleftherios P. Diamandis,
Tópico(s)Coagulation, Bradykinin, Polyphosphates, and Angioedema
Resumo7-amino-4-methyl-commarin kallikrein protein Netherton syndrome para-nitroanilide peeling skin syndrome stratum corneum serine protease inhibitor Kazal-type 5 TO THE EDITOR Peeling skin syndrome type B (PSS-type B, MIN 270300) is a congenital skin disease associated with continual skin peeling and ichthyotic erythroderma, known to display various clinical similarities with Netherton syndrome (NS, caused by genetic defects of serine protease inhibitor Kazal-type5 (SPINK5), MIN 256500) (Wile, 1924Wile U.J. Familial study of three unusual cases of congenital ichthyosiform erythroderma.Arch Dermatol Syph. 1924; 10: 487-498Crossref Scopus (22) Google Scholar; Traupe, 1989Traupe H. The ichthyoses: a guide to clinical diagnosis, genetic counseling and therapy. Springer-Verlag, New York1989: 207-210Crossref Google Scholar; Magert et al., 1999Magert H.J. Standker L. Kreutzmann P. Zucht H.D. Reinecke M. Sommerhoff C.P. et al.LEKTI, a novel 15-domain type of human serine proteinase inhibitor.J Biol Chem. 1999; 274: 21499-21502Crossref PubMed Scopus (211) Google Scholar; Chavanas et al., 2000Chavanas S. Bodemer C. Rochat A. Hamel-Teillac D. Ali M. Irvine A.D. et al.Mutations in SPINK5, encoding a serine protease inhibitor, cause Netherton syndrome.Nat Genet. 2000; 25: 141-142Crossref PubMed Scopus (658) Google Scholar). Human tissue kallikreins are a family of 15 trypsin- or chymotrypsin-like secreted serine proteases (hK1–hK15) found in a variety of tissues (Yousef and Diamandis, 2001Yousef G.M. Diamandis E.P. The new human tissue kallikrein gene family: structure, function, and association to disease.Endocr Rev. 2001; 22: 184-204Crossref PubMed Scopus (577) Google Scholar). At least eight different hKs have been identified in the stratum corneum (SC) and sweat as desquamation-related proteases (Komatsu et al., 2005Komatsu N. Saijoh K. Sidiropoulos M. Tsai B. Levesque M.A. Elliott M.B. et al.Quantification of human tissue kallikreins in the stratum corneum: dependence on age and gender.J Invest Dermatol. 2005; 125: 1182-1192Crossref PubMed Scopus (65) Google Scholar, Komatsu et al., 2006Komatsu N. Tsai B. Sidiropoulos M. Saijoh K. Levesque M.A. Takehara K. et al.Quantification of eight tissue kallikreins in the stratum corneum and sweat.J Invest Dermatol. 2006; 126: 925-929Abstract Full Text Full Text PDF PubMed Scopus (11) Google Scholar). Here, we aimed (1) to clarify the pathogenesis of PSS-type B, (2) to elucidate the relationship between PSS-type B and hK expression, and (3) to explain the reason for the clinical similarities between PSS-type B and NS. Informed consent was obtained from all patients, their parents, and normal volunteers, and our studies were performed according to the Declaration of Helsinki Principles. The Medical Ethics Committee of the Graduate School of Medical Science, School of Medicine, Kanazawa University and Juntendo University, School of Medicine, approved all described studies. Two unrelated 8-year-old female Japanese patients (Patient M and Patient K) were studied (Figure S1 and Table S1). Patients M and K were born with erythroderma accompanied by scaling and their lesions have shown no improvement to date. Patient M suffered significant growth retardation (<−2 SD) since the age of 1 and occasional herpes simplex infections in her perioral region. Both patients experience severe pruritus, temperature instability, and low sweat secretion. Patient M exhibited overabsorption of topical agents from skin, which is also observed in NS (Smith et al., 1995Smith D.L. Smith J.G. Wong S.W. deShazo R.D. Netherton's syndrome: a syndrome of elevated IgE and characteristic skin and hair findings.J Allergy Clin Immunol. 1995; 95: 116-123Abstract Full Text Full Text PDF PubMed Scopus (112) Google Scholar; Allen et al., 2001Allen A. Siegfried E. Silverman R. Williams M.L. Elias P.M. Szabo S.K. et al.Significant absorption of topical tacrolimus in 3 patients with Netherton syndrome.Arch Dermatol. 2001; 137: 747-750PubMed Google Scholar). Asthma attacks were experienced in Patient M but no allergic diseases were apparent in Patient K. Both patients displayed eosinophilia and elevated serum IgE levels. As many clinical manifestations observed in the patients are common between NS and PSS-type B (Traupe, 1989Traupe H. The ichthyoses: a guide to clinical diagnosis, genetic counseling and therapy. Springer-Verlag, New York1989: 207-210Crossref Google Scholar; Judge et al., 1994Judge M.R. Morgan G. Harper J.I. A clinical and immunological study of Netherton's syndrome.Br J Dermatol. 1994; 131: 615-621Crossref PubMed Scopus (147) Google Scholar; Griffiths et al., 1998Griffiths W.A.D. Judge M.R. Leigh I.M. Disorders of keratinization.in: Champion R.H. Burton J.L. Burns D.A. Breathnach S.M. Textbook of dermatology. 2. Blackwell Science, Oxford1998: 1486-1588Google Scholar), SPINK5 gene mutation analysis by genomic polymerase chain reaction and sequencing was performed for both patients using specific primers (Komatsu et al., 2002Komatsu N. Takata M. Otsuki N. Ohka R. Amano O. Takehara K. et al.Elevated stratum corneum hydrolytic activity in Netherton syndrome suggests an inhibitory regulation of desquamation by SPINK5-derived peptides.J Invest Dermatol. 2002; 118: 436-443Crossref PubMed Scopus (167) Google Scholar). However, no gene mutations were detected in both patients; therefore, NS was ruled out, leaving PSS-type B as the most likely diagnosis. Download .jpg (.08 MB) Help with files Figure S1Clinical features of PSS type B patients from two unrelated Japanese families. Download .pdf (.06 MB) Help with pdf files Table S1Comparison of clinical and laboratory findings between PSS type B and Netherton syndrome, including our two patients. Pathologically, the patients showed an absence of the SC or a few layers of parakeratosis, which tended to be separated from the stratum granulosum, psoriasisforme acanthosis, and perivascular infiltration with mononuclear leukocytes (Figure S2). Download .jpg (.11 MB) Help with files Figure S2Histopathological features of the skin in peeling skin syndrome type B patients. Immunohistochemistry for hK6, hK8, hK13, and SPINK5 protein showed that they were mainly expressed in the stratum granulosum and SC in normal epidermis (Figure 1). In both patients, the hKs and SPINK5 protein expressions were similarly distributed, and their stainings were deeply expanded into the lower epidermis compared with those in normal skin. It is known that the skin of NS patients shows absent or only faint staining against the same anti-SPINK5 protein antibody (Raghunath et al., 2004Raghunath M. Tontsidou L. Oji V. Aufenvenne K. Schurmeyer-Horst F. Jayakumar A. et al.SPINK5 and Netherton syndrome: novel mutations, demonstration of missing LEKTI, and differential expression of transglutaminases.J Invest Dermatol. 2004; 123: 474-483Crossref PubMed Scopus (91) Google Scholar). Consequently, Patients M and K are unlikely to suffer from NS. In the SC of both Patients M and K, all hK concentrations studied by ELISA were dramatically higher than those in the normal SC samples (Table 1a). The elevation of minor skin hKs (e.g., hK10, hK6, and hK13; <1.0 ng/mg dry weight for normal subjects) were prominent in the SC of the patients (Table 1a). In the serum, hK6, hK7, hK8, hK10, and hK13 concentrations were significantly elevated in the patients (Table 1b).Table 1aKallikrein levels in the SC of PSS-type B patients by ELISAhK (ng/mg dry weight)Normal mean±SDPatient MPatient KChymotrypsin-like hK hK710.9±6.065.8*Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05).130.7*Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05).Trypsin-like hKs hK85.8±1.862.9*Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05).63.7*Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05). hK118.7±4.137.6*Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05).56.9*Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05). hK53.1±1.48.3*Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05).13.1*Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05). hK100.67±0.4121.7*Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05).29.5*Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05). hK140.34±0.132.3*Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05).3.2*Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05). hK60.28±0.1273.2*Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05).30.0*Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05). hK130.17±0.1424.1*Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05).15.9*Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05). Total of trypsin-like hKs19.1±5.4230.1*Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05).212.3*Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05).SC, stratum corneum; SD, standard deviation; PSS, peeling skin syndrome;The values indicate the mean±SD (ng/mg dry weight). The normal subjects (<11 years) were referred from a previous study (Komatsu et al., 2005Komatsu N. Saijoh K. Sidiropoulos M. Tsai B. Levesque M.A. Elliott M.B. et al.Quantification of human tissue kallikreins in the stratum corneum: dependence on age and gender.J Invest Dermatol. 2005; 125: 1182-1192Crossref PubMed Scopus (65) Google Scholar). Kallikreins are subdivided into chymotrypsin-like hK (hK7) and trypsin-like hKs (the rest of hKs) (Yousef and Diamandis, 2001Yousef G.M. Diamandis E.P. The new human tissue kallikrein gene family: structure, function, and association to disease.Endocr Rev. 2001; 22: 184-204Crossref PubMed Scopus (577) Google Scholar).* Smirnov test showed significant differences between normal SC samples and individual patients (P<0.05). Open table in a new tab Table 1bKallikrein levels in the serum of PSS-type B patients by ELISAhK (ng/mg)Normal mean±SDPatient MPatient KChymotrypsin-like hK hK75.1±2.117.0*Smirnov test showed significant differences between the normal serum samples and individual patients (P<0.05). The amount of hK13 in normal samples usually is very low or undetectable; hK13 in normal serum was described as <0.01ng/ml without SD. The statistics for hK13 was performed considering hK13 values for normal subjects as 0.01ng/ml.35.5*Smirnov test showed significant differences between the normal serum samples and individual patients (P<0.05). The amount of hK13 in normal samples usually is very low or undetectable; hK13 in normal serum was described as <0.01ng/ml without SD. The statistics for hK13 was performed considering hK13 values for normal subjects as 0.01ng/ml.Trypsin-like hKs hK64.4±1.515.7*Smirnov test showed significant differences between the normal serum samples and individual patients (P<0.05). The amount of hK13 in normal samples usually is very low or undetectable; hK13 in normal serum was described as <0.01ng/ml without SD. The statistics for hK13 was performed considering hK13 values for normal subjects as 0.01ng/ml.10.4*Smirnov test showed significant differences between the normal serum samples and individual patients (P<0.05). The amount of hK13 in normal samples usually is very low or undetectable; hK13 in normal serum was described as <0.01ng/ml without SD. The statistics for hK13 was performed considering hK13 values for normal subjects as 0.01ng/ml. hK81.9±0.7715.2*Smirnov test showed significant differences between the normal serum samples and individual patients (P<0.05). The amount of hK13 in normal samples usually is very low or undetectable; hK13 in normal serum was described as <0.01ng/ml without SD. The statistics for hK13 was performed considering hK13 values for normal subjects as 0.01ng/ml.7.1*Smirnov test showed significant differences between the normal serum samples and individual patients (P<0.05). The amount of hK13 in normal samples usually is very low or undetectable; hK13 in normal serum was described as <0.01ng/ml without SD. The statistics for hK13 was performed considering hK13 values for normal subjects as 0.01ng/ml. hK101.2±0.565.6*Smirnov test showed significant differences between the normal serum samples and individual patients (P<0.05). The amount of hK13 in normal samples usually is very low or undetectable; hK13 in normal serum was described as <0.01ng/ml without SD. The statistics for hK13 was performed considering hK13 values for normal subjects as 0.01ng/ml.5.4*Smirnov test showed significant differences between the normal serum samples and individual patients (P<0.05). The amount of hK13 in normal samples usually is very low or undetectable; hK13 in normal serum was described as <0.01ng/ml without SD. The statistics for hK13 was performed considering hK13 values for normal subjects as 0.01ng/ml. hK50.68±0 152.0*Smirnov test showed significant differences between the normal serum samples and individual patients (P<0.05). The amount of hK13 in normal samples usually is very low or undetectable; hK13 in normal serum was described as <0.01ng/ml without SD. The statistics for hK13 was performed considering hK13 values for normal subjects as 0.01ng/ml.0.47 hK110.54±0.160.380.67 hK140.22±0.0910.432.6*Smirnov test showed significant differences between the normal serum samples and individual patients (P<0.05). The amount of hK13 in normal samples usually is very low or undetectable; hK13 in normal serum was described as <0.01ng/ml without SD. The statistics for hK13 was performed considering hK13 values for normal subjects as 0.01ng/ml. hK13<0.010.26*Smirnov test showed significant differences between the normal serum samples and individual patients (P<0.05). The amount of hK13 in normal samples usually is very low or undetectable; hK13 in normal serum was described as <0.01ng/ml without SD. The statistics for hK13 was performed considering hK13 values for normal subjects as 0.01ng/ml.0.14*Smirnov test showed significant differences between the normal serum samples and individual patients (P<0.05). The amount of hK13 in normal samples usually is very low or undetectable; hK13 in normal serum was described as <0.01ng/ml without SD. The statistics for hK13 was performed considering hK13 values for normal subjects as 0.01ng/ml. Total of trypsin-like hKs4.5±5.423.9*Smirnov test showed significant differences between the normal serum samples and individual patients (P<0.05). The amount of hK13 in normal samples usually is very low or undetectable; hK13 in normal serum was described as <0.01ng/ml without SD. The statistics for hK13 was performed considering hK13 values for normal subjects as 0.01ng/ml.16.4*Smirnov test showed significant differences between the normal serum samples and individual patients (P<0.05). The amount of hK13 in normal samples usually is very low or undetectable; hK13 in normal serum was described as <0.01ng/ml without SD. The statistics for hK13 was performed considering hK13 values for normal subjects as 0.01ng/ml.PSS, peeling skin syndrome; SD, standard deviation.The values indicate the mean±SD (ng/ml).* Smirnov test showed significant differences between the normal serum samples and individual patients (P<0.05). The amount of hK13 in normal samples usually is very low or undetectable; hK13 in normal serum was described as <0.01 ng/ml without SD. The statistics for hK13 was performed considering hK13 values for normal subjects as 0.01 ng/ml. Open table in a new tab SC, stratum corneum; SD, standard deviation; PSS, peeling skin syndrome; The values indicate the mean±SD (ng/mg dry weight). The normal subjects (<11 years) were referred from a previous study (Komatsu et al., 2005Komatsu N. Saijoh K. Sidiropoulos M. Tsai B. Levesque M.A. Elliott M.B. et al.Quantification of human tissue kallikreins in the stratum corneum: dependence on age and gender.J Invest Dermatol. 2005; 125: 1182-1192Crossref PubMed Scopus (65) Google Scholar). Kallikreins are subdivided into chymotrypsin-like hK (hK7) and trypsin-like hKs (the rest of hKs) (Yousef and Diamandis, 2001Yousef G.M. Diamandis E.P. The new human tissue kallikrein gene family: structure, function, and association to disease.Endocr Rev. 2001; 22: 184-204Crossref PubMed Scopus (577) Google Scholar). PSS, peeling skin syndrome; SD, standard deviation. The values indicate the mean±SD (ng/ml). SC trypsin-like serine protease enzymatic activity examined using Boc-Pro-Phe-Arg-AMC (PFR-) as a substrate, and plasmin-like (for Boc-Val-Leu-Lys-AMC; VLK-) and furin-like (for Pyr-Arg-Thr-Lys-Arg-AMC; R-KR-) activities were significantly elevated, whereas trypsin-like (for Boc-Phe-Ser-Arg-AMC; FSR-) and chymotrypsin-like (for MeO-Suc-Arg-Pro-Tyr-pNA-HCl; RPY-) activities showed only mild elevations in both patients (Table 1c).Table 1cSC enzymatic activities in normal subjects and PSS-type B patientsSubstrateTime (hours)NormalPSS-type BReleased AMC or pNA (nmol/mg dry weight)n=9 or 161Refer to the supplementary text for the details. According to their kinetic properties, hK5, hK6, hK8, hK13, and hK14 strongly display trypsin-like (FSR-) activity (Oka et al., 2002; Magklara et al., 2003; Kapadia et al., 2004; Felber et al., 2005; Michael et al., 2005, respectively). hK7 may be largely responsible for the chymotrypsin-like (RPY-) activity (Franzke et al., 1996; Komatsu et al., 2005b). The identity of enzymes contributing to PFR- and VLK-activities in the skin is unknown. Arg-X-Lys-Arg- (R-X-KR) sequences are repeatedly found between an inhibitory domain and other domains of SPINK5 proprotein, and the sequence is the specific proteolytic target (R-KR-↓) of furin (Seidah and Chretien, 1999; Komatsu et al., 2002; Mitsudo et al., 2003). The co-localization of SPINK5 protein and furin in normal skin (Bergeron et al., 2000; Bitoun et al., 2003) allowed us to predict the presence of a furin-like activity in normal SC. mean±SDPatient MPatient KMeanMeanTrypsin-like activity Phe-Ser-Arg-AMC215.5±1.521.015.5 Pro-Phe-Arg-AMC25.7±3.11Refer to the supplementary text for the details. According to their kinetic properties, hK5, hK6, hK8, hK13, and hK14 strongly display trypsin-like (FSR-) activity (Oka et al., 2002; Magklara et al., 2003; Kapadia et al., 2004; Felber et al., 2005; Michael et al., 2005, respectively). hK7 may be largely responsible for the chymotrypsin-like (RPY-) activity (Franzke et al., 1996; Komatsu et al., 2005b). The identity of enzymes contributing to PFR- and VLK-activities in the skin is unknown. Arg-X-Lys-Arg- (R-X-KR) sequences are repeatedly found between an inhibitory domain and other domains of SPINK5 proprotein, and the sequence is the specific proteolytic target (R-KR-↓) of furin (Seidah and Chretien, 1999; Komatsu et al., 2002; Mitsudo et al., 2003). The co-localization of SPINK5 protein and furin in normal skin (Bergeron et al., 2000; Bitoun et al., 2003) allowed us to predict the presence of a furin-like activity in normal SC.24.0*Significant differences (P<0.05) between the mean of normal samples and each patient at the specified time (Smirnov test for extreme values).18.6*Significant differences (P<0.05) between the mean of normal samples and each patient at the specified time (Smirnov test for extreme values).Chymotrypsin-like activity Arg-Pro-Tyr-pNA413.9±5.511.913.7Plasmin-like activity Val-Leu-Lys-AMC21.7±1.01Refer to the supplementary text for the details. According to their kinetic properties, hK5, hK6, hK8, hK13, and hK14 strongly display trypsin-like (FSR-) activity (Oka et al., 2002; Magklara et al., 2003; Kapadia et al., 2004; Felber et al., 2005; Michael et al., 2005, respectively). hK7 may be largely responsible for the chymotrypsin-like (RPY-) activity (Franzke et al., 1996; Komatsu et al., 2005b). The identity of enzymes contributing to PFR- and VLK-activities in the skin is unknown. Arg-X-Lys-Arg- (R-X-KR) sequences are repeatedly found between an inhibitory domain and other domains of SPINK5 proprotein, and the sequence is the specific proteolytic target (R-KR-↓) of furin (Seidah and Chretien, 1999; Komatsu et al., 2002; Mitsudo et al., 2003). The co-localization of SPINK5 protein and furin in normal skin (Bergeron et al., 2000; Bitoun et al., 2003) allowed us to predict the presence of a furin-like activity in normal SC.8.9*Significant differences (P<0.05) between the mean of normal samples and each patient at the specified time (Smirnov test for extreme values).19.6*Significant differences (P<0.05) between the mean of normal samples and each patient at the specified time (Smirnov test for extreme values).Furin-like activity Pyr-Arg-Thr-Lys-Arg-AMC23.0±1.31Refer to the supplementary text for the details. According to their kinetic properties, hK5, hK6, hK8, hK13, and hK14 strongly display trypsin-like (FSR-) activity (Oka et al., 2002; Magklara et al., 2003; Kapadia et al., 2004; Felber et al., 2005; Michael et al., 2005, respectively). hK7 may be largely responsible for the chymotrypsin-like (RPY-) activity (Franzke et al., 1996; Komatsu et al., 2005b). The identity of enzymes contributing to PFR- and VLK-activities in the skin is unknown. Arg-X-Lys-Arg- (R-X-KR) sequences are repeatedly found between an inhibitory domain and other domains of SPINK5 proprotein, and the sequence is the specific proteolytic target (R-KR-↓) of furin (Seidah and Chretien, 1999; Komatsu et al., 2002; Mitsudo et al., 2003). The co-localization of SPINK5 protein and furin in normal skin (Bergeron et al., 2000; Bitoun et al., 2003) allowed us to predict the presence of a furin-like activity in normal SC.43.0*Significant differences (P<0.05) between the mean of normal samples and each patient at the specified time (Smirnov test for extreme values).28.5*Significant differences (P<0.05) between the mean of normal samples and each patient at the specified time (Smirnov test for extreme values).AMC, 7-amino-4-methyl-commarin; pNA, para-nitroanilide; PSS, peeling skin syndrome; SC, stratum corneum; SD, standard deviation.The overall SC serine protease enzymatic activities represent released AMC or pNA from the synthetic substrates for the normal subjects (mean±SD; nmol/mg dry weight). The amount of released AMC or pNA was measured at 2 or 4 h, respectively. Each assay was performed in triplicate for the patients and the mean values are indicated.* Significant differences (P<0.05) between the mean of normal samples and each patient at the specified time (Smirnov test for extreme values).1 Refer to the supplementary text for the details. According to their kinetic properties, hK5, hK6, hK8, hK13, and hK14 strongly display trypsin-like (FSR-) activity (Oka et al., 2002Oka T. Hakoshima T. Itakura M. Yamamori S. Takahashi M. Hashimoto Y. et al.Role of loop structures of neuropsin in the activity of serine protease and regulated secretion.J Biol Chem. 2002; 277: 14724-14730Crossref PubMed Scopus (28) Google Scholar; Magklara et al., 2003Magklara A. Mellati A.A. Wasney G.A. Little S.P. Sotiropoulou G. 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Arg-X-Lys-Arg- (R-X-KR) sequences are repeatedly found between an inhibitory domain and other domains of SPINK5 proprotein, and the sequence is the specific proteolytic target (R-KR-↓) of furin (Seidah and Chretien, 1999Seidah N.G. Chretien M. Proprotein and prohormone convertases: a family of subtilases generating diverse bioactive polypeptides.Brain Res. 1999; 848: 45-62Crossref PubMed Scopus (660) Google Scholar; Komatsu et al., 2002Komatsu N. Takata M. Otsuki N. Ohka R. Amano O. Takehara K. et al.Elevated stratum corneum hydrolytic activity in Netherton syndrome suggests an inhibitory regulation of desquamation by SPINK5-derived peptides.J Invest Dermatol. 2002; 118: 436-443Crossref PubMed Scopus (167) Google Scholar; Mitsudo et al., 2003Mitsudo K. Jayakumar A. Henderson Y. Frederick M.J. Kang Y. Wang M. et al.Inhibition of serine proteinases plasmin, trypsin, subtilisin A, cathepsin G, and elastase by LEKTI: a kinetic analysis.Biochemistry. 2003; 42: 3874-3881Crossref PubMed Scopus (85) Google Scholar). The co-localization of SPINK5 protein and furin in normal skin (Bergeron et al., 2000Bergeron F. Leduc R. Day R. Subtilase-like pro-protein convertases: from molecular specificity to therapeutic applications.J Mol Endocrinol. 2000; 24: 1-22Crossref PubMed Scopus (160) Google Scholar; Bitoun et al., 2003Bitoun E. Micheloni A. Lamant L. Bonnart C. Tartaglia-Polcini A. Cobbold C. et al.LEKTI proteolytic processing in human primary keratinocytes, tissue distribution, and defective expression in Netherton syndrome.Hum Mol Genet. 2003; 12: 2417-2430Crossref PubMed Scopus (161) Google Scholar) allowed us to predict the presence of a furin-like activity in normal SC. Open table in a new tab AMC, 7-amino-4-methyl-commarin; pNA, para-nitroanilide; PSS, peeling skin syndrome; SC, stratum corneum; SD, standard deviation. The overall SC serine protease enzymatic activities represent released AMC or pNA from the synthetic substrates for the normal subjects (mean±SD; nmol/mg dry weight). The amount of released AMC or pNA was measured at 2 or 4 h, respectively. Each assay was performed in triplicate for the patients and the mean values are indicated. The overall SC trypsin-like (FSR-) and chymotrypsin-like (RPY-) activities are known as desquamation-related SC protease activities (Suzuki et al., 1996Suzuki Y. Koyama J. Moro O. Horii I. Kikuchi K. Tanida M. et al.The role of two endogenous proteases of the stratum corneum in degradation of desmoglein-1 and their reduced activity in the skin of ichthyotic patients.Br J Dermatol. 1996; 134: 460-464Crossref PubMed Scopus (70) Google Scholar). These activities are regulated in a consistent manner across different age groups, indicating that maintenance of a stable SC serine protease activity may be essential for retaining a constant number of SC layers which is known to be unaffected by aging (Ya-Xian et al., 1999Ya-Xian Z. Suetake T. Tagami H. Number of cell layers of the stratum corneum in normal skin – relationship to the anatomical location on the body, age, sex and physical parameters.Arch Dermatol Res. 1999; 291: 555-559Crossref PubMed Scopus (294) Google Scholar; Komatsu et al., 2005Komatsu N. Saijoh K. Sidiropoulos M. Tsai B. Levesque M.A. Elliott M.B. et al.Quantification of human tissue kallikreins in the stratum corneum: dependence on age and gender.J Invest Dermatol. 2005; 125: 1182-1192Crossref PubMed Scopus (65) Google Scholar). hKs may contribute to the overall SC protease activities and the degradation of intercellular adhesion molecules resulting in desquamation of corneocytes (Simon et al., 2001Simon M. Jonca N. Guerrin M. Haftek M. Bernard D. Caubet C. et al.Refined characterization of corneodesmosin proteolysis during terminal differentiation of human epidermis and its relationship to desquamation.J Biol Chem. 2001; 276: 20292-20299Crossref PubMed Scopus (146) Google Scholar; Komatsu et al., 2002Komatsu N. Takata M. Otsuki N. Ohka R. Amano O. Takehara K. et al.Elevated stratum corneum hydrolytic activity in Netherton syndrome suggests an inhibitory regulation of desquamation by SPINK5-derived peptides.J Invest Dermatol. 2002; 118: 436-443Crossref PubMed Scopus (167) Google Scholar; Caubet et al., 2004Caubet C. Jonca N. Brattsand M. Guerrin M. Bernard D. Schmidt R. et al.Degradation of corneodesmosome proteins by two serine proteases of the kallikrein family, SCTE/KLK5/hK5 and SCCE/KLK7/hK7.J Invest Dermatol. 2004; 122: 1235-1244Crossref PubMed Scopus (392) Google Scholar) (Figure 2a). SPINK5 inhibitory domains are believed to be negative regulators of desquamation-related proteases including hKs (Mitsudo et al., 2003Mitsudo K. Jayakumar A. Henderson Y. Frederick M.J. Kang Y. Wang M. et al.Inhibition of serine proteinases plasmin, trypsin, subtilisin A, cathepsin G, and elastase by LEKTI: a kinetic analysis.Biochemistry. 2003; 42: 3874-3881Crossref PubMed Scopus (85) Google Scholar; Descargues et al., 2005Descargues P. Deraison C. Bonnart C. Kreft M. Kishibe M. Ishida-Yamamoto A. et al.Spink5-deficient mice mimic Netherton syndrome through degradation of desmoglein 1 by epidermal protease hyperactivity.Nat Genet. 2005; 37: 56-65Crossref PubMed Scopus (289) Google Scholar; Egelrud et al., 2005Egelrud T. Brattsand M. Kreutzmann P. Walden M. Vitzithum K. Marx U.C. et al.hK5 and hK7, two serine proteinases abundant in human skin, are inhibited by LEKTI domain 6.Br J Dermatol. 2005; 153: 1200-1203Crossref PubMed Scopus (97) Google Scholar; Schechter et al., 2005Schechter N.M. Choi E.J. Wang Z.M. Hanakawa Y. Stanley J.R. Kang Y. et al.Inhibition of human kallikreins 5 and 7 by the serine protease inhibitor lympho-epithelial Kazal-type inhibitor (LEKTI).Biol Chem. 2005; 386: 1173-1184Crossref PubMed Scopus (84) Google Scholar) (Figure 2a). SPINK5 proprotein can be proteolytically processed at (R-KR↓) by furin-like activity to 15 individual bioactive domains (Seidah and Chretien, 1999Seidah N.G. Chretien M. Proprotein and prohormone convertases: a family of subtilases generating diverse bioactive polypeptides.Brain Res. 1999; 848: 45-62Crossref PubMed Scopus (660) Google Scholar; Komatsu et al., 2002Komatsu N. Takata M. Otsuki N. Ohka R. Amano O. Takehara K. et al.Elevated stratum corneum hydrolytic activity in Netherton syndrome suggests an inhibitory regulation of desquamation by SPINK5-derived peptides.J Invest Dermatol. 2002; 118: 436-443Crossref PubMed Scopus (167) Google Scholar; Mitsudo et al., 2003Mitsudo K. Jayakumar A. Henderson Y. Frederick M.J. Kang Y. Wang M. et al.Inhibition of serine proteinases plasmin, trypsin, subtilisin A, cathepsin G, and elastase by LEKTI: a kinetic analysis.Biochemistry. 2003; 42: 3874-3881Crossref PubMed Scopus (85) Google Scholar) (Figure 2a). SPINK5 knockout mice displayed high trypsin-like and chymotrypsin-like activities in their SC (Descargues et al., 2005Descargues P. Deraison C. Bonnart C. Kreft M. Kishibe M. Ishida-Yamamoto A. et al.Spink5-deficient mice mimic Netherton syndrome through degradation of desmoglein 1 by epidermal protease hyperactivity.Nat Genet. 2005; 37: 56-65Crossref PubMed Scopus (289) Google Scholar). Similarly, NS patients, who lack the downstream of SPINK5 products, display significantly high trypsin-like (FSR-) (Komatsu et al., 2002Komatsu N. Takata M. Otsuki N. Ohka R. Amano O. Takehara K. et al.Elevated stratum corneum hydrolytic activity in Netherton syndrome suggests an inhibitory regulation of desquamation by SPINK5-derived peptides.J Invest Dermatol. 2002; 118: 436-443Crossref PubMed Scopus (167) Google Scholar) and chymotrypsin-like (RPY-) activities (unpublished data) (Figure 2b). This indicates that SPINK5 protein possesses strong inhibitory functions towards FSR- and RPY-activities. The increased epidermal cell layers expressing SPINK5 protein in the patients' skins could lead to an enhanced expression of SPINK5 proprotein, and then the elevation of furin-like (R-KR-) activity may yield an excess amount of SPINK5 inhibitory domains by proteolysis of SPINK5 proprotein (Figure 2c). Ultimately, the mild elevation of FSR-activity and the normal RPY-activity in the two PSS-type B patients, both of whom with “intact” (and may be increased) SPINK5 protein/domain expression, could be explained by an efficient inhibitory function by SPINK5 proteins. As SPINK5 protein possesses anti-“plasmin” function (Mitsudo et al., 2003Mitsudo K. Jayakumar A. Henderson Y. Frederick M.J. Kang Y. Wang M. et al.Inhibition of serine proteinases plasmin, trypsin, subtilisin A, cathepsin G, and elastase by LEKTI: a kinetic analysis.Biochemistry. 2003; 42: 3874-3881Crossref PubMed Scopus (85) Google Scholar), we measured overall SC “plasmin-like (VLK-) activity” in the SC. However, the elevated VLK-activity in the patients suggested that SPINK5 protein may not be an adequate inhibitor for the overall SC “plasmin-like activity”. The trypsin-like (FSR-) and (PFR-) activities in the patients showed different preferences; therefore, the composition of proteases that play a role in FSR- and PFR-activities may be different. The elevated SC protease activities in the PSS-type B patients with “intact” SPINK5 implied an abnormal regulation of SC protease expressions/activities. Hence, we focused on the SC proteases, especially hKs, assuming an aberrant hK expression in these patients. As predicted, the amount of all hKs in the SC and serum of the PSS-type B patients was substantially higher than those in normal subjects. hKs may function as an enzymatic cascade pathway in many tissues (Yousef and Diamandis, 2002Yousef G.M. Diamandis E.P. Human tissue kallikreins: a new enzymatic cascade pathway.Biol Chem. 2002; 383: 1045-1057Crossref PubMed Scopus (86) Google Scholar), for example, hK5 can activate the proforms of hK7 and hK14 (Caubet et al., 2004Caubet C. Jonca N. Brattsand M. Guerrin M. Bernard D. Schmidt R. et al.Degradation of corneodesmosome proteins by two serine proteases of the kallikrein family, SCTE/KLK5/hK5 and SCCE/KLK7/hK7.J Invest Dermatol. 2004; 122: 1235-1244Crossref PubMed Scopus (392) Google Scholar), and activation of pro-hK5 is either autocatalytic or is mediated by hK14 (Brattsand et al., 2005Brattsand M. Stefansson K. Lundh C. Haasum Y. Egelrud T. A proteolytic cascade of kallikreins in the stratum corneum.J Invest Dermatol. 2005; 124: 198-203Crossref PubMed Scopus (259) Google Scholar). As hK7 exists in the SC as a mixture of the proform and the active form (Ekholm and Egelrud, 1999Ekholm E. Egelrud T. Stratum corneum chymotryptic enzyme in psoriasis.Arch Dermatol Res. 1999; 291: 195-200Crossref PubMed Scopus (74) Google Scholar), other hKs in the SC may also exist in a similar manner as mixtures. It is conceivable that the abnormally increased concentration of multiple hKs in PSS-type B patients would result in an aberrant activation of other hKs (an altered ratio of pro- and active-form in hKs) and/or activation (and possibly amplification) of an hK enzymatic cascade. This could lead to a high SC enzymatic protease activity, an overdegradation of intercellular adhesion molecules, and finally, an abnormal desquamation (Figure 2c). NS and PSS-type B may have totally different pathogenesis; however, the final stages for each disease, that is, an elevation of SC protease activities and detachment of the SC (probably owing to over-desquamation) seem to be similar (Figure 2b and c), which may explain the clinical similarities between them. NS patients exhibit severe over-absorption from their skin owing to the over-desquamation of corneocytes (Komatsu et al., 2002Komatsu N. Takata M. Otsuki N. Ohka R. Amano O. Takehara K. et al.Elevated stratum corneum hydrolytic activity in Netherton syndrome suggests an inhibitory regulation of desquamation by SPINK5-derived peptides.J Invest Dermatol. 2002; 118: 436-443Crossref PubMed Scopus (167) Google Scholar), and skin permeability barrier dysfunction, causing iatrogenic Cushing's syndrome (Smith et al., 1995Smith D.L. Smith J.G. Wong S.W. deShazo R.D. Netherton's syndrome: a syndrome of elevated IgE and characteristic skin and hair findings.J Allergy Clin Immunol. 1995; 95: 116-123Abstract Full Text Full Text PDF PubMed Scopus (112) Google Scholar) and elevated tacrolimus serum levels with a kidney disorder (Allen et al., 2001Allen A. Siegfried E. Silverman R. Williams M.L. Elias P.M. Szabo S.K. et al.Significant absorption of topical tacrolimus in 3 patients with Netherton syndrome.Arch Dermatol. 2001; 137: 747-750PubMed Google Scholar). Patient M showed over-absorption through her skin, highly suggesting that the PSS-type B patients, like NS patients, suffer from over-desquamation. In conclusion, we suggest that PSS-type B may not be an ichthyosis characterized by the retention of thick adherent scales (Frost and Van Scott, 1966Frost P. Van Scott E.J. Ichthyosiform dermatoses. Classification based on anatomic and biometric observations.Arch Dermatol. 1966; 94: 113-126Crossref PubMed Scopus (154) Google Scholar) but an over-desquamation disease owing to an over-expression of hKs and an elevation of the SC protease activities. The over-desquamation of corneocytes may explain the clinical similarities between PSS-type B and NS. The elevated SC enzymatic activities may be good therapeutic targets for PSS-type B patients. The authors state no conflict of interest. We thank the patients, their parents, and our volunteers for generously providing samples, and K. Hama, Y. Obata, Y. Yamada, C. Kuk, and CK. Cho for technical help. This work was also supported in part by the University of Texas MD Anderson Cancer Center SPORE in Head and Neck Cancer NIH-NCI P50 CA097007 (G.L.C), NIH R01 DE013954 G.L.C), Cancer Center Support Grant NIH P30 CA016672 (G.L.C), Alando J. Ballantyne Distinguished Chair in Head and Neck Surgery award (G.L.C), Michael A. O'Bannon Endowment for Cancer Research (G.L.C), Betty Berry Cancer Research Fund (G.L.C), and NIH INRS Award T32 CA060374 (G.L.C), 2004 AAO-HNSF Percy Memorial Grant (G.L.C.). Work on kallikreins in Eleftherios P. Diamandis' lab is supported by grants from the Natural Sciences and Engineering Council of Canada (NSERC) and IBEX Technologies Inc., Canada. Download .pdf (.09 MB) Help with pdf files Supplementary materials and methods, references
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