Correspondence Clinical Letter
Clinical Letter Upregulation of tumor necrosis factor alphainduced protein 3 interacting protein 1 mRNA in psoriasis vulgaris
DOI: 10.1111/ddg.12462
Dear Editors, Psoriasis vulgaris is an inflammatory skin disorder that is characterized by hyperproliferation and abnormal differentiation of keratinocytes, a mixed dermal cellular infiltrate and angiogenesis. It has been demonstrated that nuclear factor-κB (NF-κB) activity contributes to the pathogenesis of psoriasis vulgaris [1]. A tight regulation of NF-κB signaling is thus absolutely required. Two novel genes were identified as key players in the regulation of NF-κB signaling: tumor necrosis factor alpha-induced protein 3 (TNFAIP3) and TNFAIP3 interacting protein 1 (TNIP1). The proteins encoded by these two genes can bind to each other and function as a “brake” on NF-κB signaling [2]. Recent studies demonstrated that TNFAIP3 mRNA is highly expressed in patients with mild psoriasis vulgaris, but not in those with severe disease [3, 4]. Since TNIP1 may facilitate the function of TNFAIP3 in the negative feedback regulation of NF-κB activation [5], the corresponding transcript levels of TNIP1 in psoriasis vulgaris were investigated in this study. The pairwise lesional and normal-appearing skin from 20 patients with psoriasis vulgaris and control skin from ten healthy volunteers were assessed in this study with informed consent and approval from the Ethics Committee of Fudan University. Based on the clinical classification of the American National Psoriasis Foundation, body surface area (BSA) was used to stratify patients with psoriasis into two subgroups: In the “mild” group, lesions affected less than 3 % of the body surface, whereas in the “moderate-to-severe”
group, lesions covered more than 3 % of the body surface. Psoriasis area and severity index (PASI) was used to evaluate the severity of psoriasis vulgaris. Total RNA was isolated from the skin using the RNeasy Protect minikit (Qiagen, Hilden, Germany). The level of polymerase (RNA) II (DNA-directed) polypeptide A (POLR2A) mRNA was determined as an internal control for each sample. Primers used in real-time PCR were as follows: TNIP1 forward, 5′-AAAGGAGATCCAGCGGCTCAAC-3′; TNIP1 reverse, 5′-TTCTCCTCATTCATGCGCTCAC-3′; POLR2A forward, 5′-GCAGAGAAGCTGGTGCTCCGTA-3′; and POLR2A reverse, 5′-CAGCATGTTGGACTCGATGCAG -3′. SYBR Green real-time PCRs of TNIP1 and POLR2A were performed, as described previously [4]. Gene expression levels in the different groups were compared using the paired t-test and ANOVA test. Spearman’s rank test was used to analyze the correlation between different test pairs, which were performed using GraphPad Prism software (v 5.0). As indicated in Table 1, ten patients were identified with mild psoriasis vulgaris, while the other ten patients were ascribed to the “moderate-to-severe” group. TNIP1 mRNA expression levels between the groups were significantly different (p = 0.022), especially between severe lesions and normal controls (p = 0.005). Likewise, there was a statistically significant difference between lesions and normal-appearing skin in the group with “mild” psoriasis vulgaris (p = 0.017). However, no such difference was found in the “moderate-to-severe” group (p = 0.230) (Figure 1). Based on our limited number of cases, the transcript levels of TNIP1 in psoriatic lesions did not show any significant correlations with BSA, nor with PASI scores. TNIP1 encodes the A20-binding inhibitor of NF-κB (ABIN-1). In vitro studies suggested that ABIN-1 can bind NEMO/IKKγ and inhibit TNF-induced NF-κB signaling [5]. Moreover, ABIN-1 can also bind ubiquitin chains. The ubiquitin binding by ABIN-1 is important for the ability of ABIN-1 to restrict TNF and Toll-like receptor (TLR) signaling [6, 7]. Our results showed that the TNIP1 mRNA levels were significantly increased in patients with severe psoriasis
Table 1 The demographic and clinical severity of patients with psoriasis vulgaris and controls. Groups
Mild
Severe
Controls
10
10
10
50.5 (24–64)
50 (28–65)
62.5 (14–70)
Sex M/F
6/4
10/0
4/6
Mean BSA (%)
1.5
23.8
n/a
Mean PASI
4.4
18.5
n/a
No. Median age (range) (yr)
Abbr.: BSA, body skin area; PASI, psoriasis severity and area index; n/a, not available.
© 2015 Deutsche Dermatologische Gesellschaft (DDG). Published by John Wiley & Sons Ltd. | JDDG | 1610-0379/2015/1302
159
Correspondence Clinical Letter
Funding sources Nanshan Science and Technology Bureau of Shenzhen (grant 2012043), and the Research Special Fund for Public Welfare Industry of Health, Guideline-oriented Research in the Management of Some Common and Severe Skin Diseases (No. 201002016). Conflict of interest None.
Fang Wang1*, Xia Zhang2*, Ping Xia3, Li Zhang2, Zhenghua Zhang3
Figure 1 Relative tumor necrosis factor alpha-induced protein 3 interacting protein 1 mRNA expression levels in normal-appearing skin (N) and lesions (L) from patients with mild and severe levels of psoriasis vulgaris and in normal skin from healthy controls (Normal). Each symbol represents the value for an individual patient.
(1) Department of Dermatology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China (2) Department of Dermatology, Shekou People’s Hospital, Shenzhen, Guangdong, China (3) Department of Dermatology, Huashan Hospital, Fudan University, Shanghai, China *The first two authors contributed equally to the present article.
Correspondence to compared with normal controls, suggesting that TNIP1 plays a role in a negative feedback loop that down-regulates immune responses in psoriasis, especially in severe cases. Considering that no correlation between TNIP1 mRNA levels and PASI scores were found, TNIP1 is no reliable marker for the severity of psoriasis. It is worth to note that the products of the TNFAIP3 and TNIP1 genes (A20 and ABIN1, respectively) functionally bind to each other, an interaction that may synergistically impede NF-κB signaling and negatively regulate inflammation [5]. Interestingly, in our previous study, TNFAIP3 seems to be the immediate early gene for defense reactions in the pathogenesis of psoriasis vulgaris, as its mRNA expression was significantly up-regulated in lesions compared to normal-appearing skin in the “mild” psoriasis group [4]. Moreover, our results showed an upward trend of TNIP1 mRNA levels from the mild to the severe subgroup. Thus, in contrast to the “quick-acting brake” of TNFAIP3, it is conceivable that TNIP1 acts as a relatively “slow and persistent brake” on excessive inflammation [8]. Nonetheless, further studies are needed to analyze specific interactions of TNFAIP3 and TNIP1 in the immunopathogenesis of psoriasis.
Acknowledgements We thank all patients who participated in this study.
160
Zhenghua Zhang Department of Dermatology Huashan Hospital Shanghai Medical College Fudan University Shanghai, 200040, China E-mail: [email protected]
References 1 2
3
4
5
6
Doger FK, Dikicioglu E, Ergin F et al. Nature of cell kinetics in psoriatic epidermis. J Cutan Pathol 2007; 34: 257–63. Nair RP, Ding J, Duffin KC et al. Psoriasis bench to bedside: genetics meets immunology. Arch Dermatol 2009; 145: 462–4. Jiang X, Tian H, Fan Y et al. Expression of tumor necrosis factor alpha-induced protein 3 mRNA in peripheral blood mononuclear cells negatively correlates with disease severity in psoriasis vulgaris. Clin Vaccine Immunol 2012; 19: 1938–42. Zhang X, Xia P, Zhang L, Zhang Z. Upregulation of tumor necrosis factor alpha induced protein 3 mRNA in mild psoriasis vulgaris. Clin Vaccine Immunol 2013; 20: 1341. Mauro C, Pacifico F, Lavorgna A et al. ABIN-1 binds to NEMO/ IKKgamma and co-operates with A20 in inhibiting NF-kappaB. J Biol Chem 2006; 281: 18482–8. Oshima S, Turer EE, Callahan JA et al. ABIN-1 is a ubiquitin sensor that restricts cell death and sustains embryonic development. Nature 2009; 457: 906–9.
© 2015 Deutsche Dermatologische Gesellschaft (DDG). Published by John Wiley & Sons Ltd. | JDDG | 1610-0379/2015/1302
Correspondence Clinical Letter
7
Nanda SK, Venigalla RK, Ordureau A et al. Polyubiquitin binding to ABIN1 is required to prevent autoimmunity. J Exp Med 2011; 208: 1215–28.
8
Callahan JA, Hammer GE, Agelides A et al. Cutting edge: ABIN1 protects against psoriasis by restricting MyD88 signals in dendritic cells. J Immunol 2013; 191: 535–9.
© 2015 Deutsche Dermatologische Gesellschaft (DDG). Published by John Wiley & Sons Ltd. | JDDG | 1610-0379/2015/1302
161
Clinical Letter Upregulation of tumor necrosis factor alphainduced protein 3 interacting protein 1 mRNA in psoriasis vulgaris
DOI: 10.1111/ddg.12462
Dear Editors, Psoriasis vulgaris is an inflammatory skin disorder that is characterized by hyperproliferation and abnormal differentiation of keratinocytes, a mixed dermal cellular infiltrate and angiogenesis. It has been demonstrated that nuclear factor-κB (NF-κB) activity contributes to the pathogenesis of psoriasis vulgaris [1]. A tight regulation of NF-κB signaling is thus absolutely required. Two novel genes were identified as key players in the regulation of NF-κB signaling: tumor necrosis factor alpha-induced protein 3 (TNFAIP3) and TNFAIP3 interacting protein 1 (TNIP1). The proteins encoded by these two genes can bind to each other and function as a “brake” on NF-κB signaling [2]. Recent studies demonstrated that TNFAIP3 mRNA is highly expressed in patients with mild psoriasis vulgaris, but not in those with severe disease [3, 4]. Since TNIP1 may facilitate the function of TNFAIP3 in the negative feedback regulation of NF-κB activation [5], the corresponding transcript levels of TNIP1 in psoriasis vulgaris were investigated in this study. The pairwise lesional and normal-appearing skin from 20 patients with psoriasis vulgaris and control skin from ten healthy volunteers were assessed in this study with informed consent and approval from the Ethics Committee of Fudan University. Based on the clinical classification of the American National Psoriasis Foundation, body surface area (BSA) was used to stratify patients with psoriasis into two subgroups: In the “mild” group, lesions affected less than 3 % of the body surface, whereas in the “moderate-to-severe”
group, lesions covered more than 3 % of the body surface. Psoriasis area and severity index (PASI) was used to evaluate the severity of psoriasis vulgaris. Total RNA was isolated from the skin using the RNeasy Protect minikit (Qiagen, Hilden, Germany). The level of polymerase (RNA) II (DNA-directed) polypeptide A (POLR2A) mRNA was determined as an internal control for each sample. Primers used in real-time PCR were as follows: TNIP1 forward, 5′-AAAGGAGATCCAGCGGCTCAAC-3′; TNIP1 reverse, 5′-TTCTCCTCATTCATGCGCTCAC-3′; POLR2A forward, 5′-GCAGAGAAGCTGGTGCTCCGTA-3′; and POLR2A reverse, 5′-CAGCATGTTGGACTCGATGCAG -3′. SYBR Green real-time PCRs of TNIP1 and POLR2A were performed, as described previously [4]. Gene expression levels in the different groups were compared using the paired t-test and ANOVA test. Spearman’s rank test was used to analyze the correlation between different test pairs, which were performed using GraphPad Prism software (v 5.0). As indicated in Table 1, ten patients were identified with mild psoriasis vulgaris, while the other ten patients were ascribed to the “moderate-to-severe” group. TNIP1 mRNA expression levels between the groups were significantly different (p = 0.022), especially between severe lesions and normal controls (p = 0.005). Likewise, there was a statistically significant difference between lesions and normal-appearing skin in the group with “mild” psoriasis vulgaris (p = 0.017). However, no such difference was found in the “moderate-to-severe” group (p = 0.230) (Figure 1). Based on our limited number of cases, the transcript levels of TNIP1 in psoriatic lesions did not show any significant correlations with BSA, nor with PASI scores. TNIP1 encodes the A20-binding inhibitor of NF-κB (ABIN-1). In vitro studies suggested that ABIN-1 can bind NEMO/IKKγ and inhibit TNF-induced NF-κB signaling [5]. Moreover, ABIN-1 can also bind ubiquitin chains. The ubiquitin binding by ABIN-1 is important for the ability of ABIN-1 to restrict TNF and Toll-like receptor (TLR) signaling [6, 7]. Our results showed that the TNIP1 mRNA levels were significantly increased in patients with severe psoriasis
Table 1 The demographic and clinical severity of patients with psoriasis vulgaris and controls. Groups
Mild
Severe
Controls
10
10
10
50.5 (24–64)
50 (28–65)
62.5 (14–70)
Sex M/F
6/4
10/0
4/6
Mean BSA (%)
1.5
23.8
n/a
Mean PASI
4.4
18.5
n/a
No. Median age (range) (yr)
Abbr.: BSA, body skin area; PASI, psoriasis severity and area index; n/a, not available.
© 2015 Deutsche Dermatologische Gesellschaft (DDG). Published by John Wiley & Sons Ltd. | JDDG | 1610-0379/2015/1302
159
Correspondence Clinical Letter
Funding sources Nanshan Science and Technology Bureau of Shenzhen (grant 2012043), and the Research Special Fund for Public Welfare Industry of Health, Guideline-oriented Research in the Management of Some Common and Severe Skin Diseases (No. 201002016). Conflict of interest None.
Fang Wang1*, Xia Zhang2*, Ping Xia3, Li Zhang2, Zhenghua Zhang3
Figure 1 Relative tumor necrosis factor alpha-induced protein 3 interacting protein 1 mRNA expression levels in normal-appearing skin (N) and lesions (L) from patients with mild and severe levels of psoriasis vulgaris and in normal skin from healthy controls (Normal). Each symbol represents the value for an individual patient.
(1) Department of Dermatology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China (2) Department of Dermatology, Shekou People’s Hospital, Shenzhen, Guangdong, China (3) Department of Dermatology, Huashan Hospital, Fudan University, Shanghai, China *The first two authors contributed equally to the present article.
Correspondence to compared with normal controls, suggesting that TNIP1 plays a role in a negative feedback loop that down-regulates immune responses in psoriasis, especially in severe cases. Considering that no correlation between TNIP1 mRNA levels and PASI scores were found, TNIP1 is no reliable marker for the severity of psoriasis. It is worth to note that the products of the TNFAIP3 and TNIP1 genes (A20 and ABIN1, respectively) functionally bind to each other, an interaction that may synergistically impede NF-κB signaling and negatively regulate inflammation [5]. Interestingly, in our previous study, TNFAIP3 seems to be the immediate early gene for defense reactions in the pathogenesis of psoriasis vulgaris, as its mRNA expression was significantly up-regulated in lesions compared to normal-appearing skin in the “mild” psoriasis group [4]. Moreover, our results showed an upward trend of TNIP1 mRNA levels from the mild to the severe subgroup. Thus, in contrast to the “quick-acting brake” of TNFAIP3, it is conceivable that TNIP1 acts as a relatively “slow and persistent brake” on excessive inflammation [8]. Nonetheless, further studies are needed to analyze specific interactions of TNFAIP3 and TNIP1 in the immunopathogenesis of psoriasis.
Acknowledgements We thank all patients who participated in this study.
160
Zhenghua Zhang Department of Dermatology Huashan Hospital Shanghai Medical College Fudan University Shanghai, 200040, China E-mail: [email protected]
References 1 2
3
4
5
6
Doger FK, Dikicioglu E, Ergin F et al. Nature of cell kinetics in psoriatic epidermis. J Cutan Pathol 2007; 34: 257–63. Nair RP, Ding J, Duffin KC et al. Psoriasis bench to bedside: genetics meets immunology. Arch Dermatol 2009; 145: 462–4. Jiang X, Tian H, Fan Y et al. Expression of tumor necrosis factor alpha-induced protein 3 mRNA in peripheral blood mononuclear cells negatively correlates with disease severity in psoriasis vulgaris. Clin Vaccine Immunol 2012; 19: 1938–42. Zhang X, Xia P, Zhang L, Zhang Z. Upregulation of tumor necrosis factor alpha induced protein 3 mRNA in mild psoriasis vulgaris. Clin Vaccine Immunol 2013; 20: 1341. Mauro C, Pacifico F, Lavorgna A et al. ABIN-1 binds to NEMO/ IKKgamma and co-operates with A20 in inhibiting NF-kappaB. J Biol Chem 2006; 281: 18482–8. Oshima S, Turer EE, Callahan JA et al. ABIN-1 is a ubiquitin sensor that restricts cell death and sustains embryonic development. Nature 2009; 457: 906–9.
© 2015 Deutsche Dermatologische Gesellschaft (DDG). Published by John Wiley & Sons Ltd. | JDDG | 1610-0379/2015/1302
Correspondence Clinical Letter
7
Nanda SK, Venigalla RK, Ordureau A et al. Polyubiquitin binding to ABIN1 is required to prevent autoimmunity. J Exp Med 2011; 208: 1215–28.
8
Callahan JA, Hammer GE, Agelides A et al. Cutting edge: ABIN1 protects against psoriasis by restricting MyD88 signals in dendritic cells. J Immunol 2013; 191: 535–9.
© 2015 Deutsche Dermatologische Gesellschaft (DDG). Published by John Wiley & Sons Ltd. | JDDG | 1610-0379/2015/1302
161
