Gene expression profile in keratinocytes from atopic dermatitis patients

S. Pastore, L. Rogge, F. Mariotti, F. Mascia, R. La Scaleia, C. Dattilo, F. Sinigaglia, G. Girolomoni

Research output: Contribution to journalArticle

Abstract

Aim. Keratinocytes of atopic dermatitis (AD) patients show enhanced production of cytokines and chemokines, a phenomenon that could be relevant in promoting and maintaining inflammation and hence pivotal in localizing the atopic diathesis to the skin. We performed an oligonucleotide microarrray analysis to investigate the gene expression profile in keratinocyte cultures from 6 AD patients in order to search differentially expressed genes. Methods. Six well informed volunteer patients with moderate-to-severe chronic AD (age range 19-45 years) participated in this study. Skin involvement ranged from 20% to 60% of the body surface area. Six well informed volunteer healthy individuals (age range: 25-50 years) were used as controls. Results. The microarrray analysis allowed to identify 201 differentially expressed transcripts, including transcriptional regulators, signal transducers, cell cycle regulators and enzymes involved in inflammation. Ten transcripts were confirmed by real-time reverse transcription polymerase chain reaction (RT-PCR), with some diverged results from gene chip analysis. In particular, metalloproteinase-2 (MMP-2) and leupaxin were not confirmed, whereas we found heterogeneous levels for c-fos, SAP1b, acidic sphingomyelinase (SMPD1), protein phosphatase 2A (PP2A) and GTP cyclohydrolase I regulatory protein among the 6 AD patients. Still, we confirmed that cathepsin O mRNA was homogeneously up-regulated in AD keratinocytes. Finally, specific transcripts of cdc2-like PISSLRE and 15-Lipooxigenase (15-LO) were undetectable as compared to controls. Conclusion. These results indicate that the low transcript levels typical of unstimulated culture conditions may critically impair the usefulness of microarray technology in the definition of transcription profile. Furthermore, the disparate pattern of gene transcript levels in AD donors suggests that multiple and distinct molecular changes may concur to establish skin predisposition to a dysregulated response to inflammatory stimuli.

Original languageEnglish
Pages (from-to)475-483
Number of pages9
JournalGiornale Italiano di Dermatologia e Venereologia
Volume140
Issue number5
Publication statusPublished - 2005

Fingerprint

Atopic Dermatitis
Keratinocytes
Transcriptome
Skin
GTP Cyclohydrolase
Inflammation
Sphingomyelin Phosphodiesterase
Cathepsins
Protein Phosphatase 2
Body Surface Area
Matrix Metalloproteinase 2
Disease Susceptibility
Oligonucleotide Array Sequence Analysis
Transducers
Chemokines
Oligonucleotides
Genes
Reverse Transcription
Volunteers
Healthy Volunteers

Keywords

  • Atopic dermatitis
  • Gene expression profiling
  • Keratinocytes

ASJC Scopus subject areas

  • Dermatology

Cite this

Pastore, S., Rogge, L., Mariotti, F., Mascia, F., La Scaleia, R., Dattilo, C., ... Girolomoni, G. (2005). Gene expression profile in keratinocytes from atopic dermatitis patients. Giornale Italiano di Dermatologia e Venereologia, 140(5), 475-483.

Gene expression profile in keratinocytes from atopic dermatitis patients. / Pastore, S.; Rogge, L.; Mariotti, F.; Mascia, F.; La Scaleia, R.; Dattilo, C.; Sinigaglia, F.; Girolomoni, G.

In: Giornale Italiano di Dermatologia e Venereologia, Vol. 140, No. 5, 2005, p. 475-483.

Research output: Contribution to journalArticle

Pastore, S, Rogge, L, Mariotti, F, Mascia, F, La Scaleia, R, Dattilo, C, Sinigaglia, F & Girolomoni, G 2005, 'Gene expression profile in keratinocytes from atopic dermatitis patients', Giornale Italiano di Dermatologia e Venereologia, vol. 140, no. 5, pp. 475-483.
Pastore S, Rogge L, Mariotti F, Mascia F, La Scaleia R, Dattilo C et al. Gene expression profile in keratinocytes from atopic dermatitis patients. Giornale Italiano di Dermatologia e Venereologia. 2005;140(5):475-483.
Pastore, S. ; Rogge, L. ; Mariotti, F. ; Mascia, F. ; La Scaleia, R. ; Dattilo, C. ; Sinigaglia, F. ; Girolomoni, G. / Gene expression profile in keratinocytes from atopic dermatitis patients. In: Giornale Italiano di Dermatologia e Venereologia. 2005 ; Vol. 140, No. 5. pp. 475-483.
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abstract = "Aim. Keratinocytes of atopic dermatitis (AD) patients show enhanced production of cytokines and chemokines, a phenomenon that could be relevant in promoting and maintaining inflammation and hence pivotal in localizing the atopic diathesis to the skin. We performed an oligonucleotide microarrray analysis to investigate the gene expression profile in keratinocyte cultures from 6 AD patients in order to search differentially expressed genes. Methods. Six well informed volunteer patients with moderate-to-severe chronic AD (age range 19-45 years) participated in this study. Skin involvement ranged from 20{\%} to 60{\%} of the body surface area. Six well informed volunteer healthy individuals (age range: 25-50 years) were used as controls. Results. The microarrray analysis allowed to identify 201 differentially expressed transcripts, including transcriptional regulators, signal transducers, cell cycle regulators and enzymes involved in inflammation. Ten transcripts were confirmed by real-time reverse transcription polymerase chain reaction (RT-PCR), with some diverged results from gene chip analysis. In particular, metalloproteinase-2 (MMP-2) and leupaxin were not confirmed, whereas we found heterogeneous levels for c-fos, SAP1b, acidic sphingomyelinase (SMPD1), protein phosphatase 2A (PP2A) and GTP cyclohydrolase I regulatory protein among the 6 AD patients. Still, we confirmed that cathepsin O mRNA was homogeneously up-regulated in AD keratinocytes. Finally, specific transcripts of cdc2-like PISSLRE and 15-Lipooxigenase (15-LO) were undetectable as compared to controls. Conclusion. These results indicate that the low transcript levels typical of unstimulated culture conditions may critically impair the usefulness of microarray technology in the definition of transcription profile. Furthermore, the disparate pattern of gene transcript levels in AD donors suggests that multiple and distinct molecular changes may concur to establish skin predisposition to a dysregulated response to inflammatory stimuli.",
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AU - Pastore, S.

AU - Rogge, L.

AU - Mariotti, F.

AU - Mascia, F.

AU - La Scaleia, R.

AU - Dattilo, C.

AU - Sinigaglia, F.

AU - Girolomoni, G.

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N2 - Aim. Keratinocytes of atopic dermatitis (AD) patients show enhanced production of cytokines and chemokines, a phenomenon that could be relevant in promoting and maintaining inflammation and hence pivotal in localizing the atopic diathesis to the skin. We performed an oligonucleotide microarrray analysis to investigate the gene expression profile in keratinocyte cultures from 6 AD patients in order to search differentially expressed genes. Methods. Six well informed volunteer patients with moderate-to-severe chronic AD (age range 19-45 years) participated in this study. Skin involvement ranged from 20% to 60% of the body surface area. Six well informed volunteer healthy individuals (age range: 25-50 years) were used as controls. Results. The microarrray analysis allowed to identify 201 differentially expressed transcripts, including transcriptional regulators, signal transducers, cell cycle regulators and enzymes involved in inflammation. Ten transcripts were confirmed by real-time reverse transcription polymerase chain reaction (RT-PCR), with some diverged results from gene chip analysis. In particular, metalloproteinase-2 (MMP-2) and leupaxin were not confirmed, whereas we found heterogeneous levels for c-fos, SAP1b, acidic sphingomyelinase (SMPD1), protein phosphatase 2A (PP2A) and GTP cyclohydrolase I regulatory protein among the 6 AD patients. Still, we confirmed that cathepsin O mRNA was homogeneously up-regulated in AD keratinocytes. Finally, specific transcripts of cdc2-like PISSLRE and 15-Lipooxigenase (15-LO) were undetectable as compared to controls. Conclusion. These results indicate that the low transcript levels typical of unstimulated culture conditions may critically impair the usefulness of microarray technology in the definition of transcription profile. Furthermore, the disparate pattern of gene transcript levels in AD donors suggests that multiple and distinct molecular changes may concur to establish skin predisposition to a dysregulated response to inflammatory stimuli.

AB - Aim. Keratinocytes of atopic dermatitis (AD) patients show enhanced production of cytokines and chemokines, a phenomenon that could be relevant in promoting and maintaining inflammation and hence pivotal in localizing the atopic diathesis to the skin. We performed an oligonucleotide microarrray analysis to investigate the gene expression profile in keratinocyte cultures from 6 AD patients in order to search differentially expressed genes. Methods. Six well informed volunteer patients with moderate-to-severe chronic AD (age range 19-45 years) participated in this study. Skin involvement ranged from 20% to 60% of the body surface area. Six well informed volunteer healthy individuals (age range: 25-50 years) were used as controls. Results. The microarrray analysis allowed to identify 201 differentially expressed transcripts, including transcriptional regulators, signal transducers, cell cycle regulators and enzymes involved in inflammation. Ten transcripts were confirmed by real-time reverse transcription polymerase chain reaction (RT-PCR), with some diverged results from gene chip analysis. In particular, metalloproteinase-2 (MMP-2) and leupaxin were not confirmed, whereas we found heterogeneous levels for c-fos, SAP1b, acidic sphingomyelinase (SMPD1), protein phosphatase 2A (PP2A) and GTP cyclohydrolase I regulatory protein among the 6 AD patients. Still, we confirmed that cathepsin O mRNA was homogeneously up-regulated in AD keratinocytes. Finally, specific transcripts of cdc2-like PISSLRE and 15-Lipooxigenase (15-LO) were undetectable as compared to controls. Conclusion. These results indicate that the low transcript levels typical of unstimulated culture conditions may critically impair the usefulness of microarray technology in the definition of transcription profile. Furthermore, the disparate pattern of gene transcript levels in AD donors suggests that multiple and distinct molecular changes may concur to establish skin predisposition to a dysregulated response to inflammatory stimuli.

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