Molecular and Cellular Biochemistry

, Volume 392, Issue 1–2, pp 13–30 | Cite as

Forward subtractive libraries containing genes transactivated by dexamethasone in ataxia-telangiectasia lymphoblastoid cells

  • Sara Biagiotti
  • Michele Menotta
  • Elisa Giacomini
  • Lucia Radici
  • Marzia Bianchi
  • Cristina Bozzao
  • Luciana Chessa
  • Mauro Magnani
Article

Abstract

Ataxia telangiectasia (A-T) is a rare autosomal recessive disorder caused by biallelic mutations in the Ataxia Telangiectasia-mutated gene. A-T shows a complex phenotype ranging from early-onset progressive neurodegeneration to immunodeficiencies, high incidence of infections, and tumors. Unfortunately, no therapy is up to now available for treating this condition. Recently, the short term treatment of ataxia-telangiectasia patients with glucocorticoids was shown to improve their neurological symptoms and possibly reverse cerebellar atrophy. Thus, corticosteroids represent an attractive approach for the treatment of this neurodegenerative disease. However, the molecular mechanism involved in glucocorticoid action in A-T is yet unknown. The aim of our work is to construct cDNA libraries containing those genes which are transactivated by the glucocorticoid analogue, dexamethasone, in A-T human cells. For this purpose, suppression subtractive hybridization has been performed on ATM-null lymphoblastoid cell transcriptome extracted following drug administration. Annotation of whole genes contained in the libraries has been obtained by coupling subtractive hybridization with microarray analysis. Positive transcripts have been validated by quantitative PCR. Through in silico analyses, identified genes have been classified on the basis of the pathway in which they are involved, being able to address signaling required for dexamethasone action. Most of the induced transcripts are involved in metabolic processes and regulation of cellular processes. Our results can help to unravel the mechanism of glucocorticoid action in the reversion of A-T phenotype. Moreover, the induction of a specific region of the ATM transcript has been identified as putative biomarker predictive of dexamethasone efficacy on ataxic patients.

Keywords

Ataxia telangiectasia Dexamethasone Gene expression profiling Transcriptional activation Suppression subtractive hybridization 

Abbreviation

A-T

Ataxia telangiectasia

GC

Glucocorticoid

dexa

Dexamethasone

SSH

Suppression subtractive hybridization

GRE

Glucocorticoid responsive elements

LCLs

Lymphoblastoid cell lines

Notes

Acknowledgments

This work was partially supported by “Fondazione M. Vocale.” Sara Biagiotti hold a fellowship from Fondazione Cassa di Risparmio di Fano sponsorized by FANOATENEO and Associazione Nazionale A-T “Davide DeMarini.”

Supplementary material

11010_2014_2013_MOESM1_ESM.docx (49 kb)
Supplementary material 1 (DOCX 49 kb)

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Sara Biagiotti
    • 1
  • Michele Menotta
    • 1
  • Elisa Giacomini
    • 1
  • Lucia Radici
    • 1
  • Marzia Bianchi
    • 1
  • Cristina Bozzao
    • 2
  • Luciana Chessa
    • 2
  • Mauro Magnani
    • 1
  1. 1.Department of Biomolecular SciencesUniversity of Urbino “Carlo Bo”UrbinoItaly
  2. 2.Departmernt of Clinical and Molecular MedicineSapienza UniversityRomeItaly

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