Purpose. Adenosine acts as a potent anti-inflammatory autacoid; extracellular adenosine formation is generally thought to result from the sequential dephosphorylation of extracellular ATP to AMP by action of ectonuclease triphoshate diphosphohydrolase (CD39) followed by degradation to adenosine by ecto-5'-nucleotidase (CD73). The diverse cellular actions of adenosine are mediated by four specific membrane receptors (A1R, A2aR, A2bR and A3R). In animal models an up-regulation of ARs can be observed in cardiac tissue of failing heart (HF). Recently ARs mRNA expression has been assessed in human whole blood providing a useful tool to evaluate the expression of ARs in diseases characterized by a marked inflammatory component. The aim of this study was to evaluate the possible changes of transcriptomic profiling of A1R, A2aR, A2bR and A3R in human leukocytes of patients (pts) with valvular HF (Vlv) as compared to healthy subjects (C). Methods. Total RNA was extracted from leukocytes of C (n=7) and of Vlv pts (n= 6, NYHA III-IV) with PAXgene Blood RNA Kit (Qiagen, Milan, Italy). Real time PCR was performed and optimized for each ARs primer. CD39 and CD73 mRNA expression was also evaluated both in C and in Vlv pts. The experimental results were normalized with the three most stably expressed genes (SRP14, EEF1A, RPL13A). Results. In human whole blood of Vlv pts significantly higher levels of mRNA expression, for each receptors analyzed with respect to C were observed (A1R: C=1.97±0.72, Vlv pts=10.9±3.27, p=0.0098; A2aR: C=0.45±0.15, Vlv pts=3.19±0.94, p=0.0057; A2bR: C=0.43±0.16, Vlv pts=5.00±2.98, p=0.05; A3R: C=1.39±0.26, Vlv pts=2.9±0.71, p=0.04). Also CD39 and CD73 resulted upregulated in human leukocytes of Vlv pts with respect to C. Significant correlations (p<0.05) were observed between all ARs themself as well as with CD39 and CD73. Conclusions. The present study highlights, for the first time, an increase of ARs mRNA expression in the peripheral circulating cells of Vlv pts. The increase of CD39 and CD73 trascript levels is suggestive of a protective CD39-CD73-dependent adenosine production in advanced HF pts. These findings suggest that components of adenosine metabolism and signalling are altered to promote adenosine production and signalling in HF patients. Thus HF may benefit from adenosine-based drugs therapy after confirmation by clinical trials.
Adenosine receptors mRNA expression in human leukocytes of patients with valvular disease
Del Ry S;Della Latta V;Cabiati M;Sabatino L;Morales M A;
2013
Abstract
Purpose. Adenosine acts as a potent anti-inflammatory autacoid; extracellular adenosine formation is generally thought to result from the sequential dephosphorylation of extracellular ATP to AMP by action of ectonuclease triphoshate diphosphohydrolase (CD39) followed by degradation to adenosine by ecto-5'-nucleotidase (CD73). The diverse cellular actions of adenosine are mediated by four specific membrane receptors (A1R, A2aR, A2bR and A3R). In animal models an up-regulation of ARs can be observed in cardiac tissue of failing heart (HF). Recently ARs mRNA expression has been assessed in human whole blood providing a useful tool to evaluate the expression of ARs in diseases characterized by a marked inflammatory component. The aim of this study was to evaluate the possible changes of transcriptomic profiling of A1R, A2aR, A2bR and A3R in human leukocytes of patients (pts) with valvular HF (Vlv) as compared to healthy subjects (C). Methods. Total RNA was extracted from leukocytes of C (n=7) and of Vlv pts (n= 6, NYHA III-IV) with PAXgene Blood RNA Kit (Qiagen, Milan, Italy). Real time PCR was performed and optimized for each ARs primer. CD39 and CD73 mRNA expression was also evaluated both in C and in Vlv pts. The experimental results were normalized with the three most stably expressed genes (SRP14, EEF1A, RPL13A). Results. In human whole blood of Vlv pts significantly higher levels of mRNA expression, for each receptors analyzed with respect to C were observed (A1R: C=1.97±0.72, Vlv pts=10.9±3.27, p=0.0098; A2aR: C=0.45±0.15, Vlv pts=3.19±0.94, p=0.0057; A2bR: C=0.43±0.16, Vlv pts=5.00±2.98, p=0.05; A3R: C=1.39±0.26, Vlv pts=2.9±0.71, p=0.04). Also CD39 and CD73 resulted upregulated in human leukocytes of Vlv pts with respect to C. Significant correlations (p<0.05) were observed between all ARs themself as well as with CD39 and CD73. Conclusions. The present study highlights, for the first time, an increase of ARs mRNA expression in the peripheral circulating cells of Vlv pts. The increase of CD39 and CD73 trascript levels is suggestive of a protective CD39-CD73-dependent adenosine production in advanced HF pts. These findings suggest that components of adenosine metabolism and signalling are altered to promote adenosine production and signalling in HF patients. Thus HF may benefit from adenosine-based drugs therapy after confirmation by clinical trials.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
