Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by selective loss of motor neurons in the CNS. Approximately 20% of ALS cases are associated to mutations in the Cu/Zn-superoxide dismutase-1 (SOD1) gene (Rosen et al., 1993). We have recently characterized the transcriptional profiles of motor cortex samples from sporadic ALS (SALS) patients and differentiated these into two subgroups (SALS1 and SALS2), based on differentially expressed genes, encoding for 70 potential therapeutic targets (Aronica et al., 2015; Morello and Cavallaro 2015). Through a meta-analysis approach, we have identified three target genes commonly upregulated both in SOD1G93A transgenic mice and SALS2 motor cortex, including pituitary adenylate cyclase-activating polypeptide (PACAP), epidermal growth factor receptor (EGFR) and matrix metallopeptidase 2 (MMP-2). Considering that a functional association between PACAP and EGFR in lung cancer cells has been already described (Moody et al., 2011), here we investigated whether a similar PACAP/EGFR/MMP-2 axis deregulation is involved in the pathophysiology of ALS. This study has been performed in a motor neuronal cell line of mouse NSC-34, stably transfected for the inducible expression of low quantities of wild or mutant SOD1G93A. Our results have shown that treatment with 100 nm PACAP is able to rescue cells degeneration following growth factors deprivation, as previously demonstrated in a iPSC-derived model of ALS (Bonaventura et al., 2017). Conversely, cells viability is drastically decreased following treatment with PAC1 antagonist PACAP(6-38) or EGFR tyrosine kinase inhibitor gefitinib. We found that PACAP addition increased EGFR tyrosine phosphorylation through protein kinase A, but not phospholipase C. Moreover, peptide addition triggered the activation of survival signal MEK/ERK pathway, and increased the expression of MMP-2, whose levels are drastically reduced after serum starvation. Our findings suggest that a deeply characterization of the mechanism linking PACAP/EGFR/MMP-2 axis to SOD1 mutation may open a new perspective for ALS therapy
Role of PACAP/EGFR/MMP-2 axis in an in vitro model of amyotrophic lateral sclerosis
D'Amico A;
2018-01-01
Abstract
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by selective loss of motor neurons in the CNS. Approximately 20% of ALS cases are associated to mutations in the Cu/Zn-superoxide dismutase-1 (SOD1) gene (Rosen et al., 1993). We have recently characterized the transcriptional profiles of motor cortex samples from sporadic ALS (SALS) patients and differentiated these into two subgroups (SALS1 and SALS2), based on differentially expressed genes, encoding for 70 potential therapeutic targets (Aronica et al., 2015; Morello and Cavallaro 2015). Through a meta-analysis approach, we have identified three target genes commonly upregulated both in SOD1G93A transgenic mice and SALS2 motor cortex, including pituitary adenylate cyclase-activating polypeptide (PACAP), epidermal growth factor receptor (EGFR) and matrix metallopeptidase 2 (MMP-2). Considering that a functional association between PACAP and EGFR in lung cancer cells has been already described (Moody et al., 2011), here we investigated whether a similar PACAP/EGFR/MMP-2 axis deregulation is involved in the pathophysiology of ALS. This study has been performed in a motor neuronal cell line of mouse NSC-34, stably transfected for the inducible expression of low quantities of wild or mutant SOD1G93A. Our results have shown that treatment with 100 nm PACAP is able to rescue cells degeneration following growth factors deprivation, as previously demonstrated in a iPSC-derived model of ALS (Bonaventura et al., 2017). Conversely, cells viability is drastically decreased following treatment with PAC1 antagonist PACAP(6-38) or EGFR tyrosine kinase inhibitor gefitinib. We found that PACAP addition increased EGFR tyrosine phosphorylation through protein kinase A, but not phospholipase C. Moreover, peptide addition triggered the activation of survival signal MEK/ERK pathway, and increased the expression of MMP-2, whose levels are drastically reduced after serum starvation. Our findings suggest that a deeply characterization of the mechanism linking PACAP/EGFR/MMP-2 axis to SOD1 mutation may open a new perspective for ALS therapyI documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
