title: Stimulation of adenosine A2A receptors reduces intracellular cholesterol accumulation and rescues mitochondrial abnormalities in human neural cell models of Niemann-Pick C1.
abstract: Niemann Pick C 1 (NPC1) disease is an incurable, devastating lysosomal-lipid storage disorder characterized by hepatosplenomegaly, progressive neurological impairment and early death. Current treatments are very limited and the research of new therapeutic targets is thus mandatory. We recently showed that the stimulation of adenosine A2A receptors (A2ARs) rescues the abnormal phenotype of fibroblasts from NPC1 patients suggesting that A2AR agonists could represent a therapeutic option for this disease. However, since all NPC1 patients develop severe neurological symptoms which can be ascribed to the complex pathology occurring in both neurons and oligodendrocytes, in the present paper we tested the effects of the A2AR agonist CGS21680 in human neuronal and oligodendroglial NPC1 cell lines (i.e. neuroblastoma SH-SY5Y and oligodendroglial MO3.13 transiently transfected with NPC1 small interfering RNA). The down-regulation of the NPC1 protein effectively resulted in intracellular cholesterol accumulation and altered mitochondrial membrane potential. Both effects were significantly attenuated by CGS21680 (500 nM). The protective effects of CGS were prevented by the selective A2AR antagonist ZM241385 (500 nM). The involvement of calcium modulation was demonstrated by the ability of Bapta-AM (5-7 μM) in reverting the effect of CGS. The A2A-dependent activity was prevented by the PKA-inhibitor KT5720, thus showing the involvement of the cAMP/PKA signaling. These findings provide a clear in vitro proof of concept that A2AR agonists are promising potential drugs for NPC disease.
authors.affiliation: Department of Therapeutic Research and Medicine Evaluation, Istituto Superiore di Sanità, 00161, Rome, Italy. Electronic address: antonella.ferrante@iss.it.
authors.forename: A
authors.initials: A
authors.lastname: Ferrante
authors.affiliation: Department of Cell Biology and Neurosciences, Istituto Superiore di Sanità, 00161, Rome, Italy.
authors.forename: C
authors.initials: C
authors.lastname: De Nuccio
authors.affiliation: Department of Therapeutic Research and Medicine Evaluation, Istituto Superiore di Sanità, 00161, Rome, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Pepponi
authors.affiliation: Department of Cell Biology and Neurosciences, Istituto Superiore di Sanità, 00161, Rome, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Visentin
authors.affiliation: Department of Therapeutic Research and Medicine Evaluation, Istituto Superiore di Sanità, 00161, Rome, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Martire
authors.affiliation: Department of Cell Biology and Neurosciences, Istituto Superiore di Sanità, 00161, Rome, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Bernardo
authors.affiliation: Department of Cell Biology and Neurosciences, Istituto Superiore di Sanità, 00161, Rome, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Minghetti
authors.affiliation: Department of Therapeutic Research and Medicine Evaluation, Istituto Superiore di Sanità, 00161, Rome, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Popoli
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology; D058908:Adenosine A2 Receptor Agonists / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins / Q000235:genetics; D008928:Mitochondria / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D000077296:Niemann-Pick C1 Protein; D052556:Niemann-Pick Disease, Type C / Q000378:metabolism*; D009836:Oligodendroglia / Q000378:metabolism; D010627:Phenethylamines / Q000494:pharmacology; D043705:Receptor, Adenosine A2A / Q000378:metabolism*
chemical_list: D058908:Adenosine A2 Receptor Agonists; D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; C106881:NPC1 protein, human; D000077296:Niemann-Pick C1 Protein; D010627:Phenethylamines; D043705:Receptor, Adenosine A2A; C061282:2-(4-(2-carboxyethyl)phenethylamino)-5'-N-ethylcarboxamidoadenosine; D002784:Cholesterol; D017868:Cyclic AMP-Dependent Protein Kinases; D000241:Adenosine; D002118:Calcium
keywords: Adenosine A(2A) receptor; Cholesterol; Mitochondrial membrane potential; Neuroblastoma cell line; Niemann-Pick C; Oligodendroglial cell line