The formyl peptide receptor agonist FPRa14 induces differentiation of Neuro2a mouse neuroblastoma cells into multiple distinct morphologies which can be specifically inhibited with FPR antagonists and FPR knockdown using siRNA

• Published in: PloS one Vol. 14; no. 6; p. e0217815
• Main Authors: Cussell, Peter J G, Howe, Michael S, Illingworth, Thomas A, Gomez Escalada, Margarita, Milton, Nathaniel G N, Paterson, Andrew W J
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 06.06.2019; Public Library of Science (PLoS)
• Subjects: Agonists; Alzheimer's disease; Animals; Biology; Biology and Life Sciences; Cancer; Cancer research; Care and treatment; Cell Death - drug effects; Cell differentiation; Cell Differentiation - drug effects; Cell Line, Tumor; Cell Shape - drug effects; Central nervous system; Differentiation (biology); Formyl peptide receptors; G proteins; Genetic aspects; Growth factors; Humans; Invasiveness; Kinases; Ligands; Medicine and Health Sciences; Mice; Morphology; Motility; Nervous system; Neuralgia; Neuroblastoma; Neuroblastoma - pathology; Neuroblastoma cells; Neuroblasts; Neurons; Neuropathy; Neutrophils; Novels; Oxidative stress; Pain; Parkinson's disease; Peptides; Physical Sciences; Physiology; Proteins; Receptors; Receptors, Formyl Peptide - agonists; Receptors, Formyl Peptide - antagonists & inhibitors; Receptors, Formyl Peptide - metabolism; Regeneration; Research and analysis methods; Risk factors; RNA interference; RNA, Small Interfering - metabolism; siRNA; Time Factors; Toxicity; United Kingdom > UK
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0217815

The N-formyl peptide receptors (FPRs) have been identified within neuronal tissues and may serve as yet undetermined functions within the nervous system. The FPRs have been implicated in the progression and invasiveness of neuroblastoma and other cancers. In this study the effects of the synthetic FPR agonist FPRa14, FPR antagonists and FPR knockdown using siRNA on mouse neuroblastoma neuro2a (N2a) cell differentiation plus toxicity were examined. The FPRa14 (1-10μM) was found to induce a significant dose-dependent differentiation response in mouse neuroblastoma N2a cells. Interestingly, three distinct differentiated morphologies were observed, with two non-archetypal forms observed at the higher FPRa14 concentrations. These three forms were also observed in the human neuroblastoma cell-lines IMR-32 and SH-SY5Y when exposed to 100μM FPRa14. In N2a cells combined knockdown of FPR1 and FPR2 using siRNA inhibited the differentiation response to FPRa14, suggesting involvement of both receptor subtypes. Pre-incubating N2a cultures with the FPR1 antagonists Boc-MLF and cyclosporin H significantly reduced FPRa14-induced differentiation to near baseline levels. Meanwhile, the FPR2 antagonist WRW4 had no significant effect on FPRa14-induced N2a differentiation. These results suggest that the N2a differentiation response observed has an FPR1-dependent component. Toxicity of FPRa14 was only observed at higher concentrations. All three antagonists used blocked FPRa14-induced toxicity, whilst only siRNA knockdown of FPR2 reduced toxicity. This suggests that the toxicity and differentiation involve different mechanisms. The demonstration of neuronal differentiation mediated via FPRs in this study represents a significant finding and suggests a role for FPRs in the CNS. This finding could potentially lead to novel therapies for a range of neurological conditions including neuroblastoma, Alzheimer's disease, Parkinson's disease and neuropathic pain. Furthermore, this could represent a potential avenue for neuronal regeneration therapies.