title: Lamin A/C Is Required for ChAT-Dependent Neuroblastoma Differentiation.
abstract: The mouse neuroblastoma N18TG2 clone is unable to differentiate and is defective for the enzymes of the biosynthesis of neurotransmitters. The forced expression of choline acetyltransferase (ChAT) in these cells results in the synthesis and release of acetylcholine (Ach) and hence in the expression of neurospecific features and markers. To understand how the expression of ChAT triggered neuronal differentiation, we studied the differences in genome-wide transcription profiles between the N18TG2 parental cells and its ChAT-expressing 2/4 derived clone. The engagement of the 2/4 cells in the neuronal developmental program was confirmed by the increase of the expression level of several differentiation-related genes and by the reduction of the amount of transcripts of cell cycle genes. At the same time, we observed a massive reorganization of cytoskeletal proteins in terms of gene expression, with the accumulation of the nucleoskeletal lamina component Lamin A/C in differentiating cells. The increase of the Lmna transcripts induced by ChAT expression in 2/4 cells was mimicked treating the parental N18TG2 cells with the acetylcholine receptor agonist carbachol, thus demonstrating the direct role played by this receptor in neuron nuclei maturation. Conversely, a treatment of 2/4 cells with the muscarinic receptor antagonist atropine resulted in the reduction of the amount of Lmna RNA. Finally, the hypothesis that Lmna gene product might play a crucial role in the ChAT-dependent molecular differentiation cascade was strongly supported by Lmna knockdown in 2/4 cells leading to the downregulation of genes involved in differentiation and cytoskeleton formation and to the upregulation of genes known to regulate self-renewal and stemness.
authors.affiliation: CNR, Institute of Cell Biology and Neurobiology (IBCN), Rome, Italy.
authors.forename: Loredana
authors.initials: L
authors.lastname: Guglielmi
authors.affiliation: CNR, Institute of Cell Biology and Neurobiology (IBCN), Rome, Italy.
authors.forename: Marta
authors.initials: M
authors.lastname: Nardella
authors.affiliation: CNR, Institute of Cell Biology and Neurobiology (IBCN), Rome, Italy.
authors.forename: Carla
authors.initials: C
authors.lastname: Musa
authors.affiliation: CNR, Institute of Cell Biology and Neurobiology (IBCN), Rome, Italy.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Iannetti
authors.affiliation: Genomics Facility, European Brain Research Institute (EBRI), Rome, Italy.
authors.forename: Ivan
authors.initials: I
authors.lastname: Arisi
authors.affiliation: Genomics Facility, European Brain Research Institute (EBRI), Rome, Italy.
authors.forename: Mara
authors.initials: M
authors.lastname: D'Onofrio
authors.affiliation: Genomics Facility, European Brain Research Institute (EBRI), Rome, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Storti
authors.affiliation: Department of Laboratory Medicine and Department of Internal Medicine, PTV, University of Rome "Tor Vergata", Rome, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Valentini
authors.affiliation: Department of Biology and Biotechnology "Charles Darwin", Sapienza University, Rome, Italy.
authors.forename: Emanuele
authors.initials: E
authors.lastname: Cacci
authors.affiliation: Department of Biology and Biotechnology "Charles Darwin", Sapienza University, Rome, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Biagioni
authors.affiliation: Department of Biology and Biotechnology "Charles Darwin", Sapienza University, Rome, Italy.
authors.forename: Gabriella
authors.initials: G
authors.lastname: Augusti-Tocco
authors.affiliation: CNR, Institute of Cell Biology and Neurobiology (IBCN), Rome, Italy. igea.dagnano@cnr.it.
authors.forename: Igea
authors.initials: I
authors.lastname: D'Agnano
authors.affiliation: CNR, Institute of Cell Biology and Neurobiology (IBCN), Rome, Italy. armando.felsani@cnr.it.
authors.forename: Armando
authors.initials: A
authors.lastname: Felsani
mesh_terms: D000818:Animals; D002453:Cell Cycle / Q000187:drug effects / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002795:Choline O-Acetyltransferase / Q000378:metabolism*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D055785:Gene Knockdown Techniques; D063990:Gene Ontology; D034904:Lamin Type A / Q000378:metabolism*; D051379:Mice; D009447:Neuroblastoma / Q000201:enzymology* / Q000235:genetics / Q000473:pathology*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D025941:Protein Interaction Mapping; D011950:Receptors, Cholinergic / Q000378:metabolism; D011976:Receptors, Muscarinic / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects; D014212:Tretinoin / Q000494:pharmacology; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D034904:Lamin Type A; D011950:Receptors, Cholinergic; D011976:Receptors, Muscarinic; D014212:Tretinoin; D002795:Choline O-Acetyltransferase
keywords: Choline acetyltransferase; Differentiation; Gene expression; LMNA; Neuroblastoma