title: Depletion of the central metabolite NAD leads to oncosis-mediated cell death.
abstract: Depletion of the central metabolite NAD in cells results in broad metabolic defects leading to cell death and is a proposed novel therapeutic strategy in oncology. There is, however, a limited understanding of the underlying mechanisms that connect disruption of this central metabolite with cell death. Here we utilize GNE-617, a small molecule inhibitor of NAMPT, a rate-limiting enzyme required for NAD generation, to probe the pathways leading to cell death following NAD depletion. In all cell lines examined, NAD was rapidly depleted (average t½ of 8.1 h) following NAMPT inhibition. Concurrent with NAD depletion, there was a decrease in both cell proliferation and motility, which we attribute to reduced activity of NAD-dependent deacetylases because cells fail to deacetylate α-tubulin-K40 and histone H3-K9. Following depletion of NAD by >95%, cells lose the ability to regenerate ATP. Cell lines with a slower rate of ATP depletion (average t½ of 45 h) activate caspase-3 and show evidence of apoptosis and autophagy, whereas cell lines with rapid depletion ATP (average t½ of 32 h) do not activate caspase-3 or show signs of apoptosis or autophagy. However, the predominant form of cell death in all lines is oncosis, which is driven by the loss of plasma membrane homeostasis once ATP levels are depleted by >20-fold. Thus, our work illustrates the sequence of events that occurs in cells following depletion of a key metabolite and reveals that cell death caused by a loss of NAD is primarily driven by the inability of cells to regenerate ATP.
authors.affiliation: From the Departments of Translational Oncology and.
authors.forename: Christopher
authors.initials: C
authors.lastname: Del Nagro
authors.affiliation: From the Departments of Translational Oncology and.
authors.forename: Yang
authors.initials: Y
authors.lastname: Xiao
authors.affiliation: Pathology, Genentech Inc., South San Francisco, California 94080.
authors.forename: Linda
authors.initials: L
authors.lastname: Rangell
authors.affiliation: Pathology, Genentech Inc., South San Francisco, California 94080.
authors.forename: Mike
authors.initials: M
authors.lastname: Reichelt
authors.affiliation: From the Departments of Translational Oncology and Obrien.tom@gene.com.
authors.forename: Thomas
authors.initials: T
authors.lastname: O'Brien
mesh_terms: D000107:Acetylation / Q000187:drug effects; D000255:Adenosine Triphosphate / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects; D001343:Autophagy / Q000187:drug effects; D015153:Blotting, Western; D053148:Caspase 3 / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D045325:HCT116 Cells; D006574:Heterocyclic Compounds, 2-Ring / Q000737:chemistry / Q000494:pharmacology*; D006657:Histones / Q000378:metabolism; D006801:Humans; D046529:Microscopy, Electron, Transmission; D015394:Molecular Structure; D009243:NAD / Q000378:metabolism*; D054409:Nicotinamide Phosphoribosyltransferase / Q000037:antagonists & inhibitors / Q000378:metabolism; D013450:Sulfones / Q000737:chemistry / Q000494:pharmacology*; D013997:Time Factors; D014404:Tubulin / Q000378:metabolism
chemical_list: C000591022:GNE-617; D006574:Heterocyclic Compounds, 2-Ring; D006657:Histones; D013450:Sulfones; D014404:Tubulin; D009243:NAD; D000255:Adenosine Triphosphate; D054409:Nicotinamide Phosphoribosyltransferase; D053148:Caspase 3
keywords: ATP; Apoptosis; Cell Death; Cell Metabolism; GNE-617; NAMPT; Nicotinamide Adenine Dinucleotide (NAD); Oncosis