title: Macromolecular Hydrogen Sulfide Donors Trigger Spatiotemporally Confined Changes in Cell Signaling.
abstract: Hydrogen sulfide (H2S) is involved in a myriad of cell signaling processes that trigger physiological events ranging from vasodilation to cell proliferation. Moreover, disturbances to H2S signaling have been associated with numerous pathologies. As such, the ability to release H2S in a cellular environment and stimulate signaling events is of considerable interest. Herein we report the synthesis of macromolecular H2S donors capable of stimulating cell signaling pathways in both the cytosol and at the cell membrane. Specifically, copolymers having pendent oligo(ethylene glycol) and benzonitrile groups were synthesized, and the benzonitrile groups were subsequently transformed into primary aryl thioamide groups via thionation using sodium hydrosulfide. These thioamide moieties could be incorporated into a hydrophilic copolymer or a block copolymer (i.e., into either the hydrophilic or hydrophobic domain). An electrochemical sensor was used to demonstrate release of H2S under simulated physiological conditions. Subsequent treatment of HEK293 cells with a macromolecular H2S donor elicited a slow and sustained increase in cytosolic ERK signaling, as monitored using a FRET-based biosensor. The macromolecular donor was also shown to induce a small, fast and sustained increase in plasma membrane-localized PKC activity immediately following addition to cells. Studies using an H2S-selective fluorescent probe in live cells confirmed release of H2S from the macromolecular donor over physiologically relevant time scales consistent with the signaling observations. Taken together, these results demonstrate that by using macromolecular H2S donors it is possible to trigger spatiotemporally confined cell signaling events. Moreover, the localized nature of the observed signaling suggests that macromolecular donor design may provide an approach for selectively stimulating certain cellular biochemical pathways.
authors.affiliation: ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, Monash Institute of Pharmaceutical Sciences, Monash University , Parkville, VIC 3052, Australia.
authors.forename: Francesca
authors.initials: F
authors.lastname: Ercole
authors.affiliation: ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, Monash Institute of Pharmaceutical Sciences, Monash University , Parkville, VIC 3052, Australia.|Children's Cancer Institute, Lowy Cancer Research Centre , Randwick, NSW 2031, Australia.
authors.forename: Friederike M
authors.initials: FM
authors.lastname: Mansfeld
authors.affiliation: Children's Cancer Institute, Lowy Cancer Research Centre , Randwick, NSW 2031, Australia.|ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, and Australian Centre for NanoMedicine, UNSW Australia, NSW 2052, Australia.
authors.forename: Maria
authors.initials: M
authors.lastname: Kavallaris
authors.affiliation: ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, Monash Institute of Pharmaceutical Sciences, Monash University , Parkville, VIC 3052, Australia.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Whittaker
authors.affiliation: ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, Monash Institute of Pharmaceutical Sciences, Monash University , Parkville, VIC 3052, Australia.
authors.forename: John F
authors.initials: JF
authors.lastname: Quinn
authors.affiliation: Drug Discovery Biology Theme, Monash Institute of Pharmaceutical Sciences, Monash University , Parkville, VIC 3052, Australia.
authors.forename: Michelle L
authors.initials: ML
authors.lastname: Halls
authors.affiliation: ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, Monash Institute of Pharmaceutical Sciences, Monash University , Parkville, VIC 3052, Australia.|Department of Chemistry, University of Warwick , Coventry, ULCV4 7AL, United Kingdom.
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Davis
mesh_terms: D015374:Biosensing Techniques; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D049109:Cell Proliferation; D003600:Cytosol / Q000378:metabolism*; D019855:Ethylene Glycol / Q000138:chemical synthesis; D048049:Extracellular Signal-Regulated MAP Kinases / Q000187:drug effects / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D006862:Hydrogen Sulfide / Q000737:chemistry / Q000494:pharmacology*; D057927:Hydrophobic and Hydrophilic Interactions; D009570:Nitriles / Q000138:chemical synthesis; D019906:Nuclear Magnetic Resonance, Biomolecular; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry; D011493:Protein Kinase C / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*; D013440:Sulfides / Q000737:chemistry; D013854:Thioamides / Q000737:chemistry
chemical_list: D009570:Nitriles; D011108:Polymers; D013440:Sulfides; D013854:Thioamides; C014356:benzonitrile; D011493:Protein Kinase C; D048049:Extracellular Signal-Regulated MAP Kinases; D019855:Ethylene Glycol; C025451:sodium bisulfide; D006862:Hydrogen Sulfide