Neutralizing the pathological effects of extracellular histones with small polyanions
Connor H. O’ Meara, Lucy A. Coupland, Farzaneh Kordbacheh, Ben Quah, Chih‐Wei Chang, David S. Davis, Anna Bezos, Anna Browne, Craig Freeman, Dillon Hammill, Pradeep Chopra, Gergely Pipa, Paul D. Madge, Esther M. Gallant, Courtney Segovis, Angela F. Dulhunty, Leonard Arnolda, Imogen Mitchell, Levon M. Khachigian, Ross W. Stephens, Mark von Itzstein, Christopher R. Parish
Abstract
Extracellular histones in neutrophil extracellular traps (NETs) or in chromatin from injured tissues are highly pathological, particularly when liberated by DNases. We report the development of small polyanions (SPAs) (~0.9-1.4 kDa) that interact electrostatically with histones, neutralizing their pathological effects. In vitro, SPAs inhibited the cytotoxic, platelet-activating and erythrocyte-damaging effects of histones, mechanistic studies revealing that SPAs block disruption of lipid-bilayers by histones. In vivo, SPAs significantly inhibited sepsis, deep-vein thrombosis, and cardiac and tissue-flap models of ischemia-reperfusion injury (IRI), but appeared to differ in their capacity to neutralize NET-bound versus free histones. Analysis of sera from sepsis and cardiac IRI patients supported these differential findings. Further investigations revealed this effect was likely due to the ability of certain SPAs to displace histones from NETs, thus destabilising the structure. Finally, based on our work, a non-toxic SPA that inhibits both NET-bound and free histone mediated pathologies was identified for clinical development.