학술논문
An immunohistochemical atlas of necroptotic pathway expression
Document Type
Original Paper
Author
Chiou, Shene; Al-Ani, Aysha H; Pan, Yi; Patel, Komal M; Kong, Isabella Y; Whitehead, Lachlan W; Light, Amanda; Young, Samuel N; Barrios, Marilou; Sargeant, Callum; Rajasekhar, Pradeep; Zhu, Leah; Hempel, Anne; Lin, Ann; Rickard, James A; Hall, Cathrine; Gangatirkar, Pradnya; Yip, Raymond KH; Cawthorne, Wayne; Jacobsen, Annette V; Horne, Christopher R; Martin, Katherine R; Ioannidis, Lisa J; Hansen, Diana S; Day, Jessica; Wicks, Ian P; Law, Charity; Ritchie, Matthew E; Bowden, Rory; Hildebrand, Joanne M; O’Reilly, Lorraine A; Silke, John; Giulino-Roth, Lisa; Tsui, Ellen; Rogers, Kelly L; Hawkins, Edwin D; Christensen, Britt; Murphy, James M; Samson, André L
Source
EMBO Molecular Medicine. 16(7):1717-1749
Subject
Language
English
ISSN
1757-4684
Abstract
Necroptosis is a lytic form of regulated cell death reported to contribute to inflammatory diseases of the gut, skin and lung, as well as ischemic-reperfusion injuries of the kidney, heart and brain. However, precise identification of the cells and tissues that undergo necroptotic cell death in vivo has proven challenging in the absence of robust protocols for immunohistochemical detection. Here, we provide automated immunohistochemistry protocols to detect core necroptosis regulators – Caspase-8, RIPK1, RIPK3 and MLKL – in formalin-fixed mouse and human tissues. We observed surprising heterogeneity in protein expression within tissues, whereby short-lived immune barrier cells were replete with necroptotic effectors, whereas long-lived cells lacked RIPK3 or MLKL expression. Local changes in the expression of necroptotic effectors occurred in response to insults such as inflammation, dysbiosis or immune challenge, consistent with necroptosis being dysregulated in disease contexts. These methods will facilitate the precise localisation and evaluation of necroptotic signaling in vivo.
Synopsis: Necroptotic cell death is broadly linked to inflammation-associated diseases such as IBD. But, accurately identifying when and where necroptosis occurs in human disease has proven difficult. Using new methods to detect the key regulators of necroptosis - Caspase-8, RIPK1, RIPK3, MLKL - we find that:Expression of the necroptotic pathway is rare under basal conditions and largely restricted to fast-cycling barrier cells such as the intestinal epithelium.RIPK3 is a novel acute phase reactant, with levels that rapidly change in response to physiological challenges including inflammation, dysbiosis and immunisation.Exaggerated necroptosis arises in a subset of IBD patients.Subcellular relocation of Caspase-8 and other key regulators is a clinically relevant approach to pinpoint necroptosis in mouse and human tissues.
Necroptotic cell death is broadly linked to inflammation-associated diseases such as IBD. But, accurately identifying when and where necroptosis occurs in human disease has proven difficult. Using new methods to detect the key regulators of necroptosis - Caspase-8, RIPK1, RIPK3, MLKL - we find that:
Synopsis: Necroptotic cell death is broadly linked to inflammation-associated diseases such as IBD. But, accurately identifying when and where necroptosis occurs in human disease has proven difficult. Using new methods to detect the key regulators of necroptosis - Caspase-8, RIPK1, RIPK3, MLKL - we find that:Expression of the necroptotic pathway is rare under basal conditions and largely restricted to fast-cycling barrier cells such as the intestinal epithelium.RIPK3 is a novel acute phase reactant, with levels that rapidly change in response to physiological challenges including inflammation, dysbiosis and immunisation.Exaggerated necroptosis arises in a subset of IBD patients.Subcellular relocation of Caspase-8 and other key regulators is a clinically relevant approach to pinpoint necroptosis in mouse and human tissues.
Necroptotic cell death is broadly linked to inflammation-associated diseases such as IBD. But, accurately identifying when and where necroptosis occurs in human disease has proven difficult. Using new methods to detect the key regulators of necroptosis - Caspase-8, RIPK1, RIPK3, MLKL - we find that: