Transferrin Receptor Is a Specific Ferroptosis Marker.

1. Cell Rep. 2020 Mar 10;30(10):3411-3423.e7. doi: 10.1016/j.celrep.2020.02.049. Transferrin Receptor Is a Specific Ferroptosis Marker. Feng H(1), Schorpp K(2), Jin J(1), Yozwiak CE(3), Hoffstrom BG(4), Decker AM(1), Rajbhandari P(1), Stokes ME(1), Bender HG(1), Csuka JM(1), Upadhyayula PS(5), Canoll P(6), Uchida K(7), Soni RK(8), Hadian K(2), Stockwell BR(9). Author information: (1)Department of Biological Sciences, Columbia University, Northwest Corner Building, 12th Floor, MC 4846, 550 West 120(th) Street, New York, NY 10027, USA. (2)HelmholtzZentrum München, German Research Center for Environmental Health (GmbH), Assay Development and Screening Platform, Institute for Molecular Toxicology and Pharmacology, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany. (3)Department of Chemistry, Columbia University, Northwest Corner Building, 12th Floor, MC 4846, 550 West 120(th) Street, New York, NY 10027, USA. (4)Antibody Technology Resource, Fred Hutchinson Cancer Research Center, 1100 Fairview Ave. N, Seattle, WA 98109, USA. (5)Department of Neurological Surgery, Vagelos College of Physicians and Surgeons, Columbia University Irving Medical Center, New York, NY 10032, USA. (6)Department of Pathology and Cell Biology, Vagelos College of Physicians and Surgeons, Columbia University Irving Medical Center, 1130 St. Nicholas Ave., Room 1001, New York, NY 10032, USA; Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, NY 10032, USA. (7)Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan. (8)Proteomics and Macromolecular Crystallography Shared Resource, Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, NY 10032, USA. (9)Department of Biological Sciences, Columbia University, Northwest Corner Building, 12th Floor, MC 4846, 550 West 120(th) Street, New York, NY 10027, USA; Department of Chemistry, Columbia University, Northwest Corner Building, 12th Floor, MC 4846, 550 West 120(th) Street, New York, NY 10027, USA; Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, NY 10032, USA. Electronic address: bstockwell@columbia.edu. Ferroptosis is a type of regulated cell death driven by the iron-dependent accumulation of oxidized polyunsaturated fatty acid-containing phospholipids. There is no reliable way to selectively stain ferroptotic cells in tissue sections to characterize the extent of ferroptosis in animal models or patient samples. We address this gap by immunizing mice with membranes from lymphoma cells treated with the ferroptosis inducer piperazine erastin and screening ∼4,750 of the resulting monoclonal antibodies generated for their ability to selectively detect cells undergoing ferroptosis. We find that one antibody, 3F3 ferroptotic membrane antibody (3F3-FMA), is effective as a selective ferroptosis-staining reagent. The antigen of 3F3-FMA is identified as the human transferrin receptor 1 protein (TfR1). We validate this finding with several additional anti-TfR1 antibodies and compare them to other potential ferroptosis-detecting reagents. We find that anti-TfR1 and anti-malondialdehyde adduct antibodies are effective at staining ferroptotic tumor cells in multiple cell culture and tissue contexts. Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved. DOI: 10.1016/j.celrep.2020.02.049 PMCID: PMC7172030 PMID: 32160546 [Indexed for MEDLINE] Conflict of interest statement: Declaration of Interests B.R.S. is a consultant to and has equity in Inzen Therapeutics. B.R.S. also is an inventor on patents and patent applications related to ferroptosis. C.E.Y. is currently an employee of Vertex Pharmaceuticals.