NFS1 undergoes positive selection in lung tumours and protects cells from ferroptosis
Authors: Erdem M Terzi
Thales Papagiannakopoulos
Vladislav O Sviderskiy
Richard Possemato
Sylvia Adams
Samantha W Alvarez
Andre L Moreira
David M Sabatini
Kıvanç Birsoy
Journal: Nature (2017)
Discipline: Multidisciplinary
Keywords: Environmental nutrient levels; cancer cell metabolism; gene essentiality; oxygen levels; iron-sulfur cluster biosynthetic enzyme
Affiliations: Department of Pathology, New York University School of Medicine, New York, New York 10016, USA., Laura & Isaac Perlmutter Cancer Center, New York University School of Medicine, New York, New York 10016, USA., Whitehead Institute for Biomedical Research, Nine Cambridge Center, Cambridge, Massachusetts 02142, USA., Howard Hughes Medical Institute, Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA., The David H. Koch Institute for Integrative Cancer Research, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA., Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA., Broad Institute of Harvard and Massachusetts Institute of Technology, Seven Cambridge Center, Cambridge, Massachusetts 02142, USA., Laboratory of Metabolic Regulation and Genetics, The Rockefeller University, 1230 York Avenue, New York 10065, USA.

Abstract:  Environmental nutrient levels impact cancer cell metabolism, resulting in context-dependent gene essentiality. Here, using loss-of-function screening based on RNA interference, we show that environmental oxygen levels are a major driver of differential essentiality between in vitro model systems and in vivo tumours. Above the 3-8% oxygen concentration typical of most tissues, we find that cancer cells depend on high levels of the iron-sulfur cluster biosynthetic enzyme NFS1. Mammary or subcutaneous tumours grow despite suppression of NFS1, whereas metastatic or primary lung tumours do not. Consistent with a role in surviving the high oxygen environment of incipient lung tumours, NFS1 lies in a region of genomic amplification present in lung adenocarcinoma and is most highly expressed in well-differentiated adenocarcinomas. NFS1 activity is particularly important for maintaining the iron-sulfur co-factors present in multiple cell-essential proteins upon exposure to oxygen compared to other forms of oxidative damage. Furthermore, insufficient iron-sulfur cluster maintenance robustly activates the iron-starvation response and, in combination with inhibition of glutathione biosynthesis, triggers ferroptosis, a non-apoptotic form of cell death. Suppression of NFS1 cooperates with inhibition of cysteine transport to trigger ferroptosis in vitro and slow tumour growth. Therefore, lung adenocarcinomas select for expression of a pathway that confers resistance to high oxygen tension and protects cells from undergoing ferroptosis in response to oxidative damage. 

Retracted: No

PubPeer comments:
 Certainly a minor issue, but a little misleading: shNFs1 lung panels are the same picture just 180u00baC rotated. ![file] , 
 #1 We have reviewed the primary data collected for this panel in response to this comment. In this panel, we show two representative images of tumors from the lungs of mice into which mouse lung adenocarcinoma cells (KP) were instilled intratracheally. As described in Figure 2j, these cells form tumors very poorly when expressing shNfs1. Therefore, when choosing representative images we inadvertently chose images of the same tumor and did not recognize the mistake because of the rotation of the image. If you were to zoom in, you can see that the images, while of the same region, are not identical at the pixel level, reflecting the fact that the images were taken at different times. We apologize for the duplication. , 
 Raw data for Figure 3B contains a set of duplicates values between replicate 1 and 2. Could the authors check if these values are correct because it doesnu2019t sound reasonable that the authors got the same decimal values in different replicates ![file] , 
 In this experiment, we report the effects of several anti-oxidant compounds on cells in which NFS1 has been suppressed by shRNA, a treatment that blunts proliferation in standard culture conditions. In each of 3 to 5 independent experiments conducted in biological triplicate, we did not observe any effect of these compounds on the proliferation of cells in which NFS1 had been suppressed. In contrast, culturing cells in 3% oxygen lead to a complete or near complete rescue of this phenotype (detailed further in Figure 1e). These 3-5 experiments conducted in biological triplicate are aggregated in Figure 3b. In the final revision of this manuscript (to comply with Nature s new policy to include primary data and individual points on all bar graphs with fewer than ten points) when adding individual data points and compiling the Source Data files, we inadvertently included several data points twice, as indicated by the comment above. These duplicated data have been removed in the corrected source data table provided below. This change does not affect our conclusion that these compounds do not affect viability of cells in which NFS1 has been suppressed. ![file] , 
 Dear authors, are you planning to issue an Erratum to correct the duplicated histology image and the duplicated table columns? It s been two years since this discussion, but I cannot find any update on the official paper s landing page. , 
 #5 I have written to the handling Editor at Nature, who recommended issuing an Erratum. I will attend to this. , 
 Correction to: Nature https: doi.org 10.1038 nature24637Published online 22 November 2017 The authors regret to report that errors in data reporting were introduced after peer review during finalization of data for publication. In Fig. 2b, the data points in the u201cPooru201d and u201cModerateu201d groups are shifted by approximately u201330 units on the y axis. The original Fig. 2b is shown next to the corrected panel with the correct values (Supplementary Fig. 1). In the Source Data for Fig. 3b, a column is partially duplicated. These duplicated data points were erroneously reported on the panel for Fig. 3b. The original Fig. 3b is shown next to the corrected panel without the erroneous replicates (Supplementary Fig. 2). The source data for the original Fig. 3b have also been corrected and are provided. In the text, the Fig. 3 legend should have read u201cFive-day proliferation assay, MDA-MB-231 cells, 3% O2 (nu2009=u20094) or 21% O2 and with trolox (100u2009u03bcM, nu2009=u20095), Fer-1 (1u2009u03bcM, nu2009=u20093) or N-acetylcysteine (NAC, 750u2009u03bcM, nu2009=u20093).u201d Also, in Extended Data Fig. 4h, the two representative images shown on the top right, while independently acquired, are from partially overlapping areas of the same tumour. We have replaced the partially duplicated image with an independent representative image (Supplementary Fig. 3). Additional supplementary files are now provided (Supplementary Figs. 1u20133). Amended source data are included for Fig. 3b. Additional source data are now provided for all images (Figs. 2c and 4e and Extended Data Figs. 3b, 3c, 3f, 4c, 4h, 6i, 7a and 7b). https: www.nature.com articles s41586-022-05323-7 ,