PMID- 38184997 OWN - NLM STAT- MEDLINE DCOM- 20240122 LR - 20240202 IS - 2213-2317 (Electronic) IS - 2213-2317 (Linking) VI - 69 DP - 2024 Feb TI - Oncogenic KEAP1 mutations activate TRAF2-NFkappaB signaling to prevent apoptosis in lung cancer cells. PG - 103031 LID - S2213-2317(24)00007-7 [pii] LID - 10.1016/j.redox.2024.103031 [doi] LID - 103031 AB - The Kelch-like ECH-associated protein 1 (KEAP1) - Nuclear factor erythroid 2 -related factor 2 (NRF2) pathway is the major transcriptional stress response system in cells against oxidative and electrophilic stress. NRF2 is frequently constitutively active in many cancers, rendering the cells resistant to chemo- and radiotherapy. Loss-of-function (LOF) mutations in the repressor protein KEAP1 are common in non-small cell lung cancer, particularly adenocarcinoma. While the mutations can occur throughout the gene, they are enriched in certain areas, indicating that these may have unique functional importance. In this study, we show that in the GSEA analysis of TCGA lung adenocarcinoma RNA-seq data, the KEAP1 mutations in R320 and R470 were associated with enhanced Tumor Necrosis Factor alpha (TNFalpha) - Nuclear Factor kappa subunit B (NFkappaB) signaling as well as MYC and MTORC1 pathways. To address the functional role of these hotspot mutations, affinity purification and mass spectrometry (AP-MS) analysis of wild type (wt) KEAP1 and its mutation forms, R320Q and R470C were employed to interrogate differences in the protein interactome. We identified TNF receptor associated factor 2 (TRAF2) as a putative protein interaction partner. Both mutant KEAP1 forms showed increased interaction with TRAF2 and other anti-apoptotic proteins, suggesting that apoptosis signalling could be affected by the protein interactions. A549 lung adenocarcinoma cells overexpressing mutant KEAP1 showed high TRAF2-mediated NFkappaB activity and increased protection against apoptosis, XIAP being one of the key proteins involved in anti-apoptotic signalling. To conclude, KEAP1 R320Q and R470C and its interaction with TRAF2 leads to activation of NFkappaB pathway, thereby protecting against apoptosis. CI - Copyright (c) 2024 The Authors. Published by Elsevier B.V. All rights reserved. FAU - Deen, Ashik Jawahar AU - Deen AJ AD - A. I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, 70211, Finland. FAU - Adinolfi, Simone AU - Adinolfi S AD - A. I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, 70211, Finland. FAU - Harkonen, Jouni AU - Harkonen J AD - A. I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, 70211, Finland; Department of Pathology, Hospital Nova of Central Finland, Jyvaskyla, 40620, Finland. FAU - Patinen, Tommi AU - Patinen T AD - A. I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, 70211, Finland. FAU - Liu, Xiaonan AU - Liu X AD - Institute of Biotechnology, HiLIFE, University of Helsinki, Helsinki, 00014, Finland. FAU - Laitinen, Tuomo AU - Laitinen T AD - School of Pharmacy, University of Eastern Finland, Kuopio, 70211, Finland. FAU - Takabe, Piia AU - Takabe P AD - Institute of Biomedicine, University of Eastern Finland, Kuopio, 70211, Finland. FAU - Kainulainen, Kirsi AU - Kainulainen K AD - Institute of Biomedicine, University of Eastern Finland, Kuopio, 70211, Finland. FAU - Pasonen-Seppanen, Sanna AU - Pasonen-Seppanen S AD - Institute of Biomedicine, University of Eastern Finland, Kuopio, 70211, Finland. FAU - Gawriyski, Lisa M AU - Gawriyski LM AD - Institute of Biotechnology, HiLIFE, University of Helsinki, Helsinki, 00014, Finland. FAU - Arasu, Uma Thanigai AU - Arasu UT AD - A. I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, 70211, Finland. FAU - Selvarajan, Ilakya AU - Selvarajan I AD - A. I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, 70211, Finland. FAU - Makinen, Petri AU - Makinen P AD - A. I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, 70211, Finland. FAU - Laitinen, Hanna AU - Laitinen H AD - A. I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, 70211, Finland. FAU - Kansanen, Emilia AU - Kansanen E AD - A. I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, 70211, Finland; Science Service Centre, Kuopio University Hospital, Kuopio, 70211, Finland. FAU - Kaikkonen, Minna U AU - Kaikkonen MU AD - A. I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, 70211, Finland. FAU - Poso, Antti AU - Poso A AD - School of Pharmacy, University of Eastern Finland, Kuopio, 70211, Finland; Department of Internal Medicine VIII, University Hospital Tubingen, Tubingen, 72076, Germany. FAU - Varjosalo, Markku AU - Varjosalo M AD - Institute of Biotechnology, HiLIFE, University of Helsinki, Helsinki, 00014, Finland. FAU - Levonen, Anna-Liisa AU - Levonen AL AD - A. I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, 70211, Finland. Electronic address: anna-liisa.levonen@uef.fi. LA - eng PT - Journal Article DEP - 20240104 PL - Netherlands TA - Redox Biol JT - Redox biology JID - 101605639 RN - 0 (TNF Receptor-Associated Factor 2) RN - 0 (Kelch-Like ECH-Associated Protein 1) RN - 0 (NF-E2-Related Factor 2) RN - 0 (Intracellular Signaling Peptides and Proteins) RN - 0 (NF-kappa B) RN - 0 (KEAP1 protein, human) SB - IM MH - Humans MH - *Lung Neoplasms/metabolism MH - *Carcinoma, Non-Small-Cell Lung/pathology MH - TNF Receptor-Associated Factor 2/genetics/metabolism MH - Kelch-Like ECH-Associated Protein 1/genetics/metabolism MH - NF-E2-Related Factor 2/metabolism MH - Cell Line, Tumor MH - Intracellular Signaling Peptides and Proteins/metabolism MH - *Adenocarcinoma of Lung/genetics MH - Apoptosis/genetics MH - NF-kappa B/genetics/metabolism MH - Mutation PMC - PMC10808971 OTO - NOTNLM OT - Apoptosis OT - KEAP1 OT - Lung cancer OT - NFkappaB OT - TNFalpha OT - TRAF2 COIS- Declaration of competing interest The authors declare that there are no potential conflicts of interest. EDAT- 2024/01/08 00:42 MHDA- 2024/01/22 06:42 PMCR- 2024/01/04 CRDT- 2024/01/07 18:09 PHST- 2023/10/18 00:00 [received] PHST- 2023/12/22 00:00 [revised] PHST- 2024/01/02 00:00 [accepted] PHST- 2024/01/22 06:42 [medline] PHST- 2024/01/08 00:42 [pubmed] PHST- 2024/01/07 18:09 [entrez] PHST- 2024/01/04 00:00 [pmc-release] AID - S2213-2317(24)00007-7 [pii] AID - 103031 [pii] AID - 10.1016/j.redox.2024.103031 [doi] PST - ppublish SO - Redox Biol. 2024 Feb;69:103031. doi: 10.1016/j.redox.2024.103031. Epub 2024 Jan 4.