PMID- 33487631 OWN - NLM STAT- MEDLINE DCOM- 20220304 LR - 20230128 IS - 2059-3635 (Electronic) IS - 2095-9907 (Print) IS - 2059-3635 (Linking) VI - 6 IP - 1 DP - 2021 Jan 25 TI - Targeting TRPV1-mediated autophagy attenuates nitrogen mustard-induced dermal toxicity. PG - 29 LID - 10.1038/s41392-020-00389-z [doi] LID - 29 AB - Nitrogen mustard (NM) causes severe vesicating skin injury, which lacks effective targeted therapies. The major limitation is that the specific mechanism of NM-induced skin injury is not well understood. Recently, autophagy has been found to play important roles in physical and chemical exposure-caused cutaneous injuries. However, whether autophagy contributes to NM-induced dermal toxicity is unclear. Herein, we initially confirmed that NM dose-dependently caused cell death and induced autophagy in keratinocytes. Suppression of autophagy by 3-methyladenine, chloroquine, and bafilomycin A1 or ATG5 siRNA attenuated NM-induced keratinocyte cell death. Furthermore, NM increased transient receptor potential vanilloid 1 (TRPV1) expression, intracellular Ca(2+) content, and the activities of Ca(2+)/calmodulin-dependent kinase kinase beta (CaMKKbeta), AMP-activated protein kinase (AMPK), unc-51-like kinase 1 (ULK1), and mammalian target of rapamycin (mTOR). NM-induced autophagy in keratinocytes was abolished by treatment with inhibitors of TRPV1 (capsazepine), CaMKKbeta (STO-609), AMPK (compound C), and ULK1 (SBI-0206965) as well as TRPV1, CaMKKbeta, and AMPK siRNA transfection. In addition, an mTOR inhibitor (rapamycin) had no significant effect on NM-stimulated autophagy or cell death of keratinocytes. Finally, the results of the in vivo experiment in NM-treated skin tissues were consistent with the findings of the in vitro experiment. In conclusion, NM-caused dermal toxicity by overactivating autophagy partially through the activation of TRPV1-Ca(2+)-CaMKKbeta-AMPK-ULK1 signaling pathway. These results suggest that blocking TRPV1-dependent autophagy could be a potential treatment strategy for NM-caused cutaneous injury. FAU - Chen, Mingliang AU - Chen M AD - Department of Chemical Defense Medicine, School of Military Preventive Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing, 400038, China. AD - Institute of Pathology and Southwest Cancer Centre, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, 400038, China. AD - Institute of Toxicology, School of Military Preventive Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing, 400038, China. FAU - Dong, Xunhu AU - Dong X AD - Department of Chemical Defense Medicine, School of Military Preventive Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing, 400038, China. AD - Institute of Toxicology, School of Military Preventive Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing, 400038, China. FAU - Deng, Haoyue AU - Deng H AD - State Key Laboratory of Trauma, Burns and Combined Injury, Second Department of Research Institute of Surgery, Daping Hospital, Third Military Medical University (Army Medical University), Chongqing, 400042, China. FAU - Ye, Feng AU - Ye F AD - Department of Chemical Defense Medicine, School of Military Preventive Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing, 400038, China. AD - Institute of Toxicology, School of Military Preventive Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing, 400038, China. FAU - Zhao, Yuanpeng AU - Zhao Y AD - Department of Chemical Defense Medicine, School of Military Preventive Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing, 400038, China. AD - Institute of Toxicology, School of Military Preventive Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing, 400038, China. FAU - Cheng, Jin AU - Cheng J AD - Department of Chemical Defense Medicine, School of Military Preventive Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing, 400038, China. AD - Institute of Toxicology, School of Military Preventive Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing, 400038, China. FAU - Dan, Guorong AU - Dan G AD - Department of Chemical Defense Medicine, School of Military Preventive Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing, 400038, China. AD - Institute of Toxicology, School of Military Preventive Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing, 400038, China. FAU - Zhao, Jiqing AU - Zhao J AD - Department of Chemical Defense Medicine, School of Military Preventive Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing, 400038, China. AD - Institute of Toxicology, School of Military Preventive Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing, 400038, China. FAU - Sai, Yan AU - Sai Y AD - Department of Chemical Defense Medicine, School of Military Preventive Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing, 400038, China. AD - Institute of Toxicology, School of Military Preventive Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing, 400038, China. FAU - Bian, Xiuwu AU - Bian X AD - Institute of Pathology and Southwest Cancer Centre, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, 400038, China. bianxiuwu@263.net. FAU - Zou, Zhongmin AU - Zou Z AD - Department of Chemical Defense Medicine, School of Military Preventive Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing, 400038, China. zmzou@tmmu.edu.cn. AD - Institute of Toxicology, School of Military Preventive Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing, 400038, China. zmzou@tmmu.edu.cn. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20210125 PL - England TA - Signal Transduct Target Ther JT - Signal transduction and targeted therapy JID - 101676423 RN - 0 (ATG5 protein, human) RN - 0 (Autophagy-Related Protein 5) RN - 0 (Macrolides) RN - 0 (RNA, Small Interfering) RN - 0 (TRPV Cation Channels) RN - 0 (TRPV1 protein, human) RN - 50D9XSG0VR (Mechlorethamine) RN - 5142-23-4 (3-methyladenine) RN - 886U3H6UFF (Chloroquine) RN - 88899-55-2 (bafilomycin A1) RN - EC 2.7.11.1 (Autophagy-Related Protein-1 Homolog) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) RN - EC 2.7.11.17 (Calcium-Calmodulin-Dependent Protein Kinase Kinase) RN - EC 2.7.11.3 (AMP-Activated Protein Kinase Kinases) RN - JAC85A2161 (Adenine) RN - W36ZG6FT64 (Sirolimus) SB - IM MH - AMP-Activated Protein Kinase Kinases/genetics MH - Adenine/analogs & derivatives/pharmacology MH - Animals MH - Autophagy/*genetics MH - Autophagy-Related Protein 5/*genetics MH - Autophagy-Related Protein-1 Homolog/genetics MH - Blister/genetics/pathology MH - Calcium-Calmodulin-Dependent Protein Kinase Kinase/genetics MH - Cell Death/drug effects MH - Chloroquine/pharmacology MH - Humans MH - Keratinocytes/drug effects/pathology MH - Macrolides/pharmacology MH - Mechlorethamine/toxicity MH - Mice MH - RNA, Small Interfering/genetics MH - Sirolimus/pharmacology MH - Skin/drug effects/injuries/pathology MH - Skin Diseases/chemically induced/drug therapy/*genetics/pathology MH - TOR Serine-Threonine Kinases/genetics MH - TRPV Cation Channels/*genetics PMC - PMC7829253 COIS- The authors declare no competing interests. EDAT- 2021/01/26 06:00 MHDA- 2022/03/05 06:00 PMCR- 2021/01/25 CRDT- 2021/01/25 05:35 PHST- 2020/03/16 00:00 [received] PHST- 2020/10/09 00:00 [accepted] PHST- 2020/10/07 00:00 [revised] PHST- 2021/01/25 05:35 [entrez] PHST- 2021/01/26 06:00 [pubmed] PHST- 2022/03/05 06:00 [medline] PHST- 2021/01/25 00:00 [pmc-release] AID - 10.1038/s41392-020-00389-z [pii] AID - 389 [pii] AID - 10.1038/s41392-020-00389-z [doi] PST - epublish SO - Signal Transduct Target Ther. 2021 Jan 25;6(1):29. doi: 10.1038/s41392-020-00389-z.