Ammonium tetrathiomolybdate triggers autophagy-dependent NRF2 activation in vascular endothelial cells

作者全名："Zhang, Mengling; Qiu, Hongmei; Mao, Lejiao; Wang, Bin; Li, Na; Fan, Yinzhen; Weng, Ping; Hu, Siyao; Dong, Xiaomei; Qin, Xia; Chen, Chengzhi; Zou, Zhen; Yu, Chao; Zhang, Jun"

作者地址："[Zhang, Mengling; Qiu, Hongmei; Weng, Ping; Hu, Siyao; Yu, Chao] Chongqing Med Univ, Coll Pharm, Chongqing Key Lab Pharmaceut Metab Res, Chongqing 400016, Peoples R China; [Mao, Lejiao; Wang, Bin; Li, Na; Fan, Yinzhen; Dong, Xiaomei; Zou, Zhen; Zhang, Jun] Chongqing Med Univ, Inst Life Sci, Mol Biol Lab Resp Dis, Chongqing 400016, Peoples R China; [Qin, Xia] Chongqing Med Univ, Dept Pharm, Affiliated Hosp 1, Chongqing 400016, Peoples R China; [Chen, Chengzhi] Chongqing Med Univ, Sch Publ Hlth, Dept Occupat & Environm Hlth, Chongqing 400016, Peoples R China; [Chen, Chengzhi; Zou, Zhen; Zhang, Jun] Chongqing Med Univ, Res Ctr Environm & Human Hlth, Sch Publ Hlth, Chongqing 400016, Peoples R China"

通信作者："Yu, C (通讯作者)，Chongqing Med Univ, Coll Pharm, Chongqing Key Lab Pharmaceut Metab Res, Chongqing 400016, Peoples R China.; Zou, Z; Zhang, J (通讯作者)，Chongqing Med Univ, Inst Life Sci, Mol Biol Lab Resp Dis, Chongqing 400016, Peoples R China.; Zou, Z; Zhang, J (通讯作者)，Chongqing Med Univ, Res Ctr Environm & Human Hlth, Sch Publ Hlth, Chongqing 400016, Peoples R China."

来源：CELL DEATH & DISEASE

ESI学科分类：MOLECULAR BIOLOGY & GENETICS

WOS号：WOS:000844794200001

JCR分区：Q1

影响因子：9

年份：2022

卷号：13

期号：8

开始页：　

结束页：　

文献类型：Article

关键词：　

摘要："Ammonium tetrathiomolybdate (TTM) is a copper chelator in clinical trials for treatment of Wilson's disease, tumors and other diseases. In the current study, we innovatively discovered that TTM is a novel NRF2 activator and illustrated that autophagy contributed to TTM-induced NRF2 activation. We showed that TTM treatment promoted NRF2 nuclear translocation and upregulated transcription level of NRF2 target genes including HMOX1, GCLM, and SLC7A11 in vascular endothelial cells (HUVECs). Moreover, NRF2 deficiency directly hindered TTM-mediated antioxidative effects. Followingly, we revealed that overexpression of KEAP1, a negative regulator of NRF2, significantly repressed NRF2 activation induced by TTM. Further mutation analysis revealed that KEAP1 Cys151 is a major sensor responsible for TTM-initiated NRF2 signaling, suggesting that KEAP1 is involved in TTM-mediated NRF2 activation. Notably, we found that TTM can trigger autophagy as evidenced by accumulation of autophagosomes, elevation of LC3BI-II/I, increase of LC3 puncta and activation of AMPK/mTOR/ULK1 pathway. Autophagic flux assay indicated that TTM significantly enhanced autophagic flux in HUVECs. Inhibition of autophagy with knockout of autophagy key gene ATG5 resulted in suppression of TTM-induced NRF2 activation. TTM also induced phosphorylation of autophagy receptor SQSTM1 at Ser349, while SQSTM1-deficiency inhibited KEAP1 degradation and blocked NRF2 signaling pathway, suggesting that TTM-induced NRF2 activation is autophagy dependent. As the novel NRF2 activator, TTM protected against sodium arsenite (NaAsO2)-induced oxidative stress and cell death, while NRF2 deficiency weakened TTM antioxidative effects. Finally, we showed that autophagy-dependent NRF2 activation contributed to the protective effects of TTM against NaAsO2-induced oxidative injury, because of ATG5 or SQSTM1 knockout aggravated NaAsO2-induced elevation of HMOX1, cleaved PARP and gamma H2AX. Taken together, our findings highlight copper chelator TTM is a novel autophagy-dependent NRF2 activator and shed a new light on the cure for oxidative damage-related diseases."

基金机构："National Natural Science Foundation of China [81500343, 81903358]; Chongqing Talents: Exceptional Young Talents Project [CQYC2020058650]; Natural Science Foundation of Chongqing [cstc2018jcyjAX0355, cstc2021ycjh-bgzxm0105, cstc2020jcyj-msxmX0155]; Science and Technology Research Program of Chongqing Municipal Education Commission [KJQN202000423, KJCXZD2020020, KJQN202100405]; Future Medical Youth Innovation Program of Chongqing Medical University [W0038]; Chongqing Bayu Talented Young Scholar program"

基金资助正文："We appreciate Ms. Xiaoyun Dou and Aijia Song (Institute of Life Sciences, Chongqing Medical University) for the help of confocal microscopy and transmission electron microscopy. We also thank Dr. Kai Wang (Key Laboratory of Molecular Biology of Infectious Diseases, Chongqing Medical University) for providing KEAP1-Flag and NRF2-Myc plasmids. This work was supported partly by the National Natural Science Foundation of China (81500343 and 81903358), the Chongqing Talents: Exceptional Young Talents Project (CQYC2020058650), the Natural Science Foundation of Chongqing (cstc2018jcyjAX0355, cstc2021ycjh-bgzxm0105 and cstc2020jcyj-msxmX0155), the Science and Technology Research Program of Chongqing Municipal Education Commission (KJQN202000423, KJCXZD2020020, and KJQN202100405), the Future Medical Youth Innovation Program of Chongqing Medical University (W0038). J.Z., Z.Z., and C.C. are also supported by Chongqing Bayu Talented Young Scholar program."