bims-nurfca 2023-02-05 papers

bims-nurfca

Biomed News

on NRF2 and Cancer

Issue of 2023–02–05
nine papers selected by
Caner Geyik, Istinye University

FAM117B promotes gastric cancer growth and drug resistance by targeting the KEAP1/NRF2 signaling pathway.
FXR1 facilitates axitinib resistance in clear cell renal cell carcinoma via regulating KEAP1/Nrf2 signaling pathway.
Camptothecin improves sorafenib sensitivity by inhibiting Nrf2‑ARE pathway in hepatocellular carcinoma.
m6A writer WTAP targets NRF2 to accelerate bladder cancer malignancy via m6A-dependent ferroptosis regulation.
NRF2 controls iron homeostasis and ferroptosis through HERC2 and VAMP8.
Benzo[a]pyrene treatment modulates Nrf2/Keap1 axis and changes the metabolic profile in rat lung cancer.
CRISPR metabolic screen identifies ATM and KEAP1 as targetable genetic vulnerabilities in solid tumors.
MCB-613 exploits a collateral sensitivity in drug resistant EGFR -mutant non-small cell lung cancer through covalent inhibition of KEAP1.
Targeting GSTP1 as Therapeutic Strategy against Lung Adenocarcinoma Stemness and Resistance to Tyrosine Kinase Inhibitors.

J Clin Invest. 2023 Feb 01. pii: e158705. [Epub ahead of print]133(3):

FAM117B promotes gastric cancer growth and drug resistance by targeting the KEAP1/NRF2 signaling pathway.

Yunjiang Zhou, Yaxin Chen, Yongwei Shi, Leyin Wu, Yingying Tan, Tao Li, Yigang Chen, Jiazeng Xia, Rong Hu.

  Gastric cancer often shows malignant growth and insensitivity to chemotherapeutic drugs due to the regulation of complex molecular mechanisms, which results in poor prognosis for patients. However, the relevant molecular mechanisms remain unclear. In this study, we reported that family with sequence similarity 117, member B (FAM117B), promoted the growth of gastric cancer cells and reduced the sensitivity of cells to chemotherapeutic drugs. Mechanistically, FAM117B competed with nuclear factor E2-related factor 2 (NRF2) for Kelch-like ECH-associated protein 1 (KEAP1) binding, reduced the ubiquitination degradation of NRF2, and activated the KEAP1/NRF2 signaling pathway. Moreover, FAM117B-induced growth and chemoresistance of gastric cancer cells were NRF2 dependent. We found that FAM117B and NRF2 protein levels were highly expressed in tumor tissues of patients with gastric cancer and their co-overexpression represented an independent factor for poor prognosis. Collectively, our findings reveal that FAM117B is involved in promoting gastric cancer growth and drug resistance, and it could be exploited as a cancer therapeutic target.

Keywords:  Cancer; Gastric cancer; Gastroenterology; Oncology

DOI:  https://doi.org/10.1172/JCI158705
Anticancer Drugs. 2022 Nov 16.

FXR1 facilitates axitinib resistance in clear cell renal cell carcinoma via regulating KEAP1/Nrf2 signaling pathway.

Haipeng Huang, Jiange Zhang, Peng Jiang, Xiaolong Xu, Fu Huang, Binli Zhao, Xiaoming Wang, Liquan Zhou.

Axitinib is emerging as a first-line combination treatment drug for metastatic renal cell carcinoma, but the acquired resistance significantly bothers the treatment efficacy. This article is to investigate the impact of fragile X mental retardation autosomal homolog 1 (FXR1) and its mechanistic involvement with Kelch-like epoxy chloropropan-associated protein 1 (KEAP1)/NF-E2-related factor 2 (Nrf2) pathway on cell resistance to axitinib in clear cell renal cell carcinoma (ccRCC). Establishment of axitinib resistance cells (786-O, Caki-1, 786-O/axitinib, or Caki-1/axitinib) was made, and the cells were then transfected with sh-FXR1, or co-transfected with sh-FXR1 and sh-KEAP1. The quantitative real-time PCR (qRT-PCR) and western blotting assays were employed to measure the expression of FXR1, KEAP1, Nrf2, LC3 II/I, Beclin 1, p62, MDR-1, and MRP-1. In addition, the binding between FXR1 and KEAP1 was verified by RNA-immunoprecipitation and RNA pull-down assays, and FXR1-dependent KEAP1 mRNA degradation was determined. Herein, FXR1 was demonstrated to be overexpressed in ccRCC cells, and showed higher expression in 786-O/axitinib and Caki-1/axitinib cells. Mechanistically, FXR1 enriched KEAP1 mRNA, and pulled downed by biotinylated KEAP1 probes. Results of RNA stability assay reveled that KEAP mRNA stability was suppressed by FXR1. Furthermore, knockdown of FXR1 promoted cell apoptosis and showed a restrained feature on cell resistance to axitinib. Of note, KEAP1 knockdown suppressed cell autophagy, oxidative stress, resistance to axitinib, and promoted apoptosis, despite FXR1 was downregulated in ccRCC cells. In conclusion, FXR1 played an encouraging role in ccRCC cell resistance to axitinib by modulating KEAP/Nrf2 pathway.

DOI: https://doi.org/10.1097/CAD.0000000000001416
Oncol Rep. 2023 Mar;pii: 55. [Epub ahead of print]49(3):

Camptothecin improves sorafenib sensitivity by inhibiting Nrf2‑ARE pathway in hepatocellular carcinoma.

Liwei Sun, Hankang Wang, Qian Liu, Fanguang Meng, Jinliang Zhang, Xiaodong Li, Shulin Chang, Guijie Li, Feng Chen.

  Sorafenib is a targeted drug for hepatocellular carcinoma (HCC), however, its efficacy is limited. Nuclear factor erythroid 2‑related factor 2 (Nrf2) contributes to sorafenib resistance. The present study investigated camptothecin (CPT) as a Nrf2 inhibitor to sensitize HCC to sorafenib. The effect of CPT on sorafenib sensitivity in HCC was assessed in vivo using H22 mice model (n=32) and VX2 rabbit models (n=32), which were sorted into four treatment groups. The expression levels of Nrf2, its downstream genes, including heme oxygenases‑1 (HO‑1) and NAD(P)H quinone oxidoreductase 1 (NQO1), and the epithelial‑mesenchymal transition markers Snail and N‑cadherin in tumors were determined using immunohistochemical staining and western blotting. Magnetic resonance imaging was used to monitor changes in tumor microcirculation and activity before and after treatment. Mouse body weights, liver and kidney function were monitored to evaluate the safety of combined therapy. The results revealed that the mean tumor size of the combined group was significantly smaller than that of sorafenib group for both models. The expression levels of Nrf2, heme oxygenase‑1, NAD(P)H quinone oxidoreductase 1, Snail, and N‑cadherin in the sorafenib group were significantly higher than control group (P<0.05). However, the expression levels of these genes were decreased in the combined group (P<0.05). Microcirculation perfusion and tumor activity in the combined group were also lower than sorafenib group. There were no significant differences in mouse body weight or liver and kidney function among the four groups. In summary, CPT is a Nrf2 inhibitor that could enhance the efficacy of sorafenib against HCC.

Keywords:  Nrf2; camptothecin; chemoresistance; hepatocellular carcinoma; sorafenib

DOI:  https://doi.org/10.3892/or.2023.8492
Apoptosis. 2023 Jan 31.

m6A writer WTAP targets NRF2 to accelerate bladder cancer malignancy via m6A-dependent ferroptosis regulation.

Ke Wang, Gang Wang, Gang Li, Wei Zhang, Yarong Wang, Xiaofeng Lin, Chengxian Han, Hanxuan Chen, Liang Shi, Abudoula Reheman, Jingkai Li, Zhaomin Li, Xinxuan Yang.

  Recent evidence have indicated that ferroptosis, a novel iron-dependent form of non-apoptotic cell death, plays a critical role in human cancers. Besides, emerging literatures have revealed the ovel function of N6-methyladenosine (m6A) in bladder cancer physiological. However, the underlying mechanism of m6A on bladder cancer is still unclear. Here, present work revealed that m6A methyltransferase ('writer') WTAP up-regulated in bladder cancer tissue and cells, indicating the poor prognosis of bladder cancer patients. Functionally, gain/loss-of-functional experiments illustrated that WTAP promoted the viability of bladder cancer cells and inhibited the erastin-induced ferroptosis. Mechanistically, there was a remarkable m6A modification site on 3'-UTR of endogenous antioxidant factor NRF2 RNA and WTAP could install its methylation. Moreover, m6A reader YTHDF1 recognized the m6A site on NRF2 mRNA and enhanced its mRNA stability. Therefore, these findings demonstrated potential therapeutic strategyies for bladder cancer via m6A-dependent manner.

Keywords:  Bladder cancer; Ferroptosis; N6-methyladenosine; NRF2.; WTAP

DOI:  https://doi.org/10.1007/s10495-023-01817-5
Sci Adv. 2023 Feb 03. 9(5): eade9585

NRF2 controls iron homeostasis and ferroptosis through HERC2 and VAMP8.

Annadurai Anandhan, Matthew Dodson, Aryatara Shakya, Jinjing Chen, Pengfei Liu, Yongyi Wei, Hui Tan, Qian Wang, Ziyan Jiang, Kevin Yang, Joe Gn Garcia, Setsuko K Chambers, Eli Chapman, Aikseng Ooi, Yang Yang-Hartwich, Brent R Stockwell, Donna D Zhang.

Enhancing the intracellular labile iron pool (LIP) represents a powerful, yet untapped strategy for driving ferroptotic death of cancer cells. Here, we show that NRF2 maintains iron homeostasis by controlling HERC2 (E3 ubiquitin ligase for NCOA4 and FBXL5) and VAMP8 (mediates autophagosome-lysosome fusion). NFE2L2/NRF2 knockout cells have low HERC2 expression, leading to a simultaneous increase in ferritin and NCOA4 and recruitment of apoferritin into the autophagosome. NFE2L2/NRF2 knockout cells also have low VAMP8 expression, which leads to ferritinophagy blockage. Therefore, deletion of NFE2L2/NRF2 results in apoferritin accumulation in the autophagosome, an elevated LIP, and enhanced sensitivity to ferroptosis. Concordantly, NRF2 levels correlate with HERC2 and VAMP8 in human ovarian cancer tissues, as well as ferroptosis resistance in a panel of ovarian cancer cell lines. Last, the feasibility of inhibiting NRF2 to increase the LIP and kill cancer cells via ferroptosis was demonstrated in preclinical models, signifying the impact of NRF2 inhibition in cancer treatment.

DOI: https://doi.org/10.1126/sciadv.ade9585
Chem Biol Interact. 2023 Feb 01. pii: S0009-2797(23)00040-6. [Epub ahead of print] 110373

Benzo[a]pyrene treatment modulates Nrf2/Keap1 axis and changes the metabolic profile in rat lung cancer.

Kaveri R Washimkar, Manendra Singh Tomar, Sharmeen Ishteyaque, Akhilesh Kumar, Ashutosh Shrivastava, Madhav Nilakanth Mugale.

  Lung cancer is an aggressive malignancy and the leading cause of cancer-related deaths. Benzo [a]pyrene (B [a]P), a polycyclic hydrocarbon, plays a pivotal role in lung carcinogenesis. Uncovering the molecular mechanism underlying the pathophysiology of B [a]P-induced malignancy is crucial. Male Sprague Dawley rats were induced with B [a]P to generate a lung cancer model. The B [a]P-administered rats show increased body and lung weight, loss of normal pulmonary architecture, and decreased survival. This study demonstrated that B [a]P upregulates activating transcription factor-6 (ATF6) and C/EBP Homologous Protein (CHOP) and induces endoplasmic reticulum (ER) stress. B [a]P also dysregulated mitochondrial homeostasis by upregulating, PTEN-induced putative kinase-1 (PINK1) and Parkin. B [a]P affected the levels of superoxide dismutase (SOD), reduced glutathione (GSH), malondialdehyde (MDA), and increased oxidative stress. B [a]P exposure downregulated Kelch-like ECH-associated protein 1 (Keap1) and upregulated nuclear factor erythroid 2-related factor 2 (Nrf2) and Heme oxygenase-1(HO1). The metabolomic study identified that biosynthesis of nucleotide, amino acids, pentose phosphate pathway (PPP), tricarboxylic acid cycle (TCA), and glutathione metabolism were up-accumulated. On the other hand, lower accumulation of fatty acids e.g., palmitic acid, stearic acid, and oleic acid were reported in the B [a]P induced group. Overall, the results of this study indicate that B [a]P treatment affects several signaling and metabolic pathways, whose dysregulation might be involved in lung cancer induction.

Keywords:  Benzo[a]pyrene; Endoplasmic reticulum stress; Lung cancer; Metabolomics; Mitochondrial dysfunction; Oxidative stress

DOI:  https://doi.org/10.1016/j.cbi.2023.110373
Proc Natl Acad Sci U S A. 2023 Feb 07. 120(6): e2212072120

CRISPR metabolic screen identifies ATM and KEAP1 as targetable genetic vulnerabilities in solid tumors.

Haojian Li, Yue Liu, Yunjie Xiao, Crystal N Wilson, Hui Jen Bai, Maxwell D Jones, Shihchun Wang, Jennie E DeVore, Esther Y Maier, Stephen T Durant, Myriem Boufraqech, Urbain Weyemi.

  Cancer treatments targeting DNA repair deficiencies often encounter drug resistance, possibly due to alternative metabolic pathways that counteract the most damaging effects. To identify such alternative pathways, we screened for metabolic pathways exhibiting synthetic lethality with inhibition of the DNA damage response kinase Ataxia-telangiectasia-mutated (ATM) using a metabolism-centered Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 library. Our data revealed Kelch-like ECH-associated protein 1 (KEAP1) as a key factor involved in desensitizing cancer cells to ATM inhibition both in vitro and in vivo. Cells depleted of KEAP1 exhibited an aberrant overexpression of the cystine transporter SLC7A11, robustly accumulated cystine inducing disulfide stress, and became hypersensitive to ATM inhibition. These hallmarks were reversed in a reducing cellular environment indicating that disulfide stress was a crucial factor. In The Cancer Genome Atlas (TCGA) pan-cancer datasets, we found that ATM levels negatively correlated with KEAP1 levels across multiple solid malignancies. Together, our results unveil ATM and KEAP1 as new targetable vulnerabilities in solid tumors.

Keywords:  ATM; DNA repair; KEAP1; drug resistance; redox metabolism

DOI:  https://doi.org/10.1073/pnas.2212072120
bioRxiv. 2023 Jan 19. pii: 2023.01.17.524094. [Epub ahead of print]

MCB-613 exploits a collateral sensitivity in drug resistant EGFR -mutant non-small cell lung cancer through covalent inhibition of KEAP1.

Christopher F Bassil, Gray R Anderson, Benjamin Mayro, Kayleigh N Askin, Peter S Winter, Samuel Gruber, Tierney M Hall, Jacob P Hoj, Christian Cerda-Smith, Haley M Hutchinson, Shane T Killarney, Katherine R Singleton, Li Qin, Kévin Jubien-Girard, Cécile Favreau, Anthony R Martin, Guillaume Robert, Rachid Benhida, Patrick Auberger, Ann Marie Pendergast, David M Lonard, Alexandre Puissant, Kris C Wood.

Targeted therapies have revolutionized cancer chemotherapy. Unfortunately, most patients develop multifocal resistance to these drugs within a matter of months. Here, we used a high-throughput phenotypic small molecule screen to identify MCB-613 as a compound that selectively targets EGFR -mutant, EGFR inhibitor-resistant non-small cell lung cancer (NSCLC) cells harboring diverse resistance mechanisms. Subsequent proteomic and functional genomic screens involving MCB-613 identified its target in this context to be KEAP1, revealing that this gene is selectively essential in the setting of EGFR inhibitor resistance. In-depth molecular characterization demonstrated that (1) MCB-613 binds KEAP1 covalently; (2) a single molecule of MCB-613 is capable of bridging two KEAP1 monomers together; and, (3) this modification interferes with the degradation of canonical KEAP1 substrates such as NRF2. Surprisingly, NRF2 knockout sensitizes cells to MCB-613, suggesting that the drug functions through modulation of an alternative KEAP1 substrate. Together, these findings advance MCB-613 as a new tool for exploiting the selective essentiality of KEAP1 in drug-resistant, EGFR -mutant NSCLC cells.

DOI: https://doi.org/10.1101/2023.01.17.524094
Adv Sci (Weinh). 2023 Jan 29. e2205262

Targeting GSTP1 as Therapeutic Strategy against Lung Adenocarcinoma Stemness and Resistance to Tyrosine Kinase Inhibitors.

Si-Qi Wang, Jun-Jiang Chen, Yuchen Jiang, Zi-Ning Lei, Ye Chun Ruan, Yihang Pan, Judy Wai Ping Yam, Maria Pik Wong, Zhi-Jie Xiao.

  Glutathione S-transferase pi (GSTP1), a phase II detoxification enzyme, is known to be overexpressed and mediates chemotherapeutic resistance in lung cancer. However, whether GSTP1 supports cancer stem cells (CSCs) and the underlying mechanisms in lung adenocarcinoma (LUAD) remain largely unknown. This study unveiled that GSTP1 is upregulated in lung CSCs and supports tumor self-renewal, metastasis, and resistance to targeted tyrosine kinase inhibitors of LUAD both in vitro and in vivo. Mechanistically, CaMK2A (calcium/calmodulin-dependent protein kinase 2 isoform A)/NRF2 (nuclear factor erythroid 2-related factor 2)/GSTP1 is uncovered as a regulatory axis under hypoxia. CaMK2A increased GSTP1 expression through phosphorylating the Sersine558 residue of NRF2 and promoting its nuclear translocation, a novel mechanism for NRF2 activation apart from conventional oxidization-dependent activation. Upregulation of GSTP1 in turn suppressed reactive oxygen species levels and supported CSC phenotypes. Clinically, GSTP1 analyzed by immunohistochemistry is upregulated in a proportion of LUAD and serves as a prognostic marker for survival. Using patient-derived organoids from an ALK-translocated LUAD, the therapeutic potential of a specific GSTP1 inhibitor ezatiostat in combination treatment with the ALK inhibitor crizotinib is demonstrated. This study demonstrates GSTP1 to be a promising therapeutic target for long-term control of LUAD through targeting CSCs.

Keywords:  calcium/calmodulin-dependent protein kinase 2 isoform A; cancer stem cells; glutathione S-transferase pi; lung adenocarcinoma; nuclear factor erythroid 2-related factor 2

DOI:  https://doi.org/10.1002/advs.202205262