bims-nurfca 2023-10-08 papers

Issue of 2023–10–08
seven papers selected by
Caner Geyik, Istinye University

Pigment Cell Melanoma Res. 2023 Sep 30.

Nrf2 protein in melanoma progression, as a new means of treatment.

Melanoma is a potentially lethal form of skin cancer resulting from the unlimited proliferation of melanocytes. Melanocytic lineage appears to have a greater rate of reactive oxygen species (ROS) production, possibly as a result of exposure to ultraviolet (UV) light and the production of melanin. It has been established that nuclear factor erythroid 2-related factor 2 (Nrf2) serves as a master regulator of the cellular response to oxidative stresses. Recent research has shown that the Nrf2 and its critical negative regulator Kelch-like ECH-associated protein 1 (Keap1) are misregulated in melanoma, and the Keap1-Nrf2 pathway has emerged as a promising new target for treating and preventing melanoma. In melanoma, Nrf2 may either limit tumor growth or promote its development. This review covers a wide range of topics, including the dual functions played by the Keap1-Nrf2 signaling pathway in melanoma and the most recent targeting techniques of the Nrf2.

Keywords: Nrf2; antioxidant; melanocyte; melanoma; oxidative stress

DOI: https://doi.org/10.1111/pcmr.13137

Redox Biol. 2023 Sep 22. pii: S2213-2317(23)00302-6. [Epub ahead of print]67 102901

Small molecule screen identifies pyrimethamine as an inhibitor of NRF2-driven esophageal hyperplasia.

Chorlada Paiboonrungruang, Zhaohui Xiong, David Lamson, Yahui Li, Brittany Bowman, Julius Chembo, Caizhi Huang, Jianying Li, Eric W Livingston, Jon E Frank, Vivian Chen, Yong Li, Bernard Weissman, Hong Yuan, Kevin P Williams, M Ben Major, Xiaoxin Chen.

OBJECTIVE: NRF2 is a master transcription factor that regulates the stress response. NRF2 is frequently mutated and activated in human esophageal squamous cell carcinoma (ESCC), which drives resistance to chemotherapy and radiation therapy. Therefore, a great need exists for NRF2 inhibitors for targeted therapy of NRF2high ESCC.
DESIGN: We performed high-throughput screening of two compound libraries from which hit compounds were further validated in human ESCC cells and a genetically modified mouse model. The mechanism of action of one compound was explored by biochemical assays.
RESULTS: Using high-throughput screening of two small molecule compound libraries, we identified 11 hit compounds as potential NRF2 inhibitors with minimal cytotoxicity at specified concentrations. We then validated two of these compounds, pyrimethamine and mitoxantrone, by demonstrating their dose- and time-dependent inhibitory effects on the expression of NRF2 and its target genes in two NRF2Mut human ESCC cells (KYSE70 and KYSE180). RNAseq and qPCR confirmed the suppression of global NRF2 signaling by these two compounds. Mechanistically, pyrimethamine reduced NRF2 half-life by promoting NRF2 ubiquitination and degradation in KYSE70 and KYSE180 cells. Expression of an Nrf2E79Q allele in mouse esophageal epithelium (Sox2CreER;LSL-Nrf2E79Q/+) resulted in an NRF2high phenotype, which included squamous hyperplasia, hyperkeratinization, and hyperactive glycolysis. Treatment with pyrimethamine (30 mg/kg/day, p.o.) suppressed the NRF2high esophageal phenotype with no observed toxicity.
CONCLUSION: We have identified and validated pyrimethamine as an NRF2 inhibitor that may be rapidly tested in the clinic for NRF2high ESCC.

Keywords: Esophageal squamous cell carcinoma; KEAP1; Mitoxantrone; NRF2; Pyrimethamine

DOI: https://doi.org/10.1016/j.redox.2023.102901

Cancer Cell Int. 2023 Oct 04. 23(1): 229

Pan-cancer analysis of NFE2L2 mutations identifies a subset of lung cancers with distinct genomic and improved immunotherapy outcomes.

Kewei Wang, Zixi Li, Ying Xuan, Yong Zhao, Chao Deng, Meidan Wang, Chenjun Xie, Fenglai Yuan, Qingfeng Pang, Wenjun Mao, Dongyan Cai, Zhangfeng Zhong, Jie Mei.

BACKGROUND: Mutations in the KEAP1-NFE2L2 signaling pathway were linked to increased tumorigenesis and aggressiveness. Interestingly, not all hotspot mutations on NFE2L2 were damaging; some even were activating. However, there was conflicting evidence about the association between NFE2L2 mutation and Nrf2-activating mutation and responsiveness to immune checkpoint inhibitors (ICIs) in non-small cell lung cancer (NSCLC) and other multiple cancers.
METHODS: The study with the largest sample size (n = 49,533) explored the landscape of NFE2L2 mutations and their impact response/resistance to ICIs using public cohorts. In addition, the in-house WXPH cohort was used to validate the efficacy of immunotherapy in the NFE2L2 mutated patients with NSCLC.
RESULTS: In two pan-cancer cohorts, Nrf2-activating mutation was associated with higher TMB value compared to wild-type. We identified a significant association between Nrf2-activating mutation and shorter overall survival in pan-cancer patients and NSCLC patients but not in those undergoing ICIs treatment. Similar findings were obtained in cancer patients carrying the NFE2L2 mutation. Furthermore, in NSCLC and other cancer cohorts, patients with NFE2L2 mutation demonstrated more objective responses to ICIs than patients with wild type. Our in-house WXPH cohort further confirmed the efficacy of immunotherapy in the NFE2L2 mutated patients with NSCLC. Lastly, decreased inflammatory signaling pathways and immune-depleted immunological microenvironments were enriched in Nrf2-activating mutation patients with NSCLC.
CONCLUSIONS: Our study found that patients with Nrf2-activating mutation had improved immunotherapy outcomes than patients with wild type in NSCLC and other tumor cohorts, implying that Nrf2-activating mutation defined a distinct subset of pan-cancers and might have implications as a biomarker for guiding ICI treatment, especially NSCLC.

Keywords: Biomarker; Immunotherapy; NFE2L2; NGS; NSCLC

DOI: https://doi.org/10.1186/s12935-023-03056-9

Int J Radiat Oncol Biol Phys. 2023 Oct 01. pii: S0360-3016(23)05621-3. [Epub ahead of print]117(2S): e248

Radiotherapy Resistance in Prostate Cancer Cells: AKR1C3 Inhibition of Ubiquitinated Degradation of Nrf2 through Interaction with KEAP1.

F Lyu, X Gao, S Shang, S Li, X Ren, J Chen.

PURPOSE/OBJECTIVE(S): Radiotherapy is the most fundamental treatment for prostate cancer (PCa), and although radiotherapy for overall PCa patients is effective, poor prognosis and resistance to multiple treatments regimes in some highly malignant PCa, such as those with high Gleason Scores (GS) (≥9), are important bottlenecks limiting the improvement of treatment outcomes for clinical. AKR1C3 is a key PCa resistance gene that our team identified previously, but the induction of the specific mechanism of radiotherapy resistance has not been fully revealed and understand.
MATERIALS/METHODS: To analyze the correlation between its expression level and clinical radiotherapy, we used the gene expression profiles data of PCa patients in TCGA database. We generated a stepwise increase of radiotherapy dose to established PCa radiotherapy resistant cell lines and detect the AKR1C3 expression level. In addition, to explore the molecular mechanism of AKR1C3 induced prostate cancer radiation tolerance through functional enrichment analysis. Then, to treat cells with cycloheximide and the protein stability of the Nrf2 was detected. Last, the protein ubiquitination level was assayed by co-immunoprecipitation (co-IP) after treatment with MG132. Finally, protein-protein interactions were identified using co-IP to mine possible binding molecules.
RESULTS: By analyzing the expression profiles data of PCa patients in the TCGA database, we found that in the population of PCa patients treated with radiotherapy, all patients with high AKR1C3 expression died after radiotherapy, suggesting that high AKR1C3 expression may be a biomarker of resistance to radiation. Accordingly, AKR1C3 expression levels showed a positive correlation with GS score, which may be a symbol for patients with highly malignant PCa. A PCa radiotherapy resistant cell line was constructed by a stepwise increase of ionizing radiation (IR) dose, and the total IR dose of radiotherapy was 84Gy, which reached the dose of radical radiotherapy for prostate cancer. The expression of AKR1C3 was further detected by RT-qPCR and WB, and it was found that the expression of AKR1C3 was significantly up-regulated in the resistant cell line, accompanied by milder DNA damage. What's more, by GSEA functional enrichment analysis, we discovered that AKR1C3 overexpression might be related with intracellular oxidative stress damage. After CHX treatment, the protein stability of Nrf2 was significantly enhanced in AKR1C3 overexpression groups than control groups. And the administration of MG132 showed the same results, indicating that the ubiquitinated degradation of Nrf2 was inhibited in AKR1C3 overexpression groups. Further by co-IP, we found that Nrf2 was less ubiquitinated in the cytoplasm after AKR1C3 overexpression, and AKR1C3 could bind Keap1.
CONCLUSION: In sum, we found that AKR1C3 can bind with Keap1 leading to reduced ubiquitination level of Nrf2, causing upregulation of Nrf2 expression and providing new insights into PCa radiotherapy resistance.

DOI: https://doi.org/10.1016/j.ijrobp.2023.06.1187

Cancer Lett. 2023 Sep 30. pii: S0304-3835(23)00356-7. [Epub ahead of print]576 216405

Targeting LINC01607 sensitizes hepatocellular carcinoma to Lenvatinib via suppressing mitophagy.

Yuxin Zhang, Yujie Zhang, Haisu Tao, Jinghan Zhu, Yuanxiang Lu, Fangling Cheng, Yixiao Xiong, Junjie Liu, Guangzhen Cai, Zhanguo Zhang, Huifang Liang, Yifa Chen, Wanguang Zhang.

Lenvatinib is a standard therapy option for advanced hepatocellular carcinoma (HCC), but resistance limits clinical benefits. In this study, we identified inhibition of ROS levels and reduced redox status in Lenvatinib-resistant HCC. Integrating RNA-seq with unbiased whole-genome CRISPR-Cas9 screen analysis indicated LINC01607 regulated the P62 to enhance drug resistance by affecting mitophagy and antioxidant pathways. Underlying mechanisms were investigated both in vitro and in vivo. We initially confirmed that LINC01607, as a competing endogenous RNA (ceRNA) competing with mirRNA-892b, triggered protective mitophagy by upregulating P62, which reduced ROS levels and promoted drug resistance. Furthermore, LINC01607 was proved to resist oxidative stress by regulating the P62-Nrf2 axis, which transcriptionally regulated the expression of LINC01607 to form a positive feedback loop. Finally, silencing LINC01607 combined with Lenvatinib reversed resistance in animal and patient-derived organoid models. In conclusion, we proposed a novel mechanism of Lenvatinib resistance involving ROS homeostasis. This work contributed to understanding redox homeostasis-related drug resistance and provided new therapeutic targets and strategies for HCC patients.

Keywords: Hepatocellular carcinoma; LINC01607; Lenvatinib resistance; Mitophagy; Nrf2; P62

DOI: https://doi.org/10.1016/j.canlet.2023.216405

Int Urol Nephrol. 2023 Oct 03.

Alpha-pinene neutralizes cisplatin-induced reproductive toxicity in male rats through activation of Nrf2 pathway.

Selim Demir, Ahmet Mentese, Zeynep Turkmen Usta, Nihal Turkmen Alemdar, Elif Ayazoglu Demir, Yuksel Aliyazicioglu.

PURPOSE: Testicular toxicity is one of the most important side effects of cisplatin (CP) therapy. Alpha-pinene (AP) is a naturally occurring monoterpene with antioxidant character in plants. Here, we aimed to evaluate the therapeutic activity of AP against CP-induced testicular toxicity by including the nuclear factor erythroid 2-associated factor 2 (Nrf2) pathway in rats.
METHODS: Thirty male rats were divided into 5 groups: control, CP, CP + AP (5 and 10 mg/kg) and only AP (10 mg/kg). CP was administered intraperitoneally at a dose of 5 mg/kg on the first day, followed by three consecutive injections of AP. Serum reproductive hormone levels were evaluated using ELISA kits. Oxidative stress (OS), inflammation, endoplasmic reticulum stress (ERS) and apoptosis markers in testicular tissue were also determined colorimetrically. In addition, how CP affects Nrf2 pathway and the effect of AP on this situation were also addressed.
RESULTS: Treatment with CP significantly increased OS, inflammation, ERS and apoptosis in testicular tissue. Administrations of AP resulted in an amelioration of these altered parameters. The mechanism of therapeutic effect of AP appeared to involve induction of Nrf2. Furthermore, these results were also confirmed by histological data.
CONCLUSION: Results suggest that AP can exhibit therapeutic effects against CP-induced testicular toxicity. It can be concluded that AP may be a potential molecule to abolish reproductive toxicity after chemotherapy.

Keywords: Alpha-pinene; Cisplatin; Endoplasmic reticulum stress; Inflammation; Nrf2; Oxidative stress

DOI: https://doi.org/10.1007/s11255-023-03817-5

Front Genet. 2023 ;14 1266680

Insight into Nrf2: a bibliometric and visual analysis from 2000 to 2022.

Yawei Ma, Zhongqing Wang, Yuedong Hu.

Background: Nrf2 plays a pivotal role in governing the antioxidant defense system, triggering the transcription of diverse genes involved in cellular protection. Its role in mitigating oxidative damage and modulating inflammatory processes has made Nrf2 an attractive target for therapeutic interventions. Despite the growing interest in Nrf2 research, a bibliometric analysis is relatively rare. This study aimed to clarify Nrf2's role in multiple diseases, identify emerging trends and hotspots using bibliometric analysis, and provide valuable insights and potential directions for future therapeutic interventions. Methods: The Science Citation Index of Web of Science Core library from 2000 to 2022 was searched on 22 October 2022. Use Microsoft Excel, CiteSpace, Bibliometrix, and VOS viewers for data collection and visualization of research focus and trends. Results: A vast collection of 22,040 research studies on Nrf2 published between 2000 and 2022 were identified. Nrf2 research has seen significant growth globally from 2000 to 2022. China leaded in publication numbers (9,623, 43.66%), while the United States dominated in citation frequency with 261,776 citations. China Medical University was the most productive institutions (459, 2.08%). Masayuki Yamamoto topped in publications (307), while Itoh K. ranked first in citations with 3669. Free Radical Biology and Medicine was the journal with the most studies and citations on Nrf2 (613, 29,687 citations). The analysis of keyword clustering enhanced the categorization of topics and can be summarized as oxidative stress, cancer, disorders in glycolipid metabolism, inflammation, and neurological conditions. Conclusion: China and the United States are the pioneers in Nrf2 research. Recently, there has been a comprehensive exploration of Nrf2 involving both experimental and clinical aspects, as well as mechanisms and therapeutic applications. Investigating novel molecular mechanisms, including NF-κB, Ho1, and Keap1, and developing enhanced, targeted Nrf2 activators or inhibitors to uncover the interplay among cancer, glycolipid metabolic disorder, inflammation, and neurological disorders will be upcoming trends and hotspots.

Keywords: Nrf2; VOSviewer; bibliometric analysis; citespace; visualization

DOI: https://doi.org/10.3389/fgene.2023.1266680