bims-nurfca 2022-12-04 papers

bims-nurfca

Biomed News

on NRF2 and Cancer

Issue of 2022–12–04
eight papers selected by
Caner Geyik, Istinye University

Overactivated NRF2 induces pseudohypoxia in hepatocellular carcinoma by stabilizing HIF-1α.
The crosstalk between classic cell signaling pathways, non-coding RNAs and ferroptosis in drug resistance of tumors.
Triptolide promotes ferroptosis by suppressing Nrf2 to overcome leukemia cell resistance to doxorubicin.
Ryanodine receptor 2 promotes colorectal cancer metastasis by the ROS/BACH1 axis.
ARD1 stabilizes NRF2 through direct interaction and promotes colon cancer progression.
Attenuation of N-Nitrosodiethylamine -Induced Hepatocellular Carcinoma by Piceatannol and/or Cisplatin: The Interplay between Nuclear Factor (Erythroid Derived 2)-like 2 and Redox Status.
Elevated FSP1 protects KRAS-mutated cells from ferroptosis during tumor initiation.
Exosomal hsa-piR1089 promotes proliferation and migration in neuroblastoma via targeting KEAP1.

Free Radic Biol Med. 2022 Nov 29. pii: S0891-5849(22)01020-6. [Epub ahead of print]

Overactivated NRF2 induces pseudohypoxia in hepatocellular carcinoma by stabilizing HIF-1α.

Jie Zheng, Su-Jung Kim, Soma Saeidi, Seong Hoon Kim, Xizhu Fang, Yeon-Hwa Lee, Yanymee N Guillen-Quispe, Hoang Kieu Chi Ngo, Do-Hee Kim, Doojin Kim, Young-Joon Surh.

  Hypoxia-inducible factor-1α (HIF-1α) is highly expressed/activated in most hypoxic tumors including hepatocellular carcinoma (HCC). Another key transcription factor, nuclear factor erythroid 2-related factor 2 (NRF2), is also constitutively overactivated in HCC. In an attempt to determine whether HIF-1α and NRF2 could play complementary roles in HCC growth and progression, we investigated the crosstalk between these two transcription factors and underlying molecular mechanisms in cultured HCC cells and experimentally induced hepatocarcinogenesis as well as clinical settings. While silencing of HIF-1α in HepG2 human hepatoma cells did not alter the protein expression of NRF2, NRF2 knock down markedly reduced the nuclear accumulation of HIF-1α without influencing its mRNA expression. In diethylnitrosamine-induced hepatocarcinogenesis in wild type mice, there was elevated NRF2 expression with concomitant upregulation of HIF-1α. However, this was abolished in NRF2 knock out mice. NRF2 and HIF-1α co-localized and physically interacted with each other as assessed by in situ proximity ligation and immunoprecipitation assays. In addition, the interaction between NRF2 and HIF-1α as well as their overexpression was found in specimens obtained from clinical HCC patients. In normoxia, HIF-1α undergoes hydroxylation by a specific HIF-prolyl hydroxylase domain proteins (PHD), which facilitates VHL-mediated ubiquitination and proteasomal degradation of HIF-1α. NRF2 contributes to pseudohypoxia, by directly binding to the oxygen dependent degradation (ODD) domain of HIF-1α, which hampers the PHD2-mediated hydroxylation, concomitant recruitment of VHL and ubiquitination of HIF-1α.

Keywords:  HIF-1α; Hepatocellular carcinoma; Hypoxia; NRF2; Pseudohypoxia

DOI:  https://doi.org/10.1016/j.freeradbiomed.2022.11.039
Cell Signal. 2022 Nov 24. pii: S0898-6568(22)00300-X. [Epub ahead of print]102 110538

The crosstalk between classic cell signaling pathways, non-coding RNAs and ferroptosis in drug resistance of tumors.

Liangyu Yang, Lei Chen, Tingting Chen, Xinghong Gao, Yongai Xiong.

  Ferroptosis is an iron-dependent oxidative cell death characterized by the lethal accumulation of lipid-based reactive oxygen species (ROS), which is distinct from apoptosis, necrosis, and autophagy. Extensive studies suggest that ferroptosis be critical in regulating the growth and drug resistance of tumors, thus providing potential targets for cancer therapy. The development of resistance to cancer therapy remains a major challenge. Ferroptosis is associated with cancer drug resistance and inducing ferroptosis has been demonstrated to reverse drug resistance. This review focuses on a detailed account of the interplay between ferroptosis and related signaling pathways, including the Hippo signaling pathway, Keap1-Nrf2-ARE signaling pathway, Autophagy, and non-coding RNAs, which will shed light on developing the therapeutic role of regulating ferroptosis in reversing the resistance of cancer.

Keywords:  Autophagy; Ferroptosis; Hippo signaling pathway; Keap1-Nrf2-ARE signaling pathway; Non-coding RNAs; Tumor resistance

DOI:  https://doi.org/10.1016/j.cellsig.2022.110538
Mol Med Rep. 2023 Jan;pii: 17. [Epub ahead of print]27(1):

Triptolide promotes ferroptosis by suppressing Nrf2 to overcome leukemia cell resistance to doxorubicin.

Xia Wu, Shangqing Chen, Kechu Huang, Gongping Lin.

  Doxorubicin (DOX) is an extensively used chemotherapeutic drug to treat leukemia. However, there remains a pivotal clinical problem of resistance to DOX in patients with leukemia. Erythroid 2‑related factor 2 (Nrf2) is a master regulator of antioxidation response which serves a critical role in maintaining cellular oxidative homeostasis. However, whether Nrf2 is involved in DOX resistance is not totally clear. It is well‑documented that triptolide, a widely used drug to treat autoimmune disorders, possesses anti‑cancer activities, yet whether triptolide affects leukemia cell sensitivity to DOX remains to be elucidated. The present study aimed to determine the role of triptolide‑mediated downregulation of Nrf2 in regulating leukemia cell ferroptosis and resistance to DOX. For this purpose, low‑dose DOX was used to establish DOX‑resistant K562 cells and HL‑60 cells. Nrf2 mRNA and protein expression were examined by quantitative PCR and western blotting assays. The effects of triptolide on leukemia cell viability, reactive oxygen species (ROS) levels, or lipid oxidation were determined by CCK8 assay, DCFH‑DA assay, or BODIPY 581/591 C11 assay, respectively. The results show that Nrf2 expression was significantly upregulated in DOX‑resistant leukemia cells and clinical leukemia samples. Silencing of Nrf2 significantly sensitized leukemia cells to DOX. Furthermore, it was demonstrated that triptolide inhibited Nrf2 expression and induced leukemia cell ferroptosis, as evidenced by increased ROS levels and lipid oxidation as well as decreased glutathione peroxidase 4 expression. Ectopic expression of Nrf2 significantly rescued triptolide‑induced leukemia cell ferroptosis. Notably, the present study showed that triptolide re‑sensitized DOX‑resistant leukemia cells to DOX. In conclusion, the present study indicated that Nrf2 served a critical role in leukemia cell resistance to DOX and triptolide‑induced ferroptosis and suggested a potential strategy of combination therapy using triptolide and DOX in leukemia treatment.

Keywords:  Nrf2; doxorubicin resistance; ferroptosis; leukemia; triptolide

DOI:  https://doi.org/10.3892/mmr.2022.12904
Mol Oncol. 2022 Nov 30.

Ryanodine receptor 2 promotes colorectal cancer metastasis by the ROS/BACH1 axis.

Tianwei Chen, Xilin Zhang, Xufen Ding, Jing Feng, Xueli Zhang, Dong Xie, Xiang Wang.

  KRAS proto-oncogene, GTPase (KRAS)-mutant metastatic colorectal cancer (mCRC) lacks targeted therapy because the underlying mechanism remains obscure. Based on bioinformatic analysis, this study aims to elucidate a potential gene target for which an approved drug is available , and to further reveal the function as well as the underlying mechanism of the candidate gene. Here, we identified that ryanodine receptor 2 (RyR2) expression was upregulated in KRAS-mutant mCRC, and that this promoted cancer cell metastasis. S107, an approved drug to inhibit calcium release from RyR2 in the clinic, inhibited cancer cell metastasis both in vitro and in vivo. High expression of RyR2 predicts poor survival in our patient cohort. CRC patients with serosa invasion and vascular tumor thrombus are characterized by high RyR2 expression. Analysis of expression profiles upon RyR2 knockdown and inhibition reveals a set of metastasis-related molecules, and identifies BTB domain and CNC homolog 1 (BACH1) as the main transcription factor regulated by RyR2. RyR2 regulates cellular reactive oxygen species (ROS) levels, which activates nuclear factor erythroid 2-related factor 2 (Nrf2; also known as NFE2L2) and HMOX1 expression, and thus BACH1 accumulation. Collectively, this study provides evidence that the RyR2/ROS/BACH1 axis may be a potential target to intervene with CRC metastasis.

Keywords:  ROS; RyR2; colorectal cancer; metastasis

DOI:  https://doi.org/10.1002/1878-0261.13350
Life Sci. 2022 Nov 25. pii: S0024-3205(22)00917-1. [Epub ahead of print] 121217

ARD1 stabilizes NRF2 through direct interaction and promotes colon cancer progression.

Xizhu Fang, Yeon-Hwa Lee, Jeong-Hoon Jang, Su-Jung Kim, Seong Hoon Kim, Do-Hee Kim, Hye-Kyung Na, Kyung-Ok Kim, Jeong-Heum Baek, Young-Joon Surh.

   AIMS: Aberrant overactivation/overexpression of NRF2 is implicated as a driving event in tumor progression, which has been attributed to its mutation or inactivation of the inhibitory protein, KEAP1. However, alternative mechanisms responsible for sustained activation of NRF2 are less understood.
MAIN METHODS: Human colon cancer cell lines and tissues obtained from colorectal cancer patients were used. To examine the expression levels of ARD1 and NRF2, Western blot and immunofluorescence analyses were performed. To investigate the potential relevance of NRF2 and ARD1 to human colorectal cancer (CRC), NRF2 and ARD1 were individually silenced in human CRC cells (HCT-116) by transfection with their specific small interfering RNA (siRNA). To determine the functional role of ARD1 in NRF2 regulation, PLA, immunoprecipitation, nano-LC-ESI MS/MS, and in vitro acetylation assays were performed.
KEY FINDINGS: ARD1 knockdown in human colon cancer cell lines significantly reduced the protein levels of NRF2 without affecting its mRNA expression; however, silencing of NRF2 did not alter ARD1 protein expression. In addition, these two proteins were co-localized and physically interacted with each other both in human colon cancer cells (HCT-116) and human colon tumor tissues. Mechanistically, ARD1 overexpression increased the acetylation levels of NRF2. Moreover, an in vitro acetylation assay and mass spectrometric analysis demonstrated that ARD1 could directly acetylate NRF2. Ectopic expression of mutant forms of ARD1 with defective acetyltransferase activity reduced the stability of NRF2.
SIGNIFICANCE: In conclusion, ARD1 may potentiate the oncogenic function of NRF2 in human colon cancer by stabilizing this transcription factor.

Keywords:  ARD1; Acetylation; Colorectal cancer; NRF2; Posttranslational modification

DOI:  https://doi.org/10.1016/j.lfs.2022.121217
Asian Pac J Cancer Prev. 2022 Nov 01. pii: 90382. [Epub ahead of print]23(11): 3895-3903

Attenuation of N-Nitrosodiethylamine -Induced Hepatocellular Carcinoma by Piceatannol and/or Cisplatin: The Interplay between Nuclear Factor (Erythroid Derived 2)-like 2 and Redox Status.

Omnia Am Salama, Fatma Sm Moawed, Enas M Moustafa, Eman I Kandil.

   BACKGROUND: The natural compound's alternative and complementary uses have increased hopes for hepatocellular cancer treatment (HCC).
OBJECTS: The goal of this study was to see if Piceatannol (PIC) in combination with cisplatin has a synergistic effect on N, N-nitrosodiethylamine (DEN)-induced HCC in rats.
METHODS: Tissue antioxidant enzymes, malondialdehyde (MDA), and nuclear factor erythroid 2 related factors 2 (Nrf2) and tumor necrosis factor α (TNF-α) gene expression were all measured. Nuclear Factor Kabba B (NF-κB) was also tested, as well as hepatic caspase 3 and NAD (P) H quinone oxidoreductase 1 (NQO1). Liver specimens were subjected to histopathological analysis.
RESULTS: When compared to the HCC group, piceatannol and/or cisplatin caused a significant improvement in liver function tests, as well as a significant modulation in Nrf2 gene expression and antioxidant enzyme activities, as well as a significant decrease in tissue MDA, TNF-α, NF-κB levels, NQO1 activity, and prompt and caspase-3 activities. When the PIC and/or cisplatin combination was compared to each of these compounds alone, the results were substantial.
CONCLUSION: PIC in combination with cisplatin has been shown to have a synergistic anticancer impact through modulating Nrf2 and redox state. In addition, adding PIC to an HCC therapy plan that includes chemotherapeutic medicines may boost the efficacy of cisplatin while reducing its negative effects.

Keywords:  NFκ B; NQO1; Nrf2; TNF-alpha

DOI:  https://doi.org/10.31557/APJCP.2022.23.11.3895
Cell Death Differ. 2022 Nov 29.

Elevated FSP1 protects KRAS-mutated cells from ferroptosis during tumor initiation.

Fabienne Müller, Jonathan K M Lim, Christina M Bebber, Eric Seidel, Sofya Tishina, Alina Dahlhaus, Jenny Stroh, Julia Beck, Fatma Isil Yapici, Keiko Nakayama, Lucia Torres Fernández, Johannes Brägelmann, Gabriel Leprivier, Silvia von Karstedt.

Oncogenic KRAS is the key driver oncogene for several of the most aggressive human cancers. One key feature of oncogenic KRAS expression is an early increase in cellular reactive oxygen species (ROS) which promotes cellular transformation if cells manage to escape cell death, mechanisms of which remain incompletely understood. Here, we identify that expression of oncogenic as compared to WT KRAS in isogenic cellular systems renders cells more resistant to ferroptosis, a recently described type of regulated necrosis. Mechanistically, we find that cells with mutant KRAS show a specific lack of ferroptosis-induced lipid peroxidation. Interestingly, KRAS-mutant cells upregulate expression of ferroptosis suppressor protein 1 (FSP1). Indeed, elevated levels of FSP1 in KRAS-mutant cells are responsible for mediating ferroptosis resistance and FSP1 is upregulated as a consequence of MAPK and NRF2 pathway activation downstream of KRAS. Strikingly, FSP1 activity promotes cellular transformation in soft agar and its overexpression is sufficient to promote spheroid growth in 3D in KRAS WT cells. Moreover, FSP1 expression and its activity in ferroptosis inhibition accelerates tumor onset of KRAS WT cells in the absence of oncogenic KRAS in vivo. Consequently, we find that pharmacological induction of ferroptosis in pancreatic organoids derived from the LsL-KRASG12D expressing mouse model is only effective in combination with FSP1 inhibition. Lastly, FSP1 is upregulated in non-small cell lung cancer (NSCLC), colorectal cancer (CRC) and pancreatic ductal adenocarcinoma (PDAC) as compared to the respective normal tissue of origin and correlates with NRF2 expression in PDAC patient datasets. Based on these data, we propose that KRAS-mutant cells must navigate a ferroptosis checkpoint by upregulating FSP1 during tumor establishment. Consequently, ferroptosis-inducing therapy should be combined with FSP1 inhibitors for efficient therapy of KRAS-mutant cancers.

DOI: https://doi.org/10.1038/s41418-022-01096-8
Pathol Res Pract. 2022 Nov 23. pii: S0344-0338(22)00484-8. [Epub ahead of print]241 154240

Exosomal hsa-piR1089 promotes proliferation and migration in neuroblastoma via targeting KEAP1.

Hao Wang, Bozhong Shi, Xiaoyang Zhang, Ping Shen, Qiao He, Minzhi Yin, Yi Pan, Jing Ma.

  Neuroblastoma (NB) is a sympathetic nervous system tumor and one of the most common pediatric, extra-cranial, solid tumors, especially in early childhood. Its expression is heterogeneous and shows a unique clinical and prognostic feature. Due to its insidious onset, most diagnoses are accompanied by metastasis, making patient prognoses extremely poor. Novel biomarkers are urgently needed for easy diagnosis, metastasis detection, and investigation of potential mechanisms regulating NB tumor progression. Recent research highlights that circulating tumor markers could be used to diagnose and monitor prognosis in various tumors. Among them, exosomal genetic material has attracted much attention because of its tumor-secreted specificity and unique mechanism of action. In this study, we used next-generation sequencing to study PIWI-interacting RNAs (piRNAs) in exosomes derived from NB patient plasma. We found higher human piRNA 1089 (hsa-piR-1089) levels in exosomes from NB patients than from normal controls. Our receiver operating characteristic (ROC) curve analyses showed that hsa-piR-1089 had high diagnostic sensitivity and specificity. We also found that high hsa-piR-1089 expression in NB tumor tissues was associated with a high-risk Children's Oncology Group classification and metastasis. Our in vitro experiments showed that exosomal hsa-piR-1089 promoted NB cell proliferation and migration by inhibiting Kelch-like ECH-associated protein 1 (KEAP1) expression. Moreover, low KEAP1 expression was associated with NB progression in clinical samples. In conclusion, our data indicate that blood-borne exosomal hsa-piR-1089 is a diagnostic marker for NB and assessing metastasis. Our study provides a quick, simple, and noninvasive diagnostic method for NB and contributes to developing new treatment strategies.

Keywords:  Exosome; Hsa-piR1089; KEAP1; Migration; Neuroblastoma; Proliferation

DOI:  https://doi.org/10.1016/j.prp.2022.154240