Discovery of GPX4 inhibitory peptides from random peptide T7 phage display and subsequent structural analysis

Sakamoto, Kotaro; Sogabe, Satoshi; Kamada, Yusuke; Matsumoto, Shimpei; Kadotani, Akito; Sakamoto, Junichi; Tani, Akiko

doi:10.1016/j.bbrc.2016.11.035

Cited by 48 publications

(59 citation statements)

References 21 publications

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“…If these observations also apply to our results, the poor prognosis of patients with GPX4 overexpression may be owing to suppression of ferroptosis. To determine whether our result is related to ferroptosis, we would need to verify whether cell death rate increases under treatment with GPX4 inhibitors such as ferrostatin-1 and vitamin E [40] or GPX4-inhibitory peptide [41]. Though many aspects of ferroptosis remain unexplained, such as the execution factor and the substances detected downstream [40], these reports suggest that GPX4 is unique and different from other antioxidants.…”

Section: Discussionmentioning

confidence: 96%

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Glutathione peroxidase 4 overexpression inhibits ROS-induced cell death in diffuse large B-cell lymphoma

Kinowaki

Kurata

Ishibashi

et al. 2018

Laboratory Investigation

129

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Regulation of oxidative stress and redox systems has important roles in carcinogenesis and cancer progression, and for this reason has attracted much attention as a new area of cancer therapeutic targets. Glutathione peroxidase 4 (GPX4), an antioxidant enzyme, has biological important functions such as signaling cell death by suppressing peroxidation of membrane phospholipids. However, few studies exist on the expression and clinical relevance of GPX4 in malignant lymphomas such as diffuse large B-cell lymphoma. In this study, we assessed the expression of GPX4 immunohistochemically. GPX4 was expressed in 35.5% (33/93) cases of diffuse large B-cell lymphoma. The GPX4-positive group had poor overall survival (P = 0.0032) and progression-free survival (P = 0.0004) compared with those of the GPX4-negative group. In a combined analysis of GPX4 and 8-hydroxydeoxyguanosine (8-OHdG), an oxidative stress marker, there was a negative correlation between GPX4 and 8-hydroxydeoxyguanosine (P = 0.0009). The GPX4-positive and 8-hydroxydeoxyguanosine-negative groups had a significantly worse prognosis than the other groups in both overall survival (P = 0.0170) and progression-free survival (P = 0.0005). These results suggest that the overexpression of GPX4 is an independent prognostic predictor in diffuse large B-cell lymphoma. Furthermore, in vitro analysis demonstrated that GPX4-overexpressing cells were resistant to reactive oxygen species-induced cell death (P = 0.0360). Conversely, GPX4-knockdown cells were sensitive to reactive oxygen species-induced cell death (P = 0.0111). From these data, we conclude that GPX4 regulates reactive oxygen species-induced cell death. Our results suggest a novel therapeutic strategy using the mechanism of ferroptosis, as well as a novel prognostic predictor of diffuse large B-cell lymphoma.

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Section: Discussionmentioning

confidence: 96%

“…GPX4 has the potential to become a new therapeutic target as research on GPX4 advances, with the elucidation of the structure of GPX4, for example ref. [41].…”

Section: Discussionmentioning

confidence: 99%

Glutathione peroxidase 4 overexpression inhibits ROS-induced cell death in diffuse large B-cell lymphoma

Kinowaki

Kurata

Ishibashi

et al. 2018

Laboratory Investigation

129

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“…The recombinant expression of mammalian selenoproteins in bacterial protein production systems is often difficult due to low efficiency of selenocysteine incorporation [51][52][53] . Historically, structural studies of GPX4 have been possible only with U46C and U46G active-site mutants 38,52,54,55 . The structure of a selenocysteine-containing GPX4 (though not wild type) was recently reported 49 , but to date, structural studies of covalent GPX4 inhibitors have not been successful 5 .…”

Section: Extended Discussionmentioning

confidence: 99%

“…The structure of a selenocysteine-containing GPX4 (though not wild type) was recently reported 49 , but to date, structural studies of covalent GPX4 inhibitors have not been successful 5 . Co-crystal structures of GPX4 U46C in complex with reversible peptide binders have been reported, but their relevance remains unknown given that these compounds are unable to inhibit cellular GPX4 38 .…”

Section: Extended Discussionmentioning

confidence: 99%

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Targeting a Therapy-Resistant Cancer Cell State Using Masked Electrophiles as GPX4 Inhibitors

Eaton

Furst

et al. 2018

Preprint

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We recently discovered that inhibition of the lipid peroxidase GPX4 can selectively kill cancer cells in a therapy-resistant state through induction of ferroptosis. Although GPX4 lacks a conventional druggable pocket, covalent small-molecule inhibitors are able to overcome this challenge by reacting with the GPX4 catalytic selenocysteine residue to eliminate enzymatic activity. Unfortunately, all currently-reported GPX4 inhibitors achieve their activity through reactive chloroacetamide groups. We demonstrate that such chloroacetamide-containing compounds are poor starting points for further advancement given their promiscuity, instability, and low bioavailability. Development of improved GPX4 inhibitors, including those with therapeutic potential, requires the identification of new electrophilic chemotypes and mechanisms of action that do not suffer these shortcomings. Here, we report our discovery that nitrile oxide electrophiles, and a set of remarkable chemical transformations that generates them in cells from masked precursors, provide an effective strategy for selective targeting of GPX4. Our results, which include structural insights, target engagement assays, and diverse GPX4-inhibitor tool compounds, provide critical insights that may galvanize development of improved compounds that illuminate the basic biology of GPX4 and therapeutic potential of ferroptosis induction. In addition, our discovery that nitrile oxide electrophiles engage in highly selective cellular interactions and are bioavailable in their masked forms may be relevant for targeting other currently undruggable proteins, such as those revealed by recent proteome-wide ligandability studies.

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Transformable Supramolecular Self‐Assembled Peptides for Cascade Self‐Enhanced Ferroptosis Primed Cancer Immunotherapy

Wang,

Jiao,

Feng

et al. 2024

Advanced Materials

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Immunotherapy has received widespread attention for its effective and long‐term tumor‐eliminating ability. However, for immunogenic “cold” tumors, such as prostate cancer (PCa), the low immunogenicity of the tumor itself is a serious obstacle to efficacy. Here, we report a strategy to enhance PCa immunogenicity by triggering cascade self‐enhanced ferroptosis in tumor cells, turning the tumor from “cold” to “hot”. We developed a transformable self‐assembled peptide TEP‐FFG‐CRApY with alkaline phosphatase (ALP) responsiveness and GPX4 protein targeting. TEP‐FFG‐CRApY self‐assembles into nanoparticles under aqueous conditions and transforms into nanofibers in response to ALP during endosome/lysosome uptake into tumor cells, promoting lysosomal membrane permeabilization (LMP). On the one hand, the released TEP‐FFG‐CRAY nanofibers targeted GPX4 and selectively degraded the GPX4 protein under the light irradiation, inducing ferroptosis; on the other hand, the large amount of leaked Fe2+ further cascaded to amplify the ferroptosis through the Fenton reaction. TEP‐FFG‐CRApY‐induced immunogenic ferroptosis improved tumor cell immunogenicity by promoting the maturation of DC cells and increasing intra‐tumor T cell infiltration. More importantly, recovered T cells further enhanced ferroptosis by secreting large amounts of IFN‐γ. This work provides a novel strategy for the molecular design of synergistic molecularly targeted therapy for immunogenic “cold” tumors.This article is protected by copyright. All rights reserved

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Discovery of GPX4 inhibitory peptides from random peptide T7 phage display and subsequent structural analysis

Cited by 48 publications

References 21 publications

Glutathione peroxidase 4 overexpression inhibits ROS-induced cell death in diffuse large B-cell lymphoma

Glutathione peroxidase 4 overexpression inhibits ROS-induced cell death in diffuse large B-cell lymphoma

Targeting a Therapy-Resistant Cancer Cell State Using Masked Electrophiles as GPX4 Inhibitors

Transformable Supramolecular Self‐Assembled Peptides for Cascade Self‐Enhanced Ferroptosis Primed Cancer Immunotherapy

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