Systematic identification of anticancer drug targets reveals a nucleus-to-mitochondria ROS-sensing pathway

Zhang, Junbing; Simpson, Claire M.; Berner, Jacqueline; Chong, Harrison Byron; Fang, Jiafeng; Ordulu, Zehra; Weiss-Sadan, Tommy; Possemato, Anthony; Harry, Stefan; Takahashi, Mariko; Yang, Tzu-Yi; Richter, Marianne; Patel, Himani; Smith, Abby E.; Carlin, Alexander; Groot, Adriaan F. Hubertus de; Wolf, Konstantin; Shi, Lei; Wei, Ting-Yu; Dürr, Benedikt R; Chen, Nicholas J.; Vornbäumen, Tristan; Wichmann, Nina O.; Mahamdeh, Mohammed; Pooladanda, Venkatesh; Matoba, Yusuke; Kumar, Shaan; Kim, Eugene; Bouberhan, Sara; Oliva, Esther; Rueda, Bo R.; Soberman, Roy J.; Bardeesy, Nabeel; Liau, Brian B.; Lawrence, Michael S.; Stokes, Matt P.; Beausoleil, Sean A.; Bar-Peled, Liron

doi:10.1016/j.cell.2023.04.026

Cited by 77 publications

(34 citation statements)

References 133 publications

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“…Transient transfections were conducted as previously described 32 . Briefly, HEK-293T cells were plated at a confluence of 3 x 10 6 cells/10 cm dish.…”

Section: Methods Detailsmentioning

confidence: 99%

“…Loading buffer was subsequently added and samples were denatured by boiling at 95°C for 5 min. Proteins were resolved by SDS-PAGE and analyzed by immunoblotting as previously described 32 with antibodies described in Key Resource Table .…”

Section: Methods Detailsmentioning

confidence: 99%

“…In vitro binding assays were conducted as previously described 32 . In brief, HEK-293T cells expressing the indicated proteins were washed once with ice-cold PBS, snap-frozen for storage, then lysed using a chilled bath sonicator (Q700, QSonica) in CHAPS lysis buffer (40 mM HEPES pH 7.4, 10 mM KCl, 5 mM MgCl 2 , 0.3 % CHAPS).…”

Section: Methods Detailsmentioning

confidence: 99%

“…To examine the impact of SOX10 depletion on melanoma cell lines, 1,000-7,000 melanoma cells expressing the indicated sgRNAs (see above) were seeded into a 96-well plate, and proliferation was determined 6 days post-seeding by measuring relative ATP levels as previously described 32 . In brief, 50 µL Cell Titer Glo TM (Promega) was added to each sample well, and the luminescence was read on the SpectraMax M5 plate reader (Molecular Devices).…”

Section: Methods Detailsmentioning

confidence: 99%

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DrugMap: A quantitative pan-cancer analysis of cysteine ligandability

Takahashi,

Chong,

Zhang

et al. 2023

Preprint

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Cysteine-focused chemical proteomic platforms have accelerated the clinical development of covalent inhibitors of a wide-range of targets in cancer. However, how different oncogenic contexts influence cysteine targeting remains unknown. To address this question, we have developed DrugMap, an atlas of cysteine ligandability compiled across 416 cancer cell lines. We unexpectedly find that cysteine ligandability varies across cancer cell lines, and we attribute this to differences in cellular redox states, protein conformational changes, and genetic mutations. Leveraging these findings, we identify actionable cysteines in NFκB1 and SOX10 and develop corresponding covalent ligands that block the activity of these transcription factors. We demonstrate that the NFkB1 probe blocks DNA binding, whereas the SOX10 ligand increases SOX10-SOX10 interactions and disrupts melanoma transcriptional signaling. Our findings reveal heterogeneity in cysteine ligandability across cancers, pinpoint cell-intrinsic features driving cysteine targeting, and illustrate the use of covalent probes to disrupt oncogenic transcription factor activity.

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“…Transient transfections were conducted as previously described 32 . Briefly, HEK-293T cells were plated at a confluence of 3 x 10 6 cells/10 cm dish.…”

Section: Methods Detailsmentioning

confidence: 99%

Section: Methods Detailsmentioning

confidence: 99%

Section: Methods Detailsmentioning

confidence: 99%

Section: Methods Detailsmentioning

confidence: 99%

See 2 more Smart Citations

DrugMap: A quantitative pan-cancer analysis of cysteine ligandability

Takahashi,

Chong,

Zhang

et al. 2023

Preprint

Self Cite

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“…Cellular redox balance is a prerequisite for normal physiological activities of cells . The redox imbalance in cells involves the occurrence and development of many diseases, such as cancer, degenerative disease, and arthritis. − In particular, mitochondria, as producers and sinks of intracellular reactive oxygen species (ROS), have a significant impact on the redox balance of cells. − Once mitochondria are damaged, mitophagy could be initiated and elimination of dysfunctional mitochondria in a lysosome-dependent manner, maintaining healthy mitochondrial population. , Mounting evidence suggests that many pathological processes could improve the intracellular ROS level to induce mitophagy. , Among these ROS, hydrogen peroxide (H 2 O 2 ) has broad actions in oxidative damaging and redox signaling. − Excessive mitochondrial H 2 O 2 may trigger mitophagy, which in turn reduces H 2 O 2 levels to protect cells. , Therefore, real-time imaging of the H 2 O 2 dynamic in mitochondria is vital to probe the redox imbalance during mitophagy.…”

Section: Introductionmentioning

confidence: 99%

A Reversible Fluorescent Probe for In Situ Monitoring Redox Imbalance during Mitophagy

Li,

Song,

et al. 2023

Anal. Chem.

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Photoactivation Inducing Multifunctional Coupling of Fluorophore for Efficient Tumor Therapy In Situ

Zhang,

Lv,

Huo

et al. 2024

Advanced Materials

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Photoactivatable molecules, with high‐precision spatial and temporal control, have largely promoted bioimaging and phototherapy applications, particularly for fluorescent dyes. Here, we described the first photoactivatable sensor (BI) that can be triggered by broad excitation light (405‐660 nm), which further undergoes ISC (intersystem crossing) and HAT (H‐atom transfer) processes to the formation of superoxide anion radicals (O2−•) and carbon radicals. Particularly, the photoinduced gain of carbon‐centered radicals (BI•) allows for radical‐radical coupling to afford the combined crosslink product (BI‐BI), which would be oxidized in the presence of O2−• to produce an extended conjugate system with NIR emission (820 nm). Besides, the photochemically generated product (Cy‐BI) possesses ultra‐high PCE (photothermal conversion efficiency) up to 90.9%, which enables optimized potential for phototherapy. What's more, Western Blot assay revealed that both BI and the coresponding photoproduct Cy‐BI could efficiently inhibit the expression of CHK1, and the irradiation of BI and Cy‐BI further induced apoptosis and ultimately enhancing the phototherapeutic effects. Thus, the combination of cell cycle block inducing apoptosis, PDT (photodynamic therapy) and PTT (photothermal therapy) treatment significantly suppressed solid tumor in vivo antitumor efficacy explorations. This is a novel finding in developing photoactivatable molecules, as well as the broad applicability of photoimaging and phototherapy in tumor‐related areas.This article is protected by copyright. All rights reserved

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Systematic identification of anticancer drug targets reveals a nucleus-to-mitochondria ROS-sensing pathway

Cited by 77 publications

References 133 publications

DrugMap: A quantitative pan-cancer analysis of cysteine ligandability

DrugMap: A quantitative pan-cancer analysis of cysteine ligandability

A Reversible Fluorescent Probe for In Situ Monitoring Redox Imbalance during Mitophagy

Photoactivation Inducing Multifunctional Coupling of Fluorophore for Efficient Tumor Therapy In Situ

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