Covalent Cysteine Targeting of Bruton’s Tyrosine Kinase (BTK) Family by Withaferin-A Reduces Survival of Glucocorticoid-Resistant Multiple Myeloma MM1 Cells

Logie, Emilie; Chirumamilla, Chandra Sekhar; Pérez-Novo, Claudina; Shaw, Priyanka; Declerck, Ken; Palagani, Ajay; Rangarajan, Savithri; Cuypers, Bart; Neuter, Nicolas De; Fazil, Mobashar Hussain Urf Turabe; Verma, Navin Kumar; Bogaerts, Annemie; Laukens, Kris; Offner, Fritz; Vlierberghe, Pieter Van; Ostade, Xaveer Van; Berghe, Wim Vanden

doi:10.3390/cancers13071618

Cited by 11 publications

(18 citation statements)

References 108 publications

(146 reference statements)

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“…Approved; South American Origin), 1% MEM non-essential amino acids (Invitrogen, Carlsbad, CA, USA), 1% sodium pyruvate (Invitrogen, Carlsbad, CA, USA), and 1% penicillin/streptomycin solution (Invitrogen, Carlsbad, CA, USA). Cell viability was assessed by the MTT colorimetric assay (Sigma Aldrich, St. Louis, MO, USA) as previously described [108].…”

Section: Cell Culture and Cell Viability Assaysmentioning

confidence: 99%

“…Total RNA from control and RSL3-treated (3 h, 5 µM RSL3) MM1 cells was isolated using the RNeasy Mini Kit (Qiagen, Venlo, Netherlands) according to the manufacturer's protocol. Isolated RNA was quantified and qualified using the Epoch™ Microplate Spectrophotometer (BioTek, Winooski, VT, USA) and sent to BGI (BGI Group, Beijing, China) for RNA sequencing analysis as previously described [108]. Briefly, RNA integrity (RNA content > 80 ng/µL, 28 s/18 s ≥ 1.0 and RIN ≥ 7.0) was determined using the 2100 Bioanalyzer system (Agilent Technologies, Santa Clara, CA, USA), after which library preparation was initiated.…”

Section: Rna Extraction Rna Sequencing and Rna Sequencing Data Analysismentioning

confidence: 99%

“…Analysis of RNA sequencing data was performed as previously described [108]. In short, the quality of the sequencing reads was evaluated using FastQC (v0.11.5, Braham Institute, Cambridge, UK) [109] and subsequent alignment to the human reference genome build 37 (hg19) was performed with the STAR (v.2.7.3a, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, USA) tool [110].…”

Section: Rna Extraction Rna Sequencing and Rna Sequencing Data Analysismentioning

confidence: 99%

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Ferroptosis Induction in Multiple Myeloma Cells Triggers DNA Methylation and Histone Modification Changes Associated with Cellular Senescence

Logie

Puyvelde

Cuypers

et al. 2021

IJMS

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Disease relapse and therapy resistance remain key challenges in treating multiple myeloma. Underlying (epi-)mutational events can promote myelomagenesis and contribute to multi-drug and apoptosis resistance. Therefore, compounds inducing ferroptosis, a form of iron and lipid peroxidation-regulated cell death, are appealing alternative treatment strategies for multiple myeloma and other malignancies. Both ferroptosis and the epigenetic machinery are heavily influenced by oxidative stress and iron metabolism changes. Yet, only a limited number of epigenetic enzymes and modifications have been identified as ferroptosis regulators. In this study, we found that MM1 multiple myeloma cells are sensitive to ferroptosis induction and epigenetic reprogramming by RSL3, irrespective of their glucocorticoid-sensitivity status. LC-MS/MS analysis revealed the formation of non-heme iron-histone complexes and altered expression of histone modifications associated with DNA repair and cellular senescence. In line with this observation, EPIC BeadChip measurements of significant DNA methylation changes in ferroptotic myeloma cells demonstrated an enrichment of CpG probes located in genes associated with cell cycle progression and senescence, such as Nuclear Receptor Subfamily 4 Group A member 2 (NR4A2). Overall, our data show that ferroptotic cell death is associated with an epigenomic stress response that might advance the therapeutic applicability of ferroptotic compounds.

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Section: Cell Culture and Cell Viability Assaysmentioning

confidence: 99%

Section: Rna Extraction Rna Sequencing and Rna Sequencing Data Analysismentioning

confidence: 99%

Section: Rna Extraction Rna Sequencing and Rna Sequencing Data Analysismentioning

confidence: 99%

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Ferroptosis Induction in Multiple Myeloma Cells Triggers DNA Methylation and Histone Modification Changes Associated with Cellular Senescence

Logie

Puyvelde

Cuypers

et al. 2021

IJMS

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“…Once isolated, quantification was performed with the Qubit RNA BR Assay Kit (ThermoFisher, MA, USA) and RNA was stored at -80°C. Extracted RNA was used as input for RNA sequencing as previously described [90]. In brief, RNA was shipped to BGI (BGI Group, Bejing, China) where quality checks were performed using the 2100 Bioanalyzer system (Agilent Technologies, USA) and sequencing took place using the BGISE-500 platform (BGI Group, Beijing, China).…”

Section: Rna Extraction and Sequencingmentioning

confidence: 99%

Investigating the Role of Chromatin Remodeler FOXA1 in Ferroptotic Cell Death

Logie

Maes

Meenen

et al. 2021

Preprint

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Ferroptosis is a lipid peroxidation-dependent mechanism of regulated cell death known to suppress tumor proliferation and progression. Although several genetic and protein hallmarks have been identified in ferroptotic cell death, it remains challenging to fully characterize ferroptosis signaling pathways and to find suitable biomarkers. Moreover, changes taking place in the epigenome of ferroptotic cells remain poorly studied. In this context, we aimed to investigate the role of chromatin remodeler forkhead box protein A1 (FOXA1) in RSL3-treated multiple myeloma cells because, similar to ferroptosis, this transcription factor has been associated with changes in the lipid metabolism, DNA damage, and epithelial-to-mesenchymal transition (EMT). RNA sequencing and Western blot analysis revealed that FOXA1 expression is consistently upregulated upon ferroptosis induction in different in vitro and in vivo disease models. In silico motif analysis and transcription factor enrichment analysis further suggested that ferroptosis-mediated FOXA1 expression is orchestrated by specificity protein 1 (Sp1), a transcription factor known to be influenced by lipid peroxidation. Remarkably, FOXA1 upregulation in ferroptotic myeloma cells did not alter hormone signaling or EMT, two key downstream signaling pathways of FOXA1. CUT&RUN genome-wide transcriptional binding site profiling showed that GPX4-inhibition by RSL3 triggered loss of binding of FOXA1 to pericentromeric regions in multiple myeloma cells, suggesting that this transcription factor is possibly involved in genomic instability, DNA damage, or cellular senescence under ferroptotic conditions.

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“…In addition, targeting PK in various hematologic malignancies such as myeloma may be necessary when resistance to corticoid treatment develops. However, it should be kept in mind that the more specific the targeting, the more resistance mechanisms may emerge; therefore, new pharmacological approaches are required, such as the use of withaferin A, a promiscuous kinase inhibitor in glucocorticoid resistance in myeloma [ 7 ].…”

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confidence: 99%