Upregulated heme biosynthesis, an exploitable vulnerability in MYCN-driven leukemogenesis

Fukuda, Yu; Wang, Yao; Lian, Shangli; Lynch, John; Nagai, Shinjiro; Fanshawe, Bruce; Kandilci, Ayten; Janke, Laura J.; Neale, Geoffrey; Fan, Yiping; Sorrentino, Brian P.; Roussel, Martine F.; Grosveld, Gerard; Schuetz, John D.

doi:10.1172/jci.insight.92409

Cited by 38 publications

(41 citation statements)

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“…Since cancer cells frequently have elevated levels of heme and often exhibit up-regulated expression of many of the genes involved in the biosynthesis of heme (48), it is likely that our observations will be generalizable to cancer beyond B lineage acute leukemia. Indeed, inhibition of heme synthesis has even been reported to reduce tumor cell survival and proliferation in a variety of cancer types (30,49,50). The effects of inhibiting heme synthesis may be in part due to disruption of normal cellular processes including electron transport chain function, cataplerosis in the TCA cycle, p53 activity and stability, regulating trafficking of ADP and ATP, and in circadian rhythms (48).…”

Section: Discussionmentioning

confidence: 99%

“…Since sgRNAs targeting the heme synthesis pathway were associated with resistance of BCR-ABL + B-ALL cells to DHA treatment, we hypothesized that repressing heme levels pharmacologically should also render BCR-ABL + B-ALL cells resistant to DHA treatment. To test this hypothesis, we cultured BCR-ABL + B-ALL cells in succinylacetone (SA), an inhibitor of heme synthesis (30), resulting in decreased cellular heme levels and rendering the cells more resistant to death induced by culture with DHA (Sup. Fig.…”

Section: Crispr Screen Identifies Pathways Required For Dha Induced Amentioning

confidence: 99%

“…1F). Conversely, BCR-ABL + B-ALL cells were cultured in aminolevulinic acid (ALA), a precursor of heme synthesis (30), which resulted in increased cellular heme levels and made the cells more sensitive to DHA induced apoptosis (Sup. Fig.…”

Section: Crispr Screen Identifies Pathways Required For Dha Induced Amentioning

confidence: 99%

“…Cells were seeded in 96-well plates and drugs (DHA, BTdCPU, ABT-199, ABT263 solubilized in DMSO or DMSO vehicle controls) were added at the indicated concentrations. Aminolevulinic acid (ALA), a precursor of heme synthesis, or succinylacetone (SA), an inhibitor of heme synthesis, (both from Sigma) were solubilized in water and added at indicated concentrations to alter cellular heme levels (30). Cell viability was determined by staining with Annexin-V-APC and propidium iodide (BD Biosciences) and measured by flow cytometry as previously described (22).…”

Section: Introductionmentioning

confidence: 99%

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Heme-Sensing Pathway Modulates Susceptibility of Poor Prognosis B-Lineage Acute Leukemia to BH3-Mimetics

Smith

Budhraja

Lynch

et al. 2020

Preprint

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211 Words Body: 5960 Words (not counting references nor supplementary material) Heme-sensing pathway dictates apoptotic response 2 Abstract:Anti-apoptotic MCL1 is one of the most frequently amplified genes in human cancers and elevated expression confers resistance to many therapeutics including the BH3-mimetic agents dihydroartemisinin (DHA) translationally represses MCL-1 and synergizes with BH3-mimetics. To explore how DHA represses MCL-1, a genome-wide CRISPR screen identified that loss of genes in the heme synthesis pathway renders mouse BCR-ABL + B-ALL cells resistant to DHAinduced death. Mechanistically, DHA disrupts the interaction between heme and the eIF2α kinase heme regulated inhibitor (HRI) triggering the integrated stress response. Genetic ablation ofEif2ak1, which encodes HRI, blocks MCL-1 repression in response to DHA treatment and represses the synergistic killing of DHA and BH3-mimetics compared to wild-type leukemia. Furthermore, BTdCPU, a small-molecule activator of HRI, similarly triggers MCL-1 repression and synergizes with BH3-mimetics in mouse and human leukemia including both Ph + and Ph-like B-ALL. Lastly, combinatorial treatment of leukemia bearing mice with both BTdCPU and a BH3-mimetic extended survival and repressed MCL-1 in vivo. These findings reveal for the first time that the HRI-dependent cellular heme-sensing pathway can modulate apoptosis in leukemic cells by repressing MCL-1 and increasing their responsiveness to BH3-mimetics. This signaling pathway could represent a generalizable mechanism for repressing MCL-1 expression in malignant cells and sensitizing them to available therapeutics. Heme-sensing pathway dictates apoptotic response Heme-sensing pathway dictates apoptotic response 6 Materials and Methods Cells and Cell Culture. Mouse p185 + Arf-null B-ALL (hereafter referred to as BCR-ABL + B-ALL) (10) and PAX5-JAK2, RCSD1-AB1, and RCSD1-ABL2 fusion-expressing Ph-like Ba/F3 cells (23) were grown in RPMI with 10% fetal bovine serum, 55 µM 2mercaptoethanol, 2 mM glutamine, penicillin, and streptomycin (Invitrogen). Navitoclax and Venetoclax were obtained from Selleckchem. DHA was obtained from AvaChem Scientific. BTdCPU was obtained from Millipore and synthesized by the The human Ph + leukemia cell lines OP-1, TOM-1, BV-173, and SUP-B15 were cultured in RPMI with 20% fetal bovine serum, 55 µM 2-mercaptoethanol, 2 mM glutamine, penicillin, and streptomycin. Genome-wide CRISPR Screen. Cas9-expressing p185 + B-ALL cells were transduced with the Brie CRISPR KO library (Addgene #73633) at a multiplicity of infection of 0.5with an sgRNA coverage of 400x (24). One day after transduction, puromycin was added (1 µg/mL) to select for infected cells. Cells were then cultured in either DMSO or 10 µM DHA containing media for 24 hours (h), followed by a 48h culture in drug free media.As previously described, genomic DNA was extracted from the surviving cells (Qiagen DNeasy Blood and Tissue kit) and amplified by PCR for Illumina sequencing of sgRNAs by Nextseq (24). MAGeCK was used to ...

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Section: Discussionmentioning

confidence: 99%

Section: Crispr Screen Identifies Pathways Required For Dha Induced Amentioning

confidence: 99%

Section: Crispr Screen Identifies Pathways Required For Dha Induced Amentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Heme-Sensing Pathway Modulates Susceptibility of Poor Prognosis B-Lineage Acute Leukemia to BH3-Mimetics

Smith

Budhraja

Lynch

et al. 2020

Preprint

Self Cite

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“…Among the possible ways, it is tempting to speculate that heme could directly or indirectly affect many of the mitochondrial-dependent mechanisms of neurodegeneration described in the previous paragraphs. Indeed, the relationship between heme and mitochondria is based on several elements ( Figure 3 ): heme is produced through a series of reactions that occur partly in the mitochondria and partly in the cytosol [ 136 ]; heme acts as a cofactor for cytochromes c and cytochromes in complexes II-III-IV of the mitochondrial ETC [ 137 ]; heme has been reported to directly or indirectly influence ATP translocation between mitochondria and cytosol [ 138 , 139 , 140 ] mediated by adenine nucleotide translocases (ANTs); finally, heme biosynthesis is considered a cataplerotic pathway for the Kreb’s cycle due to the fact that the first step of heme production consumes succynil-CoA [ 141 , 142 ]. Thus, modulation of heme homeostasis can affect mitochondrial functions.…”

Section: Heme and Mitochondrial Dysfunction Related To Neurodegenementioning

confidence: 99%

Mitochondrial Targeting in Neurodegeneration: A Heme Perspective

2018

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Mitochondrial dysfunction has achieved an increasing interest in the field of neurodegeneration as a pathological hallmark for different disorders. The impact of mitochondria is related to a variety of mechanisms and several of them can co-exist in the same disease. The central role of mitochondria in neurodegenerative disorders has stimulated studies intended to implement therapeutic protocols based on the targeting of the distinct mitochondrial processes. The review summarizes the most relevant mechanisms by which mitochondria contribute to neurodegeneration, encompassing therapeutic approaches. Moreover, a new perspective is proposed based on the heme impact on neurodegeneration. The heme metabolism plays a central role in mitochondrial functions, and several evidences indicate that alterations of the heme metabolism are associated with neurodegenerative disorders. By reporting the body of knowledge on this topic, the review intends to stimulate future studies on the role of heme metabolism in neurodegeneration, envisioning innovative strategies in the struggle against neurodegenerative diseases.

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The heme synthesis-export system regulates the tricarboxylic acid cycle flux and oxidative phosphorylation

et al. 2021

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Upregulated heme biosynthesis, an exploitable vulnerability in MYCN-driven leukemogenesis

Cited by 38 publications

References 31 publications

Heme-Sensing Pathway Modulates Susceptibility of Poor Prognosis B-Lineage Acute Leukemia to BH3-Mimetics

Heme-Sensing Pathway Modulates Susceptibility of Poor Prognosis B-Lineage Acute Leukemia to BH3-Mimetics

Mitochondrial Targeting in Neurodegeneration: A Heme Perspective

The heme synthesis-export system regulates the tricarboxylic acid cycle flux and oxidative phosphorylation

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