HBO1 is required for the maintenance of leukaemia stem cells

MacPherson, Laura; Anokye, Juliana; Yeung, Miriam M.; Lam, Enid Y.N.; Chan, Yih Chih; Weng, Chen Fang; Yeh, Paul; Knezevic, Kathy; Butler, Miriam; Hoegl, Annabelle; Chan, Kah Lok; Burr, Marian L.; Gearing, Linden J.; Willson, Tracy A.; Liu, Joy; Choi, Jarny; Yang, Yuqing; Bilardi, Rebecca A.; Falk, Hendrik; Nguyen, Nghi; Stupple, Paul A.; Peat, T.S.; Zhang, Ming; Silva, Melanie de; Carrasco‐Pozo, Catalina; Avery, Vicky M.; Khoo, Poh Sim; Dolezal, Olan; Dennis, Matthew L.; Nuttall, Stewart D.; Surjadi, Regina; Newman, Janet; Ren, Bin; Leaver, David J.; Sun, Yuxin; Baell, Jonathan B.; Dovey, Oliver M.; Vassiliou, George S.; Grebien, Florian; Dawson, Sarah; Street, Ian P.; Monahan, Brendon J.; Burns, Christopher J.; Choudhary, Chunaram; Blewitt, Marnie E.; Voss, Anne K.; Thomas, Tim; Dawson, Mark A.

doi:10.1038/s41586-019-1835-6

Cited by 111 publications

(81 citation statements)

References 56 publications

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“…33 Similar approaches may be important in applications of CoA-based probes to study the proteome-wide selectivity of small molecule ligands. Drug-like protein acetyltransferases inhibitors have recently been developed and shown promise in oncology and other clinical settings [34][35][36][37] Methods to assess target occupancy and ligand selectivity such as those demonstrated here will likely be critical to the continued development and therapeutic validation these pre-clinical agents. Information including Figures S1-2, Supplementary Table S1, supporting materials, and methods is available online.…”

Section: Discussionmentioning

confidence: 99%

Harnessing Ionic Selectivity In Acetyltransferase Chemoproteomic Probes

Jing

Montaño

Levy

et al. 2020

Preprint

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Chemical proteomics provides a powerful strategy for the high-throughput assignment of enzyme function or inhibitor selectivity. However, identifying optimized probes for an enzyme family member of interest and differentiating signal from background remain persistent challenges in the field. To address this obstacle, here we report a physiochemical discernment strategy for optimizing chemical proteomics based on the Coenzyme A (CoA) cofactor. First, we synthesize a pair of CoA-based Sepharose pulldown resins differentiated by a single negatively charged residue, and find this change alters their capture properties in gel-based profiling experiments. Next, we integrate these probes with quantitative proteomics and benchmark analysis of ‘probe selectivity’ versus traditional ‘competitive chemical proteomics’. This reveals the former is well-suited for the identification of optimized pulldown probes for specific enzyme family members, while the latter may have advantages in discovery applications. Finally, we apply our anionic CoA pulldown probe to evaluate the selectivity of a recently reported small molecule N-terminal acetyltransferase inhibitor. These studies further validate the use of physical discriminant strategies in chemoproteomic hit identification and demonstrate how CoA-based chemoproteomic probes can be used to evaluate the selectivity of small molecule protein acetyltransferase inhibitors, an emerging class of pre-clinical therapeutic agents.

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Section: Discussionmentioning

confidence: 99%

Harnessing Ionic Selectivity In Acetyltransferase Chemoproteomic Probes

Jing

Montaño

Levy

et al. 2020

Preprint

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“…Taken together, these results explain the specific oncogenicity of the ubiquitously expressed DOT1L from another perspective. Although there is a new controversy about whether DOT1L is necessary for MLL LSC maintenance [60], the essential role of DOT1L in sustaining MLL leukemia cell self-renewal ability is undisputed [14,61]. Due to the complicated pathogenesis of MLL leukemia and the potential limitation on direct DOT1L-targeted strategies, optional approaches with more precise targeted therapies, including lncRNA and other drugs highly specific to LSC, are therefore required, particularly as combination regimens to treat different clinical responses.…”

Section: Discussionmentioning

confidence: 99%

The lncRNA LAMP5-AS1 drives leukemia cell stemness by directly modulating DOT1L methyltransferase activity in MLL leukemia

et al. 2020

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Background: Mixed-lineage leukemia (MLL) gene rearrangements trigger aberrant epigenetic modification and gene expression in hematopoietic stem and progenitor cells, which generates one of the most aggressive subtypes of leukemia with an apex self-renewal. It remains a challenge to directly inhibit rearranged MLL itself because of its multiple fusion partners and the poorly annotated downstream genes of MLL fusion proteins; therefore, novel therapeutic targets are urgently needed. Methods: qRT-PCR, receiver operating characteristic (ROC), and leukemia-free survival analysis were used to validate LAMP5-AS1 (LAMP5 antisense 1) expression and evaluate its clinical value. We performed in vitro and in vivo experiments to investigate the functional relevance of LAMP5-AS1 in MLL leukemia progression and leukemia cell stemness. RNA electrophoretic mobility shift assays (EMSA), histone methyltransferase assay, RNA pull-down assay, and RNA fluorescence in situ hybridization (FISH) were used to validate the relationship between LAMP5-AS1 and the methyltransferase activity of DOT1L. The downstream ectopic target genes of LAMP5-AS1/DOT1L were validated by the chromatin immunoprecipitation (ChIP) and western blot. Results: We discovered that a long noncoding RNA (lncRNA) LAMP5-AS1 can promote higher degrees of H3K79 methylation, followed by upregulated expression of the self-renewal genes in the HOXA cluster, which are responsible for leukemia stemness in context of MLL rearrangements. We found that LAMP5-AS1 is specifically overexpressed in MLL leukemia patients (n = 58) than that in the MLL-wt leukemia (n = 163) (p < 0.001), and the patients with a higher expression level of LAMP5-AS1 exhibited a reduced 5-year leukemia-free survival (p < 0.01). LAMP5-AS1 suppression significantly reduced colony formation and increased differentiation of primary MLL leukemia CD34+ cells. Mechanistically, LAMP5-AS1 facilitated the methyltransferase activity of DOT1L by directly binding its Lys-rich region of catalytic domain, thus promoting the global patterns of H3K79 dimethylation and trimethylation in cells. These observations supported that LAMP5-AS1 upregulated H3K79me2/me3 and the

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“…KMT2A, the translocation partner of MLL-AF4 in SEM cells, was identified in the HOXA9-MEIS1 rescue TF screen but not the original screen without ectopic expression of HOXA9. Notably, the MYST acetyltransferase HBO1 (also known as KAT7 or MYST2) and several members of the HBO1 protein complex, which were recently shown as critical regulators of leukemia stem cell maintenance, were also identified among the top hits ( MacPherson et al, 2020 ; Au et al, 2020 ). Most importantly, USF2 was enriched among the top hits in both screens ( Figure 3B ), suggesting USF2 is likely a positive regulator with less survival essentiality compared with KMT2A.…”

Section: Resultsmentioning

confidence: 99%

Functional interrogation of HOXA9 regulome in MLLr leukemia via reporter-based CRISPR/Cas9 screen

Zhang

Zhou

et al. 2020

eLife

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Aberrant HOXA9 expression is a hallmark of most aggressive acute leukemias, notably those with KMT2A (MLL) gene rearrangements. HOXA9 overexpression not only predicts poor diagnosis and outcome but also plays a critical role in leukemia transformation and maintenance. However, our current understanding of HOXA9 regulation in leukemia is limited, hindering development of therapeutic strategies. Here, we generated the HOXA9-mCherry knock-in reporter cell lines to dissect HOXA9 regulation. By utilizing the reporter and CRISPR/Cas9 screens, we identified transcription factors controlling HOXA9 expression, including a novel regulator, USF2, whose depletion significantly down-regulated HOXA9 expression and impaired MLLr leukemia cell proliferation. Ectopic expression of Hoxa9 rescued impaired leukemia cell proliferation upon USF2 loss. Cut&Run analysis revealed the direct occupancy of USF2 at HOXA9 promoter in MLLr leukemia cells. Collectively, the HOXA9 reporter facilitated the functional interrogation of the HOXA9 regulome and has advanced our understanding of the molecular regulation network in HOXA9-driven leukemia.

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HBO1 is required for the maintenance of leukaemia stem cells

Cited by 111 publications

References 56 publications

Harnessing Ionic Selectivity In Acetyltransferase Chemoproteomic Probes

Harnessing Ionic Selectivity In Acetyltransferase Chemoproteomic Probes

The lncRNA LAMP5-AS1 drives leukemia cell stemness by directly modulating DOT1L methyltransferase activity in MLL leukemia

Functional interrogation of HOXA9 regulome in MLLr leukemia via reporter-based CRISPR/Cas9 screen

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