Trabectedin Inhibits EWS-FLI1 and Evicts SWI/SNF from Chromatin in a Schedule-dependent Manner

Harlow, Matt L.; Chassé, Maggie H.; Boguslawski, Elissa A.; Sorensen, Katie M.; Gedminas, Jenna M.; Kitchen-Goosen, Susan M.; Rothbart, Scott B.; Taslim, Cenny; Lessnick, Stephen L.; Peck, Anderson; Madaj, Zachary; Bowman, Megan J.; Grohar, Patrick J.

doi:10.1158/1078-0432.ccr-18-3511

Cited by 39 publications

(53 citation statements)

References 54 publications

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“…BT12 and CHLA266 cell lines were obtained from the Children's Oncology Group, G401 cells from ATCC, TC32 cells from Dr. Lee Helman (Children's Hospital of Los Angeles) and U2OS cells from Dr. Chand Khanna (Ethos Veterinary Health LLC). Cell lines were routinely monitored for pathogens and cultured as described (16).…”

Section: Methodsmentioning

confidence: 99%

“…BT12 cells were incubated with mithramycin and the RNA was collected, reverse transcribed, and PCR amplified as previously described (16). Expression was calculated with standard ddCT methods relative to GAPDH and solvent controls.…”

Section: Quantitative Real-time Pcr (Rt-qpcr)mentioning

confidence: 99%

“…IC50s were determined by non-linear regression in duplicate for three independent experiments using MTS assay CellTiter96 and confirmed with real-time proliferation assays on the Incucyte Zoom (16).…”

Section: Cell Proliferation Assaysmentioning

confidence: 99%

“…Fractionation lysates were collected with cytoplasmic lysis buffer (25 mM HEPES pH 8.0, 50mM KCl, and 05% NP-40) and the nuclei pellet lysed with 4% LDS buffer. 30 µg of protein was resolved and transferred as described (16). The membranes were blocked with 5% milk in TBS-T and probed with Abcam (H3K27me3), EMD Millipore (SP1), and Cell Signaling (SMARCC1, SMARCE1, H3) antibodies.…”

Section: Cell Proliferation Assaysmentioning

confidence: 99%

“…Chromatin Fractionation 3 million BT12, G401, or U2OS were incubated with 100nM mithramycin or PBS control for 8 or 18-hours, washed and collected in PBS and fractionated as previously described (16).…”

Section: Cell Proliferation Assaysmentioning

confidence: 99%

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Direct therapeutic targeting of SWI/SNF induces epigenetic reprogramming and durable tumor regression in rhabdoid tumor

Chassé

et al. 2019

Preprint

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Conflict of Interest:The authors disclose no potential conflicts of interest. STATEMENT OF TRANSLATIONAL RELEVANCEThere is a tremendous need for novel therapeutic approaches for rhabdoid tumor and the more than 20% of human cancers characterized by dysregulation of the SWI/SNF chromatin remodeling complex. While approaches to target associated complexes, such as PRC2, known to be influenced by dysregulated SWI/SNF are currently being evaluated in the clinic, the direct therapeutic targeting of SWI/SNF has not been explored. Here we identify an inhibitor of SWI/SNF and thoroughly explore the therapeutic development of this compound from a mechanistic and translational perspective thus providing insight into the targeting of this complex as well as a dose, schedule, and biomarker of target inhibition that is immediately clinically translatable. ABSTRACTPurpose: Rhabdoid tumor is a pediatric cancer characterized by the biallelic inactivation of SMARCB1, a subunit of the SWI/SNF chromatin remodeling complex.SMARCB1 inactivation leads to SWI/SNF redistribution to favor a proliferative dedifferentiated cellular state. Although this deletion is the known oncogenic driver, SWI/SNF therapeutic targeting remains a challenge. Experimental Design:We define a novel epigenetic mechanism for mithramycin using biochemical fractionation, chromatin immunoprecipitation sequencing (ChIP-seq), and a dual spike-in assay for transposase accessible chromatin sequencing (ATAC-seq). We correlate epigenetic reprogramming with changes with chromatin A/B compartments and promoter accessibility with chromHMM models and RNA-seq. Finally, we demonstrate durable, marked tumor response in an intramuscular rhabdoid tumor xenograft model. Results:Here we show mithramycin and a second-generation analogue EC8042 evict mutated SWI/SNF from chromatin and are effective in rhabdoid tumor. SWI/SNF blockade triggers chromatin compartment remodeling and promoter reprogramming leading to differentiation and amplification of H3K27me3, the catalytic mark of PRC2.Treatment of rhabdoid rumor xenografts with EC8042 leads to marked, durable tumor regression and differentiation of the tumor tissue into benign mesenchymal tissue, including de novo bone formation. Conclusion:Overall, this study identifies a novel therapeutic candidate for rhabdoid tumor and an approach that may be applicable to the 20% of cancers characterized by mutated SWI/SNF.

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Section: Methodsmentioning

confidence: 99%

Section: Quantitative Real-time Pcr (Rt-qpcr)mentioning

confidence: 99%

Section: Cell Proliferation Assaysmentioning

confidence: 99%

Section: Cell Proliferation Assaysmentioning

confidence: 99%

Section: Cell Proliferation Assaysmentioning

confidence: 99%

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Direct therapeutic targeting of SWI/SNF induces epigenetic reprogramming and durable tumor regression in rhabdoid tumor

Chassé

et al. 2019

Preprint

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Ewing sarcoma and related FET family translocation‐associated round cell tumors: A century of clinical and scientific progress

Maki

Grohar

Antonescu

2022

Genes Chromosomes & Cancer

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The year 2021 marked the centenary of the first publication of a cancer termed diffuse endothelioma of bone by James Ewing. Its unique features were apparent even in the first case series he described. This new diagnosis was clearly distinct from osteogenic sarcoma and myeloma, which were already well recognized at the time. We undertake this summary to better understanding Ewing sarcoma, contrasting the logarithmic evolution of the standard of care of systemic therapy for this and related diagnoses to the exponential understanding of the molecular biology of this family of tumors. We also outline in this manuscript how the finding of genomic relatives within Ewing sarcoma itself and related tumors, first noted nearly 40 years ago, helps us appreciate the need to find therapeutic plans that are specific for each small round blue cell tumor subtype. The advent of next generation sequencing regarding previously unknown small round blue cell tumor subtypes in many ways puts us back in the shoes of James Ewing in 1921, searching anew for clues leading to better treatments for increasingly rare cancer subsets.

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Children's Oncology Group's 2023 blueprint for research: Bone tumors

Reed,

Grohar,

Rubin

et al. 2023

Pediatric Blood & Cancer

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The Children's Oncology Group (COG) Bone Tumor Committee is responsible for clinical trials and biological research on localized, metastatic, and recurrent osteosarcoma and Ewing sarcoma (EWS). Results of clinical trials in localized disease completed and published in the past 10 years have led to international standard‐of‐care chemotherapy for osteosarcoma and EWS. A recent focus on identifying disease subgroups has led to the identification of biological features associated with poor outcomes including the presence of circulating tumor DNA (ctDNA) at diagnosis, and specific genomic alterations—MYC amplification for osteosarcoma and STAG2 and TP53 mutation for EWS. Studies validating these potential biomarkers are under way. Clinical trials evaluating the addition of multitargeted kinase inhibitors, which are active in relapsed bone sarcomas, to standard chemotherapy are under way in osteosarcoma and planned in EWS. In addition, the Committee has data analyses and a clinical trial under way to evaluate approaches to local management of the primary tumor and metastatic sites. Given the rarity of bone sarcomas, we have prioritized international interactions and are in the process of forming an international data‐sharing consortium to facilitate refinement of risk stratification and study of rare disease subtypes.

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Trabectedin Inhibits EWS-FLI1 and Evicts SWI/SNF from Chromatin in a Schedule-dependent Manner

Cited by 39 publications

References 54 publications

Direct therapeutic targeting of SWI/SNF induces epigenetic reprogramming and durable tumor regression in rhabdoid tumor

Direct therapeutic targeting of SWI/SNF induces epigenetic reprogramming and durable tumor regression in rhabdoid tumor

Ewing sarcoma and related FET family translocation‐associated round cell tumors: A century of clinical and scientific progress

Children's Oncology Group's 2023 blueprint for research: Bone tumors

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