Lysine specific demethylase 1 (LSD1) plays an important role in regulating histone lysine methylation at residues K4 and K9 on H3 is attractive therapeutic target multiple malignancies. Here we report a structure-based virtual screen of compound library containing ∼2 million small molecular entities. Computational docking scoring followed by biochemical screening led to the identification novel N'-(1-phenylethylidene)-benzohydrazide series LSD1 inhibitors with hits showing IC50s 200-400 nM...

Abstract Purpose: Ewing sarcoma is a pediatric bone tumor that absolutely relies on the transcriptional activity of EWS/ETS family fusion oncoproteins. While most common fusion, EWS/FLI, utilizes lysine-specific demethylase 1 (LSD1) to repress critical suppressors, small-molecule blockade LSD1 has not yet been thoroughly explored as therapeutic approach for sarcoma. We therefore evaluated translational potential potent and specific inhibition with HCI2509 program both EWS/FLI EWS/ERG well...

Significance Ewing sarcoma is a pediatric bone malignancy driven by the fusion protein EWS/FLI. EWS/FLI mediates oncogenesis through its role as an aberrant transcription factor, but little known about molecular mechanisms underlying this function. We demonstrate in cells that activates gene targets binding at associated GGAA-microsatellites, and these repetitive sequences are necessary for cell proliferation anchorage-independent growth. Furthermore, we show previously unknown EWS portion...

Ewing sarcoma is a prototypical fusion transcription factor-associated pediatric cancer that expresses EWS/FLI or highly related FET/ETS chimera. dysregulates to induce and maintain sarcomagenesis, but the mechanisms utilized are not fully understood. We therefore sought define global effects of on chromatin conformation in cells using well-validated 'knock-down/rescue' model function combination with next generation sequencing assays evaluate how landscape changes loss, recovery,...

Multi-agent chemotherapeutic regimes remain the cornerstone treatment for Ewing sarcoma, second most common bone malignancy diagnosed in pediatric and young adolescent populations. We have reached a therapeutic ceiling with conventional cytotoxic agents, highlighting need to adopt novel approaches that specifically target drivers of sarcoma oncogenesis. As KDM1A/lysine-specific demethylase 1 (LSD1) is highly expressed cell lines tumors, elevated expression levels associated worse overall...

Endometrial cancer is the most common gynecologic malignancy. Type II endometrial carcinoma often poorly differentiated and patients diagnosed with disease (~11%) are disproportionately represented in annual deaths (48%). Recent genomic studies highlight mutations chromatin regulators as drivers tumorigenesis, suggesting use of epigenetic targeted therapies could provide clinical benefit to these patients. We investigated anti-tumor efficacy LSD1 inhibitor HCI2509 two cell lines AN3CA KLE....

Abstract Ewing sarcoma is an aggressive bone cancer of children and young adults defined by the presence a chromosomal translocation: t(11;22)(q24;q12). The encoded protein, EWS/FLI, fuses amino-terminal domain EWS to carboxyl-terminus FLI. portion intrinsically disordered transcriptional regulatory domain, while FLI contains ETS DNA-binding two flanking regions unknown function. Early studies using non-Ewing models provided conflicting information on roles each in EWS/FLI oncogenic We...

EWS/FLI is the pathognomic fusion oncoprotein that drives Ewing sarcoma. The amino-terminal EWS portion coordinates transcriptional regulation and carboxy-terminal FLI contains an ETS DNA-binding domain. acts as aberrant transcription factor, orchestrating a complex mix of gene activation repression, from both high affinity motifs repetitive GGAA-microsatellites. Our overarching hypothesis executing multi-faceted requires to use distinct molecular mechanisms at different loci. Many attempts...

Many cancers are characterized by chromosomal translocations which result in the expression of oncogenic fusion transcription factors. Typically, these proteins contain an intrinsically disordered domain (IDD) fused with DNA-binding (DBD) another protein and orchestrate widespread transcriptional changes to promote malignancy. These fusions often sole recurring genomic aberration they cause, making them attractive therapeutic targets. However, targeting factors requires a better...

Paediatric cancers commonly harbour quiet mutational landscapes and are instead characterized by single driver events such as the mutation of critical chromatin regulators, expression oncohistones, or oncogenic fusion proteins. These ultimately promote malignancy through disruption normal gene regulation development. The protein in Ewing sarcoma, EWS/FLI, is an transcription factor that reshapes enhancer landscape, resulting widespread transcriptional dysregulation. Lysine-specific...

Abstract Ewing sarcoma is an aggressive malignancy and the second most frequent bone tumor afflicting adolescent young adult population. Despite knowing oncologic driving event of for decades, outcomes treatment options this disease have not improved. A chromosomal translocation between chromosome 11 22, fusing EWSR1 gene to FLI1 (EWSR1::FLI1), cause eighty five percent cases. Due inability therapeutically target EWSR1::FLI1, field has instead attempted critical co-regulators disrupt...

Abstract Importance: This work signifies a breakthrough in our ability to study Ewing sarcoma by developing protocol purify functional WT EWSR1::FLI1. development changes the level of detail which we can EWSR1::FLI1 and potentially other difficult fusion oncoproteins. Objective: is pediatric bone soft tissue that arises adolescents young adults. The mutation causes >85% cases caused oncoprotein, Efforts recombinantly expressed have proven challenging for decades due aggregation low...

The greatest threat to human well-being in this century is climate change and related global issues.We examined the effectiveness of Modified Instrumentalism Occupational Therapy model as a framework for facilitating occupational behaviour address issues.Eleven individuals participated mixed-methods single-subject-design study. Data were gathered using Assessment Intervention Instrument Daily Inventories. Quantitative data analyzed two- three-standard deviation band methods. Qualitative...

Abstract Expression of the fusion oncoprotein EWS/FLI causes Ewing sarcoma, an aggressive pediatric tumor characterized by widespread epigenetic deregulation. These changes are targeted novel lysine-specific demethylase-1 (LSD1) inhibitors, which currently in early-phase clinical trials. Single-agent–targeted therapy often induces resistance, and successful development requires knowledge resistance mechanisms, enabling design effective combination strategies. Here, we used a genome-scale...

Abstract Ewing sarcoma is the second most common bone cancer in children and young adults. In 85% of patients, a translocation between chromosomes 11 22 results potent fusion oncoprotein, EWSR1::FLI1. EWSR1::FLI1 only genetic alteration an otherwise unaltered genome tumors. The EWSR1 portion protein intrinsically disordered domain involved transcriptional regulation by FLI contains DNA binding shown to bind core GGAA motifs repeats. A small alpha-helix FLI1, DBD-α4 helix, critical for...

Ewing sarcoma is the second most common solid bone malignancy diagnosed in pediatric and young adolescent populations. Despite aggressive multi-modal treatment strategies, 5-year event-free survival remains at 75% for patients with localized disease 20% metastases. Thus, need novel therapeutic options imperative. Recent studies have focused on epigenetic misregulation development potential new oncotargets treatment. This project study of LSD2, a flavin-dependent histone demethylase found to...

Abstract Epigenetic dysfunction is recognized as a driver in the pathology of various cancers. Promoter DNA hypermethylation, genomic hypomethylation, and aberrant histone acetylation methylation contribute to epigenetic lesions. Further, genetic mutations important regulators have been identified across solid hematological malignancies. These marks are reversible due dynamic nature regulation present attractive therapeutic targets. While therapies, like methyltransferase (DNMT) deacetylase...

Ewing sarcoma is the second most common bone cancer in children and young adults. In 85% of patients, a translocation between chromosomes 11 22 results potent fusion oncoprotein, EWS::FLI. EWS::FLI only genetic alteration an otherwise unaltered genome tumors. The EWS portion protein intrinsically disordered domain involved transcriptional regulation by FLI contains DNA binding shown to bind core GGAA motifs repeats. A small alpha-helix FLI, DBD- α 4 helix, critical for transcription function...

Ewing sarcoma is the second most common bone cancer in children and young adults. In 85% of patients, a translocation between chromosomes 11 22 results potent fusion oncoprotein, EWSR1::FLI1. EWSR1::FLI1 only genetic alteration an otherwise unaltered genome tumors. The EWSR1 portion protein intrinsically disordered domain involved transcriptional regulation by FLI contains DNA binding shown to bind core GGAA motifs repeats. A small alpha-helix FLI1, DBD-α4 helix, critical for transcription...

ABSTRACT Genes encoding the RNA-binding proteins F US, E WSR1, and T AF15 (FET proteins) are involved in chromosomal translocations rare sarcomas. FET-rearranged sarcomas often aggressive malignancies affecting patients of all ages. New therapies needed. These fuse 5’ portion FET gene with a 3’ partner transcription factor (TF). The resulting fusion oncogenic TFs protein low complexity domain (LCD) DNA binding domain. have proven stubbornly difficult to target directly promising strategies...

Abstract RNA-Seq analysis has become a routine task in numerous genomic research labs, driven by the reduced cost of bulk RNA sequencing experiments. These generate billions reads that require accurate, efficient, effective, and reproducible analysis. But time required for comprehensive remains bottleneck. Many labs rely on in-house scripts, making standardization reproducibility challenging. To address this, we developed RNA-SeqEZPZ, an automated pipeline with user-friendly point-and-click...

Abstract Lysine-specific demethylase 1 (LSD1/AOF2/KDM1A) is a flavin-dependent histone that catalyzes the post-translational oxidative demethylation of mono- and di-methylated lysines on histones. Methylation lysine residues histones can signal transcriptional activation or repression depending specific residue involved. H3K4me2 transcription-activating chromatin mark at gene promoters this by LSD1 thought to prevent expression tumor suppressor genes important in human cancer. In contrast,...