EWS-WT1 fusion isoforms establish oncogenic programs and therapeutic vulnerabilities in desmoplastic small round cell tumors.

EWS fusion oncoproteins underlie several human malignancies including Desmoplastic Small Round Cell Tumor (DSRCT), an aggressive cancer driven by EWS-WT1 fusion proteins. Here we combine chromatin occupancy and 3D profiles to identify EWS-WT1-dependent gene regulation networks and target genes. We show that EWS-WT1 is a powerful chromatin activator controlling an oncogenic gene expression program that characterizes primary tumors. Similar to wild type WT1, EWS-WT1 has two isoforms that differ in their DNA binding domain and we find that they have distinct DNA binding profiles and are both required to generate viable tumors that resemble primary DSRCT. Finally, we identify candidate EWS-WT1 target genes with potential therapeutic implications, including CCND1, whose inhibition by the clinically-approved drug Palbociclib leads to marked tumor burden decrease in DSRCT PDXs in vivo. Taken together, our studies identify gene regulation programs and therapeutic vulnerabilities in DSRCT and provide a mechanistic understanding of the complex oncogenic activity of EWS-WT1.

Preclinical spheroid models identify BMX as a therapeutic target for metastatic MYCN nonamplified neuroblastoma.

The development of targeted therapies offers new hope for patients affected by incurable cancer. However, multiple challenges persist, notably in controlling tumor cell plasticity in patients with refractory and metastatic illness. Neuroblastoma (NB) is an aggressive pediatric malignancy originating from defective differentiation of neural crest-derived progenitors with oncogenic activity due to genetic and epigenetic alterations and remains a clinical challenge for high-risk patients. To identify critical genes driving NB aggressiveness, we performed combined chromatin and transcriptome analyses on matched patient-derived xenografts (PDXs), spheroids, and differentiated adherent cultures derived from metastatic MYCN nonamplified tumors. Bone marrow kinase on chromosome X (BMX) was identified among the most differentially regulated genes in PDXs and spheroids versus adherent models. BMX expression correlated with high tumor stage and poor patient survival and was crucial to the maintenance of the self-renewal and tumorigenic potential of NB spheroids. Moreover, BMX expression positively correlated with the mesenchymal NB cell phenotype, previously associated with increased chemoresistance. Finally, BMX inhibitors readily reversed this cellular state, increased the sensitivity of NB spheroids toward chemotherapy, and partially reduced tumor growth in a preclinical NB model. Altogether, our study identifies BMX as a promising innovative therapeutic target for patients with high-risk MYCN nonamplified NB.

"The effects of non-surgical periodontal treatment plus zinc and magnesium supplementation on oxidative stress and antioxidants enzymes in type 2 diabetes patients: a quasi-experimental study".

BACKGROUND: Periodontal Disease (PD) associated with Type 2 Diabetes Mellitus (T2DM) is a chronic condition that affects the oral cavity of people living with T2DM. The mechanisms of the interaction between type 2 Diabetes Mellitus and Periodontal diseases are complex and involve multiple pathophysiological pathways related to the systemic inflammatory process and oxidative stress. Non-surgical periodontal treatment (NSTP) is considered the standard for the management of this disease; however, patients with systemic conditions such as type 2 Diabetes Mellitus do not seem to respond adequately. For this reason, the use of complementary treatments has been suggested to support non-surgical periodontal treatment to reduce the clinical consequences of the disease and improve the systemic conditions of the patient. The use of zinc gluconate and magnesium oxide as an adjunct to non-surgical periodontal treatment and its effects on periodontal clinical features and oxidative stress in patients with Periodontal diseases -type 2 Diabetes Mellitus is poorly understood. METHODS: A quasi-experimental study was performed in patients with periodontal diseases associated with T2DM. Initially, 45 subjects who met the selection criteria were included. 19 were assigned to a control group [non-surgical periodontal treatment] and 20 to the experimental group (non-surgical periodontal treatment + 500 mg of magnesium oxide and 50 mg of zinc gluconate for oral supplementation for 30 days) and the data of 6 patients were eliminated. Sociodemographic characteristics, physiological factors, biochemical parameters, and clinical features of periodontal diseases were assessed. RESULTS: In this research a change in periodontal clinical characteristics was observed, which has been associated with disease remission. Additionally, a shift in MDA levels was presented for both groups. Furthermore, the supplementation group showed an increase in antioxidant enzymes when compared to the group that only received NSPT. CONCLUSION: The use of Zinc gluconate and magnesium oxide can serve as a complementary treatment to non-surgical periodontal treatment, that supports the remission of PD as a result of regulation-reduction of oxidative biomarkers and increase in antioxidant enzymes activity. TRIAL REGISTRATION: https://www.isrctn.com ISRCTN 14,092,381. September 13º 2023. Retrospective Registration.

Relación entre la percepción de condición física y medidas antropométricas auto reportadas en estudiantes universitarios

La percepción de la condición física en estudiantes universitarios se refiere a cómo se ven a sí mismos en términos de su estado físico y salud. En esta etapa, las variables antropométricas pueden verse afectadas por estrés, cambios en hábitos de sueño y alimentación, y menor participación en actividades físicas, impactando el desempeño académico y salud general. El objetivo fue relacionar la percepción de la condición física con medidas antropométricas auto reportadas en estudiantes universitarios Esta Investigación es de tipo descriptivo-correlacional, de diseño observacional y corte transversal. Fueron evaluados 46 estudiantes universitarios de la ciudad de Talca. La condición física se midió por medio del International Fitness Scale (IFIS) y las medidas antropométricas por medio de auto reporte. La condición física general se relacionó de manera inversa con la circunferencia de cintura (r=-0,32; p=0,028), la circunferencia de cintura(r=-0,44; p=0,002) y el ICE (r=-0,43; p=0,003). Se observaron correlaciones inversas entre la condición cardiorrespiratoria con el IMC (p=0,008; r=-0,38), circunferencia de cintura e ICE (p=0,008; r=-0,38). La velocidad/agilidad se correlacionó de forma inversa con el índice de masa corporal (IMC) (p=0,001; r=-0,46), circunferencia de cintura (p=0,019; r=-0,34), cadera (p= 0,004; r= -0,40) y el ICE (p=0,004; r=-0,41). La flexibilidad se correlaciono de forma inversa con el IMC (p=0,041; r=-0,30), circunferencia de cintura (p=0,023; r=-0,33), índice cintura cadera (ICC) (p= 0,001; r=-0,45)). Por último, la fuera muscular no se correlaciono con las variables antropométricas. Se concluye que existe correlación inversa entre la percepción de la condición física y algunos autore portes de las medidas antropométricas.

The physical fitness perception of college students refers to how they see themselves in terms of their physical condition and health. In this stage, anthropometric variables may be affected by stress, changes in sleep and eating habits, and decreased participation in physical activities, impacting academic performance and general health. The objective of this study is to relate the perception of physical condition with self-reported anthropometric measures in university students. This is a descriptive-correlational, observational, cross-sectional research. Forty-six university students from the city of Talca were evaluated. Physical condition was measured by means of the International Fitness Scale (IFIS) and anthropometric measures by means of self-report. General physical condition was inversely related to waist circumference (r=-0.32; p=0.028), waist circumference (r=-0.44; p=0.002) and IFIS (r=-0.43; p=0.003). Inverse correlations were observed between cardiorespiratory fitness with BMI (p=0.008; r=-0.38), waist circumference and ECI (p=0.008; r=-0.38). Speed/agility was inversely correlated with body mass index (BMI) (p=0.001; r=-0.46), waist circumference (p=0.019; r=-0.34), hip (p= 0.004; r= -0.40) and ECI (p=0.004; r=-0.41). Flexibility was inversely correlated with BMI (p=0.041; r=-0.30), waist circumference (p=0.023; r=-0.33), waist hip index (WHI) (p= 0.001; r=-0.45)). Lastly, muscle was not correlated with the anthropometric variables. It is concluded that there is an inverse correlation between the perception of physical condition and some anthropometric measures.

A percepção da aptidão física em estudantes universitários refere-se a como eles se veem em termos de sua condição física e saúde. Nessa fase, as variáveis antropométricas podem ser afetadas pelo estresse, por mudanças nos hábitos de sono e alimentação e pela diminuição da participação em atividades físicas, o que afeta o desempenho acadêmico e a saúde em geral. O objetivo foi relacionar a percepção da aptidão física com medidas antropométricas autorreferidas em estudantes universitários. Trata-se de uma pesquisa descritiva, correlacional, transversal e observacional. Foram avaliados 46 estudantes universitários da cidade de Talca. A condição física foi medida por meio da Escala Internacional de Aptidão Física (IFIS) e as medidas antropométricas por meio de autorrelato. A aptidão física geral foi inversamente relacionada à circunferência da cintura (r=-0,32; p=0,028), circunferência da cintura (r=-0,44; p=0,002) e IFIS (r=-0,43; p=0,003). Foram observadas correlações inversas entre a aptidão cardiorrespiratória com o IMC (p=0,008; r=-0,38), a circunferência da cintura e o ECI (p=0,008; r=-0,38). A velocidade/agilidade foi inversamente correlacionada com o índice de massa corporal (IMC) (p=0,001; r=-0,46), circunferência da cintura (p=0,019; r=-0,34), quadril (p= 0,004; r= -0,40) e ECI (p=0,004; r=-0,41). A flexibilidade foi inversamente correlacionada com o IMC (p=0,041; r=-0,30), a circunferência da cintura (p=0,023; r=-0,33), o índice cintura-quadril (WHI) (p= 0,001; r=-0,45). Por fim, a musculatura não foi correlacionada com as variáveis antropométricas. Concluise que há uma correlação inversa entre a percepção da condição física e algumas medidas antropométricas.

Efectos del entrenamiento cluster sobre la hipertrofia muscular: una revisión sistemática / Effects of cluster training on muscle hypertrophy: a systematic review / Efeitos do treinamento em cluster na hipertrofia muscular: uma revisão sistemática

Resumen Objetivo: El objetivo de la presente revisión sistemática fue determinar los efectos del entrenamiento cluster sobre la hipertrofia muscular. Metodología: Se realizó una búsqueda bibliográfica en las bases de datos electrónicas Pubmed, Scopus y Web of Science, utilizando las siguientes palabras clave: 'cluster training', 'rest Interval', 'rest pause', 'hypertrophy', 'resistance training' y 'cross sectional area'. Se incluyeron ensayos clínicos que utilizaron el entrenamiento cluster como intervención en personas mayores de 18 años de ambos sexos. Resultados: La revisión sistemática obtenida durante la búsqueda de las bases de datos consultadas arrojó un total de 23 artículos, potencialmente elegibles, de los cuales se tomó una muestra de 9, con los que se podían obtener resultados que respondían al objetivo de esta revisión. La cantidad de participantes de los 9 artículos elegibles fue de 172 sujetos. Los entrenamientos cluster permiten aumentar el volumen de entrenamiento y la intensidad sin provocar elevados niveles de fatiga, favoreciendo así el desarrollo de la hipertrofia muscular. Conclusiones: Los resultados de esta revisión sistemática sugieren que los entrenamientos cluster pueden ser una herramienta eficaz para el desarrollo de la hipertrofia muscular.

Abstract Objective: The aim of this systematic review was to determine the effects of cluster training on muscle hypertrophy. Methodology: A literature search was performed in the electronic databases Pubmed, Scopus and Web of Science, using the following keywords: 'cluster training', 'rest interval', 'rest pause', 'hypertrophy', 'resistance training' and 'cross sectional area'. We included clinical trials that used cluster training as an intervention in people over 18 years of age of both sexes. Results: The systematic review obtained during the search of the databases consulted yielded a total of 23 potentially eligible articles, of which a sample of 9 was taken from which results could be obtained that responded to the objective of this review. The number of participants from the 9 eligible articles was 172 subjects. Cluster workouts allow for increased training volume and intensity without causing high levels of fatigue, thus favoring the development of muscle hypertrophy. Conclusions: The results of this systematic review suggest that cluster training can be an effective tool for the development of muscle hypertrophy.

Resumo Objetivo: O objetivo desta revisão sistemática foi determinar os efeitos do treinamento em cluster na hipertrofia muscular. Metodologia: Realizou-se uma busca na literatura nas bases de dados eletrônicas Pubmed, Scopus e Web of Science, utilizando as seguintes palavras-chave: 'cluster training', 'rest interval', 'rest pause', 'hypertrophy', 'resistance training' e 'cross sectional area'. Foram incluídos ensaios clínicos que utilizaram o treinamento em cluster como intervenção em pessoas com mais de 18 anos de ambos os sexos. Resultados: A revisão sistemática realizada durante a busca nas bases de dados consultadas resultou em um total de 23 artigos potencialmente elegíveis, dos quais uma amostra de 9 foi selecionada para obter resultados que respondessem ao objetivo desta revisão. O número de participantes nos 9 artigos elegíveis foi de 172 indivíduos. Os treinos em cluster permitem um aumento no volume e na intensidade do treinamento sem causar altos níveis de fadiga, favorecendo assim o desenvolvimento da hipertrofia muscular. Conclusões: Os resultados desta revisão sistemática sugerem que o treinamento em cluster pode ser uma ferramenta eficaz para o desenvolvimento da hipertrofia muscular.

Targeting TRIP13 in favorable histology Wilms tumor with nuclear export inhibitors synergizes with doxorubicin.

Wilms tumor (WT) is the most common renal malignancy of childhood. Despite improvements in the overall survival, relapse occurs in ~15% of patients with favorable histology WT (FHWT). Half of these patients will succumb to their disease. Identifying novel targeted therapies remains challenging in part due to the lack of faithful preclinical in vitro models. Here we establish twelve patient-derived WT cell lines and demonstrate that these models faithfully recapitulate WT biology using genomic and transcriptomic techniques. We then perform loss-of-function screens to identify the nuclear export gene, XPO1, as a vulnerability. We find that the FDA approved XPO1 inhibitor, KPT-330, suppresses TRIP13 expression, which is required for survival. We further identify synergy between KPT-330 and doxorubicin, a chemotherapy used in high-risk FHWT. Taken together, we identify XPO1 inhibition with KPT-330 as a potential therapeutic option to treat FHWTs and in combination with doxorubicin, leads to durable remissions in vivo.

A Comparison of Clear Cell Sarcoma to Jaw and Salivary Tumors Bearing EWS Fusions.

OBJECTIVE: To review tumors identified as "clear cell sarcoma" in order to determine similarities to the rare EWS fusion positive jaw and salivary gland tumors clear cell odontogenic carcinoma (CCOC) and clear cell carcinoma of the salivary gland (CCC). METHODS: PubMed was used to collect all reports of clear cell sarcoma (CCS). Search parameters were "clear cell sarcoma" and "CCS." References in the publications were screened and cross-referenced. Data extracted included demographic characteristics, presenting signs and symptoms, radiographic findings, histological and immunohistochemical features and known molecular/genetic aberrations. RESULTS: Clear cell sarcoma has several similarities to CCOC and CCC. All three tumor types have similar histologic appearances including the presence of clear cells, as well as similar genetic profiles in that all harbor an EWSR1-CREB family fusions. Additionally, these tumors appear in soft tissue as well as bone, and can have a prolonged clinical course. CCS can appear anywhere in the body, including the head and neck region. All three tumors appear to have a predilection to women, although CCS may have a slight younger age of onset as compared to CCOC and CCC (3rd vs 5th decade of life, respectively). CONCLUSION: Gaining a better understanding of the similarities and differences between these three tumors may lead to a better understanding of each one.

Sequential administration of paricalcitol followed by IL-17 blockade for progressive refractory IgA nephropathy patients.

There is no established treatment for progressive IgA nephropathy refractory to steroids and immunosuppressant drugs (r-IgAN). Interleukin 17 (IL-17) blockade has garnered interest in immune-mediated diseases involving the gut-kidney axis. However, single IL-17A inhibition induced paradoxical effects in patients with Crohn's disease and some cases of de novo glomerulonephritis, possibly due to the complete Th1 cell response, along with the concomitant downregulation of regulatory T cells (Tregs). Seven r-IgAN patients were treated with at least six months of oral paricalcitol, followed by the addition of subcutaneous anti-IL-17A (secukinumab). After a mean follow-up of 28 months, proteinuria decreased by 71% (95% CI: 56-87), P < 0.001. One patient started dialysis, while the annual eGFR decline in the remaining patients [mean (95% CI)] was reduced by 4.9 mL/min/1.73 m2 (95% CI: 0.1-9.7), P = 0.046. Circulating Th1, Th17, and Treg cells remained stable, but Th2 cells decreased, modifying the Th1/Th2 ratio. Intriguingly, accumulation of circulating Th17.1 cells was observed. This novel sequential therapy appears to optimize renal advantages in patients with r-IgAN and elicit alterations in potentially pathogenic T helper cells.

Ureteral stent after PCNL: is leaving the threads through the percutaneous tract safe and better tolerated?

OBJECTIVE: To assess safety, urinary symptoms, and feasibility of JJ stent removal with exteriorized threads through the percutaneous tract after percutaneous nephrolithotomy (PCNL). MATERIALS AND METHODS: Prospective, transversal, comparative, experimental, randomized 1-to-1 cohort study in 52 patients who underwent "tubeless" PCNL from October 2020 to November 2022. Group A with threads through the urethra and Group B through the percutaneous tract. The validated USSQ (Ureteral Stent Symptom Questionnaire) was applied in the Urology office a week after the procedure, and the JJ stent was withdrawn by pulling the threads. Hemoglobin and urine culture, and pre- and post-surgery were evaluated. RESULTS: There is a statistically significant difference in favor of group B when comparing urinary symptoms (p = 0.008), body pain (p = 0.009), and general condition (p = 0.042), mainly for non-urgency incontinence, frequency of analgesic use, and dysuria. There were significant differences between groups (p = 0.028, p = 0.026, p = 0.027, respectively). There is no association with urinary infections (p = 0.603) nor an increased risk of bleeding (p = 0.321). CONCLUSION: The removal of the JJ stent with exteriorized threads through the percutaneous tract after PCNL in the office is a feasible and safe procedure if it is removed before 8 days and has better tolerance regarding the urinary symptoms.

CIC-DUX4 Chromatin Profiling Reveals New Epigenetic Dependencies and Actionable Therapeutic Targets in CIC-Rearranged Sarcomas.

CIC-DUX4-rearranged sarcoma (CDS) is a rare and aggressive soft tissue tumor that occurs most frequently in young adults. The key oncogenic driver of this disease is the expression of the CIC-DUX4 fusion protein as a result of chromosomal rearrangements. CIC-DUX4 displays chromatin binding properties, and is therefore believed to function as an aberrant transcription factor. However, the chromatin remodeling events induced by CIC-DUX4 are not well understood, limiting our ability to identify new mechanism-based therapeutic strategies for these patients. Here, we generated a genome-wide profile of CIC-DUX4 DNA occupancy and associated chromatin states in human CDS cell models and primary tumors. Combining chromatin profiling, proximity ligation assays, as well as genetic and pharmacological perturbations, we show that CIC-DUX4 operates as a potent transcriptional activator at its binding sites. This property is in contrast with the repressive function of the wild-type CIC protein, and is mainly mediated through the direct interaction of CIC-DUX4 with the acetyltransferase p300. In keeping with this, we show p300 to be essential for CDS tumor cell proliferation; additionally, we find its pharmacological inhibition to significantly impact tumor growth in vitro and in vivo. Taken together, our study elucidates the mechanisms underpinning CIC-DUX4-mediated transcriptional regulation.

DisP-seq reveals the genome-wide functional organization of DNA-associated disordered proteins.

Intrinsically disordered regions (IDRs) in DNA-associated proteins are known to influence gene regulation, but their distribution and cooperative functions in genome-wide regulatory programs remain poorly understood. Here we describe DisP-seq (disordered protein precipitation followed by DNA sequencing), an antibody-independent chemical precipitation assay that can simultaneously map endogenous DNA-associated disordered proteins genome-wide through a combination of biotinylated isoxazole precipitation and next-generation sequencing. DisP-seq profiles are composed of thousands of peaks that are associated with diverse chromatin states, are enriched for disordered transcription factors (TFs) and are often arranged in large lineage-specific clusters with high local concentrations of disordered proteins and different combinations of histone modifications linked to regulatory potential. We use DisP-seq to analyze cancer cells and reveal how disordered protein-associated islands enable IDR-dependent mechanisms that control the binding and function of disordered TFs, including oncogene-dependent sequestration of TFs through long-range interactions and the reactivation of differentiation pathways upon loss of oncogenic stimuli in Ewing sarcoma.

Integrated Radiology, Pathology, and Pharmacy Program to Accelerate Access to Osimertinib.

PURPOSE: Targeted therapy yields superior outcomes relative to genotype-agnostic therapy for patients with epidermal growth factor receptor (EGFR)-mutant lung cancer. Workflows that facilitate timely detection of EGFR mutations and early dispensation of osimertinib can improve management of this disease. METHODS: We developed an Integrated Radiology, Pathology, and Pharmacy Program to minimize delays in initiating osimertinib. The intervention consisted of parallel workflows coupling interventional radiology, surgical pathology, and analysis of nucleic acids from frozen tissue with early pharmacy engagement. We compared time to EGFR testing results and time to treatment for participating patients with those of historical cohorts. RESULTS: Between January 2020 and December 2021, 222 patients participated in the intervention. The median turnaround time from biopsy to EGFR results was 1 workday. Forty-nine (22%) tumors harbored EGFR exon 19 deletions or EGFR L858R. Thirty-one (63%) patients were prescribed osimertinib via the intervention. The median interval between osimertinib prescription and osimertinib dispensation was 3 days; dispensation occurred within 48 hours for 42% of patients. The median interval between biopsy and osimertinib dispensation was 5 days. Three patients received osimertinib within 24 hours of EGFR results. Compared with patients with EGFR-mutant non-small-cell lung cancer who were diagnosed through routine workflows, the intervention led to a significant reduction in median time between biopsy and EGFR results (1 v 7 days; P < .01) and median time to treatment initiation (5 v 23 days; P < .01). CONCLUSION: Combining radiology and pathology workflows with early parallel pharmacy engagement leads to a significant reduction in time to initiating osimertinib. Multidisciplinary integration programs are essential to maximize clinical utility of rapid testing.

Non-canonical integrin signaling activates EGFR and RAS-MAPK-ERK signaling in small cell lung cancer.

Background: Small cell lung cancer (SCLC) is an extremely aggressive cancer type with a patient median survival of 6-12 months. Epidermal growth factor (EGF) signaling plays an important role in triggering SCLC. In addition, growth factor-dependent signals and alpha-, beta-integrin (ITGA, ITGB) heterodimer receptors functionally cooperate and integrate their signaling pathways. However, the precise role of integrins in EGF receptor (EGFR) activation in SCLC remains elusive. Methods: We analyzed human precision-cut lung slices (hPCLS), retrospectively collected human lung tissue samples and cell lines by classical methods of molecular biology and biochemistry. In addition, we performed RNA-sequencing-based transcriptomic analysis in human lung cancer cells and human lung tissue samples, as well as high-resolution mass spectrometric analysis of the protein cargo from extracellular vesicles (EVs) that were isolated from human lung cancer cells. Results: Our results demonstrate that non-canonical ITGB2 signaling activates EGFR and RAS/MAPK/ERK signaling in SCLC. Further, we identified a novel SCLC gene expression signature consisting of 93 transcripts that were induced by ITGB2, which may be used for stratification of SCLC patients and prognosis prediction of LC patients. We also found a cell-cell communication mechanism based on EVs containing ITGB2, which were secreted by SCLC cells and induced in control human lung tissue RAS/MAPK/ERK signaling and SCLC markers. Conclusions: We uncovered a mechanism of ITGB2-mediated EGFR activation in SCLC that explains EGFR-inhibitor resistance independently of EGFR mutations, suggesting the development of therapies targeting ITGB2 for patients with this extremely aggressive lung cancer type.

Highly connected 3D chromatin networks established by an oncogenic fusion protein shape tumor cell identity.

Cell fate transitions observed in embryonic development involve changes in three-dimensional genomic organization that provide proper lineage specification. Whether similar events occur within tumor cells and contribute to cancer evolution remains largely unexplored. We modeled this process in the pediatric cancer Ewing sarcoma and investigated high-resolution looping and large-scale nuclear conformation changes associated with the oncogenic fusion protein EWS-FLI1. We show that chromatin interactions in tumor cells are dominated by highly connected looping hubs centered on EWS-FLI1 binding sites, which directly control the activity of linked enhancers and promoters to establish oncogenic expression programs. Conversely, EWS-FLI1 depletion led to the disassembly of these looping networks and a widespread nuclear reorganization through the establishment of new looping patterns and large-scale compartment configuration matching those observed in mesenchymal stem cells, a candidate Ewing sarcoma progenitor. Our data demonstrate that major architectural features of nuclear organization in cancer cells can depend on single oncogenes and are readily reversed to reestablish latent differentiation programs.

The ETS transcription factor ETV6 constrains the transcriptional activity of EWS-FLI to promote Ewing sarcoma.

Transcription factors (TFs) are frequently mutated in cancer. Paediatric cancers exhibit few mutations genome-wide but frequently harbour sentinel mutations that affect TFs, which provides a context to precisely study the transcriptional circuits that support mutant TF-driven oncogenesis. A broadly relevant mechanism that has garnered intense focus involves the ability of mutant TFs to hijack wild-type lineage-specific TFs in self-reinforcing transcriptional circuits. However, it is not known whether this specific type of circuitry is equally crucial in all mutant TF-driven cancers. Here we describe an alternative yet central transcriptional mechanism that promotes Ewing sarcoma, wherein constraint, rather than reinforcement, of the activity of the fusion TF EWS-FLI supports cancer growth. We discover that ETV6 is a crucial TF dependency that is specific to this disease because it, counter-intuitively, represses the transcriptional output of EWS-FLI. This work discovers a previously undescribed transcriptional mechanism that promotes cancer.

Clinical Implications and Treatment Strategies for ESR1 Fusions in Hormone Receptor-Positive Metastatic Breast Cancer: A Case Series.

In hormone receptor-positive metastatic breast cancer (HR+ MBC), endocrine resistance is commonly due to genetic alterations of ESR1, the gene encoding estrogen receptor alpha (ERα). While ESR1 point mutations (ESR1-MUT) cause acquired resistance to aromatase inhibition (AI) through constitutive activation, far less is known about the molecular functions and clinical consequences of ESR1 fusions (ESR1-FUS). This case series discusses 4 patients with HR+ MBC with ESR1-FUS in the context of the existing ESR1-FUS literature. We consider therapeutic strategies and raise the hypothesis that CDK4/6 inhibition (CDK4/6i) may be effective against ESR1-FUS with functional ligand-binding domain swaps. These cases highlight the importance of screening for ESR1-FUS in patients with HR+ MBC while continuing investigation of precision treatments for these genomic rearrangements.

Uncommon Find: An Unusual 10 cm Bezold's Abscess Involving Dural and Mastoid Erosion.

Bezold's abscess (BA) is a rare complication of otitis media that presents as a lateral neck abscess below the mastoid tip. BA incidence has recently decreased due to early diagnosis and prompt antibiotic intervention. We present a 42-year-old male with a complicated otitis media developing a 10 cm BA. Treatment of the lesion included surgical drainage and mastoidectomy, accompanied by intravenous (IV) broad-spectrum antibiotic administration. The patient experienced no adverse events during or after surgery and was placed on postoperative observation. However, on postoperative day (POD) 2, the patient left the hospital against medical advice and did not undergo further follow-up.

Genome-wide functional perturbation of human microsatellite repeats using engineered zinc finger transcription factors.

Repeat elements can be dysregulated at a genome-wide scale in human diseases. For example, in Ewing sarcoma, hundreds of inert GGAA repeats can be converted into active enhancers when bound by EWS-FLI1. Here we show that fusions between EWS and GGAA-repeat-targeted engineered zinc finger arrays (ZFAs) can function at least as efficiently as EWS-FLI1 for converting hundreds of GGAA repeats into active enhancers in a Ewing sarcoma precursor cell model. Furthermore, a fusion of a KRAB domain to a ZFA can silence GGAA microsatellite enhancers genome wide in Ewing sarcoma cells, thereby reducing expression of EWS-FLI1-activated genes. Remarkably, this KRAB-ZFA fusion showed selective toxicity against Ewing sarcoma cells compared with non-Ewing cancer cells, consistent with its Ewing sarcoma-specific impact on the transcriptome. These findings demonstrate the value of ZFAs for functional annotation of repeats and illustrate how aberrant microsatellite activities might be regulated for potential therapeutic applications.

Chromatin-bound RB targets promoters, enhancers, and CTCF-bound loci and is redistributed by cell-cycle progression.

The interaction of RB with chromatin is key to understanding its molecular functions. Here, for first time, we identify the full spectrum of chromatin-bound RB. Rather than exclusively binding promoters, as is often described, RB targets three fundamentally different types of loci (promoters, enhancers, and insulators), which are largely distinguishable by the mutually exclusive presence of E2F1, c-Jun, and CTCF. While E2F/DP facilitates RB association with promoters, AP-1 recruits RB to enhancers. Although phosphorylation in CDK sites is often portrayed as releasing RB from chromatin, we show that the cell cycle redistributes RB so that it enriches at promoters in G1 and at non-promoter sites in cycling cells. RB-bound promoters include the classic E2F-targets and are similar between lineages, but RB-bound enhancers associate with different categories of genes and vary between cell types. Thus, RB has a well-preserved role controlling E2F in G1, and it targets cell-type-specific enhancers and CTCF sites when cells enter S-phase.

Implementation and Clinical Adoption of Precision Oncology Workflows Across a Healthcare Network.

BACKGROUND: Precision oncology relies on molecular diagnostics, and the value-proposition of modern healthcare networks promises a higher standard of care across partner sites. We present the results of a clinical pilot to standardize precision oncology workflows. METHODS: Workflows are defined as the development, roll-out, and updating of disease-specific molecular order sets. We tracked the timeline, composition, and effort of consensus meetings to define the combination of molecular tests. To assess clinical impact, we examined order set adoption over a two-year period (before and after roll-out) across all gastrointestinal and hepatopancreatobiliary (GI) malignancies, and by provider location within the network. RESULTS: Development of 12 disease center-specific order sets took ~9 months, and the average number of tests per indication changed from 2.9 to 2.8 (P = .74). After roll-out, we identified significant increases in requests for GI patients (17%; P < .001), compliance with testing recommendations (9%; P < .001), and the fraction of "abnormal" results (6%; P < .001). Of 1088 GI patients, only 3 received targeted agents based on findings derived from non-recommended orders (1 before and 2 after roll-out); indicating that our practice did not negatively affect patient treatments. Preliminary analysis showed 99% compliance by providers in network sites, confirming the adoption of the order sets across the network. CONCLUSION: Our study details the effort of establishing precision oncology workflows, the adoption pattern, and the absence of harm from the reduction of non-recommended orders. Establishing a modifiable communication tool for molecular testing is an essential component to optimize patient care via precision oncology.