Citations

Abstract Background Malignant peripheral nerve sheath tumors (MPNSTs) can arise from atypical neurofibromas (ANF). Loss of the polycomb repressor complex 2 (PRC2) is a common event. Previous studies on PRC2-regulated genes in MPNST used genetic add-back experiments in highly aneuploid MPNST cell lines which may miss PRC2-regulated genes in NF1-mutant ANF-like precursor cells. A set of PRC2-regulated genes in human Schwann cells (SCs) has not been defined. We hypothesized that PRC2 loss has direct and indirect effects on gene expression resulting in MPNST, so we sought to identify PRC2-regulated genes in immortalized human Schwann cells (iHSCs). Methods We engineered NF1-deficient iHSCs with loss of function SUZ12 or EED mutations. RNA sequencing revealed 1327 differentially expressed genes to define PRC2-regulated genes. To investigate MPNST pathogenesis, we compared genes in iHSCs to consistent gene expression differences between ANF and MPNSTs. Chromatin immunoprecipitation sequencing was used to further define targets. Methylome and proteomic analyses were performed to further identify enriched pathways. Results We identified potential PRC2-regulated drivers of MPNST progression. Pathway analysis indicates many upregulated cancer-related pathways. We found transcriptional evidence for activated Notch and Sonic Hedgehog (SHH) signaling in PRC2-deficient iHSCs. Functional studies confirm that Notch signaling is active in MPNST cell lines, patient-derived xenografts, and transient cell models of PRC2 deficiency. A combination of MEK and γ-secretase inhibition shows synergy in MPNST cell lines. Conclusions We identified PRC2-regulated genes and potential drivers of MPNSTs. Our findings support the Notch pathway as a druggable target in MPNSTs. Our identification of PRC2-regulated genes and pathways could result in more novel therapeutic approaches.

Abstract Neurofibromatosis type 1 (NF1) syndrome is an autosomal dominant cancer predisposition syndrome defined by germline deletion of one NF1 allele. Somatic loss of heterozygosity in the remaining NF1 allele in Schwann lineage cells gives rise to benign tumors known as plexiform neurofibromas. These tumors have a high risk of transforming into malignant peripheral nerve sheath tumors (MPNST) through the sequential deletion of CDKN2A/B followed by recurrent mutations in SUZ12 or EED, constituting the inactivation of polycomb repressor complex 2 (PRC2). PRC2 is a transcriptional repressor complex involved in the silencing of several development and differentiation genes. Loss of this complex leads to a depletion in trimethylation marks on Histone H3 Lysine 27 (H3K27me3) and the subsequent gain of acetylation marks on the same lysine residue (H3K27Ac). These molecular hallmarks are shared with diffuse intrinsic pontine gliomas (DIPG) harboring the H3K27M mutation which prevents effective deposition of H3K27me3, thus having similar molecular phenotypes to MPNST. However, the full range of epigenetic consequences of these events are currently not well characterized. Additionally, given that MPNSTs originate from cells of neural crest origin and DIPGs from cells of neural origin, it is not know to what degree epigenetic changes stemming from H3K27me3-loss are general across tumors or are context-dependent based on cell-of-origin. In order to contribute to this effort, the Largaespada lab has engineered Schwann cell lines to harbor deletions in NF1 and either SUZ12 or EED and assayed the H3K27ac landscape of these cells in addition to patient-derived MPNST cell lines. Utilizing the ranked ordering of super enhancers (ROSE) algorithm, we have identified common super enhancers within each MPNST cell line. These super enhancer elements are located nearby a variety of putative oncogenes responsible for the maintenance of cell identity. These elements were then compared to publicly available data from DIPG primary and patient-derived samples to understand common regulatory events driven by loss of H3K27me3 as well as molecular signatures present only in MPNST or in DIPG that may be a result of PRC2 or H3K27M alone. We further characterized these super enhancers by assessing the expression levels of their associated genes to understand their role in transcriptional activation. These efforts provide a roadmap of the super enhancer landscape of both MPNSTs and DIPGs to investigate novel mechanisms of H3K27me3-dependent tumorigenesis while also contributing to knowledge of potential effects of H3K27me3-loss that dependent on the cell-of-origin for each respective tumor. Citation Format: Christopher M. Stehn, Minu Bhunia, Kyle Williams, Alex T. Larsson, Ariel Caride, Jeong-Heon Lee, Tamas Ordog, David Largaespada. Characterizing the super enhancer landscape of H3K27me3 loss in MPNST and DIPG [abstract]. In: Proceedings of the AACR Special Conference on Brain Cancer; 2023 Oct 19-22; Minneapolis, Minnesota. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_1):Abstract nr B009.

Purpose To determine the accuracy of CT number and calcium score of a kV‐independent technique based on an artificial 120 kV reconstruction, and its potential to reduce radiation dose. Methods Anthropomorphic chest phantoms were scanned on a third‐generation dual‐source CT system equipped with the artificial 120 kV reconstruction. First, a phantom module containing a 20‐mm diameter hydroxyapatite (HA) insert was scanned inside the chest phantoms at different tube potentials (70–140 kV) to evaluate calcium CT number accuracy. Next, three small HA inserts (diameter/length = 5 mm) were inserted into a pork steak and scanned inside the phantoms to evaluate calcium score accuracy at different kVs. Finally, the same setup was scanned using automatic exposure control (AEC) at 120 kV, and then with automatic kV selection (auto‐kV). Phantoms were also scanned at 120 kV using a size‐dependent mA chart. CT numbers of soft tissue and calcium were measured from different kV images. Calcium score of each small HA insert was measured using commercial software. Results The CT number difference from 120 kV was small with tube potentials from 90 to 140 kV for both soft tissue and calcium (maximal difference of 4/5 HU, respectively). Consistent calcium scores were obtained from images of different kVs compared to 120 kV, with a relative difference <8%. Auto‐kV provided a 25–34% dose reduction compared to AEC alone. Conclusion A kV‐independent calcium scoring technique can produce artificial 120 kV images with consistent soft tissue and calcium CT numbers compared to standard 120 kV examinations. When coupled with auto‐kV, this technique can reduce radiation by 25–34% compared to that with AEC alone, while providing consistent calcium scores as that of standard 120 kV examinations.

Background: Lumbar spine injuries in National Collegiate Athletic Association (NCAA) athletes have not been well studied. Purpose: To describe the epidemiology of lumbar spine injuries in NCAA athletes during the 2009/2010 through 2013/2014 academic years utilizing the NCAA Injury Surveillance Program (ISP). Study Design: Descriptive epidemiology study. Methods: A voluntary convenience sample of NCAA varsity teams from 25 sports was examined. Mechanism of injury, injury recurrence, and time lost from sport were recorded. Injury rates were calculated as the number of injuries divided by the total number of athlete-exposures (AEs). AEs were defined as any student participation in 1 NCAA-sanctioned practice or competition. Injury rate ratios and injury proportion ratios were calculated to compare the rates within and between sports by event type, season, patient sex, mechanism, injury recurrence, and time lost from sport. Comparisons between sexes were made utilizing data that had both male and female samples. Results: An estimated 50,834 lumbar spine injuries were identified. The overall rate of injuries was 2.88 per 10,000 AEs. The rate of injuries was 2.60 per 10,000 AEs in men compared with 2.89 per 10,000 AEs in women for sex-comparable sports. Women were 1.11 times more likely than men to suffer a lumbar spine injury. Women's gymnastics (8.02 injuries per 10,000 AEs) and women's tennis (5.73 injuries per 10,000 AEs) had the highest rates of lumbar spine injuries. Athletes were 1.6 and 2.4 times more likely to sustain a lumbar spine injury during the preseason than the regular season or postseason, respectively. Noncontact was the most common mechanism of injury (45%). Injury recurrence was most common in men’s outdoor track (43%). Most injuries resulted in less than 24 hours of time loss from event participation (64%). Conclusion: The rate of lumbar spine injuries was high in NCAA athletes, and injuries commonly recurred (20.73%). In general, women were more likely to sustain a lumbar spine injury compared with men. Higher injury rates occurred during competition and via a noncontact mechanism of injury. In addition to prevention programs, reconditioning programs should be considered to prevent these injuries.