Citations

Abstract MYCN is a major driver for the childhood cancer, neuroblastoma, however, there are no inhibitors of this target. Enhanced MYCN protein stability is a key component of MYCN oncogenesis and is maintained by multiple feedforward expression loops involving MYCN transactivation target genes. Here, we reveal the oncogenic role of a novel MYCN target and binding protein, proliferation-associated 2AG4 (PA2G4). Chromatin immunoprecipitation studies demonstrated that MYCN occupies the PA2G4 gene promoter, stimulating transcription. Direct binding of PA2G4 to MYCN protein blocked proteolysis of MYCN and enhanced colony formation in a MYCN-dependent manner. Using molecular modeling, surface plasmon resonance, and mutagenesis studies, we mapped the MYCN–PA2G4 interaction site to a 14 amino acid MYCN sequence and a surface crevice of PA2G4. Competitive chemical inhibition of the MYCN–PA2G4 protein–protein interface had potent inhibitory effects on neuroblastoma tumorigenesis in vivo. Treated tumors showed reduced levels of both MYCN and PA2G4. Our findings demonstrate a critical role for PA2G4 as a cofactor in MYCN-driven neuroblastoma and highlight competitive inhibition of the PA2G4-MYCN protein binding as a novel therapeutic strategy in the disease. Significance: Competitive chemical inhibition of the PA2G4–MYCN protein interface provides a basis for drug design of small molecules targeting MYC and MYCN-binding partners in malignancies driven by MYC family oncoproteins.

Background: National Collegiate Athletic Association (NCAA) football players are at a high risk of injuries. Elbow injuries are uncommon, but there are insufficient data specifically on elbow injuries sustained in NCAA football players. Purpose: To define the epidemiology of elbow injuries in NCAA football players during the 2009-2010 to 2013-2014 seasons using data from the NCAA Injury Surveillance Program (NCAA-ISP). Study Design: Descriptive epidemiology study. Methods: Using the NCAA-ISP database, a convenience sample of NCAA football athletes was reviewed to determine the types, rates, and trends in elbow injuries. Several factors were examined, including the diagnosis, injury setting, time lost from sport, surgical necessity, and injury recurrence. Raw injury data were obtained as well as weighted totals from the NCAA-ISP to generate national estimates and adjust for underreporting. Injury rates were calculated by dividing the number of injuries by the total number of athlete-exposures (AEs). The rate ratios of injuries during competition versus practice were compared, as were the rate ratios of preseason, regular-season, and postseason injuries by type. Results: We identified 4874 total elbow injuries from the 2009-2010 to 2013-2014 seasons. The rate of injuries overall was 1.892 per 10,000 AEs. The rate for competition was 9.053 per 10,000 AEs and 1.121 per 10,000 AEs for practice. The rate ratio between competition and practice was 8.08 (95% CI, 6.04-10.80). Injury rates for the preseason, regular season, and postseason were 1.851, 1.936, and 1.406 per 10,000 AEs, respectively. Acute elbow instability was the most common injury type (65.43%). The most common mechanism was a contact injury (86.77%); 96.82% of injuries did not require surgery, and most elbow injuries required less than 24 hours of participation restriction (67.33%). Conclusion: Although elbow injuries in NCAA football players are uncommon, it is important to recognize and treat these injuries appropriately. Dislocations and ulnar collateral ligament injuries caused athletes to miss extended periods of play. Fortunately, a majority of injuries resulted in less than 24 hours of participation restriction. Particular attention should be given to preventing elbow injuries, especially ulnar collateral ligament strains, hyperextension injuries, and acute instability.

Background: It is unclear whether chondral fragments without bone have the potential to heal after fixation. Controversy exists and opinions differ regarding the optimal treatment of chondral defects after pure chondral fracture. Purpose: To determine clinical and radiographic outcomes after internal fixation of traumatic chondral fragments repaired to bone in the knee. Study Design: Case series; Level of evidence, 4. Methods: A retrospective clinical and radiographic evaluation of 10 male patients with a mean age of 14.6 years (range, 10-25 years) at the time of surgery was performed. Eight of 10 patients were skeletally immature. Patients were selected by operating surgeons per the presence of a large displaced pure chondral fragment on magnetic resonance imaging and confirmed on intraoperative inspection. All patients had a diagnosed traumatic displaced pure chondral fracture of the knee (without bone) and underwent internal fixation with minimum 1-year follow-up. Validated patient-reported and surgeon-measured outcomes were collected pre- and postoperatively. All patients were evaluated at a mean 56 months postoperatively. Results: At surgery, the mean defect size that was primarily repaired with the displaced chondral fragment was 1.9 × 2.0 cm. With minimum 1-year follow-up, there were no clinical failures. All 8 patients who had subsequent magnetic resonance imaging follow-up had radiographic evidence of complete healing of the chondral fragment back to bone. At a mean follow-up of 56 months (range, 13-171 months; median, 36 months), patients had a mean International Knee Documentation Committee score of 94.74 (range, 87.4-100), a mean Marx Activity Scale score of 14.4 (range, 8-16), and a mean Tegner Activity Scale score of 7 (range, 5-9). At final follow-up, all patients except 1 returned to sports. Conclusion: The treatment of large traumatic chondral fragments is controversial. In this select series of 10 young patients who underwent primary repair with internal fixation, there were no failures clinically. Patients demonstrated excellent short-term clinical and radiographic results after fixation of these relatively large chondral fragments in the knee.

Abstract Objective The quality of all clinical MRI is dependent on B0 homogeneity, which is optimized during the shimming part of a prescan or preparatory phase before image acquisition. The purpose of this study was to assess shimming techniques clinically employed for breast MRI across our practice, and to determine factors that correlate with higher image quality for contrast-enhanced breast MRI at 1.5T. Methods One hundred consecutive female patients were retrospectively collected with Institutional Review Board approval. Shimming-related parameters, including shim-box placement and shimming gradient offsets were extracted from prior contrast-enhanced 3D fat-suppressed T1-weighted gradient echo image acquisitions. Three breast radiologists evaluated these images for fat saturation, breast density, overall image quality, and artifacts. Technologist experience was also evaluated for variability of shimming. Generalized linear mixed models were used to compare acquisition parameters between fat saturation. P < 0.05 was considered as statistical significance. Results The percentage of soft tissue inside the field of view (FOV) (ie, Tissue/FOV) in the good fat-saturation group (0.37 ± 0.06) was significantly lower (P < 0.01) than that in the poor fat-saturation group (0.39 ± 0.06). Other shimming-related parameters were found not significantly affecting the fat-saturation outcomes. Technologists with more experience tended to have less variable shimming performance than junior technologists did. Conclusions The quality of clinical MRI and especially breast MRI is highly dependent on shimming. Decreasing Tissue/FOV was associated with good image quality (good fat saturation). Optimization of shimming may require manual shimming or higher-order field-correction strategies.

Significance Statement The lack of rapidly progressive murine models reflecting the more severe end of the spectrum of autosomal recessive polycystic kidney disease (ARPKD) inhibits progress to understanding ARPKD pathogenesis. Defects in primary cilia have been implicated in polycystic kidney disease, but their potential role is poorly understood. The authors generated and characterized new mouse and rat models of ARPKD and autosomal dominant polycystic kidney disease (ADPKD) and investigated the interaction between causative genes for these two conditions. Their digenic models demonstrated a synergistic interaction that better reflects the early-onset disease characteristic of ARPKD. Analysis of mRNA expression in the models highlighted different disrupted pathways, but with a commonality of dysregulated mechanisms associated with primary cilia. These models may improve understanding of ARPKD and preclinical testing for this disease. Background Autosomal recessive polycystic kidney disease (ARPKD) and autosomal dominant polycystic kidney disease (ADPKD) are genetically distinct, with ADPKD usually caused by the genes PKD1 or PKD2 (encoding polycystin-1 and polycystin-2, respectively) and ARPKD caused by PKHD1 (encoding fibrocystin/polyductin [FPC]). Primary cilia have been considered central to PKD pathogenesis due to protein localization and common cystic phenotypes in syndromic ciliopathies, but their relevance is questioned in the simple PKDs. ARPKD’s mild phenotype in murine models versus in humans has hampered investigating its pathogenesis. Methods To study the interaction between Pkhd1 and Pkd1 , including dosage effects on the phenotype, we generated digenic mouse and rat models and characterized and compared digenic, monogenic, and wild-type phenotypes. Results The genetic interaction was synergistic in both species, with digenic animals exhibiting phenotypes of rapidly progressive PKD and early lethality resembling classic ARPKD. Genetic interaction between Pkhd1 and Pkd1 depended on dosage in the digenic murine models, with no significant enhancement of the monogenic phenotype until a threshold of reduced expression at the second locus was breached. Pkhd1 loss did not alter expression, maturation, or localization of the ADPKD polycystin proteins, with no interaction detected between the ARPKD FPC protein and polycystins. RNA-seq analysis in the digenic and monogenic mouse models highlighted the ciliary compartment as a common dysregulated target, with enhanced ciliary expression and length changes in the digenic models. Conclusions These data indicate that FPC and the polycystins work independently, with separate disease-causing thresholds; however, a combined protein threshold triggers the synergistic, cystogenic response because of enhanced dysregulation of primary cilia. These insights into pathogenesis highlight possible common therapeutic targets.

ObjectiveType I interferon (IFN) is important to systemic lupus erythematosus (SLE) pathogenesis, but it is not clear how chronic elevations in IFN alter immune function. We compared cytokine responses after whole blood stimulation with Toll‐like receptor (TLR) agonists in high‐ and low‐IFN SLE patient subgroups.MethodsSLE patients and nonautoimmune controls were recruited, and SLE patients were categorized as either high or low IFN. Whole blood was dispensed into tubes coated with lipopolysaccharide (LPS), oligonucleotides with cytosine‐guanine repeats, Resiquimod, IFN‐α, and IFN‐α + LPS. Cytokine production in patient sera and after whole blood TLR stimulation was measured by multiplex assay, and type I IFN was assessed using a functional assay.ResultsCirculating plasmacytoid dendritic cell numbers were specifically reduced in high‐IFN SLE patients and not in low‐IFN SLE patients. In serum, we observed that the correlations between cytokines in serum differed to a much greater degree between the high‐ and low‐IFN groups (P < 0.0001) than the absolute cytokine levels differed between these same groups. In stimulated conditions, the high‐IFN patients had less cytokine production in response to TLR ligation than the low‐IFN SLE patients. LPS produced the most diverse response, and a number of interactions between type I IFN and LPS were observed.ConclusionWe find striking differences in resting and stimulated cytokine patterns in high‐ vs. low‐IFN SLE patients, which supports the biological importance of these patient subsets. These data could inform personalized treatment approaches and the pathogenesis of SLE flare following infection.

AbstractObjectiveNonalcoholic fatty liver disease can lead to hepatic inflammation/damage. Understanding the physiological mechanisms that contribute to excess hepatic lipid accumulation may help identify effective treatments.DesignWe recruited 25 nondiabetic patients with severe obesity scheduled for bariatric surgery. To evaluate liver export of triglyceride fatty acids, we measured very-low-density lipoprotein (VLDL)–triglyceride secretion rates the day prior to surgery using an infusion of autologous [1-14C]triolein-labeled VLDL particles. Ketone body response to fasting and intrahepatic long-chain acylcarnitine concentrations were used as indices of hepatic fatty acid oxidation. We measured intraoperative hepatic uptake rates of plasma free fatty acids using a continuous infusion of [U-13C]palmitate, combined with a bolus dose of [9,10-3H]palmitate and carefully timed liver biopsies. Total intrahepatic lipids were measured in liver biopsy samples to determine fatty liver status. The hepatic concentrations and enrichment from [U-13C]palmitate in diacylglycerols, sphingolipids, and acyl-carnitines were measured using liquid chromatography/tandem mass spectrometry.ResultsAmong study participants with fatty liver disease, intrahepatic lipid was negatively correlated with VLDL-triglyceride secretion rates (r = −0.92, P = 0.01) but unrelated to hepatic free fatty acid uptake or indices of hepatic fatty acid oxidation. VLDL-triglyceride secretion rates were positively correlated with hepatic concentrations of saturated diacylglycerol (r = 0.46, P = 0.02) and sphingosine-1-phosphate (r = 0.44, P = 0.03).ConclusionWe conclude that in nondiabetic humans with severe obesity, excess intrahepatic lipid is associated with limited export of triglyceride in VLDL particles rather than increased uptake of systemic free fatty acids.