Overexpression of human DNA polymerase theta is a biomarker of aggressive and DNA repair-deficient papillary thyroid cancers.

BACKGROUND: DNA polymerase theta (POLQ) is an enzyme that repairs double-strand DNA breaks. POLQ is overexpressed in several cancer types, and increased expression is associated with a poor prognosis. Ablating POLQ function in vitro increases drug sensitivity to agents that cause double-strand DNA breaks, including chemotherapies and ionizing radiation. POLQ's role in thyroid cancer remains poorly understood. METHODS: Expression of POLQ and other genes of interest were analyzed in 513 papillary thyroid cancers (505 primary tumors and 8 metastatic lesions) and 59 normal thyroid samples available in the Cancer Genome Atlas. The Cancer Genome Atlas RNA and DNA sequencing data were queried with the Xena platform. The Recombination Proficiency Score was calculated to assess DNA repair efficiency. Other signaling events associated with thyroid tumorigenesis and clinical outcomes were analyzed. Univariate and multivariate analyses were performed. Treatment with the POLQ inhibitors ART558 and Novobiocin tested the effect of POLQ inhibition on in vitro thyroid cancer growth. RESULTS: POLQ expression was increased in papillary thyroid cancers compared to normal thyroid tissue (P < .05). POLQ expression levels were inversely correlated with Recombination Proficiency Score levels (P < .05). POLQ expression was highest in tall cell papillary thyroid cancers and in metastases. Higher POLQ expression was also associated with dedifferentiation, BRAF signaling, and shorter progression-free intervals (P < .05). Treatment with POLQ inhibitors decreased in vitro thyroid cancer growth (P < .05). CONCLUSION: These findings suggest that increased POLQ expression could serve as a valuable clinical marker and a potential therapeutic target in the treatment of thyroid cancer.

Increased signal of the fibular collateral ligament of the knee on MRI, clinically significant?

INTRODUCTION: The purpose of this study was to determine the clinical significance of signal hyperintensity in the proximal fibular collateral ligament (FCL) on coronal proton density (PD) fat-saturated (FS) MRI of the knee, a common finding. This study is unique in that it characterizes the FCL of a comprehensive, large cohort of both symptomatic and asymptomatic patients, which to our knowledge represents the first study with such broad inclusion criteria. METHODS: A large case series was performed analyzing MRI of the knee of 250 patients from July 2021 through September 2021 and retrospectively reviewed. All studies were performed on 3-Tesla MRI scanners with a dedicated knee coil and in accordance with standard institutional knee MRI protocol. Signal in the proximal fibular collateral ligament was assessed on coronal PDFS and axial T2-weighted FS images. Increased signal was classified as none, mild, moderate, or severe. A corresponding chart review of clinic notes was performed to determine the presence or absence of lateral knee pain. An FCL sprain or injury was considered present if the medical record described tenderness on palpation of the lateral knee, positive finding against resistance to the leg (varus stress test) or reverse pivot shift, or any clinical suspicion for lateral complex sprain or posterolateral corner injury. RESULTS: The majority (74%) of knee MRIs demonstrated the presence of increased signal in the proximal fibular collateral ligament on coronal PD FS images. <5% of these patients had associated clinical findings of fibular collateral ligament and/or lateral supporting structure injury. DISCUSSION: Although increased signal in the proximal FCL of the knee is a common finding on coronal PDFS images, the majority are not associated with clinical symptoms. Thus, this increased signal is likely not a pathological finding in the absence of clinical findings of fibular collateral ligament sprain/injury. Our study emphasizes the importance of clinical correlation in identifying increased signal in the proximal FCL as pathologic.

Cholesterol is enriched in the sphingolipid patches on the substrate near nonpolarized MDCK cells, but not in the sphingolipid domains in their plasma membranes.

Information about the distributions of cholesterol and sphingolipids within the plasma membranes of mammalian cells provides insight into the roles of these molecules in membrane function. In this report, high-resolution secondary ion mass spectrometry was used to image the distributions of metabolically incorporated rare isotope-labeled sphingolipids and cholesterol on the surfaces of nonpolarized epithelial cells. Sphingolipid domains that were not enriched with cholesterol were detected in the plasma membranes of subconfluent Madin-Darby canine kidney cells. Surprisingly, cholesterol-enriched sphingolipid patches were observed on the substrate adjacent to these cells. Based on the shapes of these cholesterol-enriched sphingolipid patches on the substrate and their proximity to cellular projections, we hypothesize that they are deposits of membranous particles released by the cell.

Three-dimensional imaging of cholesterol and sphingolipids within a Madin-Darby canine kidney cell.

Metabolic stable isotope incorporation and secondary ion mass spectrometry (SIMS) depth profiling performed on a Cameca NanoSIMS 50 were used to image the (18)O-cholesterol and (15)N-sphingolipid distributions within a portion of a Madin-Darby canine kidney (MDCK) cell. Three-dimensional representations of the component-specific isotope distributions show clearly defined regions of (18)O-cholesterol and (15)N-sphingolipid enrichment that seem to be separate subcellular compartments. The low levels of nitrogen-containing secondary ions detected at the (18)O-enriched regions suggest that these (18)O-cholesterol-rich structures may be lipid droplets, which have a core consisting of cholesterol esters and triacylglycerides.