Clinicopathological Significance of Cyclin-Dependent Kinase 2 (CDK2) in Ductal Carcinoma In Situ and Early-Stage Invasive Breast Cancers.

Cyclin-dependent kinase 2 (CDK2) is a key cell cycle regulator, with essential roles during G1/S transition. The clinicopathological significance of CDK2 in ductal carcinomas in situ (DCIS) and early-stage invasive breast cancers (BCs) remains largely unknown. Here, we evaluated CDK2's protein expression in 479 BC samples and 216 DCIS specimens. Analysis of CDK2 transcripts was completed in the METABRIC cohort (n = 1980) and TCGA cohort (n = 1090), respectively. A high nuclear CDK2 protein expression was significantly associated with aggressive phenotypes, including a high tumour grade, lymph vascular invasion, a poor Nottingham prognostic index (all p-values < 0.0001), and shorter survival (p = 0.006), especially in luminal BC (p = 0.009). In p53-mutant BC, high nuclear CDK2 remained linked with worse survival (p = 0.01). In DCIS, high nuclear/low cytoplasmic co-expression showed significant association with a high tumour grade (p = 0.043), triple-negative and HER2-enriched molecular subtypes (p = 0.01), Comedo necrosis (p = 0.024), negative ER status (p = 0.004), negative PR status (p < 0.0001), and a high proliferation index (p < 0.0001). Tumours with high CDK2 transcripts were more likely to have higher expressions of genes involved in the cell cycle, homologous recombination, and p53 signaling. We provide compelling evidence that high CDK2 is a feature of aggressive breast cancers. The clinical evaluation of CDK2 inhibitors in early-stage BC patients will have a clinical impact.

The Clinicopathological Significance of the Cyclin D1/E1-Cyclin-Dependent Kinase (CDK2/4/6)-Retinoblastoma (RB1/pRB1) Pathway in Epithelial Ovarian Cancers.

Cyclin-dependent kinases (CDK2, CDK4, CDK6), cyclin D1, cyclin E1 and phosphorylated retinoblastoma (pRB1) are key regulators of the G1/S cell cycle checkpoint and may influence platinum response in ovarian cancers. CDK2/4/6 inhibitors are emerging targets in ovarian cancer therapeutics. In the current study, we evaluated the prognostic and predictive significance of the CDK2/4/6-cyclin D1/E1-pRB1 axis in clinical ovarian cancers (OC). The CDK2/4/6, cyclin D1/E1 and RB1/pRB1 protein expression were investigated in 300 ovarian cancers and correlated with clinicopathological parameters and patient outcomes. CDK2/4/6, cyclin D1/E1 and RB1 mRNA expression were evaluated in the publicly available ovarian TCGA dataset. We observed nuclear and cytoplasmic staining for CDK2/4/6, cyclins D1/E1 and RB1/pRB1 in OCs with varying percentages. Increased nuclear CDK2 and nuclear cyclin E1 expression was linked with poor progression-free survival (PFS) and a shorter overall survival (OS). Nuclear CDK6 was associated with poor OS. The cytoplasmic expression of CDK4, cyclin D1 and cyclin E1 also has predictive and/or prognostic significance in OCs. In the multivariate analysis, nuclear cyclin E1 was an independent predictor of poor PFS. Tumours with high nuclear cyclin E1/high nuclear CDK2 have a worse PFS and OS. Detailed bioinformatics in the TCGA cohort showed a positive correlation between cyclin E1 and CDK2. We also showed that cyclin-E1-overexpressing tumours are enriched for genes involved in insulin signalling and release. Our data not only identified the prognostic/predictive significance of these key cell cycle regulators but also demonstrate the importance of sub-cellular localisation. CDK2 targeting in cyclin-E1-amplified OCs could be a rational approach.

Immune infiltration, aggressive pathology, and poor survival outcomes in RECQL helicase deficient breast cancers.

RECQL is essential for genomic stability. Here, we evaluated RECQL in 449 pure ductal carcinomas in situ (DCIS), 152 DCIS components of mixed DCIS/invasive breast cancer (IBC) tumors, 157 IBC components of mixed DCIS/IBC and 50 normal epithelial terminal ductal lobular units (TDLUs). In 726 IBCs, CD8+, FOXP3+, IL17+, PDL1+, PD1+ T-cell infiltration (TILs) were investigated in RECQL deficient and proficient cancers. Tumor mutation burden (TMB) was evaluated in five RECQL germ-line mutation carriers with IBC by genome sequencing. Compared with normal epithelial cells, a striking reduction in nuclear RECQL in DCIS was evident with aggressive pathology and poor survival. In RECQL deficient IBCs, CD8+, FOXP3+, IL17+ or PDL1+ TILs were linked with aggressive pathology and shorter survival. In germline RECQL mutation carriers, increased TMB was observed in 4/5 tumors. We conclude that RECQL loss is an early event in breast cancer and promote immune cell infiltration.

Selective Killing of BRCA2-Deficient Ovarian Cancer Cells via MRE11 Blockade.

The MRE11 nuclease is essential during DNA damage recognition, homologous recombination, and replication. BRCA2 plays important roles during homologous recombination and replication. Here, we show that effecting an MRE11 blockade using a prototypical inhibitor (Mirin) induces synthetic lethality (SL) in BRCA2-deficient ovarian cancer cells, HeLa cells, and 3D spheroids compared to BRCA2-proficient controls. Increased cytotoxicity was associated with double-strand break accumulation, S-phase cell cycle arrest, and increased apoptosis. An in silico analysis revealed Mirin docking onto the active site of MRE11. While Mirin sensitises DT40 MRE11+/- cells to the Top1 poison SN-38, it does not sensitise nuclease-dead MRE11 cells to this compound confirming that Mirin specifically inhibits Mre11 nuclease activity. MRE11 knockdown reduced cell viability in BRCA2-deficient PEO1 cells but not in BRCA2-proficient PEO4 cells. In a Mirin-resistant model, we show the downregulation of 53BP1 and DNA repair upregulation, leading to resistance, including in in vivo xenograft models. In a clinical cohort of human ovarian tumours, low levels of BRCA2 expression with high levels of MRE11 co-expression were linked with worse progression-free survival (PFS) (p = 0.005) and overall survival (OS) (p = 0.001). We conclude that MRE11 is an attractive SL target, and the pharmaceutical development of MRE11 inhibitors for precision oncology therapeutics may be of clinical benefit.

Atypical Presentation of Erythema Nodosum Following Pfizer-BioNTech COVID-19 Vaccine.

Background: Erythema nodosum (EN) is a common form of panniculitis that could be triggered by numerous conditions including infectious and non-infectious conditions. So far, few cases of EN caused by COVID-19 vaccine had been reported. Case Report: We report a case of atypical presentation of EN mimicking cellulitis in a patient who received the first dose of the Pfizer-BioNTech COVID-19 vaccine. A 38-year-old healthy woman who developed painful swelling on the left leg one week after receiving the first dose of Pfizer-BioNTech COVID-19 vaccine. Skin biopsy was revealed septal panniculitis. Due to the temporal association and the absence of other identifiable causes, Pfizer-BioNTech COVID-19 vaccine-related EN would be the most likely explanation. Conclusion: COVID-19 vaccines could be associated with rare side effects that should be reported for a better understanding of related outcomes of COVID-19 vaccination. This case was reported to keep in mind that EN can have atypical presentation as a rare side effect of COVID-19 vaccines.

Stereomicroscopic Evaluation of Marginal Fit of E.Max Press and E.Max Computer-Aided Design and Computer-Assisted Manufacturing Lithium Disilicate Ceramic Crowns: An In vitro Study.

OBJECTIVE: The purpose of this study was to compare the marginal gap of E-max press, and E.max computer-aided design and computer-assisted manufacturing (CAD-CAM) lithium disilicate (LD) ceramic crowns fabricated by using conventional technique and CAD-CAM technique. MATERIALS AND METHODS: This was an in vitro experimental study carried out in Riyadh Elm University and King Saud University. A marginal gap of 30 LD crowns was evaluated by Stereomicroscopy. A total of 15 pressable LD (IPS E.max Press [Ivoclar Vivadent]) ceramic crowns were fabricated by using conventional lost wax pattern method (Group A). Digital impressions of the prepared dies were scanned and transferred to the milling machine. IPS E.max CAD (IPS E-max, Ivoclar, Amherst, NY, USA) LD blocks in shade Vita A2 were then milled by using DWX-50 machine for CAD-CAM crowns (Group B). Descriptive statistics of mean and standard error of marginal gaps for both groups were recorded and compared by applying Mann-Whitney U-test. All the data were analyzed by using statistical analysis software SPSS version 21.0 (Armonk, NY, USA: IBM Corp). RESULTS: The LD crowns prepared by CAD-CAM technology (26.80 ± 3.4 µm) had significantly lower (P < 0.001) marginal gap than the LD pressed crowns (38.8 ± 2.3 µm) fabricated by conventional technique. The marginal gaps between CAD-CAM versus conventional groups exhibited significant differences at (42.68 µm vs. 52.46 µm, U = 51.500, P = 0.011), Mesiobuccal (15.94 µm vs. 30.13 µm, U = 45.500, P = 0.005), distolingual (26.70 µm vs. 43.86 µm, U = 63.500, P = 0.042), and distal (12.38 µm vs 31.45 µm, U = 47.500, P = 0.006). CONCLUSIONS: Within the limitations of the study, it can be concluded that LD all ceramic crowns fabricated by using CAD-CAM techniques showed lesser marginal gap and better marginal fit compared to the conventional technique.