Sustained Disease Control in DME Patients upon Treatment Cessation with Brolucizumab.

Background: Treatment cessation due to a dry retina has not been systematically addressed in diabetic macular edema (DME). In three out of four patients receiving 6 mg of brolucizumab in the KITE study, treatment was terminated after the study ended. Methods: The KITE study was a double-masked, multicenter, active-controlled, randomized trial (NCT03481660) in DME patients. Per protocol, patients received five loading injections of Brolucizumab at 6-week intervals, with the option to adjust to 8 weeks in case of disease activity or to extend in the second year to a maximum of 16 weeks in the absence of retinal fluid. Results: After two years, one patient required eight weekly injections, while three patients reached a maximal treatment interval of 16 weeks. The severity of diabetic retinopathy improved in all patients with no dye leakage according to fluorescein angiography (FA) and no retinal fluid according to OCT in three patients. Treatment was paused in these three patients for >36 months, while the fourth patient required continuous treatment at 5-week intervals after switching to other licensed anti-VEGF agents. Conclusions: The adoption of treatment according to individual needs, including considering treatment cessation, may contribute to improved treatment adherence in many patients and be more frequently possible than expected

Targeted gene expression profile reveals CDK4 as therapeutic target for selected patients with adrenocortical carcinoma

Adrenocortical carcinomas (ACC) are aggressive tumors with a heterogeneous prognosis and limited therapeutic options for advanced stages. This study aims to identify novel drug targets for a personalized treatment in ACC. RNA was isolated from 40 formalin-fixed paraffin-embedded ACC samples. We evaluated gene expression of 84 known cancer drug targets by reverse transcriptase quantitative real time-PCR and calculated fold change using 5 normal adrenal glands as reference (overexpression by fold change >2.0). The most promising candidate cyclin-dependent kinase 4 (CDK4) was investigated at protein level in 104 ACC samples and tested by in vitro experiments in two ACC cell lines (NCI-H295R and MUC1). The most frequently overexpressed genes were TOP2A (100% of cases, median fold change = 16.5), IGF2 (95%, fold change = 52.9), CDK1 (80%, fold change = 6.7), CDK4 (62%, fold change = 2.6), PLK4 (60%, fold change = 2.8), and PLK1 (52%, fold change = 2.3). CDK4 was chosen for functional validation, as it is actionable by approved CDK4/6-inhibitors (e.g., palbociclib). Nuclear immunostaining of CDK4 significantly correlated with mRNA expression (R = 0.52, P < 0.005). We exposed both NCI-H295R and MUC1 cell lines to palbociclib and found a concentration- and time-dependent reduction of cell viability, which was more pronounced in the NCI-H295R cells in line with higher CDK4 expression. Furthermore, we tested palbociclib in combination with insulin-like growth factor 1/insulin receptor inhibitor linsitinib showing an additive effect. In conclusion, we demonstrate that RNA profiling is useful to discover potential drug targets and that CDK4/6 inhibitors are promising candidates for treatment of selected patients with ACC

Circulating cell-free DNA-based biomarkers for prognostication and disease monitoring in adrenocortical carcinoma

Adrenocortical carcinoma (ACC) is a rare aggressive cancer with heterogeneous behaviour. Disease surveillance relies on frequent imaging, which comes with significant radiation exposure. The aim of the study was to investigate the role of circulating cell-free DNA (ccfDNA)-related biomarkers (BM) for prognostication and monitoring of ACC.We investigated 34 patients with ACC and 23 healthy subjects (HS) as controls. ccfDNA was extracted by commercial kits and ccfDNA concentrations quantified by fluorimeter (BM1). Targeted sequencing was performed using a customised panel of 27 ACC-specific genes. Leucocyte DNA was used to discriminate somatic variants (BM2), while tumour DNA was sequenced in 22/34 cases for comparison. Serial ccfDNA samples were collected during follow-up in 19 ACC patients (median period 9 months) and analysed in relationship with standard radiological imaging.ccfDNA concentrations were higher in ACC than HS (mean±SD, 1.15±1.56 vs 0.05±0.05 ng/µl, P&lt;0.0001), 96% of them being above the cut-off of 0.146 ng/µl (mean HS+2SD, positive BM1). At ccfDNA sequencing, 47% of ACC showed at least one somatic mutation (positive BM2). A combined ccfDNA-BM score was strongly associated with both progression-free and overall survival (HR=2.63, 95%CI=1.13-6.13, P=0.010, and HR=5.98, 95%CI=2.29-15.6, P=0.0001, respectively). During disease monitoring, positive BM2 showed the best specificity (100%) and sensitivity (67%) to detect ACC recurrence or progress compared to BM1.In conclusion, ccfDNA-related BMs are frequently detected in ACC patients and represent a promising, minimally invasive tool to predict clinical outcome and complement surveillance imaging. Our findings will be validated in a larger cohort of ACCs with long-term follow-up

PLK1 inhibitors as a new targeted treatment for adrenocortical carcinoma

Adrenocortical carcinoma (ACC) is an aggressive malignancy with limited treatment options. Polo-like kinase 1 (PLK1) is a promising drug target; PLK1 inhibitors (PLK1i) have been investigated in solid cancers and are more effective in TP53-mutated cases. We evaluated PLK1 expression in ACC samples and the efficacy of two PLK1i in ACC cell lines with different genetic backgrounds. PLK1 protein expression was investigated by immunohistochemistry in tissue samples and correlated with clinical data. The efficacy of rigosertib (RGS), targeting RAS/PI3K, CDKs and PLKs, and poloxin (Pol), specifically targeting the PLK1 polo-box domain, was tested in TP53-mutated NCI-H295R, MUC-1, and CU-ACC2 cells and in TP53 wild-type CU-ACC1. Effects on proliferation, apoptosis, and viability were determined. PLK1 immunostaining was stronger in TP53-mutated ACC samples vs wild-type (P = 0.0017). High PLK1 expression together with TP53 mutations correlated with shorter progression-free survival (P= 0.041). NCI-H295R showed a time- and dose-dependent reduction in proliferation with both PLK1i (P< 0.05at 100 nM RGS and 30 µM Pol). In MUC-1, a less pronounced decrease was observed (P< 0.05at 1000 nM RGS and 100 µM Pol). 100 nM RGS increased apoptosis in NCI-H295R (P< 0.001), with no effect on MUC-1. CU-ACC2 apoptosis was induced only at high concentrations (P < 0.05 at 3000 nM RGS and 100 µM Pol), while proliferation decreased at 1000 nM RGS and 30 µM Pol. CU-ACC1 proliferation reduced, and apoptosis increased, only at 100 µM Pol. TP53-mutated ACC cell lines demonstrated better response to PLK1i than wild-type CU-ACC1. These data suggest PLK1i may be a promising targeted treatment of a subset of ACC patients, pre-selected according to tumour genetic signature

Targeted molecular analysis in adrenocortical carcinomas: a strategy towards improved personalized prognostication

Context: Adrenocortical carcinoma (ACC) has a heterogeneous prognosis, and current medical therapies have limited efficacy in its advanced stages. Genome-wide multiomics studies identified molecular patterns associated with clinical outcome. Objective: Here, we aimed at identifying a molecular signature useful for both personalized prognostic stratification and druggable targets, using methods applicable in clinical routine. Design: In total, 117 tumor samples from 107 patients with ACC were analyzed. Targeted next-generation sequencing of 160 genes and pyrosequencing of 4 genes were applied to formalin-fixed, paraffin-embedded (FFPE) specimens to detect point mutations, copy number alterations, and promoter region methylation. Molecular results were combined with clinical/histopathological parameters (tumor stage, age, symptoms, resection status, and Ki-67) to predict progression-free survival (PFS). Results: In addition to known driver mutations, we detected recurrent alterations in genes not previously associated with ACC (e.g., NOTCH1, CIC, KDM6A, BRCA1, BRCA2). Best prediction of PFS was obtained integrating molecular results (more than one somatic mutation, alterations in Wnt/beta-catenin and p53 pathways, high methylation pattern) and clinical/histopathological parameters into a combined score (P <0.0001, chi(2) = 68.6). Accuracy of prediction for early disease progress was 83.3% (area under the receiver operating characteristic curve: 0.872, 95% confidence interval 0.80 to 0.94). Furthermore, 17 potentially targetable alterations were found in 64 patients (e.g., inCDK4, NOTCH1, NF1, MDM2, and EGFR and in DNA repair system). Conclusions: This study demonstrates that molecular profiling of FFPE tumor samples improves prognostication of ACC beyond clinical/histopathological parameters and identifies new potential drug targets. These findings pave the way to precision medicine in this rare disease

Learning analytics to improve writing skills for young children – an holistic approach

Purpose - Due to the important role of orthography in society, the project called IDeRBlog presented in this paper created a web-based tool to motivate pupils to write text as well as to read and to comment on texts written by fellow students. In addition, IDeRBlog aims to improve student’s German orthography skills and supports teachers and parents with training materials for their students. The paper aims to discuss these issues. Design/methodology/approach - With the aid of learning analytics, the submitted text is analyzed and special feedback is given to the students so that they can try to correct the misspelled words themselves. The teachers as well as the parents are benefiting from the analysis and exercises suggested by the system. Findings - A recent study showed the efficiency of the system in form of an improvement of the students’ orthographic skills. Over a period of four months 70 percent of the students achieved a significant reduction of their spelling mistakes. Originality/value - IDeRBlog is an innovative approach to improving orthography skills combining blogging and new media with writing and practice

Emotional Speech Perception Unfolding in Time: The Role of the Basal Ganglia

The basal ganglia (BG) have repeatedly been linked to emotional speech processing in studies involving patients with neurodegenerative and structural changes of the BG. However, the majority of previous studies did not consider that (i) emotional speech processing entails multiple processing steps, and the possibility that (ii) the BG may engage in one rather than the other of these processing steps. In the present study we investigate three different stages of emotional speech processing (emotional salience detection, meaning-related processing, and identification) in the same patient group to verify whether lesions to the BG affect these stages in a qualitatively different manner. Specifically, we explore early implicit emotional speech processing (probe verification) in an ERP experiment followed by an explicit behavioral emotional recognition task. In both experiments, participants listened to emotional sentences expressing one of four emotions (anger, fear, disgust, happiness) or neutral sentences. In line with previous evidence patients and healthy controls show differentiation of emotional and neutral sentences in the P200 component (emotional salience detection) and a following negative-going brain wave (meaning-related processing). However, the behavioral recognition (identification stage) of emotional sentences was impaired in BG patients, but not in healthy controls. The current data provide further support that the BG are involved in late, explicit rather than early emotional speech processing stages

Functional Outcome After Macular Hole Surgery: Comparison of Standard Perimetry with Microperimetry.

Purpose To compare the predictive value of macular perimetry and microperimetry for visual outcomes after vitrectomy with internal limiting membrane (ILM) peeling in full-thickness macular holes (MH). Methods This retrospective, non-randomized case series refers to 100 eyes undergoing vitrectomy with ILM peeling. Best-corrected visual acuity (BCVA), standard 12° perimetry and microperimetry were perioperatively recorded. A possible predictive value of the preoperative findings on postoperative visual function (PVF) was assessed. Results Independent of the preoperative minimal MH size (range: 55-752 μm), all 100 MHs were closed. BCVA improved from 56.3 ± 12.8 to 74.8 ± 9.2 Early Treatment of Diabetic Retinopathy Study (ETDRS) letters after six months and retinal fixation stability enhanced. We found a positive correlation between BCVA and macular sensitivity 6 months postoperatively in microperimetry (r = 0.48, p < 0.010) and 12° perimetry (r = 0.45, p < 0.014), as well as with mean defect (r = 0.48, p < 0.01 and r = 0.44, p < 0.017, respectively). A correlation between preoperative visual function indices and PVF was not established. Conclusion Microperimetry and standard perimetry are equally suitable for describing perioperative retinal function in idiopathic MH. While the indices of both methods correlate comparably well with BCVA, they cannot predict PVF. This may be partially explained by the area covered by perimetry, compared to which the size of the MH is of inferior relevance

Ethylhexylglycerin impairs membrane integrity and enhances the lethal effect of phenoxyethanol

submittedVersio