A5057332386- Sarcoma Diagnosis and Treatment
- Musculoskeletal synovial abnormalities and treatments
- Soft tissue tumors and treatment
- Amyotrophic Lateral Sclerosis Research
- Neurogenetic and Muscular Disorders Research
- Healthcare and Environmental Waste Management
- Cellular transport and secretion
- Nanoparticles: synthesis and applications
- Protein Degradation and Inhibitors
- Cell Image Analysis Techniques
- Bone Metabolism and Diseases
- interferon and immune responses
- Parkinson's Disease Mechanisms and Treatments
- Epigenetics and DNA Methylation
- Endoplasmic Reticulum Stress and Disease
- Autophagy in Disease and Therapy
- Renin-Angiotensin System Studies
- Lysosomal Storage Disorders Research
- Silicon Carbide Semiconductor Technologies
- Folate and B Vitamins Research
- Connective tissue disorders research
- Neonatal Respiratory Health Research
- Immunotoxicology and immune responses
- Trace Elements in Health
- Congenital limb and hand anomalies
Institut Gustave Roussy
2024
St. Jude Children's Research Hospital
2024
University College Dublin
2017-2020
University of Chieti-Pescara
2013-2014
Mario Negri Sud Foundation
2013
Universitat de Barcelona
2002-2013
Mario Negri Institute for Pharmacological Research
2005
The Sar1 GTPase coordinates the assembly of coat protein complex-II (COPII) at specific sites endoplasmic reticulum (ER). COPII is required for ER-to-Golgi transport, as it provides a structural and functional framework to ship out cargoes produced in ER. To investigate requirement COPII-mediated transport mammalian cells, we used small interfering RNA (siRNA)-mediated depletion Sar1A Sar1B. We report that these two forms disrupts cells fail organize transitional elements coordinate...
MicroRNAs that are overexpressed in cystic fibrosis (CF) bronchial epithelial cells (BEC) negatively regulate CFTR and nullify the beneficial effects of modulators. We hypothesized it is possible to reverse microRNA-mediated inhibition using CFTR-specific target site blockers (TSBs) develop a drug-device combination inhalation therapy for CF. Lead microRNA expression was quantified series human CF non-CF samples vitro models. A panel 3′ untranslated region (UTR)-specific locked nucleic acid...
<p>EWS–WT1 drives enhanced DNA replication stress and R-loops, which contribute to DSRCT cells’ sensitivity PARPi ATRi. <b>A,</b> Assessment of fork speed (kb/minute) in JN1 cells subjected siRNA-mediated silencing EWS–WT1 or CCND1. A minimum 50 forks was analyzed per condition. Mean ± SD; each dot represents a single fork; <i>n</i> = 2, one-way ANOVA <i>post hoc</i> Dunnett test. <b>B,</b> exposed DMSO control, combination talazoparib...
<p>PARPi and ATRi have synergistic cytotoxic effects in models of DSRCT with high PARP1 expression. <b>A</b> <b>B,</b> expression (<b>A</b>) PARylation levels (<b>B</b>) as assessed by IHC a cohort 16 samples, compared those the JN1 R cell lines (PARP1 PAR are shown H-scores). Representative cases (PARP1-high vs. PARP1-low tumors; PAR-high PAR-low tumors) to right, cells. <b>C</b> <b>D,</b> Surface plots Bliss...
<p>EWS–WT1 is a determinant of DSRCT cells’ sensitivity to PARPi and ATRi. <b>A,</b> Western blot EWS–WT1 in JN1 R cells transfected with either siCNTRL or siEWS–WT1. Whole-cell lysates were generated 48 hours after transfection. <b>B–E,</b> Dose–response survival curves exposed talazoparib (<b>B</b> <b>C</b>) ATRi M4344 (<b>D</b> <b>E</b>) for 7 days the presence absence siRNA-mediated silencing EWS–WT1. Mean ±...
<div>Abstract<p>Desmoplastic small round cell tumor (DSRCT) is an aggressive sarcoma subtype that driven by the EWS–WT1 chimeric transcription factor. The prognosis for DSRCT poor, and major advances in treating have not occurred over two decades. To identify effective therapeutic approaches to target DSRCT, we conducted a high-throughput drug sensitivity screen line assessing chemosensitivity profiles 79 small-molecule inhibitors. cells were sensitive PARP inhibitors (PARPi)...
<p>The combination of PARPi and ATRi elicits a cGAS–STING–mediated cell-autonomous immune response. <b>A,</b> Western blots pTBK1, TBK, pIRF3, IRF3 in JN1 cells exposed to DMSO control, talazoparib (Tala), M4344, or both for 72 hours. <b>B</b> <b>C,</b> RT-qPCR analysis RNA isolated from talazoparib, CCL5 (<b>B</b>) CXCL10 (<b>C</b>) mRNA were analyzed separately relative RPLP0. Box whisker plots show arbitrary units gene...
<p>Supplementary File containing methods, figures and uncropped membranes.</p>
<p>PARPi and ATRi combination elicits DNA damage, replication stress, genomic instability in DSRCT cells. <b>A–D,</b> Quantification of γH2AX (<b>A</b> <b>B</b>) or RAD51 foci (<b>C</b> <b>D</b>) JN1 <b>C</b>) R (<b>B</b> cells exposed to DMSO control, PARPi talazoparib (Tala), M4344, a both for 72 hours. Cisplatin was used as the positive control. A minimum 500 nuclei analyzed per condition. Violin...
<p>A small-molecule inhibitor and drug screen identifies PARPi ATRi as candidate therapies for DSRCT. <b>A,</b> Schematic illustration of the workflow performed on JN1 cell line. <b>B,</b> Waterfall plot displaying difference in AUC between line (AUC<sub>JN1</sub>) panel 92 lines used comparison (AUC<sub>median</sub>) 79 evaluated inhibitors or drugs. Red, PARPi; blue, ATRi; green, conventional cytotoxic. <b>C–F,</b>...
There is a high demand for advanced, image-based, automated high-content screening (HCS) approaches to facilitate phenotypic in 3D cell culture models. A major challenge lies retaining the resolution of fine cellular detail but at same time imaging multicellular structures large scale. In this study, confocal microscopy-based HCS platform optical multiwell plates that enables quantitative morphological profiling populations nonuniform spheroids obtained from HT-29 human colorectal cancer...
Human amniotic fluid mesenchymal stem cells (hAFMSCs) are promising for therapeutic applications in bone damage. Calcium sensing receptor (CaSR), a G protein-coupled receptor, plays physiological role the regulation of metabolism. Thus, CaSR could be targeted by calcimimetic agonists, which may potentially helpful treating diseases. The aim our study was to characterize expression hAFMSCs and assess activity R-568 during vitro osteogenesis. Using western blotting, immunofluorescence, flow...
Abstract Desmoplastic small round cell tumor (DSRCT) is an aggressive sarcoma subtype that driven by the EWS-WT1 chimeric transcription factor. The prognosis for DSRCT poor, and major advances in treating DSCRT have not occurred over two decades. To identify effective therapeutic approaches to target DSRCT, we conducted a high-throughput drug sensitivity screen line assessing chemosensitivity profiles 79 small-molecule inhibitors. cells were sensitive PARP ATR inhibitors (PARPi, ATRi), as...
Abstract Through the nitric oxide (NO) production in vascular system, endothelial synthase (eNOS or NOS3) is a key enzyme blood pressure regulation and atherosclerosis control. Several previous studies have suggested an important role of eNOS as genetic risk factor for cardiovascular diseases. In this context, association study was carried out between two polymorphisms (the ecNOS4a/b VNTR G894T substitution) sample 101 nuclear families having one affected offspring ischemic heart disease...
Careful handling of the nanomaterials (NMs) in research labs is crucial to ensure a safe working environment. As largest university Ireland, University College Dublin (UCD) has invested significant resources update researchers with NMs. Due sizes often <100 nm, NMs including nanoparticles, harbor unprecedented materialistic properties, for example, enhanced reactivity, conductivity, fluorescence, etc. which albeit conferring an edge over bulk materials regarding applied aspects; depending on...
The effect of the C677T polymorphism in methylenetetrahydrofolate reductase (MTHFR) gene, traditionally associated with ischaemic heart disease (IHD), was assessed a Spanish population. transmission disequilibrium test (TDT) used to determine possible association sample 101 trios IHD patients. distribution MTHFR genotypes similar subjects and parental group; TT genotype present 14.9% patients, as compared 15.2% parents. frequency T allele also cases parents (39.6% vs. 42.4%; p = 0.649). TDT...
Cytochrome P450 3A5 (CYP3A5) has been proposed as a predictor of therapy response in subtypes pancreatic ductal adenocarcinoma cancer (PDAC). To validate CYP3A5 therapeutic target, we developed high-content image organoid-based screen to quantify the phenotypic responses selective inhibition enzymatic activity by clobetasol propionate (CBZ), using cohort PDAC-derived organoids (PDACOs). The chemoresistance PDACOs panel standard-of-care drugs, alone or combination with CBZ, was investigated....
In article number 1902033, Meritxell B. Cutrona and Jeremy C. Simpson describe a high-throughput automated confocal microscopy platform which allows for the generation of multiple levels quantitative information with respect to how nanoparticles interact cells. These interactions can be measured at subcellular level, cell spheroid level population level.