Project description:We use a human whole genome microarray to analyze the effects of nanosecond pulsed electric fields on Jurkat cells with the focus on early response genes to DNA damage. Keywords: nanosecond pulsed electric fields, jurkat cells, DNA damage
Project description:Glioblastoma multiforme (GBM), is the most common form of adult malignant brain tumor, highly heterogeneous and relatively resistant to therapy with a poor prognosis. As other cancers, GBM starts from a relatively small fraction of poorly differentiated and particularly aggressive cancer stem cells (CSCs), responsible for aberrant proliferation and invasion, rapidly spreading the tumor growth in the other parts of the central nervous system. Due to the extreme tumor heterogeneity, standard therapies (i.e., surgery, chemotherapy and radiotherapy) provide poor positive outcomes and cancers usually recur. Therefore, alternative approaches, possibly targeting CSCs, are necessary for searching effective therapeutic strategies against GBM. Among many emerging new therapeutic strategies, ultra-short pulsed electric fields (PEF) are considered extremely promising in cancer therapy and our previous results demonstrated the capability of a specific electric pulse protocol, characterized by a high pulse amplitude and a short duration, to selectively affect medulloblastoma CSCs preserving normal cells. Here, we tested the same exposure protocol to investigate the response of U87 GBM cells, and of U87-derived neurospheres. By analyzing different in vitro biological endpoints and taking advantage of transcriptomic and bioinformatics analyses, we found that, independently of CSCs content, PEF exposure affected cell proliferation and differentially regulated hypoxia, inflammation and P53/cell cycle checkpoints. PEF exposure also significantly reduced the clonogenic potential, reducing the ability to form new neurospheres, and inhibited the invasion potential. Importantly, exclusively in U87 neurospheres, PEF exposure changed the expression of stemness/differentiation genes. Our results, confirm this physical stimulus as a promising treatment to destabilize the extreme intra-tumoral GBM heterogeneity opening up the possibility to develop future therapeutic strategies mediated by PEF exposure.
Project description:Catheter ablation is an effective treatment to prevent recurrence of Atrial fibrillation (AF) and can be used to maintain sinus rhythm and improve symptoms of AF, but to some extent it can cause a range of adverse effects associated with catheter ablation. Pulsed electric field is a newer treatment modality to replace catheter ablation for atrial fibrillation due to its fewer side effects. Different from radiofrequency ablation, which destroys diseased myocardial tissue by thermal energy, pulsed electric field ablation achieves the purpose of atrial fibrillation ablation by inducing damage to diseased myocardial cells through irreversible electroporation. However, some experimental parameters and mechanism of pulsed electric fields remain unclear.
Project description:In the last few years, pulsed electric fields have emerged as promising clinical tools for tumor treatments. This study highlights the distinct impact of a specific pulsed electric field protocol named PEF-5 (0.3 MV/m, 40 µs, 5 pulses) on astrocytes (NHA), medulloblastoma (D283) and gli-oblastoma (U87 NS) cancer stem-like cells. We pursued this goal performing ultrastructural analyses corroborated by molecular/omics approaches to understand the vulnerability or re-sistance mechanisms triggered by PEF-5 exposure in the different cell types. Electron microscopic analyses showed that, independently of exposed cells, the main targets of PEF-5 were the cell membrane and the cytoskeleton, presenting indiscriminate filopodia disappearance on the cell surface accompanied by a rapid cell swelling. Nevertheless, cells activated different responses stemming from intracellular organelles. NHA triggered a protective mechanism to recover from the damaged membranes and eliminated the detrimental ROS to maintain cell vital functions. Conversely, PEF-5 induced a different action on mitochondria of cancer stem-like cells, more prominent on D283 compared to U87 NS cells, in correlation with a higher basal level of CD133 protein in D283 cells. In this case, a complete mitochondrial dysfunction was demonstrated, inhibiting the recovery of D283 vital processes, while a mild perturbation was observed in mito-chondria of U87 NS cells, not sufficient to impair their cell functions. Altogether, these results suggest the possibility to use PEF-based technology as a novel strategy to target selectively mi-tochondria of CSCs, preserving healthy cells. Ultrastructural analysis, supported by transcriptomic characterization in microsecond pulsed electric field exposed normal brain and brain tumor cells
Project description:In the last few years, pulsed electric fields have emerged as promising clinical tools for tumor treatments. This study highlights the distinct impact of a specific pulsed electric field protocol named PEF-5 (0.3 MV/m, 40 µs, 5 pulses) on astrocytes (NHA), medulloblastoma (D283) and gli-oblastoma (U87 NS) cancer stem-like cells. We pursued this goal performing ultrastructural analyses corroborated by molecular/omics approaches to understand the vulnerability or re-sistance mechanisms triggered by PEF-5 exposure in the different cell types. Electron microscopic analyses showed that, independently of exposed cells, the main targets of PEF-5 were the cell membrane and the cytoskeleton, presenting indiscriminate filopodia disappearance on the cell surface accompanied by a rapid cell swelling. Nevertheless, cells activated different responses stemming from intracellular organelles. NHA triggered a protective mechanism to recover from the damaged membranes and eliminated the detrimental ROS to maintain cell vital functions. Conversely, PEF-5 induced a different action on mitochondria of cancer stem-like cells, more prominent on D283 compared to U87 NS cells, in correlation with a higher basal level of CD133 protein in D283 cells. In this case, a complete mitochondrial dysfunction was demonstrated, inhibiting the recovery of D283 vital processes, while a mild perturbation was observed in mito-chondria of U87 NS cells, not sufficient to impair their cell functions. Altogether, these results suggest the possibility to use PEF-based technology as a novel strategy to target selectively mi-tochondria of CSCs, preserving healthy cells. Ultrastructural analysis, supported by transcriptomic characterization in microsecond pulsed electric field exposed normal brain and brain tumor cells.
Project description:The aim of our study was to analyze cell response to nanosecond pulsed electric field (nsPEF) at the gene expression level. TM3 Leydig cells were used as a model. Transcriptomics analysis was carried out immediately after exposure (0 h) and 4 or 24 h after treatment.
Project description:The scarcity of effective treatment options for high grade brain tumours has led to a wide ranging search for alternative means of therapy for these difficult to treat tumours. Electrical field therapy is one such area that has been considered. The OptuneTM system is an FDA approved novel anti-mitotic device that delivers continuous alternating electric fields to the patient for the treatment of primary and recurrent Glioblastoma multiforme (GBM) (tumor treating fields - TTFields). Alternative electric fields delivery systems are also being investigated for the treatment of various cancers.To further explore alternative potential mechanisms of electric fields as a whole, we ran Optune, DBS electric fields treated and control untreated KNS42, U87 and GIN-31 (primary) cell lines on Clariom S Human Assays to produce genome-wide expression data.