Current Issue : April - June Volume : 2017 Issue Number : 2 Articles : 5 Articles
Macrophage migration inhibitory factor (MIF) plays an important role in supporting the proliferation and/or survival\nof murine neural stem/progenitor cells (NSPCs); however, the downstream effectors of this factor remain unknown.\nHere, we show that MIF increases the expression of Pax6 and Chd7 in NSPCs in vitro. During neural development,\nthe chromatin remodeling factor Chd7 (chromatin helicase-DNA-binding protein 7) is expressed in the ventricular zone\nof the telencephalon of mouse brain at embryonic day 14.5, as well as in cultured NSPCs. Retroviral overexpression of\nPax6 in NSPCs increased Chd7 gene expression. Lentivirally-expressed Chd7 shRNA suppressed cell proliferation and\nneurosphere formation, and inhibited neurogenesis in vitro, while decreasing gene expression of Hes5 and N-myc. In\naddition, CHD7 overexpression increased cell proliferation in human embryonic stem cell-derived NSPCs (ES-NSPCs). In\nChd7 mutant fetal mouse brains, there were fewer intermediate progenitor cells (IPCs) compared to wildtype\nlittermates, indicating that Chd7 contributes to neurogenesis in the early developmental mouse brain. Furthermore, in\nsilico database analysis showed that, among members of the CHD family, CHD7 is highly expressed in human gliomas.\nInterestingly, high levels of CHD7 gene expression in human glioma initiating cells (GICs) compared to normal\nastrocytes were revealed and gene silencing of CHD7 decreased GIC proliferation. Collectively, our data demonstrate\nthat CHD7 is an important factor in the proliferation and stemness maintenance of NSPCs, and CHD7 is a promising\ntherapeutic target for the treatment of gliomas....
Background: The switch from oxidative phosphorylation to glycolysis in proliferating cancer cells, even under aerobic\nconditions, has been shown first in 1926 by Otto Warburg. Today this phenomenon is known as the ââ?¬Å?Warburg effectââ?¬Â and\nrecognized as\na hallmark of cancer. The metabolic shift to glycolysis is associated with the alterations in signaling pathways involved\nin energy metabolism, including glucose uptake and fermentation, and regulation of mitochondrial functions.\nHexokinases (HKs), which catalyze the first step of glycolysis, have been identified to play a role in tumorigenesis of\nhuman colorectal cancer (CRC) and melanoma. However, the mechanism of action of HKs in the promotion of tumor\ngrowth remains unclear.\nResults: The purpose of the present study was to investigate the effect of silencing of hexokinase genes (HK1, HK2, and\nHK3) in colorectal cancer (HT-29, SW 480, HCT-15, RKO, and HCT 116) and melanoma (MDA-MB-435S and SK-MEL-28) cell\nlines using short hairpin RNA (shRNA) lentiviral vectors. shRNA lentiviral plasmid vectors pLSLP-HK1, pLSLP-HK2, and\npLSLP-HK3 were constructed and then transfected separately or co-transfected into the cells. HK2 inactivation was\nassociated with increased expression of HK1 in colorectal cancer cell lines pointing to the compensation effect.\nSimultaneous attenuation of HK1 and HK2 levels led to decreased cell viability. Co-transfection with shRNA vectors against\nHK1, HK2, and HK3 mRNAs resulted in a rapid cell death via apoptosis.\nConclusions: We have demonstrated that simultaneous inactivation of HK1 and HK2 was sufficient to decrease\nproliferation and viability of melanoma and colorectal cancer cells. Our results suggest that HK1 and HK2 could be the key\ntherapeutic targets for reducing aerobic glycolysis in examined cancers....
Gene therapy holds a promising future for bridging the gap between the disciplines of\nmedicine and clinical dentistry. The dynamic treatment approaches of gene therapy have been\nadvancing by leaps and bounds. They are transforming the conventional approaches into more\nprecise and preventive ones that may limit the need of using drugs and surgery. The oral cavity is\none of the most accessible areas for the clinical applications of gene therapy for various oral tissues.\nThe idea of genetic engineering has become more exciting due to its advantages over other treatment\nmodalities. For instance, the body is neither subjected to an invasive surgery nor deep wounds,\nnor is it susceptible to systemic effects of drugs. The aim of this article is to review the gene therapy\napplications in the field of dentistry. In addition, therapeutic benefits in terms of treatment of diseases,\nminimal invasion and maximum outcomes have been discussed....
Gene transfer to airway epithelial cells is hampered by extracellular (mainly mucus) and\ncellular (tight junctions) barriers. Magnetofection has been used to increase retention time of lentiviral\nvectors (LV) on the cellular surface. In this study, magnetofection was investigated in airway epithelial\ncell models mimicking extracellular and cellular barriers. Bronchiolar epithelial cells (H441 line)\nwere evaluated for LV-mediated transduction after polarization onto filters and dexamethasone (dex)\ntreatment, which induced hemicyst formation, with or without magnetofection. Sputum from cystic\nfibrosis (CF) patients was overlaid onto cells, and LV-mediated transduction was evaluated in the\nabsence or presence of magnetofection. Magnetofection of unpolarized H441 cells increased the\ntransduction with 50 MOI (multiplicity of infection, i.e., transducing units/cell) up to the transduction\nobtained with 500 MOI in the absence of magnetofection. Magnetofection well-enhanced LV-mediated\ntransduction in mucus-layered cells by 20.3-fold. LV-mediated transduction efficiency decreased\nin dex-induced hemicysts in a time-dependent fashion. In dome-forming cells, zonula occludens-1\n(ZO-1) localization at the cell borders was increased by dex treatment. Under these experimental\nconditions, magnetofection significantly increased LV transduction by 5.3-fold. In conclusion,\nthese results show that magnetofection can enhance LV-mediated gene transfer into airway epithelial\ncells in the presence of extracellular (sputum) and cellular (tight junctions) barriers, representing\nCF-like conditions....
Haematopoietic stem cell (HSC) gene therapy has demonstrated potential to treat many\ndiseases. However, current state of the art requires sophisticated ex vivo gene transfer in a\ndedicated Good Manufacturing Practices facility, limiting availability. An automated process\nwould improve the availability and standardized manufacture of HSC gene therapy. Here, we\ndevelop a novel program for semi-automated cell isolation and culture equipment to permit\ncomplete benchtop generation of gene-modified CD34�¾ blood cell products for transplantation.\nThese cell products meet current manufacturing quality standards for both mobilized\nleukapheresis and bone marrow, and reconstitute human haematopoiesis in immunocompromised\nmice. Importantly, nonhuman primate autologous gene-modified CD34�¾ cell\nproducts are capable of stable, polyclonal multilineage reconstitution with follow-up of more\nthan 1 year. These data demonstrate proof of concept for point-of-care delivery of HSC gene\ntherapy. Given the many target diseases for gene therapy, there is enormous potential for this\napproach to treat patients on a global scale....
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