Current Issue : January - March Volume : 2018 Issue Number : 1 Articles : 5 Articles
Significant advances in gene therapy have enabled exploration of therapies for inherited\nretinal disorders, many of which are in preclinical development or clinical evaluation.\nGene therapy for retinal conditions has led the way in this growing field. The loss of\nretinal ganglion cells (RGCs) is a hallmark of a number of retinal disorders. As the\nfield matures innovations that aid in refining therapies and optimizing efficacy are in\ndemand. Gene therapies under development for RGC-related disorders, when delivered\nwith recombinant adeno associated vectors (AAV), have typically been expressed from\nubiquitous promoter sequences. Here we describe how a novel promoter from the murine\nNefh gene was selected to drive transgene expression in RGCs. The Nefh promoter, in\nan AAV2/2 vector, was shown to drive preferential EGFP expression in murine RGCs in\nvivo following intravitreal injection. In contrast, EGFP expression from a CMV promoter\nwas observed not only in RGCs, but throughout the inner nuclear layer and in amacrine\ncells located within the ganglion cell layer (GCL). Of note, the Nefh promoter sequence is\nsufficiently compact to be readily accommodated in AAV vectors, where transgene size\nrepresents a significant constraint. Moreover, this promoter should in principle provide a\nmore targeted and potentially safer alternative for RGC-directed gene therapies....
Low molecular weight polyethylenimine (1800 Da, also referred to as oligoethylenimines,\nOEI) was modified with amino acids, including two aromatic amino acids (tryptophan, phenylalanine)\nand an aliphatic amino acid (leucine). The substitution degree of amino acids could be controlled by\nadjusting the feeding mole ratio of the reactants. Fluorescence spectroscopy and circular dichroism\nexperiments demonstrated that the indole ring of tryptophan may intercalate into the DNA base\npairs and contribute to efficient DNA condensation. In vitro gene expression results revealed that\nthe modified OEIs (OEI-AAs) may provide higher transfection efficiency even than high molecular\nweight polyethylenimine (25 kDa, PEI), especially the aromatic tryptophan substituted OEI. Moreover,\nOEI-AAs exhibited excellent serum tolerance, and up to 137 times higher transfection efficiency than\nPEI 25 kDa that was obtained in the presence of serum. The cytotoxicity of OEI-AAs is much lower\nthan PEI 25 kDa. This study may afford a new method for the development of low molecular weight\noligomeric non-viral gene vectors with both high efficiency and biocompatibility....
Background: Next-generation sequencing (NGS) of cancer gene panels are widely applied to enable personalized\ncancer therapy and to identify novel oncogenic mutations.\nMethods: We performed targeted NGS on 932 clinical cases of non-small-cell lung cancers (NSCLCs) using the Ion\nAmpliSeqââ??¢ Cancer Hotspot panel v2 assay.\nResults: Actionable mutations were identified in 65% of the cases with available targeted therapeutic options,\nincluding 26% of the patients with mutations in National Comprehensive Cancer Network (NCCN) guideline genes.\nMost notably, we discovered JAK2 p.V617F somatic mutation, a hallmark of myeloproliferative neoplasms, in 1% (9/\n932) of the NSCLCs. Analysis of cancer cell line pharmacogenomic data showed that a high level of JAK2 expression\nin a panel of NSCLC cell lines is correlated with increased sensitivity to a selective JAK2 inhibitor. Further analysis of\nTCGA genomic data revealed JAK2 gain or loss due to genetic alterations in NSCLC clinical samples are associated\nwith significantly elevated or reduced PD-L1 expression, suggesting that the activating JAK2 p.V617F mutation\ncould confer sensitivity to both JAK inhibitors and anti-PD1 immunotherapy. We also detected JAK3 germline\nactivating mutations in 6.7% (62/932) of the patients who may benefit from anti-PD1 treatment, in light of recent\nfindings that JAK3 mutations upregulate PD-L1 expression.\nConclusion: Taken together, this study demonstrated the clinical utility of targeted NGS with a focused hotspot\ncancer gene panel in NSCLCs and identified activating mutations in JAK2 and JAK3 with clinical implications inferred\nthrough integrative analysis of cancer genetic, genomic, and pharmacogenomic data. The potential of JAK2 and\nJAK3 mutations as response markers for the targeted therapy against JAK kinases or anti-PD1 immunotherapy\nwarrants further investigation....
Coronary artery atherosclerosis is a chronic inflammatory disease. This study aimed to identify the key changes of\ngene expression between early and advanced carotid atherosclerotic plaque in human. Methods. Gene expression dataset GSE28829\nwas downloaded from Gene Expression Omnibus (GEO), including 16 advanced and 13 early stage atherosclerotic plaque samples\nfrom human carotid. Differentially expressed genes (DEGs) were analyzed. Results. 42,450 genes were obtained from the dataset.\nTop 100 up- and downregulated DEGs were listed. Functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes\n(KEGG) identification were performed.The result of functional and pathway enrichment analysis indicted that the immune system\nprocess played a critical role in the progression of carotid atherosclerotic plaque. Protein-protein interaction (PPI) networks\nwere performed either. Top 10 hub genes were identified from PPI network and top 6 modules were inferred. These genes were\nmainly involved in chemokine signaling pathway, cell cycle, B cell receptor signaling pathway, focal adhesion, and regulation of\nactin cytoskeleton. Conclusion. The present study indicated that analysis of DEGs would make a deeper understanding of the\nmolecular mechanisms of atherosclerosis development and they might be used as molecular targets and diagnostic biomarkers\nfor the treatment of atherosclerosis...
Epidemiological studies support an independent inverse association between high-density\nlipoprotein (HDL) cholesterol levels and heart failure incidence. The effect of selective HDL-raising\nadeno-associated viral serotype 8-human apolipoprotein (apo) A-I (AAV8-A-I) gene transfer on\ncardiac remodeling induced by transverse aortic constriction (TAC) was evaluated in C57BL/6\nlow-density lipoprotein receptor-deficient mice. Septal wall thickness and cardiomyocyte\ncross-sectional area were reduced by 16.5% (p < 0.001) and by 13.8% (p < 0.01), respectively,\neight weeks after TAC in AAV8-A-I mice (n = 24) compared to control mice (n = 39). Myocardial\ncapillary density was 1.11-fold (p < 0.05) higher and interstitial cardiac fibrosis was 45.3% (p < 0.001)\nlower in AAV8-A-I TAC mice than in control TAC mice. Lung weight and atrial weight were\nsignificantly increased in control TAC mice compared to control sham mice, but were not increased\nin AAV8-A-I TAC mice. The peak rate of isovolumetric contraction was 1.19-fold (p < 0.01) higher\nin AAV8-A-I TAC mice (n = 17) than in control TAC mice (n = 29). Diastolic function was also\nsignificantly enhanced in AAV8-A-I TAC mice compared to control TAC mice. Nitro-oxidative stress\nand apoptosis were significantly reduced in the myocardium of AAV8-A-I TAC mice compared\nto control TAC mice. In conclusion, selective HDL-raising human apo A-I gene transfer potently\ncounteracts the development of pressure overload-induced cardiomyopathy....
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