Current Issue : July - September Volume : 2017 Issue Number : 3 Articles : 6 Articles
Development of gene therapy vectors requires cellular models reflecting the genetic background of a disease thus allowing for robust preclinical vector testing. For human p47phox-deficient chronic granulomatous disease (CGD) vector testing we generated a cellular model using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 to introduce a GT-dinucleotide deletion (Ã?â?GT) mutation in p47phox encoding NCF1 gene in the human acute myeloid leukemia PLB-985 cell line. CGD is a group of hereditary immunodeficiencies characterized by impaired respiratory burst activity in phagocytes due to a defective phagocytic nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. In Western countries autosomal-recessive p47phox-subunit deficiency represents the second largest CGD patient cohort with unique genetics, as the vast majority of p47phox CGD patients carries Ã?â?GT deletion in exon two of the NCF1 gene. The established PLB-985 NCF1 Ã?â?GT cell line reflects the most frequent form of p47phox-deficient CGD genetically and functionally. It can be differentiated to granulocytes efficiently, what creates an attractive alternative to currently used iPSC models for rapid testing of novel gene therapy approaches....
Parkinson�s disease (PD) was characterized by late-onset, progressive dopamine neuron\nloss and movement disorders. The progresses of PD affected the neural function and\nintegrity. To date, most researches had largely addressed the dopamine replacement\ntherapies, but the appearance of L-dopa-induced dyskinesia hampered the use of the\ndrug. And the mechanism of PD is so complicated that it�s hard to solve the problem by\njust add drugs. Researchers began to focus on the genetic underpinnings of Parkinson�s\ndisease, searching for new method that may affect the neurodegeneration processes in\nit. In this paper, we reviewed current delivery methods used in gene therapies for PD,\nwe also summarized the primary target of the gene therapy in the treatment of PD, such\nlike neurotrophic factor (for regeneration), the synthesis of neurotransmitter (for prolong\nthe duration of L-dopa), and the potential proteins that might be a target to modulate\nvia gene therapy. Finally, we discussed RNA interference therapies used in Parkinson�s\ndisease, it might act as a new class of drug.We mainly focus on the efficiency and tooling\nfeatures of different gene therapies in the treatment of PD....
Based on the theoretical and clinical development of modern medicines, gene therapy has\nbeen a promising treatment strategy for cancer and other diseases. The practice of gene therapy\nis nearly 27 years old, since the first authorized gene transfer study took place at the National\nInstitute of Health in 1989. However, gene therapy was not readily adopted worldwide, until recently.\nSeveral gene therapy clinical trials have been carried out in China since 1998, and medical research\nin China has flourished. In this report, we review the history of gene therapy in China, focusing on\ntreatment protocol, the administration cycle, dosage calculation, and the evaluation of therapeutic\neffects, in order to provide more information for the additional development of this promising\ntreatment strategy....
Duchenne Muscular Dystrophy (DMD) is caused by a lack of dystrophin expression in patient muscle fibres. Current DMD gene therapy strategies rely on the expression of internally deleted forms of dystrophin, missing important functional domains. Viral gene transfer of full-length dystrophin could restore wild-type functionality, although this approach is restricted by the limited capacity of recombinant viral vectors. Lentiviral vectors can package larger transgenes than adeno-associated viruses, yet lentiviral vectors remain largely unexplored for full-length dystrophin delivery. In our work, we have demonstrated that lentiviral vectors can package and deliver inserts of a similar size to dystrophin. We report a novel approach for delivering large transgenes in lentiviruses, in which we demonstrate proof-of-concept for a 'template-switching' lentiviral vector that harnesses recombination events during reverse-transcription. During this work, we discovered that a standard, unmodified lentiviral vector was efficient in delivering full-length dystrophin to target cells, within a total genomic load of more than 15,000 base pairs. We have demonstrated gene therapy with this vector by restoring dystrophin expression in DMD myoblasts, where dystrophin was expressed at the sarcolemma of myotubes after myogenic differentiation. Ultimately, our work demonstrates proof-of-concept that lentiviruses can be used for permanent full-length dystrophin gene therapy, which presents a significant advancement in developing an effective treatment for DMD....
Despite improvements in drug and device therapy for heart failure, hospitalization rates and mortality have\nchanged little in the past decade. Randomized clinical trials using gene transfer to improve function of the\nfailing heart are the focus of this review. Four randomized clinical trials of gene transfer in heart failure with\nreduced ejection fraction (HFrEF) have been published. Each enrolled patients with stable symptomatic\nHFrEF and used either intracoronary delivery of a virus vector or endocardial injection of a plasmid. The\ninitial CUPID trial randomized 14 subjects to placebo and 25 subjects to escalating doses of adeno-associated\nvirus type 1 encoding sarcoplasmic reticulum calcium ATPase (AAV1.SERCA2a). AAV1.SERCA2a was well\ntolerated, and the high-dose group met a 6 month composite endpoint. In the subsequent CUPID-2 study, 243\nsubjects received either placebo or the high dose of AAV1.SERCA2a. AAV1.SERCA2a administration, while\nsafe, failed to meet the primary or any secondary endpoints. STOP-HF used plasmid endocardial injection of\nstromal cell-derived factor-1 to promote stem-cell recruitment. In a 93-subject trial of patients with ischemic\netiology heart failure, the primary endpoint (symptoms and 6min walk distance) failed, but subgroup analyses\nshowed improvements in subjects with the lowest ejection fractions. A fourth trial randomized 14\nsubjects to placebo and 42 subjects to escalating doses of adenovirus-5 encoding adenylyl cyclase 6 (Ad5.hAC6).\nThere were no safety concerns, and patients in the two highest dose groups (combined) showed improvements\nin left ventricular function (left ventricular ejection fraction and ââ?¬â??dP/dt). The safety data from four randomized\nclinical trials of gene transfer in patients with symptomatic HFrEF suggest that this approach can be\nconducted with acceptable risk, despite invasive delivery techniques in a high-risk population. Additional\ntrials are necessary before the approach can be endorsed for clinical practice....
Background: Radiogenetic therapy is a novel approach in the treatment of cancer, which employs genetic modification\nto alter the sensitivity of tumor cells to the effect of applied radiation.\nAim: To select a potent radiation inducible promoter in the context of brain tumors and to investigate if CArG radio\nresponsive motifs or other elements in the promoter nucleotide sequences can correlate to its response to radiation.\nMethods: To select initial candidates for promoter inducible elements, the levels of mRNA expression of six different\npromoters were assessed using Quantitative RTPCR in D54 MG cells before and after radiation exposure. Recombinant\nAd/reporter genes driven by five different promoters; CMV, VEGF, FLT-1, DR5 and survivin were constructed.\nGlioma cell lines were infected with different multiplicity of infection of the (promoter) Ad or CMV Ad. Cells were then\nexposed to a range of radiation (0ââ?¬â??12 Gy) at single fraction. Fluorescent microscopy, Luc assay and X-gal staining was\nused to detect the level of expression of related genes. Different glioma cell lines and normal astrocytes were infected\nwith Ad survivin and exposed to radiation. The promoters were analyzed for presence of CArG radio-responsive motifs\nand CCAAT box consensus using NCBI blast bioinformatics software.\nResults: Radiotherapy increases the expression of gene expression by 1.25ââ?¬â??2.5 fold in different promoters other\nthan survivin after 2 h of radiation. RNA analysis was done and has shown an increase in copy number of tenfold for\nsurvivin. Most importantly cells treated with RT and Ad Luc driven by survivin promoter showed a fivefold increase in\nexpression after 2 Gy of radiation in comparison to non-irradiated cells. Presence or absence of CArG motifs did not\ncorrelate with promoter response to radiation. Survivin with the best response to radiation had the lowest number of\nCCAAT box.\nConclusion: Survivin is a selective potent radiation inducible promoter for glioblastoma viral gene therapy and this\nresponse to radiation could be independent of CArG motifs....
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