Current Issue : April - June Volume : 2012 Issue Number : 2 Articles : 7 Articles
Alphavirus vectors engineered for gene delivery and expression of heterologous proteins have been considered as valuable tools for research on nurological disorders.they possess a highly efficient susceptibility for neuronal cells and can provide extreme levels of heterologous gene expression. However, they generally generate short-term\r\ntransient expression, which might limit their therapeutic use in many neurological disorders often requiring long-term\r\neven life-long presence of therapeutic agents. Recent development in gene silencing applying both RNA interference\r\nand microRNA approaches will certainly expand the application range. Moreover, alphaviruses provide interesting\r\nmodels for neurological diseases such as demyelinating and spinal motor diseases....
Endocrine disruptors (EDs) are compounds used in various industrial products,\ndrugs, and cosmetics. They can be found in the environment and disturb the endocrine and\nreproductive systems, resulting in adverse effects to humans and wildlife such as birth\ndefects and developmental disorders. Since several EDs have a structure similar to that of\nendogenous steroid hormones such as estrogens, they intend to have an affinity for steroid\nhormone receptors and alter hormone-mediated metabolism by binding to these receptors.\nEDs are therefore a global concern and assays should be developed to efficiently determine\nwhether these compounds are detrimental to biological systems. Diverse experimental\nmethods may help determine the endocrine disrupting potential of EDs and evaluate the\nadverse effects of a single and/or combination of these reagents. Currently, biomarkers\nhave been employed to objectively measure EDs potency and understand the underlying\nmechanisms. Further studies are required to develop ideal screening methods and\nbiomarkers to determine EDs potency at environmentally relevant concentrations. In this\nreview, we describe the biomarkers for estrogenicity of EDs identified both in vitro and\nin vivo, and introduce a biomarker, cabindin-D9k (CaBP-9k), that may be used to assess\nestrogenic activity of EDs....
Recombinant vectors based on a non-pathogenic human parvovirus, the adeno-associated virus (AAV), have gained attention as a potentially safe and useful alternative to the more commonly used retroviral and adenoviral vectors. AAV vectors are currently in use in Phase I/II clinical trials for gene therapy of a number of diseases such as cystic fibrosis, a-1 antitrypsin deficiency, muscular dystrophy, Batten�s disease, and Parkinson�s disease, and have shown efficacy in patients with Leber�s congenital amaurosis, and hemophilia B. For patients with hemophilia B, however, relatively large vector doses are needed to achieve therapeutic benefits. Large vector doses also trigger an immune response as a significant fraction of the vectors fails to traffic efficiently to the nucleus, and is targeted for degradation by the host cell proteasome machinery. With a better understanding of the various steps in the life cycle of AAV vectors, strategies leading to the development of novel AAV vectors that are capable of highefficiency transduction at lower doses are needed. In this review, we summarize our strategies to develop novel AAV vectors for the potential gene therapy of both hemophilia B and hemophilia A, based on our recent studies on the basic molecular biology of AAV. These strategies, including the development of novel AAV vectors by site-directed mutagenesis of critical surface-exposed tyrosine residues on AAV2 capsids to circumvent the ubiquitination step and the use of different AAV serotypes and self-complementary (sc) AAV2 vectors, and their use as helper vectors to circumvent the obstacles of second-strand DNA synthesis of single-stranded (ss) AAV, should dramatically accelerate the progress towards the potential gene therapy of both hemophilia A and hemophilia B....
Substantial progress has been made in the development of novel gene therapy strategies for central nervous system (CNS) disorders\r\nin recent years. However, unregulated transgene expression is a significant issue limiting human applications due to the potential\r\nside effects from excessive levels of transgenic protein that indiscriminately affect both diseased and nondiseased cells. Gene\r\nregulation systems are a tool by which tight tissue-specific and temporal regulation of transgene expression may be achieved.\r\nThis review covers the features of ideal regulatory systems and summarises the mechanics of current exogenous and endogenous\r\ngene regulation systems and their utility in the CNS....
Adoptive immunotherapy using TCR gene modified T cells may allow separation of beneficial Graft versus tumour responses from\r\nharmful GvHD. Improvements to this include methods to generate high avidity or high affinity TCR, improvements in vector\r\ndesign and reduction in mispairing. Following adoptive transfer, TCR transduced T cells must be able to survive and persist in\r\nvivo to give most effective antitumour responses. Central memory or naive T cells have both been shown to be more effective than\r\neffector cells at expanding and persisting in vivo. Lymphodepletion may enhance persistence of transferred T cell populations. TCR\r\ngene transfer can be used to redirect CD4 helper T cells, and these could be used in combination with CD8+ tumour specific T\r\ncells to provide help for the antitumour response. Antigen specific T regulatory T cells can also be generated by TCR gene transfer\r\nand could be used to suppress unwanted alloresponses....
Despite advances in radical surgery and chemotherapy delivery, ovarian cancer is the most lethal gynecologic malignancy.\nStandard therapy includes treatment with platinum-based combination chemotherapies yet there is no biomarker model to\npredict their responses to these agents. We here have developed and independently tested our multi-gene molecular\npredictors for forecasting patients� responses to individual drugs on a cohort of 55 ovarian cancer patients. To\nindependently validate these molecular predictors, we performed microarray profiling on FFPE tumor samples of 55 ovarian\ncancer patients (UVA-55) treated with platinum-based adjuvant chemotherapy. Genome-wide chemosensitivity biomarkers\nwere initially discovered from the in vitro drug activities and genomic expression data for carboplatin and paclitaxel,\nrespectively. Multivariate predictors were trained with the cell line data and then evaluated with a historical patient cohort.\nFor the UVA-55 cohort, the carboplatin, taxol, and combination predictors significantly stratified responder patients and\nnon-responder patients (p = 0.019, 0.04, 0.014) with sensitivity = 91%, 96%, 93 and NPV = 57%, 67%, 67% in pathologic\nclinical response. The combination predictor also demonstrated a significant survival difference between predicted\nresponders and non-responders with a median survival of 55.4 months vs. 32.1 months. Thus, COXEN single- and\ncombination-drug predictors successfully stratified platinum resistance and taxane response in an independent cohort of\novarian cancer patients based on their FFPE tumor samples....
Proteases and protease inhibitors have been identified in the ejaculates of animal taxa ranging from invertebrates to\nmammals and form a major protein class among Drosophila melanogaster seminal fluid proteins (SFPs). Other than a single\nprotease cascade in mammals that regulates seminal clot liquefaction, no proteolytic cascades (i.e. pathways with at least\ntwo proteases acting in sequence) have been identified in seminal fluids. In Drosophila, SFPs are transferred to females\nduring mating and, together with sperm, are necessary for the many post-mating responses elicited in females. Though\nseveral SFPs are proteolytically cleaved either during or after mating, virtually nothing is known about the proteases\ninvolved in these cleavage events or the physiological consequences of proteolytic activity in the seminal fluid on the\nfemale. Here, we present evidence that a protease cascade acts in the seminal fluid of Drosophila during and after mating.\nUsing RNAi to knock down expression of the SFP CG10586, a predicted serine protease, we show that it acts upstream of the\nSFP CG11864, a predicted astacin protease, to process SFPs involved in ovulation and sperm entry into storage. We also\nshow that knockdown of CG10586 leads to lower levels of egg laying, higher rates of sexual receptivity to subsequent\nmales, and abnormal sperm usage patterns, processes that are independent of CG11864. The long-term phenotypes of\nfemales mated to CG10586 knockdown males are similar to those of females that fail to store sex peptide, an important\nelicitor of long-term post-mating responses, and indicate a role for CG10586 in regulating sex peptide. These results point to\nan important role for proteolysis among insect SFPs and suggest that protease cascades may be a mechanism for precise\ntemporal regulation of multiple post-mating responses in females....
Loading....