Use Of Vectors For Gene Transfer Research Articles

Use of Gene Transfer Vectors in Lymphedema Treatment: A Systematic Review.

Different delivery mechanisms have been proposed in the literature for targeted therapies in the treatment of lymphedema. They vary from simple and direct injection to sophisticated induction of gene expression in a targeted tissue. We conducted a systematic review of publications assessing the use of viral vectors for gene transfer in lymphedema treatment. We hypothesized that viral vectors are an effective way to deliver targeted therapy in lymphedema treatment. We conducted a comprehensive systematic review of the published literature on targeted therapies associated with lymphedema surgery using the PubMed database. Eligibility criteria excluded papers that reported use of viral vectors for other medical conditions. s, presentations, reviews, meta-analyses, and non-English language articles were also excluded. From 21 potential articles found in the literature, fourteen fulfilled study eligibility criteria. Positive outcomes in terms of lymphangiogenesis were seen. The viral vectors used included adenovirus and recombinant adeno-associated virus. Most of the genes expressed were growth factors, but expression of dominant-negative transforming growth factor-β1 receptor-II or Prox1 was also proposed. Five studies targeted genetic expression on lymphedema tissue, five on transplanted lymph nodes, two on skeletal muscle, and one on adipose-derived stem cells. Publications assessing use of viral vectors for gene transfer in lymphedema treatment demonstrated that it is an effective mechanism of delivering targeted therapies. However, to date, all studies were experimental and further studies must be performed before translating these therapies into clinical practice.

EDC/NHS-mediated heparinization of small intestinal submucosa for recombinant adeno-associated virus serotype 2 binding and transduction

A major challenge in the use of gene transfer vectors as therapeutic tools is controlling vector administration at a desired tissue site. One potential solution is implanting tissue-engineering constructs loaded with gene transfer vectors such as viruses for localized transgene delivery. In this work, we conjugated recombinant adeno-associated virus serotype 2 (rAAV2) to a heparinized small intestinal submucosa (H-SIS) matrix, which resulted in vector transduction upon cellular adhesion. H-SIS was prepared by incorporating heparin, the rAAV2 receptor, into SIS through N-(3-dimethylaminopropyl)- N′-ethylcarbodiimide (EDC) and N-hydroxysuccinimide (NHS) mediated crosslinking. Incorporated heparin adsorbed rAAV2 onto the H-SIS matrix for conjugation. Using green fluorescent protein and β-galactosidase as reporters, we showed that conjugated rAAV2 was active and capable of mediating transgene delivery in cell culture. Additionally, we applied H-SIS to adsorb unpurified rAAV2 from the crude lysate of packaging cells for conjugation, avoiding the use of ultracentrifugation or chromatography in preparation of infectious rAAV2 for transduction. Our work provides a unique, modified tissue substrate H-SIS for rAAV2 binding and transduction, which can be a useful tool in developing localized gene transfer.

899. Utilizing the Vector Transport Properties of AAV-9 as a Method To Get Widespread Distribution of a Therapeutic Gene within the CNS

The use of gene therapy to treat disorders of the central nervous system has been limited due to the invasiveness of the multiple injections needed for widespread distribution of the therapeutic gene. The use of gene transfer vectors that can deliver the therapeutic gene to a larger brain region with a limited number of injections needs further exploration. Our lab and others have shown that injections of viral vectors expressing genes for lysosomal enzymes can result in retrograde and anterograde transport of the enzyme to certain afferent and efferent projections of the injected nuclei. We have also found, using adeno-associated viral (AAV) vectors packaged in AAV-9 capsid proteins, that the vector genome can be transported. Based on these findings, our hypothesis was that injecting AAV 9 into regions of the brain with widespread projections, such as the locus coeruleus (LC) or ventral tegmental area (VTA), would lead to wider distribution of vector genome and more extensive dispersal of the enzyme. For this experiment, we unilaterally injected AAV 9 expressing the human lysosomal enzyme beta-glucuronidase (hGUSB) into the LC or VTA of C3H mice, and then looked for vector and enzyme distribution 2 months post-injection. We found that a single injection of AAV 9 resulted in enzyme dispersal from the olfactory bulbs to the spinal cord, due to transport of the vector and enzyme to projection sites of the injected nucleus. This information may be useful for devising new treatment options for genetic diseases of the CNS, where multiple injections are not optimal.