RESUMO
No disponible
Assuntos
Humanos , Masculino , Adulto , Transplante de Fígado/efeitos adversos , Citomegalovirus , Citomegalovirus/isolamento & purificação , Biópsia , Paniculite/diagnóstico , Paniculite/microbiologia , Imunossupressores/uso terapêutico , Primers do DNA/uso terapêutico , Anfotericina B/uso terapêutico , Itraconazol/uso terapêutico , Voriconazol/uso terapêutico , Micoses/diagnóstico , Micoses/microbiologiaRESUMO
Gene therapy involves the transfer of genes into cells with therapeutic intent. Although several methods can accomplish this, vectors based on viruses still provide the most efficient approach. For neurosurgical purposes, preclinical and clinical applications in the areas of glioma therapy, spinal neurosurgery, and neuroprotection for treatment of Parkinson's disease and cerebral ischemia are reviewed. In general, therapies applied in the neurosurgical realm have proven relatively safe, despite occasional, well-publicized cases of morbidity and death in non-neurosurgical trials. However, continued clinical and preclinical research in this area is critical, to fully elucidate potential toxicities and to generate truly effective treatments that can be applied in neurological diseases.
Assuntos
Isquemia Encefálica/genética , Isquemia Encefálica/prevenção & controle , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/terapia , Primers do DNA/uso terapêutico , Terapia Genética , Glioma/genética , Glioma/terapia , Procedimentos Neurocirúrgicos , Doença de Parkinson/genética , Doença de Parkinson/prevenção & controle , Doenças da Medula Espinal/genética , Doenças da Medula Espinal/terapia , HumanosRESUMO
PURPOSE: Antigene radiotherapy (AR) is based on targeting localized radiodamage to specific sites in the genome by using sequence-specific triplex-forming oligonucleotides (TFO) to carry Auger-electron-emitters (A-Ettr) such as Iodine-125 (125I) to the target gene sequence. The radiodecay of an A-Ettr produces a cascade of low-energy electrons and creates a highly positively-charged daughter atom; delivered by a TFO, it should produce double-strand breaks (dsb) localized to the specific DNA target sequence. The result should be a "knock-out" of the targeted gene. METHODS AND MATERIALS: As a model, we used the MDR1 gene amplified nearly 100 times in the human KB-V1 carcinoma cell line. Chemically modified TFO complementary to the polypurine/polypyrimidine region of the MDR1 gene were synthesized and radiolabeled with 125I-dCTP by the primer extension method. Purified plasmid and genomic DNA and extracted nuclei were treated with 125I-TFO and analyzed for sequence-specific cleavage by electrophoresis in agarose gel and Southern hybridization. RESULTS: We created 125I-TFO that could effectively recognize, bind, and cleave the target sequence in plasmid and genomic DNA. We showed that these 125I-TFO in nanomolar concentrations were able to cleave the target MDR1 gene sequence in a natural environment, i.e., within the eucaryotic nucleus. CONCLUSION: 125I-TFO can effectively introduce sequence-specific dsb to a target within the MDR1 gene, both in purified DNA and inside intact nuclei. Chemically modified TFO conjugated with nuclear localization signal appear to be a promising delivery vehicle for future in vivo trials of AR.
