ABSTRACT
Published transcriptomic data from surgically removed metastatic clear cell renal cell carcinoma samples were analyzed from the genomic fabric paradigm (GFP) perspective to identify the best targets for gene therapy. GFP considers the transcriptome as a multi-dimensional mathematical object constrained by a dynamic set of expression controls and correlations among genes. Every gene in the chest wall metastasis, two distinct cancer nodules, and the surrounding normal tissue of the right kidney was characterized by three independent measures: average expression level, relative expression variation, and expression correlation with each other gene. The analyses determined the cancer-induced regulation, control, and remodeling of the chemokine and vascular endothelial growth factor (VEGF) signaling, apoptosis, basal transcription factors, cell cycle, oxidative phosphorylation, renal cell carcinoma, and RNA polymerase pathways. Interestingly, the three cancer regions exhibited different transcriptomic organization, suggesting that the gene therapy should not be personalized only for every patient but also for each major cancer nodule. The gene hierarchy was established on the basis of gene commanding height, and the gene master regulators DAPK3,TASOR, FAM27C and ALG13 were identified in each profiled region. We delineated the molecular mechanisms by which TASOR overexpression and ALG13 silencing would selectively affect the cancer cells with little consequences for the normal cells.
ABSTRACT
Radiation Therapy (RT) has been critical in cancer treatment regimens to date. However, it has been shown that ionizing radiation is also associated with increased risk of damage to healthy tissues. At high radiation doses, varied effects including inactivation of cells in treated tissue and associated functional impairment are seen. These range from direct damage to the heart; particularly, diffuse fibrosis of the pericardium and myocardium, adhesion of the pericardium, injury to the blood vessels and stenosis. Cardiac damage is mostly a late responding end-point, occurring anywhere between 1 and 10 years after radiation procedures. Cardiovascular disease following radiotherapy was more common with radiation treatments used before the late 1980s. Modern RT regimens with more focused radiation beams, allow tumors to be targeted more precisely and shield the heart and other healthy tissues for minimizing the radiation damage to normal cells. In this review, we discuss radiation therapeutic doses used and post-radiation damage to the heart muscle from published studies. We also emphasize the need for early detection of cardiotoxicity and the need for more cardio-protection approaches where feasible.
ABSTRACT
Gene therapy for Duchenne muscular dystrophy will likely require that the corrective dystrophin gene be delivered to a high fraction of muscle fibers in vivo. Because of the large size of the dystrophin cDNA, adenoviral (Ad) vectors have been developed for this application. However, Ad vectors transduce mature muscle inefficiently in part due to downregulation of Ad receptors on these cells. To circumvent this problem, we have tested fibroblast growth factor-2 (FGF) and insulin-like growth factor (IGF) as ligands for their ability to enhance Ad transduction of muscle cells. In this work, we demonstrate that covalent conjugation of FGF, but not IGF, to Ad5 vectors mediates substantial increases in transduction of skeletal muscle cells in vitro and dystrophic in vivo. Ad5 vectors expressing reporter genes were cross-linked to the ligands, using bifunctional polyethylene glycol (PEG) molecules. Ad-PEG-FGF mediated 1000- and 200-fold increases in transduction on C2C12 myoblasts and myotubes in vitro when compared with Ad5, Ad-PEG, or Ad-PEG-IGF. When tested in vivo in mdx mice, Ad-PEG-FGF mediated 6-fold higher transduction in skeletal muscle than unmodified Ad5. Similar results were seen when using lacZ as a reporter gene to observe transduction qualitatively. These data suggest that FGF may be a useful cell-binding ligand to enhance gene delivery by Ad and other vectors into skeletal muscle for the gene therapy of Duchenne muscular dystrophy and other muscle-related diseases.