| Budget Amount *help |
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2012: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2011: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2010: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| Research Abstract |
Vascular Ehlers-Danlos syndrome (vEDS), is a life-threatening dominantly inherited disorder caused by COL3A1 mutation. Although gene therapy is an important option for treatment of genetic disorders, addition of the defective gene is not applicable for dominant diseases such as vEDS. To establish a therapeutic strategy for vEDS, we examined the feasibility of two approaches in each mutation. First, we tested RNAi mediated inhibition of the mutant allele. We synthesized several small interfering RNA (siRNA) molecules targeting sequence specific for mutant mRNA and introduced into patient’s fibroblasts. Quantitative measurement of COL3A1 mRNA showed that mutant mRNA levels could be selectively decreased up to 80 % in splicing mutation and it was possible to increase the relative concentration of normal triple helix. Second, we attempted to increase theconcentration of normal COL3A1 mRNA, since half-dose of COL3A1 may not be sufficient to prevent vEDS symptoms. Lysyl oxidase(LOX)is a bifunctional protein carrying activities of both an extracellular enzyme that controls the maturation of collagen and elastin and an intracellular tranascriptional activator for the human collagen III promoter. When patient’s cells were transfected with mutant specific siRNA and LOX expression vector, specific inhibition of the mutant allele and enhancement of the normal allele wereobserved.
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