Hypoxia-modulation of EPO pDNA in Electroporated Muscle

描述(申请人提供)：使用Lchor创新和专有的TriGrid电穿孔系统将编码促红细胞生成素(EPO)的质粒DNA体内肌肉注射，已导致EPO在啮齿动物、兔子和灵长类动物中强劲的结构性表达。虽然高水平的EPO对EPO反应迟钝的患者是有益的，但这种高表达不适合对EPO快速升高敏感的慢性肾功能衰竭患者。因此，开发一种生理调节的EPO基因疗法，在患者的贫血和肌肉内传递的EPO转基因之间进行更直接的沟通，将对后一类患者群体有利。将不再需要监测血细胞比容和蛋白质的滴定剂量。毒性可能会降低。最后，非患者的虐待行为不太可能发生。 Ichor已经证明了利用与贫血相关的缺氧来调节导入贫血小鼠肌肉中的缺氧反应质粒的基因表达的可行性。然而，这种方法必须在更具临床相关性的动物模型中进行更彻底的研究。实验将在大鼠身上进行，包括在贫血动物体内筛选含有HRE的质粒，评估运动诱导的低氧对基因表达的影响，以及在疾病模型中评估低氧调节的EPO转基因表达。

DESCRIPTION (provided by applicant): In vivo intramuscular delivery of plasmid DNA encoding erythropoietin (EPO) using lchor's innovative and proprietary TriGrid electroporation system has resulted in robust constitutive expression of EPO in rodents, rabbits and primates. While high EPO levels are beneficial for patients with blunted responses to EPO, such high expression is not appropriate for chronic renal failure patients who are sensitive to rapid increases in EPO. Development of a physiologically modulated EPO gene therapy with a more direct communication between a patient's anemia and the intramuscularly delivered EPO transgene would thus be of benefit for this latter patient population. The necessities of monitoring hematocrit and titrating doses of the protein would be eliminated. Toxicities could potentially be decreased. Finally, abuse by non-patients would be unlikely. Ichor has demonstrated the feasibility of utilizing anemia-related hypoxia to modulate gene expression from hypoxia-responsive plasmids transfected into anemic mouse muscle. However, this approach must be more thoroughly investigated in a more clinically relevant animal model. Experiments will be performed using rats and will include in vivo screening of HRE-containing plasmids in anemic animals, evaluation of the effect of hypoxia induced by exercise on gene expression, and assessment of hypoxia-modulated EPO transgene expression in a disease model.