Electro-injection of IL-12 DNA to Muscle for Cancer

描述（申请人提供）：实现基因治疗作为一种治疗方法的承诺 为了治疗癌症，我们必须开发简化但有效的基因递送方法，并了解治疗性蛋白质如何杀死肿瘤。我们的建议直接涉及这两个关键因素。我们已经率先使用基于肌肉的电穿孔作为简化和有效的基因递送的新颖且廉价的方法，并且已经证明，与没有电穿孔的注射相比，伴随电穿孔的IL-12基因注射到肌肉显著增强了IL-12表达水平及其全身抗肿瘤功效，导致20%的远离基因注射部位的肿瘤被根除。我们假设，通过肌内电穿孔递送IL-12 DNA，与新的肌肉特异性表达系统协同作用，将通过上调抗肿瘤基因根除或抑制远端鳞状细胞癌（SCC）。我们的长期目标是开发一种简单有效的策略来根除手术后难以接近的小负荷转移性SCC或残留的显微镜下SCC。我们将使用称为SCCVII的SCC肿瘤模型，其允许在同基因小鼠宿主（C3 H/HeJ）中产生皮下和转移性肿瘤，以探索以下特定目的。目标1：确定电穿孔的最佳递送和表达系统，以在体内达到IL-12的治疗水平，并评估基因治疗与蛋白质治疗相比的全身毒性。目的2：确定Stat 1和原代免疫细胞在抑制IL-12引起的肿瘤生长中的作用。这项工作在根除SCC方面具有巨大的进展潜力，特别是在那些临床 小负荷转移无法触及的情况或残留显微镜下 手术后疾病仍然存在。我们提出的实验有很高的成功概率，因为我们有一个很好的模型系统，在电穿孔基因治疗领域的经验，以及完成任务所需的所有资源。

DESCRIPTION (provided by applicant): To realize the promise of gene therapy as a treatment modality for cancer, we must develop simplified but effective gene delivery approaches and understand how therapeutic proteins act to kill tumors. Our proposal directly addresses both of these critical elements. We have pioneered the use of muscle-based electroporation as a novel and inexpensive approach for simplified and effective gene delivery and have demonstrated that injection of the IL-12 gene to muscle accompanied by electroporation substantially enhances the level of IL-12 expression and its systemic antitumor efficacy, compared with injection without electroporation, leading to 20% eradication of tumors located distant to the gene injection site. We hypothesize that delivery of IL-12 DNA via intramuscular electroporation, in synergy with a novel muscle-specific expression system, will eradicate or inhibit remote squamous cell carcinoma (SCC) by upregulation of antitumor genes. Our long-term goal is to develop a simple and effective strategy for eradicating inaccessible small-load metastatic SCC or residual microscopic SCC after surgery. We will use a tumor model of SCC, known as SCCVII, which allows the generation of both subcutaneous and metastatic tumors in a syngeneic murine host (C3H/HeJ) to explore the following Specific Aims. Aim 1: Determine the optimal delivery and expression system for electroporation to achieve therapeutic levels of IL-12 in vivo, and assess the systemic toxicity of gene therapy in comparison to protein therapy. Aim 2: Determine the role of Stat 1 and primary immune cells in inhibition of tumor growth elicited by IL-12. This work holds enormous potential for progress in eradicating SCC, particularly in those clinical situations where small-load metastases are inaccessible or in cases where residual microscopic disease persists after surgery. The experiments that we propose have a high probability of success because we have an excellent model system in place, experience in the electroporation gene therapy field, and all the resources necessary to complete the task.