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的前景 对于癌症的治疗模式，我们必须开发简单但有效的基因传递方法，并了解治疗性蛋白质如何作用于杀死肿瘤。我们的建议直接涉及这两个关键要素。我们已经率先使用基于肌肉的电穿孔作为一种新的、廉价的简化和有效的基因传递方法，并且已经证明，与没有电穿孔的注射相比，肌肉注射IL-12基因和电穿孔显著提高了IL-12的表达水平和系统的抗肿瘤效果，导致远离基因注射部位的肿瘤被根除了20%。我们假设，通过肌肉内电穿孔传递IL-12DNA，与一种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来根除或抑制远程鳞状细胞癌(SCC)。我们的长期目标是开发一种简单而有效的策略来根除难以接近的小负荷转移性鳞癌或术后残留的微小鳞癌。我们将使用一种被称为SCCVII的SCC肿瘤模型，该模型允许在同基因小鼠宿主(C3H/HeJ)中产生皮下和转移肿瘤，以探索以下特定目的。目的1：确定电穿孔的最佳传递和表达系统，以达到体内IL-12的治疗水平，并与蛋白质治疗比较，评价基因治疗的全身毒性。目的：探讨STAT-1和原代免疫细胞在抑制IL-12诱导的肿瘤生长中的作用。这项工作在根除鳞状细胞癌方面具有巨大的潜力，特别是在临床上 无法接触到小负荷转移瘤的情况或残留显微镜下的情况 手术后，疾病仍然存在。我们提出的实验有很高的成功几率，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.