Non-viral gene therapy for cancer pain

项目摘要/摘要 口腔癌疼痛强度高于其他癌症。口腔癌患者的生活质量是 是所有癌症患者中最低的，因为无法控制的疼痛会干扰必要的口腔功能，包括进食， 说话和吞咽。外源性阿片类药物对这种类型的疼痛最不有效，而且有显著的副作用 效果。我们的长期目标是开发一种有效且安全的口腔癌疼痛治疗方法。我们最近 证实OPRM1(µ-阿片受体基因)在口服中甲基化和下调 将同一患者的癌症与匹配的正常组织进行比较；这些患者报告在 癌症。我们进一步证明，病毒转导后OPRM1的再表达显著降低 小鼠模型中的癌症疼痛。肿瘤表面阿片受体的表达导致阿片类物质的分泌 进入癌症的微环境。 病毒转导方法的缺点是病毒基因传递具有 安全问题和有限的承载能力。为了克服这些障碍，我们开发了两种非病毒杂交 向量。第一种是由细胞渗透肽(组氨酸修饰的HIV-1 TAT肽序列)组成 和半胱氨酸残基)与阳离子脂结合。第二个载体用阳离子聚合物取代 脂类。该载体具有良好的转染率，在体内外具有最小的细胞毒性。 此外， 与正常细胞相比，非病毒载体优先转染口腔癌细胞。基于我们的 初步工作我们假设OPRM1基因在口腔癌中使用我们的非病毒 载体将减轻癌症疼痛和恢复口腔面部功能，而不会有过多毒性。 在具体目标1中，我们 会死吗？ 非病毒载体体外转导OPRM1基因抑制肿瘤效应的实验研究 疼痛。我们的目标是将我们的非病毒转基因方法应用于临床。我们直接预见临床医生 将我们的非病毒载体接种到口腔癌中。因此，在具体目标2中，我们将确定可行性 非病毒载体体内OPRM1基因转移(即直接进入舌癌)的疗效 减轻癌症引起的疼痛。因为我们进行临床试验的先决条件是毒性和安全性研究 在特定的目标3中，我们将分析非病毒治疗的肿瘤小鼠的毒性和免疫反应。 OPRM1基因的传递。拟议的研究具有重要意义，因为我们将使用本地交付技术 直接进入癌症，以减少全身药物的潜在副作用。我们的方法是创新的 因为我们将转导癌细胞来治疗癌症疼痛。 促进开发一种治疗癌症疼痛的有效疗法。 最终，这些研究可能会

Project Summary/Abstract The intensity of oral cancer pain is higher than other cancers. Quality of life for oral cancer patients is the lowest of all cancer patients because uncontrolled pain interferes with necessary oral functions including eating, talking and swallowing. Exogenous opioids are minimally effective for this type of pain and have significant side effects. Our long-term goal is to develop an effective and safe treatment for oral cancer pain. We recently demonstrated that OPRM1 (the gene for the µ-opioid receptor) is methylated and down regulated in oral cancer compared to matched normal tissues in the same patients; these patients reported pain at the site of cancer. We further demonstrated that OPRM1 re-expression with viral transduction significantly reduced cancer pain in a mouse model. Expression of the µ-opioid receptor on the cancer led to the secretion of opioids into the cancer microenvironment. Drawbacks of a viral transduction approach are that viral gene delivery has safety concerns and limited carrying capacity. To overcome these barriers we developed two non-viral hybrid vectors. The first is composed of a cell-permeable peptide (HIV-1 Tat peptide sequence modified with histidine and cysteine residues) combined with a cationic lipid. The second vector substitutes cationic polymer for the lipid. The vectors have excellent transfection efficiency with minimal cytotoxicity in vitro and in vivo. Moreover, the non-viral vectors preferentially transfects oral cancer cells compared to normal cells. Based on our preliminary work we hypothesize that re-expression of the OPRM1 gene within oral cancer using our non-viral vectors will attenuate cancer pain and restore orofacial function without excessive toxicity. In Specific Aim 1 we will d etermine the efficacy of ex vivo OPRM1 gene transfer with non-viral vectors to attenuate cancer-induced pain. Our goal is to move our method of non-viral transfection to the clinic. We foresee clinicians directly inoculating our non-viral vector into an oral cancer. Therefore, in Specific Aim 2 we will determine the feasibility and efficacy of in vivo OPRM1 gene transfer (i.e., directly into the tongue cancer) with non-viral vectors for attenuation of cancer-induced pain. Because our prerequisites for a clinical trial are toxicity and safety studies in Specific Aim 3 we will analyze toxicity and immune response in the cancer mice treated with non-viral OPRM1 gene delivery. The proposed research is significant because we will use a local delivery technique directly into the cancer to reduce the potential side effects of systemic drugs. Our approach is innovative because we will transduce the cancer cells for the treatment of cancer pain. facilitate the development of an effective therapy to treat cancer pain. Ultimately, these studies might