Non-viral gene therapy for cancer pain.

 描述：口腔癌疼痛强度高于其他癌症。口腔癌患者的生活质量是所有癌症患者中最低的，因为无法控制的疼痛干扰了 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，并且有显著的副作用。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。我们最近证明，与同一患者的匹配正常组织相比，口腔癌中OPRM1(阿片受体基因)甲基化和下调；这些患者报告癌症部位疼痛。我们进一步证明，在小鼠模型中，OPRM1通过病毒转导重新表达显著减少了癌症疼痛。肿瘤表面阿片受体的表达导致阿片类物质分泌到肿瘤微环境中。病毒转导方法的缺点是病毒基因传递有安全问题和有限的携带能力。为了克服这些障碍，我们开发了两种新型的非病毒杂交载体：细胞渗透性多肽与阳离子脂类或阳离子聚合物结合。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。根据我们的初步工作，我们假设，使用我们的非病毒载体在口腔癌中重新表达OPRM1基因将减轻癌症疼痛，恢复口腔面部功能，而不会有过多的毒性。在特定的目标1中，我们将确定非病毒载体的体外OPRM1基因转移对减轻癌症诱导的疼痛的效果。我们的目标是将我们的非病毒转基因方法应用于临床。我们预见临床医生会直接将我们的非病毒载体接种到口腔癌中。因此，在特定的目标2中，我们将确定体内OPRM1基因转移(即直接进入舌癌)与非病毒载体一起用于减轻癌症引起的疼痛的可行性和有效性。因为我们进行临床试验的先决条件是特定目的的毒性和安全性研究3，所以我们将分析非病毒OPRM1基因治疗的癌症小鼠的毒性和免疫反应。这项拟议的研究意义重大，因为我们将使用一种直接进入癌症的局部给药技术，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用于治疗癌症疼痛，并且我们的非病毒载体比正常细胞更有效地靶向口腔癌细胞。最终，这些研究可能有助于开发一种有效的治疗癌症疼痛的方法。

 DESCRIPTION: 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 o 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 novel non-viral hybrid vectors: a cell-permeable peptide combined with either a cationic lipid or a cationic polymer. The vectors have excellent transfection efficiency with minimal cytotoxicity in vitro and in vivo. Moreover, the non viral vectors preferentially transfected 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 determine 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 and our non-viral vector more efficiently targets oral cance cells relative to normal cells. Ultimately, these studies might facilitate the development of an effective therapy to treat cancer pain.