Intratumor co-delivery of DNA and RNA to relieve cancer pain

肿瘤内共同递送 DNA 和 RNA 以缓解癌症疼痛

• 批准号: 10327329
• 负责人: Brian L Schmidt
• 金额: $ 57.37万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-08 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10327329
• 关键词: Absence of pain sensation; Afferent Neurons; Agonist; Animal Model; Attenuated; Biological Assay; CRISPR/Cas technology; Cancer Etiology; Cancer Patient; Cancer cell line; Cations; Cells; Chronic; Clinical Trials; DNA; DNA delivery; Data; Development; Down-Regulation; Drug usage; Eating; Future; Generations; Genes; Genomics; Goals; Head and Neck Cancer; Heterogeneity; Human; Knowledge; Lead; Legal patent; Life; Lipids; Malignant Neoplasms; Measures; Mechanics; Methodology; Mission; Mus; Nerve; Nociception; Non-Viral Vector; Normal tissue morphology; Opioid; Oral; Orofacial Pain; Outcome; PAR-2 Receptor; Pain; Pain Measurement; Pain management; Partner in relationship; Pathway interactions; Patient Self-Report; Patients; Peptides; Peripheral; Permeability; Pharmaceutical Preparations; Phase I Clinical Trials; Pre-Clinical Model; Public Health; Quality of life; RNA; RNA Interference; RNA delivery; Receptor Gene; Regulation; Research; Salvelinus; Signal Pathway; Signal Transduction; Simplexvirus; Specificity; Testing; Therapeutic; Transfection; Translating; Treatment Efficacy; United States National Institutes of Health; Variant; Viral Genes; Work; Xenograft procedure; antagonist; attenuation; base; beta-Endorphin; cancer cell; cancer pain; cancer therapy; chemical carcinogen; chronic neuropathic pain; clinically relevant; cytotoxicity; debilitating pain; disability; drinking; evidence base; functional restoration; gene therapy; in vivo; innovation; knock-down; malignant mouth neoplasm; mouse model; mu opioid receptors; non-opioid analgesic; non-viral gene therapy; novel; orofacial; pain signal; side effect; tumor microenvironment

Oral cancer causes more prevalent and severe pain than any other cancer. Oral cancer patients suffer from severe, chronic, mechanically-induced pain; talking and eating are profoundly disrupted. Opioids are plagued with side effects and ineffective as tolerance develops; there is no satisfactory treatment for oral cancer pain. Our long-term goal is to develop cancer-targeting non-viral gene therapy to disrupt nociceptive signaling in the cancer microenvironment with minimal off-target effects and translate our approach to patients. We created two non-viral vectors with excellent transfection efficiency and no cytotoxicity: a cell-permeable peptide com- bined with a cationic lipid for DNA; and, a lipopolymer for RNA. Non-viral transfection with OPRM1 (μ-opioid receptor gene, MOR) DNA led to re-expression of MOR and partial attenuation of nociception (i.e., pain) in cancer mouse models. Protease-activated receptor-2 (PAR2) is elevated in the primary afferent neurons that innervate the cancer and drive pain. Knockdown of F2rl1 (PAR2 gene) partially attenuated nociception. While both approaches partially reduce cancer pain, our goal is elimination. Complete analgesia poses a challenge; there are multiple and redundant pain pathways, and genomic heterogeneity in oral cancer produces variations in the pathways. As a strategy to obstruct these multiple and varied pathways, we posit that a combination of OPRM1 re-expression and F2RL1 downregulation in the cancer could eliminate pain. The central hypothesis is that a non-viral vector approach to gain OPRM1 re-expression and F2RL1 downregulation within the oral can- cer will eliminate cancer pain and fully restore function. The rationale for this project is that demonstration of therapeutic efficacy of co-delivery of OPRM1 DNA and F2RL1 RNAi or CRISPR/Cas9 will yield a scientifically rigorous framework for development of non-opioid therapies that can be translated to patients. The central hy- pothesis will be tested in two specific aims: 1) Determine the feasibility and efficacy of ex vivo transfection (i.e., transfection of oral cancer cells with dual genes prior to generation of a xenograft mouse model); and, in vivo transfection (i.e., direct inoculation of genes into the oral cancer in the chemical carcinogen mouse model) with OPRM1 DNA and/or F2RL1 RNAi or CRISPR/Cas9 with the non-viral vectors to eliminate cancer-induced pain in mouse models; 2) Measure pain in oral cancer patients, analyze OPRM1 and F2RL1 expression in their can- cers relative to matched normal tissue, and analyze the correlation between dysregulation of these genes and patients’ pain. The results will give us data regarding the proportion of oral cancer patients who might benefit from co-delivery of DNA and RNA and set the stage for a clinical trial. The research proposed in this applica- tion is innovative because co-delivery of DNA and RNA into a cancer with non-viral vectors for the manage- ment of pain has not been done. The proposed research is significant because it is expected to prove strong scientific justification for the continued development and future clinical trials of novel dual gene therapy. Ulti- mately, such knowledge might lead to development of novel non-opioid therapies for cancer pain.

口腔癌比任何其他癌症引起的疼痛更普遍和更严重。口腔癌患者患有 严重的、慢性的、机械引起的疼痛；说话和进食都被严重干扰。阿片类药物深受困扰 随着耐受性的发展，副作用和无效；口腔癌疼痛没有令人满意的治疗方法。 我们的长期目标是开发以癌症为靶点的非病毒基因疗法，以干扰 癌症微环境，最大限度地减少非目标影响，并将我们的方法转化为患者。我们创造了 两种高效、无细胞毒性的非病毒载体：一种细胞通透性多肽。 与用于DNA的阳离子脂结合；以及用于RNA的脂聚合物。OPRM_1(μ-阿片)的非病毒转染法 受体基因，MOR)DNA导致MOR的重新表达和痛觉(即痛觉)的部分减弱 癌症小鼠模型。蛋白水解酶激活受体-2(PAR2)在初级传入神经元中升高。 会刺激癌症并引发疼痛。敲除F2RL1(PAR2基因)可部分减弱伤害性感受。而当 这两种方法都部分地减轻了癌症的痛苦，我们的目标是消除疼痛。完全止痛是一个挑战； 口腔癌有多条多余的疼痛通路，基因组的异质性产生了变异。 在小路上。作为一种阻止这些多种多样的途径的策略，我们假设 癌组织中OPRM1的重新表达和F2RL1的下调可以消除疼痛。中心假设是 一种非病毒载体方法在口腔中获得OPRM1的重新表达和F2RL1的下调可以- CER将消除癌症疼痛，并完全恢复功能。该项目基本原理是演示 OPRM1 DNA和F2RL1 RNAi或CRISPR/Cas9联合治疗的疗效将产生科学的 开发可转化为患者的非阿片类药物治疗的严格框架。中央高铁- 假体将在两个特定目的中进行测试：1)确定体外转染法的可行性和有效性(即， 在建立异种小鼠模型之前用双基因转染口腔癌细胞)；以及，在体内 转基因(即直接将基因接种到化学致癌小鼠模型的口腔癌中) OPRM1 DNA和/或F2RL1 RNAi或CRISPR/Cas9与非病毒载体结合以消除癌症引起的疼痛 在小鼠模型中；2)测量口腔癌患者的疼痛，分析OPRM1和F2RL1在他们的大脑中的表达。 与匹配的正常组织的CER，并分析这些基因的失调与 病人的痛苦。这一结果将为我们提供口腔癌患者可能受益的比例的数据 从DNA和RNA的联合传递中脱颖而出，为临床试验奠定了基础。本申请中提出的研究-- TION是创新的，因为将DNA和RNA与非病毒载体共同传递到癌症中，以管理- 痛苦的治疗还没有做好。这项拟议的研究意义重大，因为预计它将被证明是强有力的 为新的双基因疗法的持续开发和未来临床试验提供科学依据。多- 更重要的是，这些知识可能会导致新的非阿片类药物治疗癌症疼痛的开发。