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: 10543510
• 负责人: Brian L Schmidt
• 金额: $ 58.38万
• 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/10543510
• 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; 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; Legal patent; Life; Lipids; Malignant Neoplasms; Measures; Mechanics; Methodology; Methylation; Mission; Mus; Nerve; Nociception; Non-Viral Vector; Normal tissue morphology; Opioid; Opioid Receptor; Oral; Orofacial Pain; Outcome; PAR-2 Receptor; Pain; Pain Measurement; Pain management; 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; Receptor Gene; Regulation; Research; 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; beta-Endorphin; cancer cell; cancer pain; cancer therapy; carcinogenesis; chemical carcinogen; chronic neuropathic pain; clinically relevant; cytotoxicity; debilitating pain; demethylation; disability; drinking; evidence base; functional restoration; gene therapy; in vivo; innovation; invention; 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.

口腔癌引起的疼痛比任何其他癌症患者都更为普遍和严重。 严重的、慢性的、机械性疼痛；说话和进食受到阿片类药物的严重干扰。 具有副作用，并且随着耐受性的发展而无效；对于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针对癌症的非病毒基因疗法，以破坏癌症中的伤害性信号传导 我们创建了具有最小脱靶效应的癌症微环境，并将我们的方法转化为患者。 两种具有优异转染效率且无细胞毒性的非病毒载体：一种细胞渗透性肽组合物 与用于 DNA 的阳离子脂质结合；以及用于 RNA 的非病毒转染（μ-阿片类药物）。 受体基因，MOR）DNA导致MOR的重新表达和伤害感受（即疼痛）的部分减弱 癌症小鼠模型中，初级传入神经元中的蛋白酶激活受体 2 (PAR2) 升高。 神经支配癌症并驱动疼痛，而 F2rl1（PAR2 基因）的敲低则部分减弱了伤害感受。 这两种方法都可以部分减轻癌症疼痛，但我们的目标是完全消除疼痛； 疼痛通路有多种且冗余，口腔癌的基因组异质性会产生变异 作为阻碍这些多种途径的策略，我们假设将以下方法结合起来： 癌症中 OPRM1 重新表达和 F2RL1 下调可以消除疼痛。 一种非病毒载体方法可在口腔内重新表达 OPRM1 并下调 F2RL1 cer 将消除癌症疼痛并完全恢复功能。 OPRM1 DNA 和 F2RL1 RNAi 或 CRISPR/Cas9 共同递送的治疗效果将产生科学的结果 开发可转化为患者的非阿片类药物疗法的严格框架。 假设将在两个具体目标上进行测试：1）确定离体转染的可行性和有效性（即， 在生成异种移植小鼠模型之前用双基因转染口腔癌细胞）； 转染（即将基因直接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 与非病毒载体一起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，分析其口腔中的 OPRM1 和 F2RL1 表达 cer 相对于匹配的正常组织，并分析这些基因的失调与 研究结果将为我们提供有关可能受益的口腔癌患者比例的数据。 DNA 和 RNA 的共同传递，并为本申请中提出的研究奠定了基础。 化是创新的，因为将 DNA 和 RNA 与非病毒载体共同递送到癌症中以进行管理 所提出的研究意义重大，因为它预计会被证明是强有力的。 新型双基因疗法的持续开发和未来临床试验的科学依据。 顺便说一句，这些知识可能会导致开发出针对癌症疼痛的新型非阿片类药物疗法。