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都更普遍和严重。口腔癌患者患 严重的、慢性的、机械引起的疼痛;说话和进食严重中断。阿片类药物困扰着 随着耐受性的发展，具有副作用和无效;对于口腔癌疼痛没有令人满意的治疗。 我们的长期目标是开发靶向癌症的非病毒基因疗法，以破坏肿瘤细胞中的伤害性信号传导。 癌症微环境与最小的脱靶效应，并将我们的方法转化为患者。我们创建 两种具有优异转染效率和无细胞毒性的非病毒载体：细胞可渗透肽复合物， 与用于DNA的阳离子脂质结合;和，用于RNA的脂质聚合物。用OPRM 1（μ-阿片样物质）非病毒转染 受体基因，莫尔）DNA导致莫尔的再表达和伤害感受的部分减弱（即，疼痛） 癌症小鼠模型。蛋白酶激活受体-2（PAR 2）在初级传入神经元中升高， 使癌细胞神经化并引发疼痛。F2 rl 1（PAR 2基因）的敲低部分减弱伤害感受。而 这两种方法都能部分减轻癌症疼痛，我们的目标是消除。完全镇痛是一个挑战; 存在多种和冗余的疼痛通路，并且口腔癌的基因组异质性产生变异， 在路径上。作为一种阻止这些多种多样途径的策略，我们认为， OPRM 1的重新表达和F2 RL 1的下调可以消除疼痛。核心假设是 一种非病毒载体方法可以在口腔内获得OPRM 1的重新表达和F2 RL 1的下调， cer将消除癌症疼痛并完全恢复功能。该项目的理由是， OPRM 1 DNA和F2 RL 1 RNAi或CRISPR/Cas9共同递送的治疗功效将产生科学的结果 为开发可用于患者的非阿片类药物疗法提供了严格的框架。中央卫生- 假设将在两个特定目的中进行测试：1）确定离体转染的可行性和功效（即， 在产生异种移植小鼠模型之前用双基因转染口腔癌细胞）;以及，在体内 转染（即，将基因直接接种到化学致癌物小鼠模型中的口腔癌中）， OPRM 1 DNA和/或F2 RL 1 RNAi或CRISPR/Cas9与非病毒载体联合使用以消除癌症诱导的疼痛 2）测量口腔癌患者的疼痛，分析其can中OPRM 1和F2 RL 1的表达， cers相对于匹配的正常组织，并分析这些基因的失调与 病人的痛苦。结果将为我们提供有关口腔癌患者可能受益的比例的数据 从DNA和RNA的共同输送，并为临床试验奠定了基础。本申请中提出的研究- 这种方法是创新的，因为用非病毒载体将DNA和RNA共递送到癌症中， 疼痛的部分还没有完成。这项拟议中的研究意义重大，因为预计它将被证明是强有力的 为新型双基因疗法的持续开发和未来临床试验提供科学依据。Ulti 然而，这些知识可能会导致癌症疼痛的新型非阿片类药物疗法的发展。