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.

与任何其他癌症相比，口腔癌会引起更普遍和严重的疼痛。口腔癌患者患有 严重，慢性，机械诱发的疼痛；说话和饮食受到严重破坏。阿片类药物困扰 副作用和无效的耐受性发展；口腔癌疼痛没有满意因素治疗。 我们的长期目标是开发针对癌症的非病毒基因疗法，以破坏伤害性信号传导 癌症微环境具有最小的脱靶效应，并将我们的方法转化为患者。我们创建了 两个非病毒载体具有出色的翻译效率，没有细胞毒性：一种可渗透的肽毒素 - 用阳离子脂质结合DNA；和RNA的脂聚合物。用OPRM1（μ-阿片类药物）非病毒转染 受体基因，MOR）DNA导致了MOR的重新表达，并且在 癌小鼠模型。蛋白酶激活的受体2（PAR2）在主要传入神经元中升高 支配癌症并加剧疼痛。 F2RL1（PAR2基因）的敲低部分减弱了伤害感受。尽管 两种方法都部分减轻了癌症的疼痛，我们的目标是消除。完全镇痛面临挑战； 有多种和冗余的疼痛途径，口腔癌中的基因组异质性会产生变化 在路径中。作为阻碍这些多种和多样化途径的策略，我们认为 OPRM1的重新表达和癌症中的F2RL1下调可以消除疼痛。中心假设是 一种非病毒矢量方法来获得OPRM1重新表达和口服范围内的F2RL1下调 CER将消除癌症疼痛并完全恢复功能。该项目的理由是 OPRM1 DNA和F2RL1 RNAI或CRISPR/CAS9的共递送的治疗效率将科学产生 可转化为患者的非阿片类疗法开发的严格框架。中央 将在两个具体的目的中测试了放大器：1）确定离体转染的可行性和效率（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）；并且，体内 转染（即，在化学致癌小鼠模型中直接接种基因到口腔癌中） OPRM1 DNA和/或F2RL1 RNAi或CRISPR/CAS9与非病毒载体消除癌症诱导的疼痛 在鼠标模型中； 2）测量口腔癌患者的疼痛，分析OPRM1和F2RL1的表达 CER相对于匹配的正常组织，并分析这些基因失调和 患者的疼痛。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 来自DNA和RNA的共递送，并为临床试验奠定了基础。该应用程序提出的研究 - 由于DNA和RNA的共递送为癌症，因此具有创新性 疼痛尚未完成。拟议的研究很重要，因为预计将证明强大 新型双基因疗法的持续发展和未来临床试验的科学理由。终极 偶然，这种知识可能导致新的非阿片类药物疗法的癌症疼痛发展。