Ligand-mediated, vehicle-free delivery of small RNAs

配体介导的无载体小 RNA 递送

• 批准号: 10737260
• 负责人: Andrea L Kasinski
• 金额: $ 38.63万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10737260
• 关键词: 2019-nCoV; Affinity; Albumins; Antineoplastic Agents; Antitumor Response; Binding; Biodistribution; Carcinoma; Cells; Circulation; Clinic; Clinical; Colon Carcinoma; Coupling; Data; Diagnostic; Disease; Dose; Drug Kinetics; Drug Targeting; FDA approved; FOLH1 gene; Folic Acid; Funding; Gene Targeting; Generations; Goals; Half-Life; Hour; Implant; Kinetics; Lead; Life; Ligands; Liver; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of ovary; Malignant neoplasm of prostate; Mediating; Methods; MicroRNAs; Mission; Mus; Non-Small-Cell Lung Carcinoma; Oncology; Phase; Property; Public Health; RNA; RNA Interference; RNA vaccine; Renal carcinoma; Renal clearance function; Safety; Serum; Serum Albumin; Small RNA; Specificity; Testing; Therapeutic Agents; Therapeutic antibodies; Time; Treatment Protocols; United States National Institutes of Health; Work; Xenograft procedure; cancer cell; cancer therapy; delivery vehicle; folate-binding protein; improved; in vivo; in vivo Model; malignant breast neoplasm; microRNA delivery; nuclease; overexpression; prevent; prostate cancer cell; risk mitigation; safety study; success; therapeutic RNA; therapeutic miRNA; tumor; uptake

PROJECT SUMMARY Like the challenges and skepticism that faced the antibody therapeutics field over a decade ago, RNA therapeutics is facing the same. And, like the antibody therapeutics field, we are beginning to realize the clinical impact of RNA therapeutics amiss these challenges. This is most clearly highlighted with the recent approval of two mRNA vaccines to prevent against SARS-CoV-2 and the first three FDA approved RNAi drugs targeted to the liver. Unfortunately, RNA-based drugs targeted to cancer cells is lagging behind, even with countless years of work that has revealed the power of using RNAi for treating oncological diseases. Lack of success in this space is attributed to inability to deliver RNAi safely and effectively. We previously developed a method that can safely deliver therapeutic microRNAs (miRNAs) to tumors that overexpress the folate receptor. However, the anti-tumor response was short-lived due to instability of the miRNA and poor pharmacokinetics, necessitating frequent dosing. To overcome these insufficiencies requires a stabilized miRNA that retains targeting activity. Recently we screened a panel of fully modified versions of miR-34a (FM-miR-34a) and identified one with >400- fold increased stability and outstanding in vivo efficacy when conjugated to folate. Treatment of mice implanted with breast cancer xenografts with folate-FM-miR-34a resulted in complete cures in two out of six mice and significant tumor regression in the remaining four. Based on this exciting data, here we propose to advance FM- miR-34 forward in two ways. In Aim 1 we will evaluate the activity, efficacy, and safety profile of FM-miR-34a in in vivo models of lung and prostate cancer. FM-miR-34a will be conjugated to: i) folate for delivery to lung cancer, and ii) PSMA-617, a ligand that targets prostate specific membrane antigen (PSMA) for delivery to prostate cancer. In Aim 2 we propose to capitalize on the stability afforded by FM-miR-34 to increase the circulation ½ time of folate-FM-miR-34 and PSMA-617-FM-miR-34a though incorporating an albumin binding moiety (ABM) into the ligands. Using these ligands we will evaluate serum albumin binding and stability of the new ligands. We will also verify that conjugation to ABM does not alter the activity of miR-34a nor cell binding and internalization kinetics. Finally, we will assess in vivo distribution of ligand-ABM-miR-34a conjugates. At the completion of this work we expect to have an all-encompassing miRNA delivery vehicle that can target a stabilized tumor suppressive RNAs specifically to NSCLC and prostate cancer. We will also have new ligands with increased circulation ½ life. The data obtained will ultimately have a significant impact in cancer treatment by providing new opportunities to advance the next phase of miRNA-based therapeutics. While proposed for NSCLC and prostate cancer, based on the utility of miR-34a for treating other cancers and overexpression of the folate receptor alone on many epithelial cancers, including ovary, kidney, and colon cancers, successful completion of this study could have far-reaching positive consequences.

项目总结 就像十多年前抗体治疗领域面临的挑战和怀疑一样，RNA 治疗公司也面临着同样的问题。而且，就像抗体治疗领域一样，我们开始意识到临床 RNA疗法的影响忽略了这些挑战。这一点最明显的体现在最近对 两种用于预防SARS-CoV-2的mRNA疫苗和FDA批准的前三种针对以下目标的RNAi药物 肝脏。不幸的是，以RNA为基础的针对癌细胞的药物却落后了，甚至已经有无数年了 揭示了使用RNAi治疗肿瘤疾病的力量的工作。在这方面缺乏成功 太空被归因于无法安全有效地运送RNAi。我们之前开发了一种方法，可以 安全地将治疗性MicroRNAs(MiRNAs)输送到叶酸受体过度表达的肿瘤。然而， 由于miRNA的不稳定和不良的药代动力学，抗肿瘤反应是短暂的，因此有必要 频繁服药。为了克服这些不足，需要一个稳定的、保持靶向活性的miRNA。 最近，我们筛选了miR-34a(FM-miR-34a)的完全修改版本的小组，并确定了其中一个带有&gt；400- 当与叶酸结合时，折叠增加了稳定性和突出的体内疗效。移植小鼠的治疗 对于乳腺癌，带有叶酸-FM-miR-34a的异种移植导致6只小鼠中有2只完全治愈， 其余4例肿瘤明显消退。基于这一令人振奋的数据，我们在此建议推进FM- 和平号-34在两个方面向前推进。在目标1中，我们将评估FM-miR-34a的活性、有效性和安全性 肺癌和前列腺癌的体内模型。FM-miR-34a将偶联到：i)用于输送到肺癌的叶酸， 和ii)PSMA-617，一种靶向前列腺特异性膜抗原(PSMA)的配体，可输送到前列腺 癌症。在目标2中，我们建议利用FM-miR-34提供的稳定性来增加发行量1/2。 加入白蛋白结合部分的叶酸-FM-miR-34和PSMA-617-FM-miR-34a的时间 进入配位体。使用这些配体，我们将评估血清白蛋白的结合和新配体的稳定性。我们 还将验证与ABM的结合不会改变miR-34a的活性，也不会改变细胞结合和内化 运动学。最后，我们将评估配体-ABM-miR-34a结合物的体内分布。 在这项工作完成后，我们希望拥有一种包罗万象的miRNA递送工具，它可以 靶向稳定的肿瘤抑制RNA，特异性针对非小细胞肺癌和前列腺癌。我们还将有新的 具有更长循环寿命的配体。获得的数据最终将对癌症产生重大影响 通过提供新的机会推进基于miRNA的治疗的下一阶段。而当 建议用于非小细胞肺癌和前列腺癌，基于miR-34a用于治疗其他癌症和 叶酸受体单独在许多上皮性癌中的过度表达，包括卵巢、肾和结肠癌 对于癌症，这项研究的成功完成可能会产生深远的积极影响。