TNBC Ligand-displaying Exosomes Using RNA Nanotechnology for Targeted Cytosol Delivery of RNAi without Endosome Entrapment

TNBC 配体展示外泌体，利用 RNA 纳米技术实现 RNAi 的靶向胞质溶胶递送，无需内体截留

• 批准号: 10404055
• 负责人: Daniel W Binzel
• 金额: $ 32.46万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-11 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10404055
• 关键词: Antibodies; Apoptosis; Biodistribution; Breast Cancer Cell; Breast Cancer Patient; Breast Cancer Treatment; Breast Cancer therapy; Caveolae; Cell Proliferation; Cells; Chemicals; Clathrin; Clinic; Clinical Treatment; Clinical Trials; Complex; Cytosol; Data; Drug Delivery Systems; Drug resistance; Dyes; ERBB2 gene; Endocytosis; Endosomes; Estrogen Receptors; Evaluation; Gene Silencing; Genes; Goals; Growth; Homing; Hydrophobicity; Image; In Vitro; Investigation; Investigational New Drug Application; KB Cells; Label; Lead; Ligand Binding; Ligands; Malignant Neoplasms; Membrane; Membrane Fusion; Methods; Methotrexate; MicroRNAs; Nanotechnology; Oncogenic; Organ; Organelles; Pathway interactions; Patient-derived xenograft models of breast cancer; Phagocytosis; Progesterone Receptors; Prognosis; Proteins; RNA; RNA Interference; RNA Interference Therapy; Receptor Cell; Recurrence; Reporting; Safety; Site; Small Interfering RNA; Surface; System; Technology; Testing; Therapeutic; Toxic effect; Translations; Treatment Efficacy; Tumor Burden; Yin; aggressive breast cancer; aptamer; cancer cell; cancer subtypes; cancer therapy; chemotherapy; clinical translation; exosome; hormone therapy; imaging agent; immunogenicity; in vitro Model; in vivo; inhibitor; knock-down; lead candidate; malignant breast neoplasm; mortality; mouse model; nano; nanocarrier; nanoparticle; novel; patient derived xenograft model; pharmacokinetics and pharmacodynamics; pre-clinical; preclinical study; siRNA delivery; side effect; survivin; targeted delivery; therapeutic RNA; therapeutic candidate; therapeutic miRNA; therapeutic siRNA; trafficking; triple-negative invasive breast carcinoma; tumor; tumor growth; uptake

SUMMARY Triple negative breast cancer (TNBC) is a heterogeneous, complex, and aggressive breast cancer subtype. TNBC patients respond poorly to chemotherapy, leading to high mortality rates and a worsening prognosis. RNAi therapeutics, including siRNA and miRNA, have shown tremendous potential for TNBC cancer therapy. Developing a safe targeted delivery system with endosomal avoidance of the payload is crucial in terms of realizing the full potential of RNAi therapeutics and could revolutionize clinical treatment of TNBC. Our group has demonstrated that delivery of anti-miR21 to TNBC can efficiently inhibit TNBC cell proliferation (Shu D, et al. ACS Nano. 2015, 27:9731; Yin H, et al. Shu D. Mol Therapy. 2019, 27:1252). We have also recently reported the use of RNA nanoparticle orientation for decorating exosome surfaces with targeting ligands to deliver siRNA loaded exosomes for TNBC treatment. We proved that exosomes can be utilized as nanocarriers to deliver siRNA to TNBC cancer cells very efficiently to inhibit cancer growth (Pi F, Binzel D et al. Nat. Nanotechnol. 2018;13:82). Preliminary data from the investigation with KB cell models in vitro has shown the mechanism behind the high efficiency of cancer inhibition to be the cytosolic delivery of siRNA via exosomes without endosomal trapping (Zhen Z, et al. J Control Release. 2019,311:43). In this study, we will investigate the targeting and delivery mechanisms of RNAi to TNBC cells by exosomes displayed with TNBC specific ligands for targeted delivery. It’s our goal to select a lead TNBC therapeutic candidate to move towards potential clinical translation. We will combine the targeting and drug delivery capabilities of RNA nanotechnology and exosomes for targeted delivery of RNAi to cell cytosols without endosomal entrapment. We will construct and evaluate the multi-functional exosomal RNA nanoparticle complexes harboring targeting ligands including RNA aptamers (EGFRapt, or alternatively, CD133apt, CD44apt, and nAHRsapt) or chemical ligand (Methotrexate) and tumor suppressing RNAi therapeutics (siRNA, suppressor miRNAs and anti- oncogenic miRNA). We will further investigate the pathways of internalization and intracellular trafficking in addition to investigating whether the payloads in the exosome are delivered to cytosol via fusion or to the endosome via endocytosis. In depth studies of the release and cellular processing of RNAi cargo loaded into exosomes will be completed. We will investigate the PK/PD parameters, delivery mechanisms, antitumor efficacy, and safety of the therapeutic RNA nanoparticles in order to select a lead candidate for preclinical translation. The preclinical studies will give a clear understanding of the viability of exosome-RNA nanoparticle complexes for TNBC therapy and provide data to move towards an Investigational New Drug (IND) application that will facilitate advancement to clinical trials.

概括 三重阴性乳腺癌（TNBC）是一种异质，复杂和侵略性的乳腺癌亚型。 TNBC患者对化学疗法的反应较差，导致高死亡率和令人担忧的预后。 包括siRNA和miRNA在内的RNAi疗法对TNBC癌症治疗显示出巨大的潜力。 从内体避免有效载荷的情况下，开发安全的目标交付系统至关重要 意识到RNAi疗法的全部潜力，并可能彻底改变TNBC的临床治疗。 我们的小组已经证明，抗MIR21向TNBC的递送可以有效抑制TNBC细胞增殖 （Shu D等人ACS Nano。2015，27：9731; Yin H等人D. Mol Therapy。2019，27：1252）。我们也有 最近报道了使用RNA纳米颗粒方向用靶向配体装饰外泌体表面 输送siRNA负载外泌体进行TNBC处理。我们证明外泌体可以用作 纳米载体非常有效地向TNBC癌细胞运送siRNA以抑制癌症的生长（Pi F，Binzel D等。 纳特。纳米技术。 2018; 13：82）。在体外使用KB细胞模型研究的初步数据已显示 癌症抑制效率高的机制是通过外泌体的siRNA的胞质递送 没有内体捕获（Zhen Z等人Jontrol版本。2019,311：43）。在这项研究中，我们将调查 用TNBC特异性展示的外泌体对RNAi对TNBC细胞的靶向和递送机制 用于靶向输送的配体。我们的目标是选择铅TNBC治疗候选者以朝着潜在的 临床翻译。我们将结合RNA纳米技术的靶向和药物输送能力和 将RNAi靶向递送至无内体夹层的细胞细胞质的外泌体。 我们将构建和评估携带的多功能外泌体RNA纳米颗粒复合体 靶向包括RNA适体（Egfrapt或替代的CD133APT，CD44APT和NAHRSAPT）或NAHRSAPT）的靶向配体 化学配体（甲氨蝶呤）和肿瘤抑制RNAI疗法（siRNA，抑制miRNA和抗 - 致癌miRNA）。我们将进一步研究内部化和细胞内贩运的途径 除了调查外泌体中的有效载荷是通过融合传递到细胞质的还是 内吞作用的内体。在深入研究RNAi货物的释放和细胞加工中 外泌体将完成。我们将研究PK/PD参数，输送机制，抗肿瘤 治疗性RNA纳米颗粒的功效和安全性，以便选择临床前候选者 翻译。临床前研究将清楚地了解外泌体-RNA纳米颗粒的生存能力 用于TNBC治疗的复合物，并提供数据以朝向研究新药（IND）应用 这将有助于临床试验的进步。