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，导致高死亡率和程序性恶化。 RNAi疗法包括siRNA和miRNA，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的临床信任。 我们的小组已经证明，抗MIR21向TNBC的递送可以有效抑制TNBC细胞的繁殖力 （Shu d等，ACS Nano。 最近报道了使用RNA纳米属方向用靶向配体装饰外泌体表面 为了提供siRNA的外泌体进行TNBC处理。 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。 NANOT。 癌症抑制效率高效率的机制是通过外泌体的siRNA的毛毛虫 没有内体诱捕（Zhen Z等人J控制释放。 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 有针对性交付的配体。 临床翻译。我们将结合RNA纳米技术的靶向和药物能力 针对目标的外泌体将RNAi递送到无内体夹层的细胞细胞质中。 我们将构建和评估携带的多功能外泌体RNA纳米复合物 靶向配体，包括RNA适体（Egfrapt或替代性，CD133APT，CD44APT和NAHRSAPT）或 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）。 除了调查外泌体中的有效载荷是通过融合还是通过融合输送到细胞质的 内吞作用的内体。 外泌体将完成。 治疗性RNA纳米颗粒的功效和安全性，以便选择Proclinical的铅候选者 翻译。 用于TNBC疗法的复合物，并提供数据以移动朝向调查的新药（IND）应用 这将促进临床试验的进步。