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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

基本信息

批准号：
10616514
负责人：
Daniel W Binzel
金额：
$ 33.57万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
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

项目摘要

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.

摘要三阴性乳腺癌是一种异质性、复杂性和侵袭性的乳腺癌亚型。 TNBC患者对化疗的反应很差，导致高死亡率和预后恶化。 RNAi疗法，包括siRNA和miRNA，已经显示出在TNBC癌症治疗方面的巨大潜力。就以下方面而言，开发一种安全的靶向递送系统并避免有效载荷是至关重要的实现RNAi疗法的全部潜力，并可能彻底改变TNBC的临床治疗。本课题组已经证实，将miR21抗体导入TNBC可以有效地抑制TNBC细胞的增殖 (Shu D等人)ACS Nano。2015，27：9731；尹浩等人。舒丁·摩尔疗法。2019年27：1252)。我们还有最近报道了利用RNA纳米颗粒取向来修饰具有靶向配体的外切体表面目的：为TNBC治疗提供携带siRNA的外切体。我们证明了外切体可以被用作将siRNA非常有效地输送到TNBC癌细胞以抑制癌症生长的纳米载体(PI F，Binzel D等人)。纳特。纳米替卡诺(Nanotechnol)。2018年；13：82)。用体外KB细胞模型进行研究的初步数据显示高效抑癌背后的机制是通过外切体胞内递送siRNA 没有内体诱捕(甄志等人)J控制释放。2019,311：43)。在这项研究中，我们将调查具有TNBC特异性的外切体对TNBC细胞的RNAi靶向和递送机制靶向递送的配体。我们的目标是选择一位领先的TNBC治疗候选者，以迈向潜在的临床翻译。我们将把RNA纳米技术的靶向和药物输送能力与外切体将RNAi靶向传递到细胞胞浆，而不需要内吞体包埋。我们将构建和鉴定含有多功能外体RNA纳米颗粒的复合体。靶向配体包括RNA适配子(EGFRapt或CD133apt、CD44apt和nAHRap)或化学配体(甲氨蝶呤)和肿瘤抑制RNAi疗法(siRNA、抑制子miRNAs和抗- 致癌miRNA)。我们将进一步研究内化和细胞内转运的途径。除了调查外切体中的有效载荷是通过融合传递到细胞质还是通过融合传递到细胞质通过内吞作用形成的内吞体。RNAi货物的释放和细胞加工的深入研究外显体将被完成。我们将研究PK/PD参数，传递机制，抗肿瘤治疗性RNA纳米粒的有效性和安全性，以选择临床前的主要候选者翻译。临床前研究将使人们清楚地了解外切体-rna纳米颗粒的可行性。用于TNBC治疗的复合体，并为迈向研究性新药(IND)应用提供数据这将促进临床试验的进展。