Development of novel nanoparticle formulations for therapeutic in vivo lncRNA knockdown and circRNA-mediated oncomiR inhibition

除了编码蛋白质的mRNA外，还存在各种非编码RNA（小微RNA（miRNA）或长非编码RNA（lncRNA））。在癌症中，它们的水平可能异常高或低。病理性上调的致癌lncRNA是治疗性抑制的非常有趣的候选物。将在结直肠癌和胃癌细胞中探索它们通过lncRNA敲除的可药性。上调的致癌miRNA（oncomiR）也可以是抑制的靶标。在这方面，所谓的环状RNA（circRNA）代表有吸引力的候选药物，因为作为所谓的oncomiR诱饵，它们结合并因此抑制oncomiR。这两种策略都严重依赖于RNA作为药物的有效递送。基于艾格纳小组提供的用于RNA递送的聚合物纳米颗粒平台技术，该项目将开发新的改性纳米颗粒。这旨在进一步增强siRNA和oncomiR诱饵递送的功效和生物相容性、生物降解性和改进。 更具体地说，基于现有的聚乙烯亚胺（PEI）聚合物和衍生物，该项目将产生更大的一组定义的新型化学修饰，也旨在开发可生物降解的聚合物。此外，还将探索上述最佳聚合物系统与脂质纳米颗粒（脂质体）的组合。这些脂聚复合物的目的是两种系统的有利性质的组合。为此，将对各种已确定的脂质制剂进行检测。将直接将脂质聚合复合物与相应的脂质体和聚合物进行比较，以确定与单一组分相比，哪些组合将显示出改善的特性。将分析新型纳米颗粒的物理/物理化学、生物学、药理学和毒理学特性。它们将在各种二维和三维细胞和组织培养模型中被广泛研究。因此，该项目的主要重点是开发、表征和测试优化的纳米颗粒，作为潜在RNA治疗的基础，特别是用于敲低癌-lncRNA和递送环状oncomiR诱饵。大多数有前途的聚合物和RNA药物将在携带来自肿瘤细胞或来自患者肿瘤的结肠直肠癌或胃癌的小鼠中进行临床前体内治疗研究（所谓的“异种”（AVATAR）模型）。纳米颗粒和输送技术也将提供给联盟内的其他项目，用于研究和临床前开发新的治疗策略。

Besides protein-encoding mRNAs, various non-coding RNAs (small microRNAs (miRNAs) or long non-coding RNAs (lncRNAs)) exist. In cancer, their levels can be aberrantly high or low. Pathologically upregulated, oncogenic lncRNAs are very interesting candidates for therapeutic inhibition. Their druggability through lncRNA knockdown will be explored in colorectal and gastric carcinoma cells. Upregulated oncogenic miRNAs (oncomiRs) may be targets for inhibition as well. So-called circular RNAs (circRNAs) represent attractive drug candidates in this regard, because as so-called oncomiR decoys, they bind and thus inhibit oncomiRs. Both strategies heavily rely on the efficient delivery of RNAs as drugs. Based on polymeric nanoparticle platform technologies for RNA delivery available in the Aigner group, this project will develop new modified nanoparticles. This aims at further enhancement of efficacy and biocompatibility, biodegradability and refinement for siRNA and oncomiR decoy delivery. More specifically, based on available polyethylenimine (PEI) polymers and derivatives, this project will generate a larger set of defined novel chemical modifications, also aiming at the development of biodegradable polymers. Furthermore, the combination of optimal polymeric systems from the above with lipid nanoparticles (liposomes) will be explored as well. These lipopolyplexes aim at the combination of favorable properties of both systems. For this, various established lipid formulations will be tested. Lipopolyplexes will be directly compared with the corresponding liposomes and polymers to identify which combinations would show improved properties in comparison to the single components. The novel nanoparticles will be analyzed regarding their physical/physicochemical, biological, pharmacological and toxicological properties. They will be extensively studied in various 2-dimensional and 3-dimensional cell and tissue culture models. The major focus of this project is thus on the development, characterization and testing of optimized nanoparticles as basis of potential RNA therapeutics, especially for the knockdown of onco-lncRNAs and the delivery of circular oncomiR decoys. Most promising polymers and RNA drugs will proceed towards pre-clinical in vivo therapy studies in mice bearing colorectal carcinomas or gastric carcinomas from tumor cells or from patients’ tumors (so-called “xenopatient” (AVATAR) models). Nanoparticles and delivery technologies will also be provided to other projects within the consortium for research and pre-clinical development of novel therapeutic strategies.