Center for Targeted Drug Delivery, Department of Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, Harry and Diane Rinker Health Science Campus, Irvine, California 92618, United States.
Mol Pharm. 2021 Mar 1;18(3):986-1002. doi: 10.1021/acs.molpharmaceut.0c00969. Epub 2021 Jan 26.
RNAi is a biological process that utilizes small interfering RNA (siRNA) to prevent the translation of mRNA to protein. This mechanism could be beneficial in preventing the overexpression of proteins in cancer. However, the cellular delivery of siRNA has proven to be challenging due to its inherent negative charge and relative instability. Here, we designed a multicomponent delivery system composed of a specifically designed peptide (linear or cyclic fatty acyl peptide conjugates and hybrid cyclic/linear peptides) and several lipids (DOTAP, DOPE, cholesterol, and phosphatidylcholine) to form a nanoparticle, which we have termed as peptide lipid-associated nucleic acids (PLANAs). Five formulations were prepared (a formulation with no peptide, which was named lipid-associated nucleic acid or LANA, and PLANA formulations A-D) using a mini extruder to form uniform nanoparticles around 100 nm in size with a slightly positive charge (less than +10 mv). Formulations were evaluated for peptide incorporation, siRNA encapsulation efficiency, release profile, toxicity, cellular uptake, and protein silencing. Our experiments showed effective encapsulation of siRNA (>95%), a controlled release profile, and negligible toxicity in formulations that did not contain a positively charged lipid. The results also revealed that PLANAs C and D exhibited optimum cellular uptake (with 80-90% siRNA-positive cells for most of the formulations). PLANA D formulation was selected to silence two model proteins (Src and RPS6KA5) in the triple-negative human breast cancer cell line MDA-MB-231, with promising silencing efficiency, which diminished the expression of RPS6KA5 and Src to approximately 29 and 38% compared to naïve cells, respectively. Many approaches have been investigated for safe and efficient delivery of nucleic acids in the last 20 years; however, many have failed due to the multifaceted challenges to overcome. Our results show a promising potential for a multicomponent design that incorporates different components for a variety of delivery tasks, which warrants further investigation of PLANAs .
RNAi 是一种利用小干扰 RNA（siRNA）防止 mRNA 翻译为蛋白质的生物过程。该机制可用于预防癌症中蛋白质的过度表达。然而，siRNA 的细胞递送已被证明具有挑战性，因为其固有的负电荷和相对不稳定性。在这里，我们设计了一种由特定设计的肽（线性或环状脂肪酸肽缀合物和混合环/线性肽）和几种脂质（DOTAP、DOPE、胆固醇和磷脂酰胆碱）组成的多组分递送系统，形成纳米颗粒，我们将其命名为肽脂质相关核酸（PLANA）。使用迷你挤出机制备了五种制剂（一种没有肽的制剂，命名为脂质相关核酸或 LANA，以及 PLANA 制剂 A-D），以形成粒径约为 100nm 的均匀纳米颗粒，带轻微正电荷（小于+10mv）。评估制剂的肽掺入、siRNA 包封效率、释放曲线、毒性、细胞摄取和蛋白质沉默。我们的实验表明，在不包含带正电荷脂质的制剂中，siRNA 的有效包封率>95%，释放曲线可控，毒性可忽略不计。结果还表明，PLANA C 和 D 表现出最佳的细胞摄取（对于大多数制剂，有 80-90%的 siRNA 阳性细胞）。选择 PLANA D 制剂来沉默三阴性人乳腺癌细胞系 MDA-MB-231 中的两种模型蛋白（Src 和 RPS6KA5），沉默效率高，与原始细胞相比，RPS6KA5 和 Src 的表达分别降低到约 29%和 38%。在过去的 20 年中，已经研究了许多安全有效的核酸传递方法；然而，由于需要克服多方面的挑战，许多方法都失败了。我们的结果表明，对于包含多种成分的多组分设计具有很大的潜力，可用于各种传递任务，这需要进一步研究 PLANAs。
