School of Biotechnology, Kalinga Institute of Industrial Technology Deemed to be University, Campus 11, Patia, Bhubaneswar 751024, Odisha, India; Biopioneer Private Limited, Patia, Bhubaneswar, Odisha, India.
School of Biotechnology, Kalinga Institute of Industrial Technology Deemed to be University, Campus 11, Patia, Bhubaneswar 751024, Odisha, India.
Eur J Pharm Sci. 2022 Apr 1;171:106125. doi: 10.1016/j.ejps.2022.106125. Epub 2022 Jan 14.
Vascular endothelial growth factor (VEGF) is considered as one of the vital growth factors for angiogenesis, which is primarily responsible for the progress and maintenance of new vascular network in tumor. Numerous studies report that inhibition of VEGF-induced angiogenesis is a potent technique for cancer suppression. Recently, RNA interference, especially small interfering RNA (siRNA) signified a promising approach to suppress the gene expression. However, the clinical implementation of biological macromolecules such as siRNA is significantly limited because of stability and bioavailability issues. Herein, self-assembled peptide nanospheres have been generated from L,L-cyclic peptides using hydrophobic (Trp), positively charged (Arg) and cysteine (Cys) amino acid residues and demonstrated as vehicles for intracellular delivery of VEGF siRNA and VEGF antisense oligonucleotide. Formation of peptide nanostructures is confirmed by HR-TEM, AFM, SEM and DLS analysis. Possible mechanism of self-assembly of the cyclic peptides and their binding with macromolecules are demonstrated by in-silico analysis. Gel electrophoresis reveals that the newly generated peptide based organic materials exhibit strong binding affinity toward siRNAs / antisense oligonucleotides (ASOs) at optimum concentration. Flow cytometry and confocal microscopy results confirm the efficiency of the new biomaterials toward the intracellular delivery of fluorescent labeled siRNA / ASOs. Furthermore, VEGF expression evaluated by western blot and RT-PCR upon the delivery of functional VEGF siRNA/ASOs suggests that very low concentrations of VEGF siRNA/ASOs cause significant gene knockdown at protein and mRNA levels, respectively.
血管内皮生长因子（VEGF）被认为是血管生成的重要生长因子之一，主要负责肿瘤中新血管网络的进展和维持。大量研究报告表明，抑制 VEGF 诱导的血管生成是抑制癌症的有效技术。最近，RNA 干扰，特别是小干扰 RNA（siRNA），成为抑制基因表达的一种很有前途的方法。然而，由于稳定性和生物利用度问题，生物大分子如 siRNA 的临床实施受到了显著限制。在此，使用疏水性（Trp）、正电荷（Arg）和半胱氨酸（Cys）氨基酸残基的 L,L-环肽生成了自组装肽纳米球，并将其证明为 VEGF siRNA 和 VEGF 反义寡核苷酸的细胞内递送载体。通过高分辨率 TEM、AFM、SEM 和 DLS 分析证实了肽纳米结构的形成。通过计算机模拟分析证明了环肽的自组装和与大分子结合的可能机制。凝胶电泳显示，新生成的基于肽的有机材料在最佳浓度下对 siRNAs/反义寡核苷酸（ASOs）表现出很强的结合亲和力。流式细胞术和共聚焦显微镜结果证实了新型生物材料对荧光标记的 siRNA/ASOs 细胞内递送的效率。此外，通过 Western blot 和 RT-PCR 评估功能性 VEGF siRNA/ASOs 的递送后 VEGF 表达表明，非常低浓度的 VEGF siRNA/ASOs 分别在蛋白和 mRNA 水平上导致显著的基因敲低。
