Afonso Inês, Neves Ana R, Eusébio Dalinda, Albuquerque Tânia, Vivès Eric, Boisguérin Prisca, Santos Adriana O, Sousa Ângela, Costa Diana
CICS-UBI-Health Sciences Research Centre, University of Beira Interior, 6200-506 Covilhã, Portugal.
PhyMedExp, Université de Montpellier, INSERM, CNRS, 34295 Montpellier, France.
Pharmaceutics. 2024 Oct 29;16(11):1389. doi: 10.3390/pharmaceutics16111389.
Gene therapy can be viewed as a promising/valuable therapeutic approach directed to cancer treatment, including glioblastoma. Concretely, the combination of gene therapy with chemotherapy could increase its therapeutic index due to a synergistic effect. In this context, bovine serum albumin (BSA)-coated temozolomide (TMZ)-peptide (WRAP5)/p53 gene-based plasmid DNA complexes were developed to promote payload co-delivery. Design of experiments (DoE) was employed to unravel the BSA-coated TMZ-WRAP5/p53 nanocomplexes with the highest potential by considering the nitrogen to phosphate groups ratio (N/P), and the BSA concentration as inputs and the size, polydispersity index, surface charge and p53-based plasmid complexation capacity (CC) as DoE outputs. The obtained quadratic models were statistically significant (-value < 0.05) with an adequate coefficient of determination, and the correspondent optimal points were successfully validated. The optimal complex formulation had N/P of 1.03, a BSA concentration of 0.08%, a size of approximately 182 nm, a zeta potential of +9.8 mV, and a pDNA CC of 96.5%. The optimal nanocomplexes are approximately spherical. A cytotoxicity assay showed that these BSA-coated TMZ-WRAP5/p53 complexes did not elicit toxicity in normal brain cells, and a hemolysis study demonstrated the hemocompatibility of the complexes. The complexes were stable in cell culture medium and fetal bovine serum and assured pDNA protection and release. Moreover, the optimal BSA-coated complexes were able of gene transcription and promoted a significant inhibition of glioblastoma cell viability. The reported findings instigate the development of future research to evaluate their potential utility to TMZ/p53 co-delivery. The DoE tool proved to be a powerful approach to explore and tailor the composition of BSA-coated TMZ-WRAP5/p53 complexes, which are expected to contribute to the progress toward a more efficient therapy against cancer and, more specifically, against glioblastoma.
基因治疗可被视为一种有前景/有价值的癌症治疗方法，包括胶质母细胞瘤。具体而言，由于协同效应，基因治疗与化疗的联合应用可提高其治疗指数。在此背景下，开发了牛血清白蛋白（BSA）包被的替莫唑胺（TMZ）-肽（WRAP5）/p53基因质粒DNA复合物，以促进有效载荷的共递送。采用实验设计（DoE）方法，通过将氮磷比（N/P）和BSA浓度作为输入参数，将粒径、多分散指数、表面电荷和基于p53的质粒络合能力（CC）作为DoE输出参数，来解析具有最高潜力的BSA包被的TMZ-WRAP5/p53纳米复合物。所获得的二次模型具有统计学意义（p值<0.05）且决定系数合适，相应的最优点也得到了成功验证。最佳复合物配方的N/P为1.03，BSA浓度为0.08%，粒径约为182nm，zeta电位为+9.8mV，pDNA CC为96.5%。最佳纳米复合物近似球形。细胞毒性试验表明，这些BSA包被的TMZ-WRAP5/p53复合物在正常脑细胞中不引起毒性，溶血研究证明了复合物的血液相容性。复合物在细胞培养基和胎牛血清中稳定，确保了pDNA的保护和释放。此外，最佳的BSA包被复合物能够进行基因转录并显著抑制胶质母细胞瘤细胞的活力。所报道的研究结果促使开展未来研究以评估它们在TMZ/p53共递送方面的潜在效用。DoE工具被证明是一种探索和定制BSA包被的TMZ-WRAP5/p53复合物组成的有效方法，有望推动针对癌症，尤其是针对胶质母细胞瘤的更有效治疗的进展。
