In vivo Evaluation of Non-viral NICD Plasmid-Loaded PLGA Nanoparticles in Developing Zebrafish to Improve Cardiac Functions

Abstract

In the era of the advanced nanomaterials, use of nanoparticles has been highlighted in biomedical research. However, the demonstration of DNA plasmid delivery with nanoparticles for in vivo gene delivery experiments must be carefully tested due to many possible issues, including toxicity. The purpose of the current study was to deliver a Notch Intracellular Domain (NICD)-encoded plasmid via poly(lactic-co-glycolic acid) (PLGA) nanoparticles and to investigate the toxic environmental side effects for an in vivo experiment. In addition, we demonstrated the target delivery to the endothelium, including the endocardial layer, which is challenging to manipulate gene expression for cardiac functions due to the beating heart and rapid blood pumping. For this study, we used a zebrafish animal model and exposed it to nanoparticles at varying concentrations to observe for specific malformations over time for toxic effects of PLGA nanoparticles as a delivery vehicle. Our nanoparticles caused significantly less malformations than the positive control, ZnO nanoparticles. Additionally, the NICD plasmid was successfully delivered by PLGA nanoparticles and significantly increased Notch signaling related genes. Furthermore, our image based deep-learning analysis approach evaluated that the antibody conjugated nanoparticles were successfully bound to the endocardium to overexpress Notch related genes and improve cardiac function such as ejection fraction, fractional shortening, and cardiac output. This research demonstrates that PLGA nanoparticle-mediated target delivery to upregulate Notch related genes which can be a potential therapeutic approach with minimum toxic effects.

Nanotechnology-based non-viral vectors for gene delivery in cardiovascular diseases

“…Another strategy is to attach targeting motifs to increase selective cell binding and internalization through receptor-mediated endocytosis. Messerschmidt et al (Messerschmidt et al, 2022) Coupled anti-Tie2+Tie1 antibody to the surface of PLGA and successfully delivered the plasmid encoding Notch intracellular domain (NICD) to the zebrafish endocardial layer via antibody binding to the endocardium, which resulted in the overexpression of Notch-related genes and significant improvement in cardiac function; T. Wang et al (T. Wang et al, 2022) used platelet vesicle (PMV) camouflaged PLGA nanoparticles (PMVs@PLGA) as a carrier for miRNA inhibitors, and miRNA-targeted transport into cardiomyocytes indirectly increased the expression of Nuclear factor (erythroid-derived 2)-related factor 2 (Nrf2) by competitively binding to miR-155-5p, which protectsed myocardium in the occurrence of myocardial ischemia-reperfusion injury (MIRI), and provided a new potential pathway for the targeted treatment of MIRI.…”

Immunomodulation in age‐related disorders and nanotechnology interventions

“…Although this technology is still in the nascent stage, keeping in mind declined immune responses in the elderly, care should be taken in assessing cell membrane mediated immune responses in nanomedicine for drug delivery. Efficient models such as zebra fish can be employed for faster screening of toxicity from nano‐formulations before entering large animal models and eventually making way to clinical trials (Haque & Ward, 2018; Messerschmidt et al, 2022). Overall, most of the recent nanotechnology for treating age‐associated diseases as discussed in this review are largely restricted to a laboratory setting with very few entering clinical trials.…”