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Fabrication of self-assembled peptide nanoparticles for in vitro assessment of cell apoptosis pathway and in vivo therapeutic efficacy

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Abstract

Near-infrared fluorescent (NIRF) dye-coupled self-assembled RGD-linked proapoptotic peptide nanoparticles have been synthesized with spherical shape and size ~ 30–40 nm diameters. The peptide sequence was coupled with cyanine 5.5 probe as NIRF-dye to introduce optical imaging properties and pH-dependent method was used to design Cy5.5 coupled self-assembled peptide nanoparticles (f-SAPNs). This nanoprobe has the ability to target αvβ3-integrin receptor overexpressed on cancer cell’s surface with improved internalization capabilities into the mitochondria. The in situ study showed that this peptide sequence has potential to disrupt the mitochondrial membrane efficiently, activating the Caspase-3 enzyme, and ultimately induces cell apoptosis. It has been observed from in vitro study that the degree of apoptosis for f-SAPNs was increased from 25.6% to 96.3%, while decreased degree of necrosis from 51.7% to 0.2% compared with its parent peptide analog (Cy5.5-c[RGDKLAK]; f-CP) occurs. Further investigations revealed that these f-SAPNs showed high uptake in U87MG glioblastoma cells in comparison with PC-3 prostate cancer cells. Moreover, in vivo therapeutic studies represented the prominent decrease in the size of tumor tissue treated with f-CP and f-SAPNs (201 ± 13 mm3 and 104 ± 6 mm3, respectively) compared with untreated tumor tissues (366 ± 18 mm3). These outcomes highlighted the specificity, and efficacy of f-SAPNs toward αvβ3-integrin expressing tumor tissue in vivo and suggested that these novel designed f-SAPNs may serve as a potential theranostic drug for brain tumor glioblastoma multiforme.

Graphical abstract

The pH-sensitive method gives NIRF dye-coupled self-assembled peptide nanoparticle (f-SAPNs), enables the tunable synthesis of spherical nanoparticles with high stability towards proteolysis, improved biocompatibility, and promising therapeutic efficacy.

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Funding

This research work was financially supported by the National Natural Science Foundation of China (No. 21575055).

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Authors and Affiliations

  1. State Key Laboratory of Applied Organic Chemistry, College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou, 730000, Gansu, People’s Republic of China

    Syed Faheem Askari Rizvi, Shuai Mu & Haixia Zhang

  2. School of Pharmacy, Lanzhou University, Lanzhou, 730000, Gansu, People’s Republic of China

    Chunyan Zhao

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  4. Haixia Zhang
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Correspondence to Haixia Zhang.

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All animal studies performed in this study were in accordance with the compliance of animal ethical committee of the National Institute of Health and National Regulation of China for Care and Use of Laboratory Animals (Lanzhou University, Gansu, China).

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Supplementary file1 (DOCX 5099 KB) The online version of supplementary material contains description about characterizations, protocols for in vitro cell studies, in silico study, and therapeutic study available (Pdf).

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Rizvi, S.F.A., Mu, S., Zhao, C. et al. Fabrication of self-assembled peptide nanoparticles for in vitro assessment of cell apoptosis pathway and in vivo therapeutic efficacy. Microchim Acta 189, 53 (2022). https://doi.org/10.1007/s00604-021-05148-7

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