Study on the cellular internalization mechanisms and in vivo anti-bone metastasis prostate cancer efficiency of the peptide T7-modified polypeptide nanoparticles

Abstract

Bone-metastasis prostate cancer (BMPCa)-targeting gene therapy is gaining increasing concern in recent years. The peptide T7-modified polypeptide nanoparticles for delivery DNA (CRD-PEG-T7/ pPMEPA1) was prepared as our previous study. However, the feasibility of CRD-PEG-T7/pPMEPA1 for BMPCa treatment, the mechanisms underlying cellular uptake, anti-BMPCa effect, and administration safety requires further research. LNCaP cells treated with endocytosis inhibitors and excessive T7 under different culture condition were carried out to investigate the mechanisms of cellular uptake of the CRD-PEG-T7-pPMEPA1. A transwell assay was applied to evaluate the cell migration ability. Besides, the tumor volume and survival rates of the PCa xenograft mice model were recorded to estimate the antitumor effect. In addition, the weight profiles of the PCa tumor-bearing mice, the blood chemistry, and the HE analysis of visceral organs and tumor was conducted to investigate the administration safety of CRD-PEG-T7/pPMEPA1. The results showed that PCa cellular uptake was decreased after treating with excessive free T7, endocytosis inhibitors and lower incubation temperature. Besides, CRD-PEG-T7/ pPMEPA1 could inhibit the LNCaP cells chemotaxis and tumor growth. In addition, the survival duration of the PCa tumor-bearing mice treating with CRD-PEG-T7/pPMEPA1 was significantly prolonged with any systemic toxicity or damage to the organs. In conclusion, this research proposes a promising stratagem for treatment BMPCa by providing the biocompatible and effective carrier for delivery DNA therapeutic agents.

αvβ3 integrin-specific exosomes engineered with cyclopeptide for targeted delivery of triptolide against malignant melanoma

“…The results indicated that internalization was associated with clathrin-mediated and caveolin-dependent endocytosis. However, there was no significant difference in cellular uptake between the amiloride and control groups (p > 0.05), implying that macropinocytosis-mediated endocytosis is not involved in A375 cell uptake of cRGD-Exo/TP, which is consistent with previous studies [ 44 , 45 ]. Upon reducing the incubation temperature or pretreatment with inhibitors, the internalization of cRGD-Exo/TP was markedly decreased to a certain extent compared with the control (at 37 °C), strongly indicating that multiple internalization pathways are involved.…”

“…Therefore, we also studied the anti-invasion effect of cRGD-Exo/TP via a Transwell assay. Serum (10%) added to the bottom chamber for stimulated haptotaxis motility was introduced to evaluate the anti-invasion ability of cRGD-Exo/TP on A375 cells [ 45 ]. Microscopic photographs (Fig.…”

“…A375 cells were pretreated with various endocytic inhibitors or cultured at low temperature to probe the internalization mechanisms of cRGD-Exo/TP [ 45 ]. A375 cells incubated in 12-well plates were pretreated with serum-free DMEM containing sodium azide (5 mg/mL), nystatin (50 μg/mL), chlorpromazine (5 μg/mL) and amiloride (50 μg/mL) for 1 h each.…”

Non-viral Vectors in Gene Therapy: Recent Development, Challenges, and Prospects

“…applied T7-modified polypeptide nanoparticles, CRD-PEG-T7, to deliver the pDNA pPMEPA1 for bone metastatic prostate cancer treatment. They found the incorporation of T7 inhibited tumor growth and extended survival time of tumor-bearing mice ( 53 ).…”

Recent Advances in Nanomedicine for the Diagnosis and Treatment of Prostate Cancer Bone Metastasis

“…In PCa patients, low expression levels of prostrate transmembrane protein androgen induced 1 (PMEPA1) have been linked to a higher incidence of bone metastasis [ 198 , 199 ]. To evaluate the merits of regulating PMEPA1 to elicit a therapeutic benefit in PCa, Gu et al designed a nanoparticle delivery system for pPMEPA1 plasmid DNA [ 200 ]. First, they covalently tethered the polypeptides R7D6 (CRRRRRRRCDDDDDD; CRD) and T7 (HAIYPRH) to the N-hydroxysuccinimide and maleimide functional groups of an α-maleimide-ω-N-hydroxysuccinimide-PEG polymer, respectively.…”