Enhanced Caspase-Mediated Abrogation of Autophagy by Temozolomide-Loaded and Panitumumab-Conjugated Poly(lactic-co-glycolic acid) Nanoparticles in Epidermal Growth Factor Receptor Overexpressing Glioblastoma Cells

Abstract

The mechanism of cell death has attracted a great deal of research interest in the design of antitumor therapy in recent days. Several attempts have been carried out in this direction and in our study also, we studied this phenomenon with the design of panitumumab (PmAb)-conjugated and temozolomide (TMZ)-loaded poly(lactic-co-glycolic acid) nanoparticles (PLGA-NPs), termed PmAb-TMZ-PLGA-NPs. First, PmAb was functionalized on the surface of TMZ-PLGA-NPs using ethyl(dimethylaminopropyl)carbodiimide (EDC)−N-hydroxysuccinimide (NHS) chemistry. Targeted PLGA-NPs significantly enhanced the cellular uptake of nanoparticles in the U-87 MG cell line as a result of the high epidermal growth factor receptor (EGFR) expression, compared to the LN229 cell line. Our study demonstrated that following the treatment of PmAb-TMZ-PLGA-NPs, a more pronounced anticancer effect was noticed in comparison with free TMZ and TMZ-PLGA-NPs. Further, a more pronounced cytotoxic effect of PmAb-TMZ-PLGA-NPs was observed in the high EGFR-overexpressed glioblastoma multiforme (GBM) model (U-87 MG) cell line compared to the low EGFR GBM model (LN229). Our study demonstrated that the treatment of PmAb-TMZ-PLGA-NPs in GBM tried to adopt the autophagic pathway of the cell survival mechanism with the elevated level of autophagic marker (Beclin-1 and LC3B) at 24 h time point, thereby suppressing the expression of caspase-9 and PARP. However, at the 48 h time point, the elevated expression of caspase-9 and PARP with the downregulation of Beclin-1 and LC3B, following the treatment of PmAb-TMZ-PLGA-NPs in the GBM model, suggested that apoptotic cell death was superior over autophagic cell survival. It was also noteworthy the activation of caspase-9 was correlated with the continuous overproduction of reactive oxygen species up to a 48 h time point after the treatment of PmAb-TMZ-PLGA-NPs. This result sheds light on the biological effect of targeted chemotherapy and illustrates that PmAb-TMZ-PLGA-NPs could be applied for EGFR-overexpressed different cancer models.

Construction of Surface-Modified Polydopamine Nanoparticles for Sequential Drug Release and Combined Chemo-Photothermal Cancer Therapy

“…Cell death caused by ROS is a frequent target for tumor treatment . Upgraded ROS levels damage the cell nucleus, membrane structure, and organelles and eventually lead to apoptosis . DOX is known to activate nicotinamide adenine dinucleotide phosphate oxidase and further induce the production of ROS to accelerate cell death .…”

“…42 Upgraded ROS levels damage the cell nucleus, membrane structure, and organelles and eventually lead to apoptosis. 43 DOX is known to activate nicotinamide adenine dinucleotide phosphate oxidase and further induce the production of ROS to accelerate cell death. 44 As shown in Figure 5E, compared to the PBS group and free DOX group, the ROS level of the DOX-PLGA@PDA−DOX/PEG/FA group was much greater and exerted a concentrationdependent manner.…”

Hypoxia-Mediated ROS Amplification Triggers Mitochondria-Mediated Apoptotic Cell Death via PD-L1/ROS-Responsive, Dual-Targeted, Drug-Laden Thioketal Nanoparticles

“…The synthesized TKNPs underwent ROS-responsive degradation after incubation with H 2 O 2 because the thioketal groups were converted to hydrophilic sulfoxides and sulfones . To immobilize antibodies on the surfaces of polymeric nanoparticles, glutaraldehyde cross-linking, carbodiimide coupling, and other strategies have been implemented . However, CE is low as additional activation and conjugation steps in the buffers are required .…”

“…Chen et al successfully demonstrated that B16F10 tumors were hypoxic and produced abundant ROS . Targeted drug therapy localizes the agent to the tumor site and minimizes its distribution to nontarget organs. , We then evaluated the tumor accumulation profile for our targeted system. Our biodistribution study suggested that after systemic targeted therapy injection, the nanoparticles were concentrated and retained at the tumor site and minimally penetrated the heart, lung, spleen, and other nontarget organs (Figure ).…”

Nanotherapeutics Overcoming the Blood-Brain Barrier for Glioblastoma Treatment

“…However, it is worth noting that the accumulation of TFR-T12 peptide-modified micelles in liver tissue is more than that of unmodified micelles, which might be due to the more presence of TFR in the liver tissue. Banstola et al (2020) coupled Panitumumab to the surface of polyglycolic acid nanoparticles loaded with temozolomide (PmAb-TMZ-PLGA-NPs), where the panitumumab targets the epidermal growth factor receptor (EGFR), which are overexpressed in glioma. Their approach has been shown to improve the tumor-targeting efficiency while exerting the therapeutic effect of the monoclonal antibody, at the same time increasing the tumor internalization of the nanoparticles and the toxic effect of temozolomide.…”