Polymeric nanoparticles as cancer-specific DNA delivery vectors to human hepatocellular carcinoma

Abstract

Hepatocellular carcinoma (HCC) is the third most deadly cancer in the US, with a meager 5-year survival rate of less than 20%. Such unfavorable numbers are closely related to the heterogeneity of the disease and the unsatisfactory therapies currently used to manage patients with invasive HCC. Outside of the clinic, gene therapy research is evolving to overcome the poor responses and toxicity associated with standard treatments. The inadequacy of gene delivery vectors, including poor intracellular delivery and cell specificity, are major barriers in the gene therapy field. Herein, we described a non-viral strategy for effective and cancer-specific DNA delivery to human HCC using biodegradable Poly(Beta-Amino Ester) (PBAE) nanoparticles (NPs). Varied PBAE NP formulations were evaluated for transfection efficacy and cytotoxicity to a range of human HCC cells as well as healthy human hepatocytes. To address HCC heterogeneity, nine different sources of human HCC cells were utilized. The polymeric NPs composed of 2-((3-aminopropyl)amino)ethanol end-modified poly(1,5-pentanediol diacrylate-co-3-amino-1-propanol) (‘536’) at a 25 polymer-to-DNA weight-to-weight ratio led to high transfection efficacy to all of the liver cancer lines, but not to hepatocytes. Each individual HCC line had a significantly higher percentage of exogenous gene expression than the healthy liver cells (P < 0.01). Notably, this biodegradable end-modified PBAE gene delivery vector was not cytotoxic and maintained the viability of hepatocytes above 80%. In a HCC/hepatocyte co-culture model, in which cancerous and healthy cells share the same micro-environment, 536 25 w/w NPs specifically transfected cancer cells. PBAE NP administration to a subcutaneous HCC mouse model, established with one of the human lines tested in vitro, confirmed effective DNA transfection in vivo. PBAE-based NPs enabled high and preferential DNA delivery to HCC cells, sparing healthy hepatocytes. These biodegradable and liver cancer-selective NPs are a promising technology to deliver therapeutic genes to liver cancer.

Assessment of Gold Nanoparticles-Inhibited Cytochrome P450 3A4 Activity and Molecular Mechanisms Underlying Its Cellular Toxicity in Human Hepatocellular Carcinoma Cell Line C3A

“…Thus, it becomes very important to identify the molecular mechanism and biological pathway underlying cellular disturbance and toxicity of NP in target cells and tissues. Recent in vitro studies demonstrated that gene expression profiling combined with cellular and biochemical responses has provided a direct assessment of cellular perturbation and potential NP toxicity [ 7 – 10 ].…”

Polymeric Nonviral Gene Delivery Systems for Cancer Immunotherapy

“…The presence of acid lysosomes has been proved to achieve the gene release from nanoparticles within a few hours during immune cell‐mediated cell uptake process ( Figure a). [ 46 ] Due to these advantages, Tsai et al. first synthesized PBAE by Michael addition reaction to form complex adjuvants with CpG, which showed the targeting effect and immune stimulation on tumor cells (Figure 3b).…”

“…Reproduced with permission. [ 46 ] Copyright 2017, Elsevier Inc. b) Synthetic schematic illustration of PBAE/CpG DNA polyplex. Reproduced with permission.…”

Gene Therapy for Liver Cancers: Current Status from Basic to Clinics

“…Taking advantage of membrane receptors on hepatic stellate cells, liver-targeted gene delivery for hepatic fibrosis has been attempted using the mannose 6-phosphate/insulin-like growth factor-II receptor [132][133][134], integrins [135], high-affinity membrane receptor for retinol-binding protein, and the galactosyl receptor as targets [101]. Target-specific gene delivery is a desirable feature, and polymeric nanoparticles [131] targeting cancer-specific DNA have shown promising anti-tumor effects in vitro and in vivo [136]. A cationic solid lipid nanoparticle also showed effective inhibition of HCC growth by delivering shRNA for the NURP gene [137], and similarly, siRNA to PLK1 gene delivered by chitosan nanoparticles efficiently suppressed HCC cell growth both in vitro and in vivo [138].…”