Targeted delivery of polo-like kinase 1 siRNA nanoparticles using an EGFR-PEG bispecific antibody inhibits proliferation of high-risk neuroblastoma

J Control Release. 2024 Mar:367:806-820. doi: 10.1016/j.jconrel.2024.02.007. Epub 2024 Feb 15.

Abstract

High-risk neuroblastoma has poor survival due to treatment failure and off-target side effects of therapy. Small molecule inhibitors have shown therapeutic efficacy at targeting oncogenic cell cycle dysregulators, such as polo-like kinase 1 (PLK1). However, their clinical success is limited by a lack of efficacy and specificity, causing off-target toxicity. Herein, we investigate a new treatment strategy whereby a bispecific antibody (BsAb) with dual recognition of methoxy polyethylene glycol (PEG) and a neuroblastoma cell-surface receptor, epidermal growth factor receptor (EGFR), is combined with a PEGylated small interfering RNA (siRNA) lipid nanoparticle, forming BsAb-nanoparticle RNA-interference complexes for targeted PLK1 inhibition against high-risk neuroblastoma. Therapeutic efficacy of this strategy was explored in neuroblastoma cell lines and a tumor xenograft model. Using ionizable lipid-based nanoparticles as a low-toxicity and clinically safe approach for siRNA delivery, we identified that their complexing with EGFR-PEG BsAb resulted in increases in cell targeting (1.2 to >4.5-fold) and PLK1 gene silencing (>2-fold) against EGFR+ high-risk neuroblastoma cells, and enhancements correlated with EGFR expression on the cells (r > 0.94). Through formulating nanoparticles with PEG-lipids ranging in diffusivity, we further identified a highly diffusible PEG-lipid which provided the most pronounced neuroblastoma cell binding, PLK1 silencing, and significantly reduced cancer growth in vitro in high-risk neuroblastoma cell cultures and in vivo in a tumor-xenograft mouse model of the disease. Together, this work provides an insight on the role of PEG-lipid diffusivity and EGFR targeting as potentially relevant variables influencing the therapeutic efficacy of siRNA nanoparticles in high-risk neuroblastoma.

Keywords: Bispecific antibodies; EGFR targeting; Lipid nanoparticles; Neuroblastoma therapy; Polo-like kinase 1 silencing; Targeted siRNA delivery.

MeSH terms

  • Animals
  • Cell Cycle Proteins / genetics
  • Cell Line, Tumor
  • Cell Proliferation
  • ErbB Receptors / genetics
  • Humans
  • Lipids / pharmacology
  • Mice
  • Nanoparticles* / chemistry
  • Neuroblastoma* / drug therapy
  • Polo-Like Kinase 1
  • Polyethylene Glycols / chemistry
  • Protein Serine-Threonine Kinases
  • Proto-Oncogene Proteins
  • RNA, Small Interfering

Substances

  • RNA, Small Interfering
  • Protein Serine-Threonine Kinases
  • Cell Cycle Proteins
  • Polo-Like Kinase 1
  • Polyethylene Glycols
  • Proto-Oncogene Proteins
  • ErbB Receptors
  • Lipids
  • EGFR protein, human