Penumbra-targeted CircOGDH siRNA-loaded nanoparticles alleviate neuronal apoptosis in focal brain ischaemia

Stroke Vasc Neurol. 2024 Apr 30;9(2):134-144. doi: 10.1136/svn-2022-002009.

Abstract

Background: Nanoparticles (NPs) are a class of substances that can be loaded with therapeutic agents delivered to specific areas. In our earlier research, we identified a neuron-derived circular RNA (circRNA), circular oxoglutarate dehydrogenase (CircOGDH), as a promising therapeutic target for acute ischaemic stroke. This study dedicated to explore a prospective preliminary strategy of CircOGDH-based NP delivered to the ischaemic penumbra region in middle cerebral artery occlusion/reperfusion (MCAO/R) mice.

Methods: Immunofluorescence in primary cortex neurons and in vivo fluorescence imaging revealed endocytosis of Poly(lactide-co-glycolide) (PLGA) poly amidoamine(PAMAM)@CircOGDH small interfering RNA (siRNA) NPs. Western blotting analysis and CCK8 assay were performed to evaluate the apoptotic level in ischaemic neurons treated with PLGA-PAMAM@CircOGDH siRNA NPs. Quantitative reverse transcription PCR experiments, mice behaviour test, T2 MRI analysis, Nissl and TdT-mediated dUTP nick end labeling (TUNEL) co-staining were performed to evaluate the apoptosis level of ischaemic penumbra neurons in MCAO/R mice. Biosafety evaluation of NPs in MCAO/R mice was detected by blood routine examination, liver and kidney function examination and HE staining.

Results: PLGA-PAMAM@CircOGDH siRNA NPs were successfully assembled. Endocytosis of PLGA-PAMAM@CircOGDH siRNA NPs in ischaemic neurons alleviated neuronal apoptotic level in vitro and in vivo. Furthermore, mice behaviour test showed that the neurological defects of MCAO/R mice were significantly alleviated after the tail injection of PLGA-PAMAM@CircOGDH siRNA NPs, and no toxic effects were observed.

Conclusion: In conclusion, our results suggest that PLGA-PAMAM@CircOGDH siRNA NPs can be delivered to the ischaemic penumbra region and alleviate neuron apoptosis in MCAO/R mice and in ischaemic neurons; therefore, our study provides a desirable approach for using circRNA-based NPs for the treatment of ischaemic stroke.

Keywords: Brain; Cerebral Infarction; Material; Stroke.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis* / drug effects
  • Cells, Cultured
  • Dendrimers*
  • Disease Models, Animal*
  • Drug Carriers / chemistry
  • Endocytosis
  • Infarction, Middle Cerebral Artery* / genetics
  • Infarction, Middle Cerebral Artery* / metabolism
  • Infarction, Middle Cerebral Artery* / pathology
  • Ischemic Stroke / metabolism
  • Ischemic Stroke / pathology
  • Ischemic Stroke / therapy
  • Male
  • Mice
  • Mice, Inbred C57BL*
  • Nanoparticle Drug Delivery System
  • Nanoparticles
  • Neurons* / drug effects
  • Neurons* / metabolism
  • Neurons* / pathology
  • Polylactic Acid-Polyglycolic Acid Copolymer / chemistry
  • RNA Interference
  • RNA, Circular* / genetics
  • RNA, Circular* / metabolism
  • RNA, Small Interfering* / metabolism
  • RNAi Therapeutics

Substances

  • RNA, Circular
  • RNA, Small Interfering
  • Drug Carriers
  • Nanoparticle Drug Delivery System
  • PAMAM Starburst
  • Polylactic Acid-Polyglycolic Acid Copolymer
  • Dendrimers