Targeting CD146 using folic acid–conjugated nanoparticles and suppression of tumor growth in a mouse glioma model

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  • 1 Departments of Neurosurgery,
  • 2 Pharmacology, and
  • 3 Biochemistry, Kochi Medical School, Kochi University, Nankoku, Kochi; and
  • 4 Japan Society for the Promotion of Science, Tokyo, Japan
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OBJECTIVE

Glioma stem cells (GSCs) are responsible for tumor initiation, therapeutic resistance, and recurrence. CD146 is mainly expressed in dividing GSCs and regulates cell cycle progression. However, the evaluation of the efficacy of targeted therapy against CD146 in vivo remains to be investigated. In this study, the authors aimed to develop gene therapy targeting GSCs using chitosan oligosaccharide lactate (COL) nanoparticles (NPs) conjugated with folic acid–polyethylene glycol (FA-PEG-COL NPs) for in vitro and in vivo delivery of CD146 small-interfering RNA (siCD146) and to determine the effect of CD146 knockdown on tumor growth.

METHODS

To examine the uptake of NPs by tumor cells, immunofluorescence staining, flow cytometry, and in vivo imaging were performed. The knockdown effect of siCD146 was measured by western blot and water-soluble tetrazolium salt–8 assay in mouse glioma cells. The efficacy of siRNA therapy–targeted GSCs was evaluated by monitoring tumor growth through in vivo imaging and histological analysis.

RESULTS

In vivo accumulation of the FA-PEG-COL NPs in subcutaneous and intracranial gliomas following NP administration via a mouse tail vein was observed. Additionally, in vitro delivery of siCD146 ionically cross-linked NPs, reduced CD146 levels, and suppressed growth in the glioma tumor sphere. Evaluation of the in vivo therapeutic effects of siCD146–cross-linked NPs in a mouse glioma model revealed significant suppression of intracranial tumor growth, with complete removal of the tumor observed in some mice on histological examination. Furthermore, delivery of siCD146 significantly reduced the Ki-67 index in residual tumor tissues relative to that in control mice.

CONCLUSIONS

CD146 is a potential therapeutic target, and folic acid–conjugated NPs delivering siRNA may facilitate gene therapy in malignant gliomas.

ABBREVIATIONS BBB = blood-brain barrier; COL = chitosan oligosaccharide lactate; EPR = enhanced permeability and retention; FA = folic acid; FA-PEG-COL = FA–polyethylene glycol COL; FITC = fluorescein isothiocyanate; FOLR1 = folate receptor 1; GBM = glioblastoma; GSC = glioma stem cell; NP = nanoparticle; PBS = phosphate-buffered saline; RT-PCR = reverse transcription polymerase chain reaction; siCD146 = CD146 siRNA; siRNA = small-interfering RNA; TS = tumor sphere; WST-8 = water-soluble tetrazolium salt–8.

Supplementary Materials

    • Supplemental Materials and Methods (PDF 830 KB)

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Contributor Notes

Correspondence Toshio Yawata: Kochi University, Kochi, Japan. yawatat@kochi-u.ac.jp.

INCLUDE WHEN CITING Published online July 24, 2020; DOI: 10.3171/2020.4.JNS193078.

Disclosures The authors report no conflict of interest concerning the materials or methods used in this study or the findings specified in this paper.

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