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Self-assembling HA/PEI/dsRNA-p21 ternary complexes for CD44 mediated small active RNA delivery to colorectal cancer

Chen-Lin Feng1, Yan-Xing Han1, Hui-Hui Guo1

  • 1a State Key Laboratory of Bioactive Substance and Function of Natural Medicines , Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College , Beijing , PR China.

Drug Delivery
|October 11, 2017
PubMed

Insights

A novel hyaluronic acid/polyethyleneimine/double-strand RNA (HA/PEI/dsRNA-p21) complex effectively delivers dsRNA-p21 to colorectal cancer cells. This targeted delivery suppresses tumor growth by activating p21 gene expression with reduced toxicity.

Area of Science:

  • Biomedical Engineering
  • Cancer Therapy
  • Nanotechnology

Background:

  • Sequence-specific double-strand RNA (dsRNA-p21) activates p21 gene expression, suppressing colorectal cancer (CRC) growth.
  • Efficient delivery systems for dsRNA-p21 remain a significant challenge for effective CRC therapy.

Purpose of the Study:

  • To develop a self-assembled hyaluronic acid/polyethyleneimine/dsRNA-p21 ternary complex (TC-dsRNA-p21) for targeted delivery of dsRNA-p21 into CRC cells.
  • To enhance tumor targeting and reduce the cytotoxicity of polyethyleneimine (PEI) through hyaluronic acid (HA) shielding.
  • To evaluate the efficacy of TC-dsRNA-p21 in vitro and in vivo for CRC treatment.

Main Methods:

  • Formation of HA/PEI/dsRNA-p21 ternary complexes (TC-dsRNA-p21) for dsRNA-p21 delivery.
  • Characterization of TC-dsRNA-p21 properties, including size, zeta potential, cytotoxicity, and stability.
  • Assessment of cellular uptake and transfection efficiency in CD44-positive CRC cells using confocal microscopy and flow cytometry.
  • In vitro evaluation of p21 gene and protein expression, cell cycle arrest, proliferation, and colony formation.
  • In vivo biodistribution studies following in situ application of TC-dsRNA-p21.

Main Results:

  • TC-dsRNA-p21 exhibited increased size, decreased zeta potential, low cytotoxicity, and high stability compared to binary complexes (BC-dsRNA-p21).
  • High transfection efficiency of TC-dsRNA-p21 was observed in CD44-abundant Lovo cells.
  • TC-dsRNA-p21 effectively activated p21 mRNA and protein expression, leading to G0/G1 cell cycle arrest and suppressed CRC cell proliferation and colony formation.
  • In vivo studies showed effective accumulation of TC-dsRNA-p21 in the rectal wall for up to 10 hours after in situ application.

Conclusions:

  • The developed TC-dsRNA-p21 system demonstrates enhanced tumor-targeting capabilities and reduced cytotoxicity.
  • TC-dsRNA-p21 effectively delivers dsRNA-p21, activating p21 expression and inhibiting CRC cell growth both in vitro and in vivo.
  • This targeted delivery system holds significant potential for the development of novel dsRNA-p21-based therapies for colorectal cancer.
Keywords:
Colorectal cancerhyaluronic acidsmall active RNAternary complexestumor targeted

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