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Tumor targeted genome editing mediated by a multi-functional gene vector for regulating cell behaviors

Bo-Ya Liu1, Xiao-Yan He1, Ren-Xi Zhuo1

  • 1Key Laboratory of Biomedical Polymers of Ministry of Education, Department of Chemistry, Wuhan University, Wuhan 430072, People's Republic of China.

Journal of Controlled Release : Official Journal of the Controlled Release Society
|October 20, 2018
PubMed

Insights

Novel nanoparticles deliver CRISPR-Cas9 gene editing to cancer cells, significantly reducing CDK11 expression and reversing tumor-induced immunosuppression for cancer prevention.

Area of Science:

  • Biotechnology
  • Cancer Research
  • Nanomedicine

Background:

  • Effective gene delivery is crucial for cancer therapy and prevention.
  • Genome editing offers precise control over cellular behavior.
  • Developing targeted delivery systems for CRISPR-Cas9 is a key challenge.

Purpose of the Study:

  • To construct multifunctional nanoparticles for targeted delivery of CRISPR-Cas9 to tumor cells.
  • To evaluate the efficacy of these nanoparticles in gene editing and cancer modulation.
  • To assess the impact of genome editing on tumor-associated proteins and the tumor immune microenvironment.

Main Methods:

  • Construction of self-assembled nanoparticles using natural polymers (carboxymethyl chitosan).
  • Incorporation of CRISPR-Cas9 plasmid, protamine sulfate, endosomolytic peptide (KALA), and AS1411 aptamer.
  • In vitro evaluation of cellular uptake, endosomal escape, and gene delivery efficiency.
  • Assessment of CDK11 gene knockout and modulation of downstream proteins (MMP-9, VEGF, p53).
  • Analysis of immune-related protein expression changes post-genome editing.

Main Results:

  • Nanoparticles demonstrated enhanced delivery efficiency and specific targeting of tumor cell nuclei.
  • CRISPR-Cas9 mediated by nanoparticles achieved >75% decrease in CDK11 expression.
  • Significant down-regulation of MMP-9 and VEGF, and up-regulation of p53 observed.
  • Genome editing led to increased expression of immune-related proteins (Fas, CD80, MICA, MICB, HLA-1) and decreased expression of immunosuppressive proteins (CD47, MUC1).

Conclusions:

  • Multifunctional nanoparticles effectively deliver CRISPR-Cas9 for targeted gene editing in cancer cells.
  • Genome editing via these nanoparticles modulates key proteins involved in tumor development, metastasis, and immune evasion.
  • The approach shows promise for reversing tumor-induced immunosuppression and preventing tumor development.

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