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Intronic Polyadenylation in Acquired Cancer Drug Resistance Circumvented by Utilizing CRISPR/Cas9 with

Terry S Elton1, Victor A Hernandez1, Jessika Carvajal-Moreno1

  • 1Division of Pharmaceutics and Pharmacology, College of Pharmacy, The Ohio State University, Columbus, OH 43210, USA.

Cancers
|July 9, 2022
PubMed

Insights

Intronic polyadenylation (IPA) causes drug resistance by altering TOP2α/170 expression in leukemia. CRISPR/Cas9 gene editing offers a potential strategy to overcome this resistance by correcting aberrant splicing and polyadenylation.

Area of Science:

  • Molecular Biology
  • Genetics
  • Cancer Research

Background:

  • Intronic polyadenylation (IPA) contributes to cancer development and chemoresistance through transcriptome alterations.
  • DNA topoisomerase IIα (TOP2α/170) is a key anticancer target, but its efficacy is limited by drug resistance, often linked to reduced nuclear levels.
  • Acquired resistance to TOP2α-targeted drugs in leukemia involves variant TOP2α mRNA transcripts due to IPA, leading to truncated isoforms.

Purpose of the Study:

  • To review mechanisms of pre-mRNA processing and alternative polyadenylation.
  • To explore the use of CRISPR/Cas9 gene editing with homology directed repair (HDR) to reduce IPA.
  • To investigate the therapeutic potential of targeting aberrant IPA in acquired drug resistance.

Main Methods:

  • Review of pre-mRNA processing and alternative polyadenylation mechanisms.
  • Application of CRISPR/Cas9 gene editing via HDR to target specific splice sites.
  • Case study of TOP2α exon 19/intron 19 splice site editing in etoposide-resistant K562 leukemia cells.

Main Results:

  • Aberrant IPA is implicated in acquired drug resistance to TOP2α-targeted therapies.
  • CRISPR/Cas9/HDR-mediated editing of the TOP2α exon 19/intron 19 splice site can decrease IPA.
  • This editing strategy shows potential for circumventing acquired TOP2α-mediated drug resistance in leukemia.

Conclusions:

  • Aberrant intronic polyadenylation is a significant factor in acquired drug resistance to TOP2α-targeted agents.
  • CRISPR/Cas9/HDR presents a promising therapeutic approach to address drug resistance driven by aberrant splicing and polyadenylation.
  • Targeting IPA offers a novel strategy to enhance the efficacy of anticancer therapies.
Keywords:
CRISPR/Cas9RNA splicingalternative polyadenylationdrug resistancegene editinghomology directed repairintronic polyadenylationtopoisomerase IIα

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