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In vivo engineering of oncogenic chromosomal rearrangements with the CRISPR/Cas9 system

Danilo Maddalo1, Eusebio Manchado1, Carla P Concepcion2

  • 1Memorial Sloan Kettering Cancer Center, Cancer Biology and Genetics Program, 1275 York Avenue, New York, New York 10065, USA.

Nature
|October 23, 2014
PubMed

Insights

Researchers developed a new CRISPR/Cas9 method to create mouse models of cancer by inducing specific chromosomal rearrangements. This technique successfully generated a model for echinoderm microtubule-associated protein like 4 (EML4)-anaplastic lymphoma kinase (ALK) lung cancer, which responded to ALK inhibitors.

Area of Science:

  • Oncology
  • Genetics
  • Molecular Biology

Background:

  • Chromosomal rearrangements are key drivers in human cancer pathogenesis, frequently leading to therapeutically targetable gene fusions.
  • The echinoderm microtubule-associated protein like 4 (EML4)-anaplastic lymphoma kinase (ALK) fusion gene, resulting from an inversion on chromosome 2, is a significant oncogene in a subset of non-small cell lung cancers (NSCLC).
  • Existing methods for modeling such genetic events in mice are complex and require extensive germline manipulation.

Purpose of the Study:

  • To establish an efficient in vivo method for inducing specific chromosomal rearrangements using CRISPR/Cas9 technology.
  • To generate a novel mouse model for EML4-ALK-driven lung cancer.
  • To validate the utility of this model for studying cancer pathogenesis and therapeutic response.

Main Methods:

  • Utilized viral-mediated delivery of the CRISPR/Cas9 system to induce targeted chromosomal inversions in somatic cells of adult mice.
  • Generated a mouse model specifically harboring the EML4-ALK fusion gene.
  • Analyzed the resulting tumors for histopathological and molecular characteristics, and assessed their response to ALK inhibitors.

Main Results:

  • The CRISPR/Cas9 method efficiently induced the specific EML4-ALK inversion in vivo.
  • The generated mouse model accurately recapitulated key features of human ALK-positive NSCLC, including tumor formation and expression of the fusion gene.
  • Tumors in the mouse model exhibited sensitivity to ALK inhibitors, mirroring clinical observations in human patients.

Conclusions:

  • Viral-mediated CRISPR/Cas9 delivery provides an efficient strategy for modeling chromosomal rearrangements and associated cancers in vivo.
  • This approach significantly advances the ability to create genetically engineered mouse models for human cancers.
  • The developed EML4-ALK lung cancer model serves as a valuable platform for preclinical research and drug development targeting ALK-driven NSCLC.

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