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@ARTICLE{Khan:302840,
      author       = {R. Khan and L. Phely and S. Ehrenfeld$^*$ and T. Schmitz
                      and P. Veratti and J. Wolfes and K. Shoumariyeh$^*$ and G.
                      Andrieux and U. S. Martens and S. d. Bra and M. Auer and O.
                      Schilling and M. Börries$^*$ and M. Speicher and A. L.
                      Illert and J. Duyster$^*$ and C. Miething$^*$},
      title        = {{M}odeling the t(2;5) {T}ranslocation of {A}naplastic
                      {L}arge {C}ell {L}ymphoma {U}sing {CRISPR}-{M}ediated
                      {C}hromosomal {E}ngineering.},
      journal      = {Cancers},
      volume       = {17},
      number       = {13},
      issn         = {2072-6694},
      address      = {Basel},
      publisher    = {MDPI},
      reportid     = {DKFZ-2025-01380},
      pages        = {2226},
      year         = {2025},
      abstract     = {ALK+ Anaplastic Large Cell Lymphoma (ALCL) is an aggressive
                      T-cell lymphoma that is characterized by expression of the
                      Anaplastic Lymphoma Kinase (ALK), which is induced by the
                      t(2;5) chromosomal rearrangement, leading to the expression
                      of the NPM-ALK fusion oncogene. Most previous preclinical
                      models of ALK+ ALCL were based on overexpression of the
                      NPM-ALK cDNA from heterologous promoters. Due to the
                      enforced expression, this approach is prone to artifacts
                      arising from synthetic overexpression, promoter competition
                      and insertional variation.To improve the existing ALCL
                      models and more closely recapitulate the oncogenic events in
                      ALK+ ALCL, we employed CRISPR/Cas-based chromosomal
                      engineering to selectively introduce translocations between
                      the Npm1 and Alk gene loci in murine cells.By inducing
                      precise DNA cleavage at the syntenic loci on chromosome 11
                      and 17 in a murine IL-3-dependent Ba/F3 reporter cell line,
                      we generated de novo Npm-Alk translocations in vivo, leading
                      to IL-3-independent cell growth. To verify efficient
                      recombination, we analyzed the expression of the NPM-ALK
                      fusion protein in the recombined cells and could also show
                      the t(11;17) in the IL-3 independent Ba/F3 cells. Subsequent
                      functional testing of these cells using an Alk-inhibitor
                      showed exquisite responsiveness towards Crizotinib,
                      demonstrating strong dependence on the newly generated ALK
                      fusion oncoprotein. Furthermore, a comparison of the gene
                      expression pattern between Ba/F3 cells overexpressing the
                      Npm-Alk cDNA with Ba/F3 cells transformed by CRISPR-mediated
                      Npm-Alk translocation indicated that, while broadly
                      overlapping, a set of pathways including the unfolded
                      protein response pathway was increased in the Npm-Alk
                      overexpression model, suggesting increased reactive changes
                      induced by exogenous overexpression of Npm-Alk. Furthermore,
                      we observed clustered expression changes in genes located in
                      chromosomal regions close to the breakpoint in the new
                      CRISPR-based model, indicating positional effects on gene
                      expression mediated by the translocation event, which are
                      not part of the older models.Thus, CRISPR-mediated
                      recombination provides a novel and more faithful approach to
                      model oncogenic translocations, which may lead to an
                      improved understanding of the molecular pathogenesis of ALCL
                      and enable more accurate therapeutic models of malignancies
                      driven by oncogenic fusion proteins.},
      keywords     = {ALCL (Other) / CRISPR/Cas (Other) / Npm-Alk (Other) /
                      chromosomal engineering (Other) / chromosomal translocation
                      (Other) / oncogenic fusion protein (Other)},
      cin          = {FR01},
      ddc          = {610},
      cid          = {I:(DE-He78)FR01-20160331},
      pnm          = {899 - ohne Topic (POF4-899)},
      pid          = {G:(DE-HGF)POF4-899},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:40647524},
      pmc          = {pmc:PMC12249153},
      doi          = {10.3390/cancers17132226},
      url          = {https://inrepo02.dkfz.de/record/302840},
}