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  • Brief Communication
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Spontaneous expression of the CIC::DUX4 fusion oncoprotein from a conditional allele potently drives sarcoma formation in genetically engineered mice

Oncogene volume 43, pages 1223–1230 (2024)Cite this article

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Abstract

CIC::DUX4 sarcoma (CDS) is a rare but highly aggressive undifferentiated small round cell sarcoma driven by a fusion between the tumor suppressor Capicua (CIC) and DUX4. Currently, there are no effective treatments and efforts to identify and translate better therapies are limited by the scarcity of patient tumor samples and cell lines. To address this limitation, we generated three genetically engineered mouse models of CDS (Ch7CDS, Ai9CDS, and TOPCDS). Remarkably, chimeric mice from all three conditional models developed spontaneous soft tissue tumors and disseminated disease in the absence of Cre-recombinase. The penetrance of spontaneous (Cre-independent) tumor formation was complete irrespective of bi-allelic Cic function and the distance between adjacent loxP sites. Characterization of soft tissue and presumed metastatic tumors showed that they consistently expressed the CIC::DUX4 fusion protein and many downstream markers of the disease credentialing the models as CDS. In addition, tumor-derived cell lines were generated and ChIP-seq was preformed to map fusion-gene specific binding using an N-terminal HA epitope tag. These datasets, along with paired H3K27ac ChIP-sequencing maps, validate CIC::DUX4 as a neomorphic transcriptional activator. Moreover, they are consistent with a model where ETS family transcription factors are cooperative and redundant drivers of the core regulatory circuitry in CDS.

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Fig. 1: Fusion of a human DUX4 C-terminal domain to endogenous Cic is sufficient to generate small round cell sarcomas.
Fig. 2: Cic haploinsufficiency is not required for CIC::DUX4 sarcomagenesis.
Fig. 3: Mouse tumors express CIC::DUX4 and a transcriptional signature consistent with CDS.
Fig. 4: CIC::DUX4 behaves as a neomorphic transcriptional activator.

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Data availability

All RNA-sequencing and ChIP-sequencing data is freely accessible through the Gene Expression Omnibus (GEO) under the series ID: GSE241371.

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Author information

Author notes

  1. These authors contributed equally: Peter G. Hendrickson, Kristianne M. Oristian.

Authors and Affiliations

  1. Department of Radiation Oncology, Duke University Medical Center, Durham, NC, USA

    Peter G. Hendrickson, Kristianne M. Oristian, MaKenna R. Browne, Lixia Luo, Yan Ma & David G. Kirsch

  2. Developmental and Stem Cell Biology Program, Duke University Medical Center, Durham, NC, USA

    MaKenna R. Browne

  3. Department of Pathology, Duke University Medical Center, Durham, NC, USA

    Diana M. Cardona

  4. Program in Genetics and Genome Biology, The Hospital for Sick Children (SickKids), University of Toronto, Toronto, ON, Canada

    Joshua O. Nash & Adam Shlien

  5. Laboratory of Medicine and Pathobiology, University of Toronto, Toronto, ON, Canada

    Joshua O. Nash, Pedro L. Ballester, Scott Davidson & Adam Shlien

  6. Department of Pediatrics, Duke University Medical Center, Durham, NC, USA

    Corinne M. Linardic

  7. Department of Pharmacology and Cancer Biology, Duke University Medical Center, Durham, NC, USA

    Corinne M. Linardic & David G. Kirsch

  8. Radiation Medicine Program, Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada

    David G. Kirsch

  9. Department of Radiation Oncology, University of Toronto, Toronto, ON, Canada

    David G. Kirsch

  10. Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada

    David G. Kirsch

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Contributions

The study was conceived and designed by PGH, KMO, and DGK. Animal data was collected and analyzed by PGH and KMO with help from MRB, LL, YM, DMC, and CML. RNA-sequencing analysis and neural network classification was performed by JON, PLB, SD, and AS. The paper with written by PGH, KMO, and DGK. This work was supported by grants from the Department of Defense (W81XWH-22-1-0454) to DGK, Alex’s Lemonade Stand Foundation to CML, the National Cancer Institute (7R35CA197616) to DGK and (1R38CA245204) PGH, and the American Society of Radiation Oncology (852948) to PGH.

Corresponding author

Correspondence to David G. Kirsch.

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Competing interests

DGK is a cofounder of and stockholder in XRAD Therapeutics, which is developing radiosensitizers. DGK is a member of the scientific advisory board and owns stock in Lumicell Inc, a company commercializing intraoperative imaging technology. None of these affiliations represents a conflict of interest with respect to the work described in this manuscript. DGK is a coinventor on a patent for a handheld imaging device and is a coinventor on a patent for radiosensitizers. XRAD Therapeutics, Merck, Bristol Myers Squibb, and Varian Medical Systems have provided research support to DGK, but this did not support the research described in this manuscript. The other authors have no conflicting financial interests.

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Hendrickson, P.G., Oristian, K.M., Browne, M.R. et al. Spontaneous expression of the CIC::DUX4 fusion oncoprotein from a conditional allele potently drives sarcoma formation in genetically engineered mice. Oncogene 43, 1223–1230 (2024). https://doi.org/10.1038/s41388-024-02984-8

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