Hijacking of normal developmental programs is a common mechanism of tumorigenesis and epigenetic deregulation of developmental transcription programs is central to the genesis of most, if not all, pediatric cancers. Ewing sarcomas, aggressive bone and soft tissue tumors that predominately arise in adolescence, continue to be associated with high rates of mortality and novel approaches to therapy are needed. Ewing sarcomas are characterized by the presence of pathognomonic driver fusion oncogenes, most commonly EWS/FLI1, and their likely cellular origin is mesenchymal stem/progenitor cells (MSC). EWS/FLI1 initiates tumorigenesis by inducing widespread disruption of transcriptional regulation as a consequence of altered recruitment of chromatin remodelers to target gene enhancers and promoters. We have reported that posterior HOXD genes, in particular HOXD13, are aberrantly over-expressed by Ewing sarcoma and that posterior HOXD genes contribute to maintenance of the tumorigenic phenotype. Our preliminary studies suggest that EWS/FLI1 can induce expression of HOXD13, in a cell context dependent fashion, and that this activation is mediated by aberrant activation of developmental enhancers that are otherwise active in only very discrete spatiotemporal developmental windows. In addition, chromatin immunoprecipitation (ChIP) studies have demonstrated that in EWS/FLI1+ cells the HOXD13 promoter is preferentially enriched with the MLL- dependent activating histone modification H3K4me3 and bound by both MLL and menin proteins. Significantly, exposure of Ewing sarcoma cells to novel small molecule inhibitors of MLL:menin interaction, that are currently in preclinical development for MLL-fusion positive leukemias, leads to a dramatic loss tumorigenicity and concomitant loss posterior HOXD gene expression. Thus, these studies demonstrate that that, like MLL-fusion positive leukemias, maintenance of the oncogenic phenotype of Ewing sarcoma is critically dependent on MLL:menin-dependent activation of developmental HOX genes. In this proposal, we will test the hypothesis that HOXD13 functions an obligate cooperative oncogene in Ewing sarcoma and that dependency on its continued expression presents a previously unrecognized tumor-specific vulnerability that can be therapeutically exploited.
In Aim 1 will use innovative Bru-seq technologies and functional validation studies to define the downstream transcriptional targets of HOXD13 that promote tumorigenicity.
In Aim 2 we will use targeted ChIP assays and chromatin conformation studies to determine how EWS/FLI1 leads to epigenetic activation of HOXD13.
In Aim 3 we will test the therapeutic potential of MI-503, a small molecule inhibitor of MLL:menin protein:protein interaction, in in vivo tumor models. These studies will together elucidate the contribution of HOXD13 to Ewing sarcoma pathogenesis and test the potential of a new class of epigenetic modifying agents for Ewing sarcoma-targeted therapies.

Public Health Relevance

Genes that are essential for normal embryonic and post-natal development of organs and tissues are frequently hijacked in disease, in particular in the context of pediatric cancers. We have discovered that HOXD13, a gene that is normally expressed only in early limb development, is aberrantly activated in Ewing sarcoma, a bone and soft tissue tumor that peaks in adolescence. In this proposal we investigate the mechanistic basis of HOXD13 activation in Ewing sarcoma and define the downstream targets of HOXD13 that contribute to tumor growth and progression. In addition, we will test whether targeting HOXD13 using epigenetic drug therapy presents a novel therapeutic opportunity for Ewing sarcoma patients.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA215981-03
Application #
9829546
Study Section
Cancer Molecular Pathobiology Study Section (CAMP)
Program Officer
Mietz, Judy
Project Start
2017-12-18
Project End
2022-11-30
Budget Start
2019-12-01
Budget End
2020-11-30
Support Year
3
Fiscal Year
2020
Total Cost
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Pediatrics
Type
Schools of Medicine
DUNS #
073133571
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109