The function of Ewing sarcoma proteins in mitosis.
Ewing sarcoma is the second most common bone tumor in children. The common molecular abnormality in Ewing sarcoma is the t(11; 22) chromosomal translocation, which results in the expression of a chimeric fusion protein containing EWS-derived sequences at the amino terminus fused to the carboxyl-terminus of the ETS transcription factor FLI1. My research goal is to elucidate the molecular mechanism whereby the Ewing sarcoma fusion protein EWS/FLI1 leads to malignant transformation.
In recent studies we reported that both the knockdown of EWS, and expression of the EWS/FLI1 fusion protein, in zebrafish embryos and HeLa cells leads to mitotic defects. We subsequently demonstrated a biochemical interaction between EWS/FLI1 and wildtype EWS, leads to inhibition of EWS activity by EWS/FLI1 in a dominant negative manner.
The hypothesis of my research is that the EWS/FLI1-EWS interaction in Ewing sarcoma induces mitotic defects leading to malignant transformation. Chromosomal instability is a hallmark of malignancy. Therefore, the functional loss of EWS due to its interaction with EWS/FLI may cause mitotic defects leading to chromosome instability, and ultimately to Ewing Sarcoma. We will analyze both EWS/FLI1 and EWS in human cells and in zebrafish. Experiments will be carried out in human cell lines to understand the molecular basis of EWS/FLI1 more closely, and to take advantage of the availability of reagents to study the mitotic spindle. These findings will then be assessed in zebrafish by generating transgenic fish expressing EWS/FLI1. The zebrafish will provide an in vivo whole animal model amenable to genetic screening for second hit mutations or suppressor genes.
We will directly address the molecular basis for this interaction and assess the activity of the EWS/FLI1 fusion in vivo in the zebrafish. Understanding how the fusion proteins resulting from these translocations lead to malignant transformation remains an important biological and clinical problem.
Park H, Kim H, Hassebroek V, Azuma Y, Slawson C, and Azuma M: Chromosomal localization of Ewing sarcoma EWSR1/FLI1 protein promotes the induction of aneuploidy. J Biol Chem. 2021, Vol 296, 100164, https://doi.org/10.1074/jbc.RA120.014328
So J, Kim M, Lee, SH, Ko S, Lee D, Park H, Azuma M, Parsons M, Prober D and Shin D: Attenuating the EGFR-ERK-SOX9 axis promotes liver progenitor cell-mediated liver regeneration in zebrafish. Hepatology. 2020. Vol 73, No4, p1494-1508
Huang W, Beer RL, Delaspre F, Wang G, Edelman HE, Park H, Azuma M, and Parsons MJ: Sox9b is a mediator of retinoic acid signaling restricting endocrine progenitor differentiation. Dev Biol. 2016. 418(1):28-39.
Park H, Richard Galbraith and Azuma M: Loss of Ewing sarcoma EWS allele promotes tumorigenesis by inducing chromosomal instability in zebrafish. Scientific Rep. 2016. 6: 32297. 10.1038/srep32297.
Lanza C, Tan EP, Zhang Z, Machacek M, Brinker AE, Azuma M, and Slawson C: Reduced O-GlcNAcase Expression Promotes Mitotic Errors and Spindle Defects. Cell Cycle. 2016. 15 (10): 1363-1375
*Wiley EOIII, Fuiten AM, Doosey MH, Lohman BK, Merkes C and Azuma M (*Corresponding author): The development and morphology of the caudal skeleton of zebrafish, Danio rerio from a phylogenetic perspective : Reassessment of Homologies from a Polyural Perspective. Copeia. 2015, 103 (4), 740-750
Merkes C, Turkalo TK, Wilder N, Park H and Azuma M: Ewing sarcoma Ewsa protein regulates chondrogenesis of Meckel’s cartilage through modulation of Sox9 in zebrafish. PLoS ONE, 2015, 10(1), e0116627
Park H, Turkalo TK, Nelson K, Folmsbee SS, Robb C, Roper B, and Azuma M (* equal contribution) : Ewing sarcoma EWS protein regulates midzone formation by recruiting Aurora B kinase to the midzone. Cell Cycle. 2014 Jun19; 13 (15) : p2391 – 2399
English MA, Lei L, Blake T, Wincovitch SM Sr, Sood R, Azuma M, Hickstein DD, and Liu PP : Incomplete splicing, cell division defects, and hematopoietic blockage in dhx8 mutant zebrafish. Developmental Dynamics. 2012 241(5): p879 - p889
Shive HR, West RR, Embree LJ, Azuma M, Sood R, Liu P, and Hickstein DD: brca2 in zebrafish ovarian development, Spermatogenesis, and tumorigenesis. Proc Natl Acad Sci USA, 2010 107(45) p19350
#Embree LJ, #Azuma M, and Hickstein DD: Ewing sarcoma fusion protein EWSR1/FLI1 interacts with EWSR1 leading to mitotic defects in zebrafish embryos and human cell lines. Cancer Res. 2009 69(10): 4363-71 (# equal contribution)
*Azuma M, Embree LJ, Sabaawy H and Hickstein DD (*Principle Investigator): Ewing sarcoma protein Ewsr1 maintains mitotic integrity and proneural cell survival in the zebrafish embryo. PLoS ONE 2(10): e979, 2007
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