Prostate adenocarcinoma (PCa) is a leading cause of death from cancer and there is no effective treatment for "castration-resistant" disease. This project is based on our recent findings that an actin-nucleating protein, Diaphanous related formin-3 (DRF3), is an inhibitor of the "amoeboid" phenotype in human PCa cells, and that chromosomal loss at the DRF3 coding locus occurs at high frequency in metastatic but not primary disease. Functional studies have demonstrated that DRF3 is a member of the EGF receptor (EGFR) network and is positioned at a signaling node that controls the transition from mesenchymal to amoeboid cancer cell phenotypes. The mesenchymal-amoeboid transition (MAT) has recently been identified as a dynamic phenotypic shift that can affect migration of tumor cells through matrices and reduce their sensitivity to protease inhibitors. DRF3 also inhibits the secretion of bioactive microvesicles capable of altering the tumor microenvironment and associates with cholesterol-rich lipid raft membrane microdomains. These data suggest that the DRF3 network may be sensitive to cholesterol- or fatty acid-targeting therapies and/or chemoprevention, which have shown promise in human studies and animal models. We will test the hypothesis that amoeboid properties arising from DRF3 loss results in a more aggressive phenotype. We further hypothesize that amoeboid behavior coincides with upregulation of lipid anabolism and that amoeboid tumor cells may become addicted to lipid-dependent pathways. Thirdly, we hypothesize that a distinct amoeboid signaling network exists and that components of this network may represent novel tumor biomarkers.
The specific aims are:
Aim 1. Determine the biological consequences of DRF3 loss in prostate cancer. The role of androgen, the androgen receptor, and ERBB receptor tyrosine kinase signaling in the amoeboid phenotype will be determined in the context of DRF3 silencing. We will assess whether DRF3 actively mediates the transition to the mesenchymal phenotype. The effect of DRF3 silencing on tumor growth, metastasis, and sensitivity to ERBB- and cholesterol-targeting therapy, and androgen ablation will be assessed. Patterns of DRF3 expression in human prostate cancers will be evaluated and correlated with clinical parameters.
Aim 2. Identify the signaling network that mediates the mesenchymal-amoeboid transition in prostate cancer. A series of directed as well as unbiased experiments, evaluating perturbations in RNA, protein, and palmitoyl-protein networks under conditions of stable DRF3 silencing and EGFR activation will be employed to uncover the signaling network controlling the MAT in prostate cancer cells. Models will be constructed from these data and validated to identify a small series of informative indicators of the MAT. These markers will be used to determine whether the amoeboid phenotype can be detected in vivo using a state-of-the-art platform relevant to prognostic evaluation. These studies will provide new insight into the relationship between membrane dynamics, lipid metabolism and PCa progression to lethal disease.
This proposal originates from the recent discoveries by the Freeman laboratory that an actin-nucleating protein, Diaphanous related formin-3 (DRF3), is an inhibitor of the amoeboid tumor phenotype and that chromosomal loss at the gene encoding DRF3 occurs with high frequency in patients with metastatic prostate cancer. DRF3 protein expression also declines with disease progression. The mesenchymal-amoeboid transition (MAT) has recently been identified as a dynamic phenotypic transition that can affect migration of tumor cells through matrices and reduce their sensitivity to protease inhibitors. Our data indicate that DRF3 is positioned at a signal transduction node that controls the MAT. Thus, DRF3 has the characteristics of a novel type of tumor- or metastasis-suppressor protein. Our data suggest that this network is linked to anabolic lipogenesis, a feature of aggressive tumor cells. The metabolic phenotype of tumor cells with amoeboid properties may make them vulnerable to therapeutic interventions that target cholesterol or lipid metabolic pathways. Our goals are to determine the biological consequences of DRF3 downregulation in prostate cancer, and to uncover the MAT signaling network that responds to DRF3 silencing. These studies will provide new insight into the relationship between membrane dynamics, lipid metabolism and castrate-resistant prostate cancer. DRF3 loss and the MAT may also be functionally important in other solid tumor systems. Consequently, these studies may provide information of more general significance to human cancer.
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|Freeman, Michael R (2013) The ubiquitin ligase Siah2 is revealed as an accomplice of the androgen receptor in castration resistant prostate cancer. Asian J Androl 15:447-8|
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|Ayala, Gustavo; Morello, Matteo; Frolov, Anna et al. (2013) Loss of caveolin-1 in prostate cancer stroma correlates with reduced relapse-free survival and is functionally relevant to tumour progression. J Pathol 231:77-87|
|Li, Xiangyan; Chen, Yi-Ting; Josson, Sajni et al. (2013) MicroRNA-185 and 342 inhibit tumorigenicity and induce apoptosis through blockade of the SREBP metabolic pathway in prostate cancer cells. PLoS One 8:e70987|
|Solomon, Keith R; Allott, Emma H; Freeman, Michael R et al. (2013) Words of wisdom. Re: Dysregulation of cholesterol homeostasis in human prostate cancer through loss of ABCA1. Eur Urol 63:1128-9|
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