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Changmeng Cai,
Hongyun Wang,
Housheng Hansen He,
Sen Chen,
Lingfeng He,
Fen Ma,
Lorelei Mucci,
Qianben Wang, Christopher Fiore,
Adam G Sowalsky,
Massimo Loda,
X Shirley Liu,
Myles Brown,
Steven P Balk,
Xin Yuan
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Changmeng Cai,
Hongyun Wang,
Housheng Hansen He,
Sen Chen,
Lingfeng He,
Fen Ma,
Lorelei Mucci,
Qianben Wang, Christopher Fiore,
Adam G Sowalsky,
Massimo Loda,
X Shirley Liu,
Myles Brown,
Steven P Balk,
Xin Yuan
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Changmeng Cai,
Hongyun Wang,
Housheng Hansen He,
Sen Chen,
Lingfeng He,
Fen Ma,
Lorelei Mucci,
Qianben Wang, Christopher Fiore,
Adam G Sowalsky,
Massimo Loda,
X Shirley Liu,
Myles Brown,
Steven P Balk,
Xin Yuan
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Changmeng Cai,
Hongyun Wang,
Housheng Hansen He,
Sen Chen,
Lingfeng He,
Fen Ma,
Lorelei Mucci,
Qianben Wang, Christopher Fiore,
Adam G Sowalsky,
Massimo Loda,
X Shirley Liu,
Myles Brown,
Steven P Balk,
Xin Yuan
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Changmeng Cai,
Hongyun Wang,
Housheng Hansen He,
Sen Chen,
Lingfeng He,
Fen Ma,
Lorelei Mucci,
Qianben Wang, Christopher Fiore,
Adam G Sowalsky,
Massimo Loda,
X Shirley Liu,
Myles Brown,
Steven P Balk,
Xin Yuan
[show abstract]
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ABSTRACT: Fusion of the androgen receptor-regulated (AR-regulated) TMPRSS2 gene with ERG in prostate cancer (PCa) causes androgen-stimulated overexpression of ERG, an ETS transcription factor, but critical downstream effectors of ERG-mediating PCa development remain to be established. Expression of the SOX9 transcription factor correlated with TMPRSS2:ERG fusion in 3 independent PCa cohorts, and ERG-dependent expression of SOX9 was confirmed by RNAi in the fusion-positive VCaP cell line. SOX9 has been shown to mediate ductal morphogenesis in fetal prostate and maintain stem/progenitor cell pools in multiple adult tissues, and has also been linked to PCa and other cancers. SOX9 overexpression resulted in neoplasia in murine prostate and stimulated tumor invasion, similarly to ERG. Moreover, SOX9 depletion in VCaP cells markedly impaired invasion and growth in vitro and in vivo, establishing SOX9 as a critical downstream effector of ERG. Finally, we found that ERG regulated SOX9 indirectly by opening a cryptic AR-regulated enhancer in the SOX9 gene. Together, these results demonstrate that ERG redirects AR to a set of genes including SOX9 that are not normally androgen stimulated, and identify SOX9 as a critical downstream effector of ERG in TMPRSS2:ERG fusion-positive PCa.
The Journal of clinical investigation 03/2013; 123(3):1109-22. · 15.39 Impact Factor
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Christopher Fiore,
Dyane Bailey,
Niamh Conlon,
Xiaoqiu Wu,
Neil Martin,
Michelangelo Fiorentino,
Stephen Finn,
Katja Fall,
Swen-Olof Andersson,
Ove Andren,
Massimo Loda,
Richard Flavin
[show abstract]
[hide abstract]
ABSTRACT: Automated scanning devices and image analysis software provide a means to overcome the limitations of manual semiquantitative scoring of immunohistochemistry. Common drawbacks to automated imaging systems include an inability to classify tissue type and an inability to segregate cytoplasmic and nuclear staining.
Immunohistochemistry for the membranous marker α-catenin, the cytoplasmic marker stathmin and the nuclear marker Ki-67 was performed on tissue microarrays (TMA) of archival formalin-fixed paraffin-embedded tissue comprising 471 (α-catenin and stathmin) and 511 (Ki-67) cases of prostate adenocarcinoma. These TMA were quantitatively analysed using two commercially available automated image analysers, the Ariol SL-50 system and the Nuance system from CRi. Both systems use brightfield microscopy for automated, unbiased and standardised quantification of immunohistochemistry, while the Nuance system has spectral deconvolution capabilities.
Overall concordance between scores from both systems was excellent (r=0.90; 0.83-0.95). The software associated with the multispectral imager allowed accurate automated classification of tissue type into epithelial glandular structures and stroma, and a single-step segmentation of staining into cytoplasmic or nuclear compartments allowing independent evaluation of these areas. The Nuance system, however, was not able to distinguish reliably between tumour and non-tumour tissue. In addition, variance in the labour and time required for analysis between the two systems was also noted.
Despite limitations, this study suggests some beneficial role for the use of a multispectral imaging system in automated analysis of immunohistochemistry.
Journal of clinical pathology 03/2012; 65(6):496-502. · 2.43 Impact Factor
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Whitney K Hendrickson,
Richard Flavin,
Julie L Kasperzyk,
Michelangelo Fiorentino,
Fang Fang,
Rosina Lis, Christopher Fiore,
Kathryn L Penney,
Jing Ma,
Philip W Kantoff,
Meir J Stampfer,
Massimo Loda,
Lorelei A Mucci,
Edward Giovannucci
[show abstract]
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ABSTRACT: Data suggest that circulating 25-hydroxyvitamin D [25(OH)D] interacts with the vitamin D receptor (VDR) to decrease proliferation and increase apoptosis for some malignancies, although evidence for prostate cancer is less clear. How VDR expression in tumor tissue may influence prostate cancer progression has not been evaluated in large studies.
We examined protein expression of VDR in tumor tissue among 841 patients with prostate cancer in relation to risk of lethal prostate cancer within two prospective cohorts, the Physicians' Health Study and Health Professionals Follow-Up Study. We also examined the association of VDR expression with prediagnostic circulating 25(OH)D and 1,25-dihydroxyvitamin D levels and with two VDR single nucleotide polymorphisms, FokI and BsmI.
Men whose tumors had high VDR expression had significantly lower prostate-specific antigen (PSA) at diagnosis (P for trend < .001), lower Gleason score (P for trend < .001), and less advanced tumor stage (P for trend < .001) and were more likely to have tumors harboring the TMPRSS2:ERG fusion (P for trend = .009). Compared with the lowest quartile, men whose tumors had the highest VDR expression had significantly reduced risk of lethal prostate cancer (hazard ratio [HR], 0.17; 95% CI, 0.07 to 0.41). This association was only slightly attenuated after adjustment for Gleason score and PSA at diagnosis (HR, 0.33; 95% CI, 0.13 to 0.83) or, additionally, for tumor stage (HR, 0.37; 95% CI, 0.14 to 0.94). Neither prediagnostic plasma vitamin D levels nor VDR polymorphisms were associated with VDR expression.
High VDR expression in prostate tumors is associated with a reduced risk of lethal cancer, suggesting a role of the vitamin D pathway in prostate cancer progression.
Journal of Clinical Oncology 06/2011; 29(17):2378-85. · 18.37 Impact Factor
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Paul L Nguyen,
Jing Ma,
Jorge E Chavarro,
Matthew L Freedman,
Rosina Lis,
Giuseppe Fedele, Christopher Fiore,
Weiliang Qiu,
Michelangelo Fiorentino,
Stephen Finn,
Kathryn L Penney,
Anna Eisenstein,
Fredrick R Schumacher,
Lorelei A Mucci,
Meir J Stampfer,
Edward Giovannucci,
Massimo Loda
[show abstract]
[hide abstract]
ABSTRACT: Fatty acid synthase (FASN) regulates de novo lipogenesis, body weight, and tumor growth. We examined whether common germline single nucleotide polymorphisms (SNPs) in the FASN gene affect prostate cancer (PCa) risk or PCa-specific mortality and whether these effects vary by body mass index (BMI).
In a prospective nested case-control study of 1,331 white patients with PCa and 1,267 age-matched controls, we examined associations of five common SNPs within FASN (and 5 kb upstream/downstream, R(2) > 0.8) with PCa incidence and, among patients, PCa-specific death and tested for an interaction with BMI. Survival analyses were repeated for tumor FASN expression (n = 909).
Four of the five SNPs were associated with lethal PCa. SNP rs1127678 was significantly related to higher BMI and interacted with BMI for both PCa risk (P(interaction) = .004) and PCa mortality (P(interaction) = .056). Among overweight men (BMI > or = 25 kg/m(2)), but not leaner men, the homozygous variant allele carried a relative risk of advanced PCa of 2.49 (95% CI, 1.00 to 6.23) compared with lean men with the wild type. Overweight patients carrying the variant allele had a 2.04 (95% CI, 1.31 to 3.17) times higher risk of PCa mortality. Similarly, overweight patients with elevated tumor FASN expression had a 2.73 (95% CI, 1.05 to 7.08) times higher risk of lethal PCa (P(interaction) = .02).
FASN germline polymorphisms were significantly associated with risk of lethal PCa. Significant interactions of BMI with FASN polymorphisms and FASN tumor expression suggest FASN as a potential link between obesity and poor PCa outcome and raise the possibility that FASN inhibition could reduce PCa-specific mortality, particularly in overweight men.
Journal of Clinical Oncology 09/2010; 28(25):3958-64. · 18.37 Impact Factor
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Michelangelo Fiorentino,
Gregory Judson,
Kathryn Penney,
Richard Flavin,
Jennifer Stark, Christopher Fiore,
Katja Fall,
Neil Martin,
Jing Ma,
Jennifer Sinnott,
Edward Giovannucci,
Meir Stampfer,
Howard D Sesso,
Philip W Kantoff,
Stephen Finn,
Massimo Loda,
Lorelei Mucci
[show abstract]
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ABSTRACT: BRCA1 functions as a tumor suppressor; recent work suggests that BRCA1 may also induce cell cycle arrest to allow for DNA repair. We hypothesized that BRCA1 expression in prostate tumor tissue may be associated with prostate cancer progression through regulation of the cell cycle. We used immunohistochemistry to evaluate BRCA1 protein expression in archival tumor samples from 393 prostate cancer cases in the Physicians' Health Study. The men were followed prospectively from diagnosis to development of metastases and mortality. Fifteen percent of tumors stained positive for BRCA1. BRCA1-positive tumors had substantially increased tumor proliferation index compared with negative tumors (47.0 Ki67-positive nuclei versus 10.3, P = 0.0016) and were more likely to develop lethal cancer compared with BRCA1-negative tumors (hazard ratio, 4.6; 95% confidence interval, 2.4-8.7). These findings strengthen the hypothesis that BRCA1 plays a role in cell cycle control and show that BRCA1 is a marker of clinical prostate cancer prognosis. Cancer Res; 70(8); 3136-9. (c)2010 AACR.
Cancer Research 04/2010; 70(8):3136-9. · 7.86 Impact Factor
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Qianben Wang,
Wei Li,
Yong Zhang,
Xin Yuan,
Kexin Xu,
Jindan Yu,
Zhong Chen,
Rameen Beroukhim,
Hongyun Wang,
Mathieu Lupien, [......],
Bo Han,
Arul M Chinnaiyan,
Mark A Rubin,
Lawrence True,
Michelangelo Fiorentino, Christopher Fiore,
Massimo Loda,
Philip W Kantoff,
X Shirley Liu,
Myles Brown
[show abstract]
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ABSTRACT: The evolution of prostate cancer from an androgen-dependent state to one that is androgen-independent marks its lethal progression. The androgen receptor (AR) is essential in both, though its function in androgen-independent cancers is poorly understood. We have defined the direct AR-dependent target genes in both androgen-dependent and -independent cancer cells by generating AR-dependent gene expression profiles and AR cistromes. In contrast to what is found in androgen-dependent cells, AR selectively upregulates M-phase cell-cycle genes in androgen-independent cells, including UBE2C, a gene that inactivates the M-phase checkpoint. We find that epigenetic marks at the UBE2C enhancer, notably histone H3K4 methylation and FoxA1 transcription factor binding, are present in androgen-independent cells and direct AR-enhancer binding and UBE2C activation. Thus, the role of AR in androgen-independent cancer cells is not to direct the androgen-dependent gene expression program without androgen, but rather to execute a distinct program resulting in androgen-independent growth.
Cell 08/2009; 138(2):245-56. · 32.40 Impact Factor
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Eric L Snyder,
Deborah J Sandstrom,
Kenneth Law, Christopher Fiore,
Ewa Sicinska,
Joseph Brito,
Dyane Bailey,
Jonathan A Fletcher,
Massimo Loda,
Scott J Rodig,
Paola Dal Cin,
Christopher D M Fletcher
[show abstract]
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ABSTRACT: Genomic amplification of c-Jun and its upstream kinases have been implicated as a mechanism of progression from well-differentiated to dedifferentiated liposarcoma. To further define the role of c-Jun in liposarcoma progression, we performed immunohistochemistry for c-Jun and its activating kinase ASK1 on a series of liposarcomas (n = 81). We correlated the results with fluorescence in situ hybridization to detect c-Jun amplification. We also derived new cell lines from dedifferentiated liposarcomas with c-Jun amplification. c-Jun protein is expressed in the majority of dedifferentiated liposarcomas (91%) and their well-differentiated components (59%), but only in the minority of pure well-differentiated liposarcomas (27%). When c-Jun is amplified in dedifferentiated liposarcoma, it is interspersed with amplified MDM2 on ring and giant marker chromosomes. MDM2 amplification is one of the earliest events in liposarcoma development, and these results suggest that c-Jun was amplified at a similar time in the evolution of the tumour. In addition, shRNA to c-Jun in c-Jun-amplified liposarcoma cells reduces cell number in vitro and inhibits tumour formation in vivo without an observable effect on the differentiation state of the liposarcoma cells. Thus, c-Jun amplification is oncogenic in liposarcomas but not always sufficient for inhibition of adipocytic differentiation.
The Journal of Pathology 05/2009; 218(3):292-300. · 6.32 Impact Factor
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Toshiro Migita,
Stacey Ruiz,
Alessandro Fornari,
Michelangelo Fiorentino,
Carmen Priolo,
Giorgia Zadra,
Fumika Inazuka,
Chiara Grisanzio,
Emanuele Palescandolo,
Eyoung Shin, [......],
Andrew L Kung,
Phillip G Febbo,
Aravind Subramanian,
Lorelei Mucci,
Jing Ma,
Sabina Signoretti,
Meir Stampfer,
William C Hahn,
Stephen Finn,
Massimo Loda
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ABSTRACT: Overexpression of the fatty acid synthase (FASN) gene has been implicated in prostate carcinogenesis. We sought to directly assess the oncogenic potential of FASN.
We used immortalized human prostate epithelial cells (iPrECs), androgen receptor-overexpressing iPrECs (AR-iPrEC), and human prostate adenocarcinoma LNCaP cells that stably overexpressed FASN for cell proliferation assays, soft agar assays, and tests of tumor formation in immunodeficient mice. Transgenic mice expressing FASN in the prostate were generated to assess the effects of FASN on prostate histology. Apoptosis was evaluated by Hoechst 33342 staining and by fluorescence-activated cell sorting in iPrEC-FASN cells treated with stimulators of the intrinsic and extrinsic pathways of apoptosis (ie, camptothecin and anti-Fas antibody, respectively) or with a small interfering RNA (siRNA) targeting FASN. FASN expression was compared with the apoptotic index assessed by the terminal deoxynucleotidyltransferase-mediated UTP end-labeling method in 745 human prostate cancer samples by using the least squares means procedure. All statistical tests were two-sided.
Forced expression of FASN in iPrECs, AR-iPrECs, and LNCaP cells increased cell proliferation and soft agar growth. iPrECs that expressed both FASN and androgen receptor (AR) formed invasive adenocarcinomas in immunodeficient mice (12 of 14 mice injected formed tumors vs 0 of 14 mice injected with AR-iPrEC expressing empty vector (P < .001, Fisher exact test); however, iPrECs that expressed only FASN did not. Transgenic expression of FASN in mice resulted in prostate intraepithelial neoplasia, the incidence of which increased from 10% in 8- to 16-week-old mice to 44% in mice aged 7 months or more (P = .0028, Fisher exact test), but not in invasive tumors. In LNCaP cells, siRNA-mediated silencing of FASN resulted in apoptosis. FASN overexpression protected iPrECs from apoptosis induced by camptothecin but did not protect iPrECs from Fas receptor-induced apoptosis. In human prostate cancer specimens, FASN expression was inversely associated with the apoptotic rate (mean percentage of apoptotic cells, lowest vs highest quartile of FASN expression: 2.76 vs 1.34, difference = 1.41, 95% confidence interval = 0.45 to 2.39, Ptrend = .0046).
These observations suggest that FASN can act as a prostate cancer oncogene in the presence of AR and that FASN exerts its oncogenic effect by inhibiting the intrinsic pathway of apoptosis.
CancerSpectrum Knowledge Environment 05/2009; 101(7):519-32. · 14.07 Impact Factor
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Michelangelo Fiorentino,
Giorgia Zadra,
Emanuele Palescandolo,
Giuseppe Fedele,
Dyane Bailey, Christopher Fiore,
Paul L Nguyen,
Toshiro Migita,
Raffaella Zamponi,
Dolores Di Vizio,
Carmen Priolo,
Chandan Sharma,
Wanling Xie,
Martin E Hemler,
Lorelei Mucci,
Edward Giovannucci,
Stephen Finn,
Massimo Loda
[show abstract]
[hide abstract]
ABSTRACT: Fatty acid synthase (FASN), a key metabolic enzyme for liponeogenesis highly expressed in several human cancers, displays oncogenic properties such as resistance to apoptosis and induction of proliferation when overexpressed. To date, no mechanism has been identified to explain the oncogenicity of FASN in prostate cancer. We generated immortalized prostate epithelial cells (iPrECs) overexpressing FASN, and found that 14C-acetate incorporation into palmitate synthesized de novo by FASN was significantly elevated in immunoprecipitated Wnt-1 when compared to isogenic cells not overexpressing FASN. Overexpression of FASN caused membranous and cytoplasmic β-catenin protein accumulation and activation, whereas FASN knockdown by short-hairpin RNA resulted in a reduction in the extent of β-catenin activation. Orthotopic transplantation of iPrECs overexpressing FASN in nude mice resulted in invasive tumors that overexpressed β-catenin. A strong significant association between FASN and cytoplasmic (stabilized) β-catenin immunostaining was found in 862 cases of human prostate cancer after computerized subtraction of the membranous β-catenin signal (P<0.001, Spearman's ρ=0.33). We propose that cytoplasmic stabilization of β-catenin through palmitoylation of Wnt-1 and subsequent activation of the pathway is a potential mechanism of FASN oncogenicity in prostate cancer.Keywords: β-catenin, fatty acid synthase, prostate cancer, immunohistochemistry, palmitoylation, Wnt1
Laboratory Investigation 10/2008; 88(12):1340-1348. · 3.64 Impact Factor
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Michelangelo Fiorentino,
Giorgia Zadra,
Emanuele Palescandolo,
Giuseppe Fedele,
Dyane Bailey, Christopher Fiore,
Paul L Nguyen,
Toshiro Migita,
Raffaella Zamponi,
Dolores Di Vizio,
Carmen Priolo,
Chandan Sharma,
Wanling Xie,
Martin E Hemler,
Lorelei Mucci,
Edward Giovannucci,
Stephen Finn,
Massimo Loda
[show abstract]
[hide abstract]
ABSTRACT: Fatty acid synthase (FASN), a key metabolic enzyme for liponeogenesis highly expressed in several human cancers, displays oncogenic properties such as resistance to apoptosis and induction of proliferation when overexpressed. To date, no mechanism has been identified to explain the oncogenicity of FASN in prostate cancer. We generated immortalized prostate epithelial cells (iPrECs) overexpressing FASN, and found that (14)C-acetate incorporation into palmitate synthesized de novo by FASN was significantly elevated in immunoprecipitated Wnt-1 when compared to isogenic cells not overexpressing FASN. Overexpression of FASN caused membranous and cytoplasmic beta-catenin protein accumulation and activation, whereas FASN knockdown by short-hairpin RNA resulted in a reduction in the extent of beta-catenin activation. Orthotopic transplantation of iPrECs overexpressing FASN in nude mice resulted in invasive tumors that overexpressed beta-catenin. A strong significant association between FASN and cytoplasmic (stabilized) beta-catenin immunostaining was found in 862 cases of human prostate cancer after computerized subtraction of the membranous beta-catenin signal (P<0.001, Spearman's rho=0.33). We propose that cytoplasmic stabilization of beta-catenin through palmitoylation of Wnt-1 and subsequent activation of the pathway is a potential mechanism of FASN oncogenicity in prostate cancer.
Laboratory Investigation 10/2008; 88(12):1340-8. · 3.64 Impact Factor
