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GENETIC VARIATION ON 8q24 AND SUSCEPTIBILITY TO PROSTATE CANCER IN CAUCASIAN MEN
226
THE JOURNAL OF UROLOGY®
PC-3 cells. WL-276 inhibited in vivo tumor growth in a dose-dependent manner which correlated with serum levels of the agent. CONCLUSIONS: From a translational prospective, WL-276 provides a proof of principle that Bcl-2/microtubule dual antagonists can be developed and such dual antagonists are promising treatments against HRPC. Moreover, as the communication between proliferation (microtubule) and apoptosis (Bcl-2 proteins) remains unclear, WL-276 also represents a unique pharmacological probe to investigate this potential network. Source of Funding: NCI/NIH - 5R01CA114294.
643 ELIMINATION OF DIETARY CARBOHYDRATE EXTENDS SURVIVAL OF MICE BEARING LNCAP XENOGRAFTS W Cooper Buschemeyer*, Alok Tewari, David Hsu, John Mavropoulos, Dmitriy Rokhfeld, Tracy Johnson, Salvatore Pizzo, Phillip G Febbo, Stephen J Freedland. Durham, NC. INTRODUCTION AND OBJECTIVE: We previously showed a no-carbohydrate ketogenic diet (NCKD) slowed prostate tumor growth relative to a Western-type diet in a murine xenograft model using a hormone-sensitive cell line (LAPC-4). Given our concerns that the high fat content of the NCKD may nonetheless accelerate tumor growth in some prostate cancer (PC) models, we conducted a second study with a different cell line (LNCaP) in order to assess the generality of our LQLWLDO¿QGLQJV METHODS: 130 individually caged male SCID mice were fed either a NCKD(84% fat, 0% carb, 16% protein), a low-fat diet (LFD)(12% fat, 72% carb, 16% protein), or a Western diet (WD )(40% fat, 44% carb, 16% protein). Mice on the LFD were fed ad libitum, WD mice received identical calories, and NCKD were fed 12.5% extra calories relative to LFD mice in order to maintain similar average body weights. After six weeks of feeding, all mice were injected with 1x106 LNCaP cells in 0DWULJHO GD\ DQGVDFUL¿FHGZKHQWXPRUVZHUHPP51$ was isolated from tumors in each diet group and placed on Affymetrix HTA-HG-133A arrays. Gene set enrichment analysis (GSEA) was performed to identify pathways differentially regulated by diet. RESULTS: At day 36, the NCKD and LFD groups had identical average body weights and both were heavier than the WD group by 1.5g (6.4%) (ANOVA, p=0.0391). Mice fed the WD experienced VLJQL¿FDQWO\ZRUVHVXUYLYDOWKDQPLFHLQWKH/)'JURXS S DQG PLFH LQ WKH 1&.' JURXS S ZLWK QR VLJQL¿FDQW GLIIHUHQFH LQ survival between LFD and NCKD groups (p=0.67). Pathways found to EHVLJQL¿FDQWO\HQULFKHG S LQWKH:'JURXSRYHU1&.'LQFOXGH WKRVHLPSOLFDWHGLQREHVLW\LQVXOLQUHVLVWDQFHDQGLQÀDPPDWLRQ,QWKH WD group compared to the LFD group, pathways involved in histone GHDFHW\ODWLRQ SUROLIHUDWLRQ DQG LQVXOLQ VHQVLWLYLW\ ZHUH VLJQL¿FDQWO\ enriched. Across all three diets, as carbohydrate content increased, LQÀDPPDWRU\SDWKZD\HQULFKPHQWZDVREVHUYHG CONCLUSIONS: As with LAPC-4, the NCKD slows tumor growth relative to the WD. While the mechanisms through which diet LPSDFWV3&JURZWKUHPDLQVXQFOHDURXUH[SUHVVLRQDQDO\VLVLGHQWL¿HG multiple biological processes that were associated with diet, and VSHFL¿FDOO\IRXQGHYLGHQFHIRULQFUHDVHGLQÀDPPDWRU\DFWLYDWLRQZLWKLQ the tumor with increased dietary carbohydrate. Given the previously HVWDEOLVKHG OLQN EHWZHHQ LQÀDPPDWLRQ DQG SURVWDWH FDQFHU LQLWLDWLRQ DQG SURJUHVVLRQ IXUWKHU IXQFWLRQDO FKDUDFWHUL]DWLRQ PD\ SURYLGH D mechanistic explanation for the observed impact of diet on tumor growth. Source of Funding: None
644 DIFFERENTIAL GENE EXPRESSION IN HUMAN PROSTATE CANCER BASED ON ETHNIC OR GEOGRAPHIC ORIGIN Wilmer B Roberts*, Thomas A Dunn, Chiledum A Ahaghotu, Rick Kittles, Stephen J Freedland, William B Isaacs, Jun Luo. Baltimore, MD, Washington, DC, Chicago, IL, and Durham, NC. INTRODUCTION AND OBJECTIVE: Although prostate cancer is a common disease among men of both African and European descent, both the incidence and mortality rate are substantially higher in African $PHULFDQ0HQ:HVRXJKWWRDQDO\]HWKHJHQHWLFDQGRWKHUPROHFXODU
Vol. 179, No. 4, Supplement, Monday, May 19, 2008
differences that underlie the differential risk for prostate cancer initiation and progression among the ethnically diverse populations using whole genome approaches such as DNA microarrays. METHODS: Paired samples of normal and cancerous prostate tissues obtained from African American (AA) and European American (EA) men undergoing radical prostatectomy were processed for RNA and then evaluated via microarrays containing over 20,000 human genes. Candidate markers representing differentially expressed genes were then evaluated using reverse-transcriptase polymerase chain reaction (RT-PCR) in an independent set of specimens. 5(68/76 7KUHHJHQHVZHUHUHDGLO\LGHQWL¿HGZLWKFKDQJHV in expression in tumor tissue relative to normal tissue and with differential expression in AA men relative to EA men. Ephrin-B2 is overexpressed LQ$$ WXPRU WLVVXH ZKLOH 3/$* ZDV VSHFL¿FDOO\ RYHUH[SUHVVHG LQ EA tumors. HMGCS2, a gene that appeared to be underexpressed in more aggressive prostate cancers, is also underexpressed in AA tumors. Studies are ongoing to validate these expression changes in H[SDQGHGVDPSOHVHWVDQGWRGH¿QHWKHLUIXQFWLRQDOFRQVHTXHQFHVLQ men with prostate cancer. CONCLUSIONS: These differential gene expression changes may provide insight into the underlying biology of prostate cancer in ethnically diverse populations. It may at least partially explain the increased incidence of and/or mortality from prostate cancer experienced by AA relative to EA men. Upon further validation and functional analysis, the individual markers may be useful for differential diagnosis and LQGLYLGXDOL]HGWKHUDS\ Source of Funding: None
645 GENETIC VARIATION ON 8q24 AND SUSCEPTIBILITY TO PROSTATE CANCER IN CAUCASIAN MEN Prodipto Pal, Huifeng Xi, Saurav Guha, Guangyun Sun, Brian T Helfand, Joshua J Meeks, Brian K Suarez, William J Catalona, Ranjan Deka*. Cincinnati, OH, Chicago, IL, and St. Louis, MO. INTRODUCTION AND OBJECTIVE: Several recent whole JHQRPH DVVRFLDWLRQ VWXGLHV KDYH LQGHSHQGHQWO\ LGHQWL¿HG PXOWLSOH prostate cancer (PCa) susceptibility variants on 8q24. We have previously reported results of case-control association studies in research participants from the Chicago metropolitan area. In this study, we evaluated the risk associated variants in this chromosomal region in a sample of European Americans derived from the St. Louis metropolitan area. METHODS: A case-control association study was performed RQDVDPSOHRI KLVWRORJLFDOO\YHUL¿HG3&DFDVHVDQGHWKQLFDOO\ matched controls. We employed a tagging approach to select a set of maximally informative SNPs (tagSNPs) distributed in three previously defined regions on 8q24 associated with PCa susceptibility (Nat Genet volume 39, 2007). Forty-four tagSNPs, including the three most VLJQL¿FDQWO\ DVVRFLDWHG 613V UV UV UV spanning a region of 430kb were genotyped using the SNPlex platform. Association was tested using likelihood ratio test, and multiple corrections were performed using permutations. RESULTS: We observed three distinct regions of linkage GLVHTXLOLEULXPFRQVLVWHQWZLWKSUHYLRXVUHSRUWV$VLJQL¿FDQWDVVRFLDWLRQ was observed for ten of the 44 tagSNPs after adjusting for multiple corrections (P~1.0E-3~1.0E-4). We did not find association with rs1447295 and nine other SNPs in region 1. Although rs16901979 (region 2) did not show association, six variants surrounding this SNP VKRZHGVLJQL¿FDQWDVVRFLDWLRQ7KHUHPDLQLQJIRXUVLJQL¿FDQW613VDUH located in region 3, that include rs6983267. &21&/86,216 $OWKRXJK WKLV VWXG\ GRHV QRW FRQ¿UP WKH association of rs1447295 and region 1, we corroborate reported signals LQUHJLRQVDQG7KLVVWXG\UHDI¿UPVWKDWJHQHWLFYDULDWLRQRQT LQÀXHQFHVXVFHSWLELOLW\WR3&DLQPHQRI(XURSHDQDQFHVWU\ Source of Funding: Urological Research Foundation.
THE JOURNAL OF UROLOGY®
PC-3 cells. WL-276 inhibited in vivo tumor growth in a dose-dependent manner which correlated with serum levels of the agent. CONCLUSIONS: From a translational prospective, WL-276 provides a proof of principle that Bcl-2/microtubule dual antagonists can be developed and such dual antagonists are promising treatments against HRPC. Moreover, as the communication between proliferation (microtubule) and apoptosis (Bcl-2 proteins) remains unclear, WL-276 also represents a unique pharmacological probe to investigate this potential network. Source of Funding: NCI/NIH - 5R01CA114294.
643 ELIMINATION OF DIETARY CARBOHYDRATE EXTENDS SURVIVAL OF MICE BEARING LNCAP XENOGRAFTS W Cooper Buschemeyer*, Alok Tewari, David Hsu, John Mavropoulos, Dmitriy Rokhfeld, Tracy Johnson, Salvatore Pizzo, Phillip G Febbo, Stephen J Freedland. Durham, NC. INTRODUCTION AND OBJECTIVE: We previously showed a no-carbohydrate ketogenic diet (NCKD) slowed prostate tumor growth relative to a Western-type diet in a murine xenograft model using a hormone-sensitive cell line (LAPC-4). Given our concerns that the high fat content of the NCKD may nonetheless accelerate tumor growth in some prostate cancer (PC) models, we conducted a second study with a different cell line (LNCaP) in order to assess the generality of our LQLWLDO¿QGLQJV METHODS: 130 individually caged male SCID mice were fed either a NCKD(84% fat, 0% carb, 16% protein), a low-fat diet (LFD)(12% fat, 72% carb, 16% protein), or a Western diet (WD )(40% fat, 44% carb, 16% protein). Mice on the LFD were fed ad libitum, WD mice received identical calories, and NCKD were fed 12.5% extra calories relative to LFD mice in order to maintain similar average body weights. After six weeks of feeding, all mice were injected with 1x106 LNCaP cells in 0DWULJHO GD\ DQGVDFUL¿FHGZKHQWXPRUVZHUHPP51$ was isolated from tumors in each diet group and placed on Affymetrix HTA-HG-133A arrays. Gene set enrichment analysis (GSEA) was performed to identify pathways differentially regulated by diet. RESULTS: At day 36, the NCKD and LFD groups had identical average body weights and both were heavier than the WD group by 1.5g (6.4%) (ANOVA, p=0.0391). Mice fed the WD experienced VLJQL¿FDQWO\ZRUVHVXUYLYDOWKDQPLFHLQWKH/)'JURXS S DQG PLFH LQ WKH 1&.' JURXS S ZLWK QR VLJQL¿FDQW GLIIHUHQFH LQ survival between LFD and NCKD groups (p=0.67). Pathways found to EHVLJQL¿FDQWO\HQULFKHG S LQWKH:'JURXSRYHU1&.'LQFOXGH WKRVHLPSOLFDWHGLQREHVLW\LQVXOLQUHVLVWDQFHDQGLQÀDPPDWLRQ,QWKH WD group compared to the LFD group, pathways involved in histone GHDFHW\ODWLRQ SUROLIHUDWLRQ DQG LQVXOLQ VHQVLWLYLW\ ZHUH VLJQL¿FDQWO\ enriched. Across all three diets, as carbohydrate content increased, LQÀDPPDWRU\SDWKZD\HQULFKPHQWZDVREVHUYHG CONCLUSIONS: As with LAPC-4, the NCKD slows tumor growth relative to the WD. While the mechanisms through which diet LPSDFWV3&JURZWKUHPDLQVXQFOHDURXUH[SUHVVLRQDQDO\VLVLGHQWL¿HG multiple biological processes that were associated with diet, and VSHFL¿FDOO\IRXQGHYLGHQFHIRULQFUHDVHGLQÀDPPDWRU\DFWLYDWLRQZLWKLQ the tumor with increased dietary carbohydrate. Given the previously HVWDEOLVKHG OLQN EHWZHHQ LQÀDPPDWLRQ DQG SURVWDWH FDQFHU LQLWLDWLRQ DQG SURJUHVVLRQ IXUWKHU IXQFWLRQDO FKDUDFWHUL]DWLRQ PD\ SURYLGH D mechanistic explanation for the observed impact of diet on tumor growth. Source of Funding: None
644 DIFFERENTIAL GENE EXPRESSION IN HUMAN PROSTATE CANCER BASED ON ETHNIC OR GEOGRAPHIC ORIGIN Wilmer B Roberts*, Thomas A Dunn, Chiledum A Ahaghotu, Rick Kittles, Stephen J Freedland, William B Isaacs, Jun Luo. Baltimore, MD, Washington, DC, Chicago, IL, and Durham, NC. INTRODUCTION AND OBJECTIVE: Although prostate cancer is a common disease among men of both African and European descent, both the incidence and mortality rate are substantially higher in African $PHULFDQ0HQ:HVRXJKWWRDQDO\]HWKHJHQHWLFDQGRWKHUPROHFXODU
Vol. 179, No. 4, Supplement, Monday, May 19, 2008
differences that underlie the differential risk for prostate cancer initiation and progression among the ethnically diverse populations using whole genome approaches such as DNA microarrays. METHODS: Paired samples of normal and cancerous prostate tissues obtained from African American (AA) and European American (EA) men undergoing radical prostatectomy were processed for RNA and then evaluated via microarrays containing over 20,000 human genes. Candidate markers representing differentially expressed genes were then evaluated using reverse-transcriptase polymerase chain reaction (RT-PCR) in an independent set of specimens. 5(68/76 7KUHHJHQHVZHUHUHDGLO\LGHQWL¿HGZLWKFKDQJHV in expression in tumor tissue relative to normal tissue and with differential expression in AA men relative to EA men. Ephrin-B2 is overexpressed LQ$$ WXPRU WLVVXH ZKLOH 3/$* ZDV VSHFL¿FDOO\ RYHUH[SUHVVHG LQ EA tumors. HMGCS2, a gene that appeared to be underexpressed in more aggressive prostate cancers, is also underexpressed in AA tumors. Studies are ongoing to validate these expression changes in H[SDQGHGVDPSOHVHWVDQGWRGH¿QHWKHLUIXQFWLRQDOFRQVHTXHQFHVLQ men with prostate cancer. CONCLUSIONS: These differential gene expression changes may provide insight into the underlying biology of prostate cancer in ethnically diverse populations. It may at least partially explain the increased incidence of and/or mortality from prostate cancer experienced by AA relative to EA men. Upon further validation and functional analysis, the individual markers may be useful for differential diagnosis and LQGLYLGXDOL]HGWKHUDS\ Source of Funding: None
645 GENETIC VARIATION ON 8q24 AND SUSCEPTIBILITY TO PROSTATE CANCER IN CAUCASIAN MEN Prodipto Pal, Huifeng Xi, Saurav Guha, Guangyun Sun, Brian T Helfand, Joshua J Meeks, Brian K Suarez, William J Catalona, Ranjan Deka*. Cincinnati, OH, Chicago, IL, and St. Louis, MO. INTRODUCTION AND OBJECTIVE: Several recent whole JHQRPH DVVRFLDWLRQ VWXGLHV KDYH LQGHSHQGHQWO\ LGHQWL¿HG PXOWLSOH prostate cancer (PCa) susceptibility variants on 8q24. We have previously reported results of case-control association studies in research participants from the Chicago metropolitan area. In this study, we evaluated the risk associated variants in this chromosomal region in a sample of European Americans derived from the St. Louis metropolitan area. METHODS: A case-control association study was performed RQDVDPSOHRI KLVWRORJLFDOO\YHUL¿HG3&DFDVHVDQGHWKQLFDOO\ matched controls. We employed a tagging approach to select a set of maximally informative SNPs (tagSNPs) distributed in three previously defined regions on 8q24 associated with PCa susceptibility (Nat Genet volume 39, 2007). Forty-four tagSNPs, including the three most VLJQL¿FDQWO\ DVVRFLDWHG 613V UV UV UV spanning a region of 430kb were genotyped using the SNPlex platform. Association was tested using likelihood ratio test, and multiple corrections were performed using permutations. RESULTS: We observed three distinct regions of linkage GLVHTXLOLEULXPFRQVLVWHQWZLWKSUHYLRXVUHSRUWV$VLJQL¿FDQWDVVRFLDWLRQ was observed for ten of the 44 tagSNPs after adjusting for multiple corrections (P~1.0E-3~1.0E-4). We did not find association with rs1447295 and nine other SNPs in region 1. Although rs16901979 (region 2) did not show association, six variants surrounding this SNP VKRZHGVLJQL¿FDQWDVVRFLDWLRQ7KHUHPDLQLQJIRXUVLJQL¿FDQW613VDUH located in region 3, that include rs6983267. &21&/86,216 $OWKRXJK WKLV VWXG\ GRHV QRW FRQ¿UP WKH association of rs1447295 and region 1, we corroborate reported signals LQUHJLRQVDQG7KLVVWXG\UHDI¿UPVWKDWJHQHWLFYDULDWLRQRQT LQÀXHQFHVXVFHSWLELOLW\WR3&DLQPHQRI(XURSHDQDQFHVWU\ Source of Funding: Urological Research Foundation.