- Email: [email protected]
1427 BCAR1 VARIANTS IN HEREDITARY PROSTATE CANCER FAMILIES WITH LINKAGE TO 16Q23
e550
THE JOURNAL OF UROLOGY姞
1426 TPD52 AS “8Q AMPLIFICATION TARGET“ WITH STRONG PROGNOSTIC RELEVANCE IN PROSTATE CANCER Sarah Minner*, Julia Rohwer, Pierre Tennstedt, Stefan Kurtz, Malte Mader, Lia Burkhardt, Markus Graefen, Hartwig Huland, Guido Sauter, Ronald Simon, Thorsten Schlomm, Hamburg, Germany INTRODUCTION AND OBJECTIVES: Tumor protein D52 (TPD52) is localized on 8q21.1, a region most frequently altered in prostate cancer. Increased expression of TPD52 as a result of gene amplification has been suggested as a marker for prostate cancer progression and metastasis. The aim of this study was to further clarify the potential relevance of TPD52 gene alterations with respect to tumor phenotype and clinical outcome. METHODS: A preexisting prostate cancer tissue microarray (TMA) consisting of over 2000 prostate cancer samples with clinical follow-up data was utilized in this study. Dual-labeling FISH (fluorescence in situ hybridization) with a centromeric probe for chromosome 8 (CEP8) and a probe for the TPD52 gene was performed to analyse TPD52 copy number in relation to chromosome 8 copy number. RESULTS: Gains of TPD52 were found to be significantly associated with Gleason grade, tumor stage and an increased risk for biochemical tumor recurrence (p⬍0.0001 each). Increased TPD52 gene copy number was found in 8.4% Gleason 3⫹3 tumors, in 18.7% Gleason 3⫹4 tumors, in 36.5% Gleason 4⫹3 tumors and in 59.0% Gleason ’4⫹4 tumors. Accordingly, TPD52 gains were found in 11.2% pT2 tumors, in 27.7% pT3 tumors and in 28.6% pT4 tumors. Gene amplifications, defined as TPD52 to CEP8 signal ratio ⱖ 2.0, were found in 1.8% of cases. However, high-level amplifications, defined as a TPD52 copy number of at least 10 and a TPD52 to CEP8 signal ratio of ⱖ2.0, were only found in 2 cases. CONCLUSIONS: These data suggest a strong impact of TPD52 gene copy number on tumor phenotype and clinical outcome. These data are especially interesting in the light of recent reports suggesting TPD52 protein as a target for vaccination in tumors. Source of Funding: This project was supported by the German Federal Ministry of Education and Science in the framework of the program for medical genome research (FKZ:01GS08189).
1427 BCAR1 VARIANTS IN HEREDITARY PROSTATE CANCER FAMILIES WITH LINKAGE TO 16Q23 Emilie Johnson*, Anna Ray, Kimberly Zuhlke, Kathleen Cooney, Ann Arbor, MI INTRODUCTION AND OBJECTIVES: In a genome-wide linkage scan of multiplex prostate cancer (PCa) families from the University of Michigan Prostate Cancer Genetics Project (PCGP), linkage was observed to markers on chromosome 16q23. This region has previously been implicated in hereditary PCa and has been shown to be deleted in approximately 50% of PCa cases suggesting a tumor suppressor mechanism. We set out to analyze BCAR1, which maps to 16q23 and has been implicated in both breast cancer and PCa cancer progression, to assess its potential role in hereditary PCa. METHODS: PCGP families displaying linkage (non-parametric linkage score ⬎ 0) to chromosome 16q23 markers were identified. All BCAR1 exons and intron/exon boundaries were directly sequenced using DNA from the youngest available individual with PCa from each of 96 linked families. We also abstracted relevant clinical data for each of the individuals who underwent sequencing of BCAR1. RESULTS: Of the 96 individuals sequenced, the majority (75.8%) had localized PCa, and median age at diagnosis was 56 years. A total of 38 SNPs were identified, including 10 missense variants. Three of the individuals sequenced were wild type at all 38 single nucleotide polymorphisms (SNPs), and 62 individuals (65%) had at least one nonsynonomous SNP. No truncating mutations were identified in BCAR1. Of particular interest was S76P, which demonstrated a minor allele frequency (MAF) of 0.34 in our study population, which is significantly greater than the previously published MAF of 0.13.
Vol. 183, No. 4, Supplement, Tuesday, June 1, 2010
CONCLUSIONS: Although there were no truncating mutations identified in BCAR1 within our population of hereditary PCa patients, multiple missense variants were identified. The most common nonsynonomous SNP identified was S76P, which had a MAF 2.65 times that of a previously published population. BCAR1 variations likely do not completely explain the 16q23 linkage peak seen in our population. However, BCAR1 is an interesting target for further genotyping and functional studies in both sporadic PCa and hereditary PCa populations. Source of Funding: NIH RO1 CA79596
1428 SIGNIFICANCE OF CCR9-CCL25 AXIS IN PROSTATE CANCER METASTASIS. Praveen Sharma, Louisville, KY; Rajesh Singh, Atlanta, GA; Kristian Novakovic, Louisville, KY; William Grizzle, Birmingham, AL; Shailesh Singh*, Louisville, KY INTRODUCTION AND OBJECTIVES: Prostate cancer (PCa) remains the second leading cause of cancer-related deaths among men in the US. Bone metastasis is major cause of PCa associated deaths and handful treatment alternatives are available to treat this condition. Therefore, better understating of metastatic process and identification of molecular signature on metastatic cells are urgently needed to develop therapeutic for metastatic PCa. We have shown that CCR9, a CC chemokine receptor is highly expressed by the PCa tissue and cells and play an important role in PCa cell migration and invasion in vitro under the chemotactic gradient of CCL25, which is the only natural ligand for CCR9. CCL25 is also known as a thymus-expressed chemokine (TECK) and primarily expressed by the thymus, which involutes with age. Interestingly, we have observed higher serum CCL25 in PCa patients compared to normal healthy donors. Therefore, major goal of this study was to identify the source of CCL25 other than thymus and determine the role of CCR9-CCL25 axis in PCa metastasis. METHODS: Immunohistochemisrty was performed to determine the expression of CCL25 by the primary prostate tumor. Bone marrow cells from mice with or without xenograft tumor was isolated and analyzed by flow cytometry for CCL25 expression. Mice were challenged with Luciferase expressing PC3 cell by intracardiac injection or directly into the tibia of nude mice. After tumor challenge mice were treated with CCL25 neutralization antibodies every second day. Xenogen/Caliper imaging system was used to monitor metastasis and tumor growth/regression non-invasively. RESULTS: For the first time, we show secretion of CCL25 within the primary prostate tumor in a paracrine manner. Interestingly, CCL25 was highly produced by the bone marrow cells isolated from tumor bearing mice as compared to non-tumor bearing mice. Furthermore, inhibition of CCR9-CCL25 interaction in vivo using CCL25 neutralization antibodies, showed significant inhibition of bone metastasis after intra-cardiac injection of PC3 cells and tumor establishment after intra-tibial injection. CONCLUSIONS: These results provide clinical and biological significance of CCR9-CCL25 axis in prostate cancer progression. Higher production of CCL25 by the bone marrow cells of the tumor bearing mouse confirms that CCL25 produced the bone marrow stromal cells are induced by the factors produced, by the tumor cell to facilitate metastasis to the bone. Therefore, developing CCR9 directed therapies, may improve the therapeutic outcome of prostate cancer either as a single agent or in combination with other therapeutic modalities. Source of Funding: Partially supported by DOD-PCRP-NIA (PC051123- W81XWH-06-1-0521)
THE JOURNAL OF UROLOGY姞
1426 TPD52 AS “8Q AMPLIFICATION TARGET“ WITH STRONG PROGNOSTIC RELEVANCE IN PROSTATE CANCER Sarah Minner*, Julia Rohwer, Pierre Tennstedt, Stefan Kurtz, Malte Mader, Lia Burkhardt, Markus Graefen, Hartwig Huland, Guido Sauter, Ronald Simon, Thorsten Schlomm, Hamburg, Germany INTRODUCTION AND OBJECTIVES: Tumor protein D52 (TPD52) is localized on 8q21.1, a region most frequently altered in prostate cancer. Increased expression of TPD52 as a result of gene amplification has been suggested as a marker for prostate cancer progression and metastasis. The aim of this study was to further clarify the potential relevance of TPD52 gene alterations with respect to tumor phenotype and clinical outcome. METHODS: A preexisting prostate cancer tissue microarray (TMA) consisting of over 2000 prostate cancer samples with clinical follow-up data was utilized in this study. Dual-labeling FISH (fluorescence in situ hybridization) with a centromeric probe for chromosome 8 (CEP8) and a probe for the TPD52 gene was performed to analyse TPD52 copy number in relation to chromosome 8 copy number. RESULTS: Gains of TPD52 were found to be significantly associated with Gleason grade, tumor stage and an increased risk for biochemical tumor recurrence (p⬍0.0001 each). Increased TPD52 gene copy number was found in 8.4% Gleason 3⫹3 tumors, in 18.7% Gleason 3⫹4 tumors, in 36.5% Gleason 4⫹3 tumors and in 59.0% Gleason ’4⫹4 tumors. Accordingly, TPD52 gains were found in 11.2% pT2 tumors, in 27.7% pT3 tumors and in 28.6% pT4 tumors. Gene amplifications, defined as TPD52 to CEP8 signal ratio ⱖ 2.0, were found in 1.8% of cases. However, high-level amplifications, defined as a TPD52 copy number of at least 10 and a TPD52 to CEP8 signal ratio of ⱖ2.0, were only found in 2 cases. CONCLUSIONS: These data suggest a strong impact of TPD52 gene copy number on tumor phenotype and clinical outcome. These data are especially interesting in the light of recent reports suggesting TPD52 protein as a target for vaccination in tumors. Source of Funding: This project was supported by the German Federal Ministry of Education and Science in the framework of the program for medical genome research (FKZ:01GS08189).
1427 BCAR1 VARIANTS IN HEREDITARY PROSTATE CANCER FAMILIES WITH LINKAGE TO 16Q23 Emilie Johnson*, Anna Ray, Kimberly Zuhlke, Kathleen Cooney, Ann Arbor, MI INTRODUCTION AND OBJECTIVES: In a genome-wide linkage scan of multiplex prostate cancer (PCa) families from the University of Michigan Prostate Cancer Genetics Project (PCGP), linkage was observed to markers on chromosome 16q23. This region has previously been implicated in hereditary PCa and has been shown to be deleted in approximately 50% of PCa cases suggesting a tumor suppressor mechanism. We set out to analyze BCAR1, which maps to 16q23 and has been implicated in both breast cancer and PCa cancer progression, to assess its potential role in hereditary PCa. METHODS: PCGP families displaying linkage (non-parametric linkage score ⬎ 0) to chromosome 16q23 markers were identified. All BCAR1 exons and intron/exon boundaries were directly sequenced using DNA from the youngest available individual with PCa from each of 96 linked families. We also abstracted relevant clinical data for each of the individuals who underwent sequencing of BCAR1. RESULTS: Of the 96 individuals sequenced, the majority (75.8%) had localized PCa, and median age at diagnosis was 56 years. A total of 38 SNPs were identified, including 10 missense variants. Three of the individuals sequenced were wild type at all 38 single nucleotide polymorphisms (SNPs), and 62 individuals (65%) had at least one nonsynonomous SNP. No truncating mutations were identified in BCAR1. Of particular interest was S76P, which demonstrated a minor allele frequency (MAF) of 0.34 in our study population, which is significantly greater than the previously published MAF of 0.13.
Vol. 183, No. 4, Supplement, Tuesday, June 1, 2010
CONCLUSIONS: Although there were no truncating mutations identified in BCAR1 within our population of hereditary PCa patients, multiple missense variants were identified. The most common nonsynonomous SNP identified was S76P, which had a MAF 2.65 times that of a previously published population. BCAR1 variations likely do not completely explain the 16q23 linkage peak seen in our population. However, BCAR1 is an interesting target for further genotyping and functional studies in both sporadic PCa and hereditary PCa populations. Source of Funding: NIH RO1 CA79596
1428 SIGNIFICANCE OF CCR9-CCL25 AXIS IN PROSTATE CANCER METASTASIS. Praveen Sharma, Louisville, KY; Rajesh Singh, Atlanta, GA; Kristian Novakovic, Louisville, KY; William Grizzle, Birmingham, AL; Shailesh Singh*, Louisville, KY INTRODUCTION AND OBJECTIVES: Prostate cancer (PCa) remains the second leading cause of cancer-related deaths among men in the US. Bone metastasis is major cause of PCa associated deaths and handful treatment alternatives are available to treat this condition. Therefore, better understating of metastatic process and identification of molecular signature on metastatic cells are urgently needed to develop therapeutic for metastatic PCa. We have shown that CCR9, a CC chemokine receptor is highly expressed by the PCa tissue and cells and play an important role in PCa cell migration and invasion in vitro under the chemotactic gradient of CCL25, which is the only natural ligand for CCR9. CCL25 is also known as a thymus-expressed chemokine (TECK) and primarily expressed by the thymus, which involutes with age. Interestingly, we have observed higher serum CCL25 in PCa patients compared to normal healthy donors. Therefore, major goal of this study was to identify the source of CCL25 other than thymus and determine the role of CCR9-CCL25 axis in PCa metastasis. METHODS: Immunohistochemisrty was performed to determine the expression of CCL25 by the primary prostate tumor. Bone marrow cells from mice with or without xenograft tumor was isolated and analyzed by flow cytometry for CCL25 expression. Mice were challenged with Luciferase expressing PC3 cell by intracardiac injection or directly into the tibia of nude mice. After tumor challenge mice were treated with CCL25 neutralization antibodies every second day. Xenogen/Caliper imaging system was used to monitor metastasis and tumor growth/regression non-invasively. RESULTS: For the first time, we show secretion of CCL25 within the primary prostate tumor in a paracrine manner. Interestingly, CCL25 was highly produced by the bone marrow cells isolated from tumor bearing mice as compared to non-tumor bearing mice. Furthermore, inhibition of CCR9-CCL25 interaction in vivo using CCL25 neutralization antibodies, showed significant inhibition of bone metastasis after intra-cardiac injection of PC3 cells and tumor establishment after intra-tibial injection. CONCLUSIONS: These results provide clinical and biological significance of CCR9-CCL25 axis in prostate cancer progression. Higher production of CCL25 by the bone marrow cells of the tumor bearing mouse confirms that CCL25 produced the bone marrow stromal cells are induced by the factors produced, by the tumor cell to facilitate metastasis to the bone. Therefore, developing CCR9 directed therapies, may improve the therapeutic outcome of prostate cancer either as a single agent or in combination with other therapeutic modalities. Source of Funding: Partially supported by DOD-PCRP-NIA (PC051123- W81XWH-06-1-0521)