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MP90-05 PROSTATE CANCER GENE EXPRESSION SIGNATURES ASSOCIATED WITH CAPSULE AND SEMINAL VESICLE INVASION AND BIOCHEMICAL RECURRENCE AFTER RADICAL PROSTATECTOMY
THE JOURNAL OF UROLOGYâ
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MP90-04 A ROLE FOR HISTONE DEMETHYALASE JMJD1A IN THE DNA DAMAGE RESPONSE OF PROSTATE CANCER CELLS Lingling Fan, Guihong Peng, Baltimore, MD; Natasha Sahgal, London, United Kingdom; Ladan Fazli, Vancouver, Canada; Martin Gleave, vancouver, Canada; Yuji Zhang, Baltimroe, MD; Arif Hussain, Feyruz Rassool, jianfei qi*, Baltimore, MD INTRODUCTION AND OBJECTIVES: The major therapies for prostate cancer (PCa) such as radio-, chemo- or androgen deprivation therapy induce DNA damage. Thus the DNA damage response (DDR) and expression of DDR genes are key factors in determining outcome of PCa therapies. Histone demethylase JMJD1A regulates the gene expression by demethylating H3K9. Our previous work shows that JMJD1A is essential for the long-term proliferation and survival of PCa cells in vitro or in the xenograft prostate tumor model, partly via its effect on the activities of androgen receptor (AR) and c-Myc transcription factors. Here, we find a new role for JMJD1A in the expression of DDR genes and sensitivity of PCa cells to irradiation and androgen deprivation. METHODS: Real-time PCR was used to measure the mRNA levels of some DDR genes in the PCa cells (control or siRNA knockdown (KD) of JMJD1A, AR or c-Myc). Chromatin immunoprecipitation (ChIP) was used to evaluate the binding of JMJD1A to the AR or MYC binding sites on these DDR genes. JMJD1A-KD PCa cells were treated with or without ionizing radiation (IR), and analyzed by the r-H2AX staining (a widely used marker for the DNA double strand breaks (DSBs)), western blot analysis of DNA damage signaling proteins, flow cytometry analysis of cell cycle, and GFP reporter assays for the DSB repair via the homologous recombination (HR) or non-homologous end joining (NHEJ) pathways. The growth of these PCa cells was also measured under the IR and androgen deprivation conditions. RESULTS: JMJD1A was found to promote the expression of some DDR genes by co-activating AR and/or c-Myc. The DDR genes regulated by JMJD1A encode factors mediating the DSB repair by both HR and NHEJ pathways. JMJD1A KD led to increased r-H2AX foci in the non-irradiated cells, and the delayed clearance of r-H2AX foci in the irradiated cells. JMJD1A KD also increased the levels of phospho-DNAPK or phospho-CHEK2 in the western blot analysis and caused G2 cell cycle arrest. Consistently, JMJD1A KD reduced the percentage of GFPpositive cells that reflects the DSB repair by both HR and NHEJ pathways. JMJD1A-KD cells showed a reduced growth under IR and androgen deprivation conditions. CONCLUSIONS: JMJD1A promotes the DSB repair via regulation of DDR gene expression through AR and/or c-Myc. The role of JMJD1A in the DSB repair may contribute to the resistance of PCa cells to the radio- and androgen deprivation therapies. Source of Funding: NCI R00 grant CA154888
MP90-05 PROSTATE CANCER GENE EXPRESSION SIGNATURES ASSOCIATED WITH CAPSULE AND SEMINAL VESICLE INVASION AND BIOCHEMICAL RECURRENCE AFTER RADICAL PROSTATECTOMY Travis Allemang*, Norfolk, VA; Michael Williams, Virginia Beach, VA; Shilpa Katta, Gyorgy Petrovics, Rockville, MD; Inger Rosner, Rockville, MD; Albert Dobi, Rockville, MD INTRODUCTION AND OBJECTIVES: Major strides in oncologic treatment and prognostication have occurred by insights into cancer gene expression signatures. The goal of this study was to identify prostate cancer gene expression signatures associated with extra capsular extension, seminal vesicle invasion, and biochemical recurrence (BCR). METHODS: Gene expression microarray data from 40 Caucasian men who underwent radical prostatectomy was analyzed from the publicly available NCBI Gene Expression Omnibus GSE32448
Vol. 195, No. 4S, Supplement, Tuesday, May 10, 2016
developed by the CPDR. Groups analyzed were based on final pathologic stage pT3a and pT3b disease, as well as those with negative surgical margins which developed BCR following radical prostatectomy. Gene expression signatures for each group were identified using bioinformatics analysis. Signatures were then applied to GenomatixÓ software for defining key regulatory nodes. Further comparison was made to the gene panels utilized in the commercially available ProlarisTM, Oncotype DX Genomic Prostate ScoreTM, and DecipherTM. RESULTS: There was a median follow-up of 15.5 years post prostatectomy. Those with pT3a disease were found to have 182 unique gene signatures of which 83 of the genes were found to have biological functions to include cell migration, locomotion or regulation of epithelial cell proliferation (p <0.0001). Those with pT3b disease were found to have 1569 unique gene signatures compared to subjects with pT3a disease. These genes were found to be associated with cellular functions to include protein targeting and localization, RNA processing and cell cycle (p<0.001). Genes unique to the development of BCR include down regulation of VEGF-A and the HLA gene family as key regulatory nodes. The genes unique to the absence of BCR include upregulation of ERG and downregulation of ETS. Compared to the commercially available gene panels, subjects with pT3b disease shared 13 unique genes with these gene panels. Among those with BCR only 1 gene was uniquely shared with the commercially available gene panels. CONCLUSIONS: These results suggest that unique gene expression signatures exist at the time of prostatectomy which are associated with pT3a, pT3b disease, and BCR. Among the commercially available gene panels, these products were found to have genes which correlated predominantly with advanced pathologic disease and had little correlation with BCR among this cohort. Future work is warranted to validate the expression of these unique genes as they could serve to compliment current prognostic approaches. Source of Funding: None
MP90-06 FULL EXOME SEQUENCING, COPY NUMBER VARIATION AND TRANSCRIPTOME ANALYSES REVEALS DIVERSITY AND MUTATIONAL EVOLUTION IN A LONGITUDINAL SERIES OF SURGICAL PROSTATE CANCER BONE METASTASES FROM A PATIENT WITH PROGRESSIVELY THERAPY-RESISTANT CANCER. Christina Jamieson*, Michelle Muldong, La Jolla, CA; Michael A. Liss, San Antonio, TX; Seung Chol Park, Iksan, Korea, Republic of; Theresa Mendoza, Abigail Gallegos, Lee Edsall, Unwanaobong Nseyo, Olga Miakicheva, Brittany Burton, Danielle Burner, Amy Strasner, Christopher J. Kane, Anna A. Kulidjian, Theresa Gaasterland, La Jolla, CA INTRODUCTION AND OBJECTIVES: Prostate cancer metastasizes to bone in 50-90% of patients with advanced disease yet relatively little is known about genome-wide genetic alterations in prostate cancer bone metastases. Recent whole genome, exome and transcriptome sequencing studies on prostate cancer have identified recurrent mutations and gene rearrangements such as the TMPRSS2ERG fusion and variants of the androgen receptor (AR). Here we determined bone metastatic prostate cancer genomic and transcriptomic variation in a longitudinal series of surgical bone metastasis samples from the same patient. METHODS: We collected surgical prostate cancer bone metastases samples at the time of orthopaedic repair surgery with matched peripheral blood and prostatectomy tissue and established a new boneniche xenograft model with a primary patient surgical prostate cancer bone metastasis, PCSD1 (Prostate Cancer San Diego 1). Next generation whole exome sequencing was performed on genomic DNA extracted from flash frozen samples or single cell suspensions of surgical bone metastasis specimens, peripheral blood mononuclear cells and from primary FFPE archived prostatectomy tissue. DNA libraries were prepared using the NimbleGen DNA Sample Preparation kit and sequencing was performed on the Illumina HiSeq 2000 sequencing
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MP90-04 A ROLE FOR HISTONE DEMETHYALASE JMJD1A IN THE DNA DAMAGE RESPONSE OF PROSTATE CANCER CELLS Lingling Fan, Guihong Peng, Baltimore, MD; Natasha Sahgal, London, United Kingdom; Ladan Fazli, Vancouver, Canada; Martin Gleave, vancouver, Canada; Yuji Zhang, Baltimroe, MD; Arif Hussain, Feyruz Rassool, jianfei qi*, Baltimore, MD INTRODUCTION AND OBJECTIVES: The major therapies for prostate cancer (PCa) such as radio-, chemo- or androgen deprivation therapy induce DNA damage. Thus the DNA damage response (DDR) and expression of DDR genes are key factors in determining outcome of PCa therapies. Histone demethylase JMJD1A regulates the gene expression by demethylating H3K9. Our previous work shows that JMJD1A is essential for the long-term proliferation and survival of PCa cells in vitro or in the xenograft prostate tumor model, partly via its effect on the activities of androgen receptor (AR) and c-Myc transcription factors. Here, we find a new role for JMJD1A in the expression of DDR genes and sensitivity of PCa cells to irradiation and androgen deprivation. METHODS: Real-time PCR was used to measure the mRNA levels of some DDR genes in the PCa cells (control or siRNA knockdown (KD) of JMJD1A, AR or c-Myc). Chromatin immunoprecipitation (ChIP) was used to evaluate the binding of JMJD1A to the AR or MYC binding sites on these DDR genes. JMJD1A-KD PCa cells were treated with or without ionizing radiation (IR), and analyzed by the r-H2AX staining (a widely used marker for the DNA double strand breaks (DSBs)), western blot analysis of DNA damage signaling proteins, flow cytometry analysis of cell cycle, and GFP reporter assays for the DSB repair via the homologous recombination (HR) or non-homologous end joining (NHEJ) pathways. The growth of these PCa cells was also measured under the IR and androgen deprivation conditions. RESULTS: JMJD1A was found to promote the expression of some DDR genes by co-activating AR and/or c-Myc. The DDR genes regulated by JMJD1A encode factors mediating the DSB repair by both HR and NHEJ pathways. JMJD1A KD led to increased r-H2AX foci in the non-irradiated cells, and the delayed clearance of r-H2AX foci in the irradiated cells. JMJD1A KD also increased the levels of phospho-DNAPK or phospho-CHEK2 in the western blot analysis and caused G2 cell cycle arrest. Consistently, JMJD1A KD reduced the percentage of GFPpositive cells that reflects the DSB repair by both HR and NHEJ pathways. JMJD1A-KD cells showed a reduced growth under IR and androgen deprivation conditions. CONCLUSIONS: JMJD1A promotes the DSB repair via regulation of DDR gene expression through AR and/or c-Myc. The role of JMJD1A in the DSB repair may contribute to the resistance of PCa cells to the radio- and androgen deprivation therapies. Source of Funding: NCI R00 grant CA154888
MP90-05 PROSTATE CANCER GENE EXPRESSION SIGNATURES ASSOCIATED WITH CAPSULE AND SEMINAL VESICLE INVASION AND BIOCHEMICAL RECURRENCE AFTER RADICAL PROSTATECTOMY Travis Allemang*, Norfolk, VA; Michael Williams, Virginia Beach, VA; Shilpa Katta, Gyorgy Petrovics, Rockville, MD; Inger Rosner, Rockville, MD; Albert Dobi, Rockville, MD INTRODUCTION AND OBJECTIVES: Major strides in oncologic treatment and prognostication have occurred by insights into cancer gene expression signatures. The goal of this study was to identify prostate cancer gene expression signatures associated with extra capsular extension, seminal vesicle invasion, and biochemical recurrence (BCR). METHODS: Gene expression microarray data from 40 Caucasian men who underwent radical prostatectomy was analyzed from the publicly available NCBI Gene Expression Omnibus GSE32448
Vol. 195, No. 4S, Supplement, Tuesday, May 10, 2016
developed by the CPDR. Groups analyzed were based on final pathologic stage pT3a and pT3b disease, as well as those with negative surgical margins which developed BCR following radical prostatectomy. Gene expression signatures for each group were identified using bioinformatics analysis. Signatures were then applied to GenomatixÓ software for defining key regulatory nodes. Further comparison was made to the gene panels utilized in the commercially available ProlarisTM, Oncotype DX Genomic Prostate ScoreTM, and DecipherTM. RESULTS: There was a median follow-up of 15.5 years post prostatectomy. Those with pT3a disease were found to have 182 unique gene signatures of which 83 of the genes were found to have biological functions to include cell migration, locomotion or regulation of epithelial cell proliferation (p <0.0001). Those with pT3b disease were found to have 1569 unique gene signatures compared to subjects with pT3a disease. These genes were found to be associated with cellular functions to include protein targeting and localization, RNA processing and cell cycle (p<0.001). Genes unique to the development of BCR include down regulation of VEGF-A and the HLA gene family as key regulatory nodes. The genes unique to the absence of BCR include upregulation of ERG and downregulation of ETS. Compared to the commercially available gene panels, subjects with pT3b disease shared 13 unique genes with these gene panels. Among those with BCR only 1 gene was uniquely shared with the commercially available gene panels. CONCLUSIONS: These results suggest that unique gene expression signatures exist at the time of prostatectomy which are associated with pT3a, pT3b disease, and BCR. Among the commercially available gene panels, these products were found to have genes which correlated predominantly with advanced pathologic disease and had little correlation with BCR among this cohort. Future work is warranted to validate the expression of these unique genes as they could serve to compliment current prognostic approaches. Source of Funding: None
MP90-06 FULL EXOME SEQUENCING, COPY NUMBER VARIATION AND TRANSCRIPTOME ANALYSES REVEALS DIVERSITY AND MUTATIONAL EVOLUTION IN A LONGITUDINAL SERIES OF SURGICAL PROSTATE CANCER BONE METASTASES FROM A PATIENT WITH PROGRESSIVELY THERAPY-RESISTANT CANCER. Christina Jamieson*, Michelle Muldong, La Jolla, CA; Michael A. Liss, San Antonio, TX; Seung Chol Park, Iksan, Korea, Republic of; Theresa Mendoza, Abigail Gallegos, Lee Edsall, Unwanaobong Nseyo, Olga Miakicheva, Brittany Burton, Danielle Burner, Amy Strasner, Christopher J. Kane, Anna A. Kulidjian, Theresa Gaasterland, La Jolla, CA INTRODUCTION AND OBJECTIVES: Prostate cancer metastasizes to bone in 50-90% of patients with advanced disease yet relatively little is known about genome-wide genetic alterations in prostate cancer bone metastases. Recent whole genome, exome and transcriptome sequencing studies on prostate cancer have identified recurrent mutations and gene rearrangements such as the TMPRSS2ERG fusion and variants of the androgen receptor (AR). Here we determined bone metastatic prostate cancer genomic and transcriptomic variation in a longitudinal series of surgical bone metastasis samples from the same patient. METHODS: We collected surgical prostate cancer bone metastases samples at the time of orthopaedic repair surgery with matched peripheral blood and prostatectomy tissue and established a new boneniche xenograft model with a primary patient surgical prostate cancer bone metastasis, PCSD1 (Prostate Cancer San Diego 1). Next generation whole exome sequencing was performed on genomic DNA extracted from flash frozen samples or single cell suspensions of surgical bone metastasis specimens, peripheral blood mononuclear cells and from primary FFPE archived prostatectomy tissue. DNA libraries were prepared using the NimbleGen DNA Sample Preparation kit and sequencing was performed on the Illumina HiSeq 2000 sequencing