THE HIGH-RESOLUTION GENOMIC PROFILING USING WHOLEGENOME SINGLE NUCLEOTIDE POLYMORPHISM GENOTYPING IN CLEAR CELL RENAL CELL CARCINOMA

Vol. 179, No. 4, Supplement, Sunday, May 18, 2008

263 THE HIGH-RESOLUTION GENOMIC PROFILING USING WHOLEGENOME SINGLE NUCLEOTIDE POLYMORPHISM GENOTYPING IN CLEAR CELL RENAL CELL CARCINOMA Akira Yokomizo*, Ken Yamamoto, Yasuhiro Tada, Seiji Naito. Fukuoka, Japan. INTRODUCTION AND OBJECTIVE: A variety of chromosomal aberrations has been reported in clear cell renal cell carcinoma (ccRCC), however, these data were derived from comparative genomic K\EULGL]DWLRQ&*+ DQGORVVRIKHWHUR]\JRVLW\/2+ DQDO\VLVZKLFK is inadequate to detect the microdeletions, microduplications and copy neutral LOH for whole genome scanning. In this study, Human CNV370Duo DNAAnalysis BeadChip, which covers the 370,000 single nucleotide polymorphism (SNP) markers in median spacing of 4.9kb in human genome, was applied to detect the genomic alterations in ccRCC. METHODS: DNA isolated from 18 primary ccRCC and PDWFKHGQRUPDO'1$IURPSHULSKHUDOZKLWHEORRGFHOOVZDVDQDO\]HG using Human CNV370-Duo DNAAnalysis BeadChip. The obtained data was evaluated by the BeadStudio software provided by illumine Inc. RESULTS: The CNV370 DNA chip detected genetic aberrations in all tumors. Most common loss including the copy neutral /2+ZDVLGHQWL¿HGLQSZKLFKZDVIRXQGLQRIWKHWXPRUV2WKHU frequent changes were losses of 1p (14%), 2q (28%), 4p (22%), 4q (22%), 8p (33%), 8q (22%) and gain of 2q (11%), 5q (28%). The copy QHXWUDO/2+WKDWZDVPLVVHGLQSULRU/2+VWXG\ZDVLGHQWL¿HGLQ DUHDRIDOOFKURPRVRPHVDQDO\]HG )XUWKHUPRUHPLFURGHOHWLRQV DQG PLFURDPSOL¿FDWLRQV OHVV WKDQ 0E ZHUH GHWHFWHG LQ  UHJLRQV The candidate genes were easily accessed by the SNP tag number, and some of them were not previously reported as the RCC associated genes. However, none of them were commonly changed in these VSHFLPHQVDQDO\]HGVXJJHVWLQJWKDWWKHQRYHOJHQHVDUHOHVVH[SHFWHG to be involved commonly in ccRCC carcinogenesis. CONCLUSIONS: This technology provided not only an overview of chromosomal aberrations but also high resolution DNA changes. In this study, short arm of chromosome 3, where the VHL gene located, were most commonly deleted and other candidate tumor suppressor genes/oncogenes were not detected in this genome-wide screening. It suggests that 3p deletions are expected to be the initial event to develop the ccRCC. Source of Funding: 21 Century COE Program.

264 PROTEOMIC PROFILING IDENTIFIES KEY REGULATORY METABOLIC ENZYMES AND STRESS PROTEINS DIFFERENTIALLY EXPRESSED IN RENAL CELL CARCINOMAS Vladimir A Valera*, Beatriz A Walter, W Marston Linehan, Maria J Merino. Bethesda, MD. INTRODUCTION AND OBJECTIVE: Several genetic and epigenetic alterations have been described in renal cell carcinomas (RCC) related to tumor initiation and progression. However, particular FKDQJHV LQ WKH SURWHLQ H[SUHVVLRQ SUR¿OHV RI WKHVH WXPRUV KDYH QRW been described yet. Accurate knowledge of the repertoire of proteins associated with renal cancers will not only provide insights into these mechanisms but will provide new markers for early detection and potential therapeutic targets. METHODS: After microdissection, enriched cell populations IURPIUR]HQWLVVXHVHFWLRQVGHULYHGIURPDVSHFWUXPRILQKHULWHGDQG sporadic RCC patients, and a set of benign tumors and normal kidney ZHUH DQDO\]HG E\ GLPHQVLRQDO SRO\DFU\ODPLGH JHO HOHFWURSKRUHVLV '3$*( *HOVZHUHGLJLWDOO\VFDQQHGFRPSDUHGDQGVLJQL¿FDQW GLIIHUHQWLDOO\ H[SUHVVHG SURWHLQ VSRWV ZHUH H[FLVHG DQG DQDO\]HG E\ 0$/',72) 06 ,GHQWL¿HG SURWHLQ H[SUHVVLRQ ZDV IXUWKHU YDOLGDWHG E\ LPPXQRKLVWRFKHPLVWU\ LQ DQ LQGHSHQGHQW JURXS RI IRUPDOLQ¿[HG paraffin embedded tumor samples and by western blot analysis. Proteomic and immunohistochemical results were correlated to standard clinicopatologic variables including tumor grade and stage. RESULTS: On agerage, 900 protein spots from both RCC VDPSOHVDQGFRUUHVSRQGLQJQRUPDONLGQH\VDPSOHVZHUHYLVXDOL]HGLQ a single gel. Sixty-six protein spots were found statistically (p<0.05) up or down regulated between cancer and normal tissues, with changes ranging from 2 up to 13.5-fold. From these spots, eight different

THE JOURNAL OF UROLOGY®

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SRO\SHSWLGHVZHUHVXFFHVVIXOO\LGHQWL¿HGPDLQO\UHSUHVHQWLQJHQ]\PHV involved in metabolic pathways. Spot pattern allowed unsupervised FODVVL¿FDWLRQ DOJRULWKPV WR VXFFHVVIXOO\ FOXVWHU WXPRU VDPSOHV LQWR ZHOOGH¿QHGJURXSV3URWHLQH[SUHVVLRQYDOLGDWLRQGHPRQVWUDWHGWKDW among the identified polypeptides, Triosephosphate Isomerase 1 (TPI-1) and Heat-shock protein 27 (HSP27) were the most related to clinicopathological features such as tumor type and grade. &21&/86,2163URWHRPLFSUR¿OLQJRI5&&E\'3$*( LGHQWL¿HGDVHULHVRIGLIIHUHQWLDOO\H[SUHVVHGSURWHLQVEHWZHHQQHRSODVWLF and normal tissue. Most of these proteins represent key metabolic UHJXODWRU\HQ]\PHVDQGVWUHVVUHVSRQVHSURWHLQV3URWHLQH[SUHVVLRQ ZDVFRQVLVWHQWDPRQJFDVHVDOORZLQJFODVVL¿FDWLRQRIWXPRUVEDVHG RQ VSRW SDWWHUQ 7KH DQDO\VLV DOVR KLJKOLJKWV WKDW WKH SURWHLQ SUR¿OH may eventually prove to be useful in identifying disease biomarkers and therapeutic targets. Source of Funding: NCI.

Pediatrics: Congenital Anomalies-Lower Urinary Tract & Genitalia (I) Moderated Poster Session 10 Sunday, May 18, 2008

10:30 am - 12:30 pm

265 EARLY FEMINIZING GENITOPLASTY: SEXUAL FUNCTION IN ADULT LIFE Arianna Lesma*, Tommaso C Camerota, Carmen Maccagnano, (OHQD6WUDGD0DQXHOD7XWROR1LFROR%XI¿8PEHUWR&DSLWDQLR$OGR Bocciardi, Patrizio Rigatti, Francesco Montorsi. Milan, Italy. INTRODUCTION AND OBJECTIVE: Reductive clitoroplasty LVDQLQWHJUDOVWHSRIIHPLQL]LQJJHQLWRSODVW\IRUWKHWUHDWPHQWRIFKLOGUHQ with ambiguous genitalia. The aim of the study is to evaluate the sensitivity of external genitalia and to assess sexual function and orgasm LQDJURXSRIDGXOWSDWLHQWVVXEPLWWHGWRHDUO\IHPLQL]LQJJHQLWRSODVW\ during childhood METHODS: From 1980 to 2006, 90 consecutive patients XQGHUZHQW IHPLQL]LQJ JHQLWRSODVW\ ZLWK UHGXFWLYH FOLWRULGRSODVW\ :H included patients affected by congenital adrenal hyperplasia, who were surgically treated in their infancy, and now ageing 18 years or more. Thermal (TS) and vibratory sensibility (VS) of clitoris, vagina and labia PLQRUD ZHUH DQDO\]HG XVLQJ WKH *HQLWR6HQVRU\$QDOD\]HU *6$  Psychosexual outcome was assessed with the following validated TXHVWLRQQDLUHV %HFN¶V TXHVWLRQQDLUH =XQJ 6HOIUDWLQJ$Q[LHW\ 6FDOH (SAS), Female Sexual Function Index (FSFI), and Female Sexual Distress Scale (FSDS). RESULTS: Sixty patients (60/90=67%) were younger of 18 years old and were not eligible for the study. Only 24 of the remaining 30 patients (24/90=27%) referred complete sexual life. Fifteen of them (15/90=17%) agreed to participate the study. Masturbation and complete sexual intercourses were reported by all patients but in 4 cases (4/15=27%), who had sexual intercourse without penetration. TS and VS of clitoris and labia minora were investigated in all cases, whereas YDJLQDO 76 DQG 96 FRXOG EH DQDO\]HG RQO\ LQ     DQG  (6/15=40%) patients respectively, because of modest vaginal stenosis. At GSA evaluation TS appeared decreased both at clitoris and at the vagina, whereas VS resulted decreased at clitoris but within normal limits at the vagina in all cases. Beck’s test showed slight depression in 3 (3/15=20%) patients. SAS was positive for slight or moderate anxiety in all patients. At FSFI all patients reported active sexual desire, good arousal, adequate OXEUL¿FDWLRQDQGSUHVHQFHRIRUJDVP$W)6'6  SDWLHQWV described stable satisfactory heterosexual relationship, 3 (3/15=20%) patients reported stable unsatisfactory heterosexual relationship without penetration, 3 (3/15=20%) patients refused the test, and 1 (1/15=7%) patient described satisfactory homosexual relationship. CONCLUSIONS: Our study suggests that vaginal VS was conserved in all patients, whereas clitoral VS, vaginal TS and clitoris TS were altered. All patients reported orgasm even in absence of penetration. Decreased but conserved clitoral sensibility does not seem to have clinical relevance. Source of Funding: None