e162 THE JOURNAL OF UROLOGY� Vol. 191, No. 4S, Supplement, Saturday, May 17, 2014

Outstanding Posters: Research

Moderated Poster

Saturday, May 17, 2014 1:00 PM-3:00 PM

OP1-01BLADDER REINNERVATION USING A PRIMARILY MOTOR DONORNERVE (FEMORAL NERVE BRANCHES) IS FUNCTIONALLYSUPERIOR TO USING A PRIMARILY SENSORY DONOR NERVE(GENITOFEMORAL NERVE)

Sandra Gomez-Amaya*, Mary Barbe, Alan Braverman, Philadelphia,PA; Justin Brown, San Diego, CA; Neil Lamarre, Vicky Massicotte,Michael Ruggieri, Philadelphia, PA

INTRODUCTION AND OBJECTIVES: To determine if transferof a primarily motor somatic nerve (Femoral, F) to the anterior vesicalbranch of the pelvic nerve (PN) allows better reinnervation of thedetrusor muscle after spinal root injury, compared to a primarily sensorynerve (genitofemoral, GF). Return of function was determined usingfunctional electrical stimulation (FES) and urodynamic observations.We hypothesized that transferring a nerve with more motor axons to theanterior vesical branch of the PN (F nerve) would more effectivelyreinnervate the detrusor muscle and restore function.

METHODS: Four surgerieswereperformed in27 femalemongrelhounds: 1) genitofemoral nerve transfer with vesicostomy (GF-NT-V:n¼12); 2) genitofemoral nerve transfer without vesicostomy (GF-NT:n¼5); 3) femoral nerve transfer with vesicostomy (F-NT-V: n¼5); and 4)femoral nerve transfer without vesicostomy (F-NT: n¼5). Bladders weredecentralized by bilateral transection of all sacral roots that inducebladder contractions by intraoperative FES.GFnerves or two branches ofthe left F nerve were transferred to PN vesical branches, bilaterally.Bladder emptying in animalswithout vesicostomieswasaccomplished byCred�e maneuver during the 6 month recovery period (242�6.2days).

RESULTS: Return of bladder functionwas observed as follows: 1)in 8 of 12 GF-NT-V dogs after direct stimulation of transferred GF nerve orlumbar spinal roots (meandetrusor pressure¼ 1.9�0.6 cmH2O); 2) in4of5GF-NT dogs after direct stimulation of transferred GF nerve or FES oflumbar spinal cord segment contributing to GF nerve origin (L2-3; meandetrusor pressure ¼ 4.7�1.7 cmH2O); 3) in all 5 F-NT-V dogs after trans-cutaneous stimulation of transferred F nerve, or direct stimulation of lumbarcord segment or spinal roots contributing to F nerve origin (L2-5; meandetrusor pressure ¼ 11.2�2.5 cmH2O); and 4) in 4 of 5 F-NT dogs afterdirect stimulation of transferred F nerve or FES of lumbar cord segmentcontributing to F nerve origin (mean detrusor pressure¼ 4.8�1.3).

CONCLUSIONS: These results indicate that although thebladder can be reinnervated by nerve transfer using either the GF or Fdonor nerve, the F-NT-V was the most efficient. This surgical approachmay be useful for patients with lower motor spinal cord injury toaccomplish bladder emptying, improving their quality of life.

Source of Funding: The project described was supported byAward Number R01NS070267 from the National Institute OfNeurological Disorders And Stroke. The content is solely theresponsibility of the authors and does not necessarily representthe official views of the National Institute Of NeurologicalDisorders And Stroke or the National Institutes of Health.

OP1-02THE ASSOCIATION BETWEEN MIF-173 G>C POLYMORPHISMAND PROSTATE CANCER IN SOUTHERN CHINESE.

Ninghong Song*, Jiangsu Province, China, People’s Republic of;Huang Su, Nanjing, China, People’s Republic of

INTRODUCTION AND OBJECTIVES: Accumulating epidemi-ological and molecular evidence suggests that inflammation is an

important component in the etiology of PCa. Macrophage migrationinhibitory factor (MIF) plays an important role in the pro- and anti-in-flammatory response to infection. This study is aimed at investigatingthe potential association between MIF-173 G>C polymorphism,Gleason score, clinical stage, and prostate-specific antigen (PSA)value with respect to PCa incidence among the Han nationality inSouthern China.

METHODS: Genotyping was performed by using tetraprimerpolymerase chain reaction (PCR) on 259 PCa patients and 301 cancer-free controls.

RESULTS: We found that the MIF-173*C variant allele wassignificantly associated with an increased risk of PCa [adjusted oddratio (OR) ¼ 2.99, 95% confident interval (CI): 1.94-4.60] and higherGleason scores from the PCa subjects (adjusted OR ¼ 10.72, 95% CI:5.35-21.49). In addition, we noted that the MIF -173*C variant allele wasrelated to higher clinical stages and PSA values in PCa patients(adjusted OR ¼ 15.68, 95% CI: 7.40-33.23; adjusted OR ¼ 4.37, 95%CI: 2.41-7.92, respectively).

CONCLUSIONS: Our data suggest that MIF-173 poly-morphisms may be associated with a higher incidence of prostatecancer compared to controls, and appears to be associated with higherGleason scores, higher clinical stages, and PSA values in those withprostate cancer.

Source of Funding: none

OP1-03INTRAVESICAL BACILLUS CALMETTE-GUERIN (BCG)EFFICIENTLY TACKLES S6K1 BUT NOT 4E-BP1PHOSPHORYLATION IN NON-MUSCLE INVASIVE BLADDERCANCER

Karen Ferrari, Juliana A. de Camargo, Guilherme Z. Rocha,Jos�e B. C. Carvalheira, Athanase Billis, M�ario J. A. Saad,Leonardo O. Reis*, Campinas, Brazil

INTRODUCTION AND OBJECTIVES: Bacillus Calmette-Gu�erin (BCG) is the gold standard treatment of non-muscle invasivebladder cancer (NMIBC). We aimed to characterize the functionalconsequences of intravesical BCG on the molecular mechanism ofthe PI3K/AKT/mTOR signaling pathway in NMIBC, never reportedto date.

METHODS: Female (Fischer 344) rats, 6 weeks old, received1.5 mg/kg MNU (N-methyl-n-nitrosourea) intravesically every otherweek for 6 weeks and were randomly divided into 2 groups: - MNUtumor group (n¼10) received 0.3 ml saline and - BCG treatment group(n¼10) received 106 cfu of BCG intravesically for 6 weeks. At week 15all bladders were collected for histopathology, immunohistochemistry,proliferation index (Ki-67), and immunoblotting (pAKT, pAMPK, pp53,pp70S6K, RICTOR, RAPTOR, mTOR, p4EBP1). Student’s t-test wasused to compare groups. Two tailed p<0.05 was considered statisticallysignificant.

RESULTS: Papillary carcinoma (pTa) and high-grade intra-epithelial neoplasia (carcinoma in situ) predominated in the MNU groupwhile normal urothelium, papillary and flat hyperplasia were morecommon in BCG treatment. NMIBC treated with BCG exhibited sup-pression of S6K1 (Figure 1 B) but not 4E-BP1 phosphorylation (Figure 1A), suggesting that 4E-BP1 is regulated differently that S6K1, escapingthe BCG action. The cellular proliferation index decreased in the BCGtreatment compared to MNU (Figure 1 C). Table 1 shows themammalian target of rapamycin pathway phosphorylation status ofBCG treatment compared to MNU.

CONCLUSIONS: 4E-BP1 might be a worthwhile new target forBCG refractory NMIBC. In the future, activation status of S6K1 and4E-BP1 might stratify patients that could benefit from targeting suchmolecular elements. This is the first study to explore the mammaliantarget of rapamycin pathway in the intravesical BCG treatmentof NMIBC.

Vol. 191, No. 4S, Supplement, Saturday, May 17, 2014 THE JOURNAL OF UROLOGY� e163

Table 1: Mammalian target of rapamycin pathway phosphorylation status

of BCG treatment compared to MNU

Immunoblotting

BCG/MNU (%) p values

mTOR

66.84 0.5299

pAKT

56.74 0.3278

pAMPK

121.33 0.6399

pp53

73.40 0.5166

RICTOR

80.28 0.6448

RAPTOR

88.62 0.8867

p4EBP1

140.16 0.5069

pp70S6K

17.08 0.0034

Source of Funding: Premio CAPES de Teses e Medicina III2012 (Leonardo Oliveira Reis) e Coordenaç~ao deAperfeiçoamento de Pessoal de Nível Superior.

OP1-04IDENTIFICATION AND VALIDATION OF PROTEIN BIOMARKERSOF RESPONSE TO NEOADJUVANT CHEMOTHERAPY IN MUSCLEINVASIVE UROTHELIAL CARCINOMA

Alexander Baras*, Nilay Gandhi, Enrico Munari, Sheila Faraj,Mohammad Hoque, Mark Schoenberg, Trinity Bivalacqua,George Netto, Baltimore, MD

INTRODUCTION AND OBJECTIVES: Neoadjuvant chemo-therapy (NAC) results in 5-10% increase in 5-year survival rate inmuscle invasive bladder cancer (MIBC) patients. More importantly,those who achieve a complete response (ypT0) have a 5 year survivalrate of 80% as opposed to 40% for those who do not. However, NAC isnot widely utilized due to concerns related to delay of cystectomy,potential side-effects, and inability to predict effectiveness. Recentlysuggested molecular signatures of chemoresponsiveness, whichcould prove useful in this setting, are yet to be translated into clin-ical practice.

METHODS: mRNA expression data from a prior report on aNAC-treated MIBC cohort (Kato et. al) were analyzed in combinationwith the antibody database of the Human Protein Atlas (HPA). Weidentified classifier candidate biomarkers that can be detected byimmunohistochemistry (IHC) in TURB biopsy material prior to NAC. Thecandidate biomarkers were subsequently validated in an independentcohort of 52 MIBC specimens. Response to NAC was defined as thelack of residual MIBC at cystectomy.

RESULTS: By filtering based on large amplitude mRNAchanges and the area under the receiver operator curve (AUROC), weidentified 21 genes whose mRNA expression profiles differentiateresponse to NAC (n¼33), Figure 1A. Using the HPA, we found thatcommercially available antibiodies to the protein products of 8 of thesegenes exhibited differential staining across a set of urothelial carci-nomas cataloged in the HPA. In addition, 12 normal tissues wereidentified from the HPA that can serve as positive and negative controlsfor the 8 proteins. Our initial studies with 2 of these markers in amultivariate logistic regression model have yielded strong performance(80% accuracy) in identifying patients in our independent validationcohort that are not likely to benefit from NAC due to chemoresistance(Fischer’s exact test p-value 0.02), Figure 1B & 1C.

CONCLUSIONS: We illustrate the feasibility of translating geneexpression signature data on NAC response into an IHC classifierapplicable to TURB specimens. The performance achieved by the initialtwo markers in our study is being further characterized and the otherputative protein markers are being assessed in our TMA cohort.

Source of Funding: none.

OP1-05INTEGRATIVE DNA METHYLATION AND GENE EXPRESSIONANALYSES IDENTIFIES DISCOIDIN DOMAIN RECEPTOR 1 (DDR1)ASSOCIATION WITH IDIOPATHIC NONOBSTRUCTIVEAZOOSPERMIA

Ranjith Ramasamy*, Alex Ridgeway, Josephine Addai, Larry Lipshultz,Dolores Lamb, Houston, TX

INTRODUCTION AND OBJECTIVES: Spermatogenesis is acomplex process that involves proliferation, differentiation, and celladhesion. Spermatogenic failure or non�obstructive azoospermia(NOA) results from mechanisms involved are incompletely understood.DDR1 is a member of a small subfamily of receptor tyrosine kinasesthat is involved in adhesion, migration, proliferation, apoptosis, cellmorphogenesis and differentiation. Since, DDR1 is expressed in humanpost�meiotic germ cells of testis, we hypothesized that abnormal DDR1expression could be a possible mechanism that can compromisespermatogenesis in a subset of men with idiopathic NOA.

METHODS: We used the high resolution Infinium 450Kmethylation array and compared fibrobalsts cultured from testicular bi-opsies of 19 NOA men and 4 fertile controls. Microarray data wasanalyzed using Minfi (R software package) utilizing subset�quantilewithin array normalization. We investigated the functional role of