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Purpose Mismatch repair (MMR) deficiency and microsatellite instability (MSI) are associated with cisplatin resistance in human germ cell tumors (GCTs). BRAF mutation (V600E) is found in MSI colorectal cancers. The role of RAS/RAF pathway mutations in GCT treatment response is unknown. Patients and Methods Two patient cohorts were investigated: 100 control GCTs (50 seminomas and 50 nonseminomas) and 35 cisplatin-based chemotherapy-resistant GCTs. MMR proteins were analyzed by immunohistochemistry, and eight microsatellite loci were examined for MSI. Tumors were assessed for specific BRAF and KRAS mutations. Results Resistant tumors showed a higher incidence of MSI than controls: 26% versus 0% in two or more loci (P < .0001). All resistant tumors were wild-type KRAS, and two controls (2%) contained a KRAS mutation. There was a significantly higher incidence of BRAF V600E mutation in resistant tumors compared with controls: 26% versus 1% (P < .0001). BRAF mutations were highly correlated with MSI (P = .006), and MSI and mutated BRAF were correlated with weak or absent staining for hMLH1 (P = .017 and P = .008). Low or absent staining of hMLH1 was correlated with promoter hypermethylation (P < .001). Tumors lacking expression of hMLH1 or MSH6 were significantly more frequent in resistant GCTs than in controls (P = .001 and 0.0036, respectively). Within the subgroup of resistant tumors, patients with MSI showed a trend to longer progression-free survival (P = .068). Conclusion We report for the first time a correlation between a gene mutation—BRAF V600E—and cisplatin resistance in nonseminomatous GCTs. Furthermore, a correlation between MMR deficiency, MSI, and treatment failure is confirmed.

9547 Background: Elderly cancer patients (pts) are a heterogenous population. A CGA helps to describe the heterogeneity in a structured way, and can help to detect changes occurring during cancer t...

9553 Background: Elderly cancer patients (pts) are a heterogeneous population. As chronological age does not provide sufficient information on individual limitations and resources, a CGA helps to describe the heterogeneity in a structured way. Assessment instruments might be sensitive to gender differences. We therefore analyzed the IN-GHO registry for gender associated differences in results of CGA. Methods: The internet based IN-GHO registry prospectively collects data of elderly cancer pts from > 100 centres in Germany and Austria. Data from 1,580 pts aged 70+ years were analyzed comparing results between female (n=883) and male (n=697) pts by Chi-square or Mann-Whitney U test. Analyses included results from CGA, physicians´ rating (fit vs. compromised vs. frail) and pts´ self rating of fitness for treatment (Likert scale 1 = very fit to 6 = very unfit), type (combination vs. monotherapy vs. no), and dosage (full vs. adapted) of treatment. Results: Mean age was 76.7 years (range 70-97), 71.5% had a solid tumor, and 28.5% a hematological malignancy. Mean age was slightly but significantly higher in women than in men (76.9 vs. 76.4; p=0.02), reflected in a higher number of pts. aged 80+ (26.5% vs. 21.1%; p=0.013). No gender differences were observed in physicians´ rating and pts´ self rating of fitness for therapy, or in type or dosage of treatment. Furthermore, no gender associated differences were noted in Karnofsky-Performance-Scale, comorbidity (Charlson-Comorbidity-Score), polypharmacy, and cognition (Mini-Mental-Status-Examination). Gender associated differences, however, were noted in body mass index (more women in categories 35), mean ADL (91.7 vs. 93.4, p=0.03), mean IADL (6.9 vs. 6.7 (p=0.049), and timed-up-and-go test (< 10 seconds 36.1% vs. 43.6%, p=0.005). The main differences between women and men in ADL and IADL scores were noted in the item bathing (ADL) and food preparation, housekeeping, and laundry (IADL). Conclusions: The internet based registry is a valuable tool to gain data on prognostic factors, treatment, and outcome in old cancer pts. In several items of CGA, significant differences exist between women and men.

4559 Background: About 20-30% of patients (pts) with advanced germ-cell cancer (GCC) relapse after cisplatin-based chemotherapy. This database evaluates first salvage treatment and the prognostic categories at first relapse according to the International Prognostic Factors Study Group (= IPFSG; JCO 2010). Methods: A total of 144 pts (78% nonseminoma) with relapsed or refractory GCC undergoing 1st salvage treatment with either conventional (CD-CX) or high-dose chemotherapy with autologous stem cell support (HD-CX) from 16 German centers were retrospectively analysed. Results: Subgroups according to the IPFSG prognostic categories, were: very low risk in 9/144 (6%), low risk in 26/144 (18%), intermediate risk in 78/144 (54%), high risk in 27/144 (19%), and very high risk in 4/144 pts (3%). 1st salvage treatment consisted of HD-CX in 96 (67%) and CD-CX in 48 pts (33%). Treatment response was CR/PR- in 60%, PR+/SD in 33%, and PD in 7%. After a median follow-up (mFU) of 21 months (mos) (range, 0 – 193), 53% of all pts had relapsed and 30% had died resulting in a median progression-free survival (PFS) of 7 mos (95%CI 0-16) and overall survival (OS) of 47 mos (95%CI 21-73). At subsequent relapses, 25/48 pts (52%) received HD-CX as > 2nd-salvage treatment. For the total cohort, PFS rate after 2 years was 35%, and OS rate after 5 years was 53%. Stratification according to IPFSG prognostic categories significantly correlated with PFS (p=0.001) and OS (p=0.004) after 1st salvage treatment. Even among high-risk and very high risk (n=31, mFU 12 mos) a PFS of 37 % at 2 years and an OS of 60 % at 2 years was observed, after 1st or 2nd salvage treatment, which might be due to patient selection and short follow-up. Conclusions: IPFSG prognostic categories highly correlated with observed PFS and OS after first salvage treatment in this cohort of pts with refractory or relapsed GCC. First salvage treatment with CD- or HD-CX resulted in overall 5 year-OS rates of about 50% across all prognostic categories. Even pts with high and very high risk achieved 2 year-OS rates of about 60% with salvage HD-CX at first or subsequent relapses.

4590 Background: Efficacy of second-line treatment after cisplatinum failure in patients (pts) with advanced urothelial cancer is limited. Based on encouraging preclinical data and results from several phase I trials in solid cancers, we evaluated the safety and efficacy of the combination of paclitaxel and RAD001 (everolimus) in these pts. Methods: In this single-arm, multicenter phase II trial, pts failing prior cisplatinum based combination treatment for advanced urothelial cancer were treated with paclitaxel (175 mg/m² i.v., three-weekly) and the mTOR-inhibitor RAD001 (10 mg p.o., once daily). Pts were treated until tumor progression by RECIST criteria or until a maximum of 6 cycles was completed. A one-stage design was used to evaluate the overall response rate (ORR) as primary endpoint. Results: 27 pts (18 men, 9 women; median age 63 (35-76) years; ECOG ≥ 1: 11/27pts, hepatic metastases: 10 pts, Hb

4597 Background: Bone metastases (mets) are rare events in GCT patients (pts) and data on characteristics, risk factors, treatment and outcome in these pts are largely missing. Methods: A subgroup of 104 ptswith bone metsfrom the IPFSG database was analyzed. All patients experienced unequivocal relapse/progression after > 3 cycles of cisplatin-based chemotherapy and received conventional-dose (CDCT) or high-dose salvage chemotherapy (HDCT). Results: 104 / 1594 pts presented with bone mets (6,5%) at first relapse. Histology was pure seminoma in 26%, and non-seminoma in 74%. At initial diagnosis, 54% of pts had presented with "poor risk" according to IGCCCG. Overall response rate (ORR; CR+PR) to 1st-line chemotherapy was 89%. Median PFS after primary treatment was 3 months (range 0 - 140). Bone mets at first relapse were accompanied by abdominal, lung, mediastinal, liver and brain mets in 54%, 48%, 28%, 36%, and 14%, respectively. Tumor marker profiles were heterogeneous (elevation of AFP / ß-HCG: 51% / 37%). Salvage treatment was CDCT in 34%, and HDCT in 66% of pts. ORR to salvage chemotherapy was 68% for the total cohort, and 43% vs. 81% for CDCT vs. HDCT (p

9114 Background: Older cancer patients (pts) compose a heterogeneous population. Data from clinical trials are limited, and little is known about treatment decisions and outcomes. The internet based IN-GHO registry prospectively collects data of cancer pts ≥ 70 years from >100 centers in Germany and Austria. Methods: Data from 1.563 cancer pts were analysed, comparing results between pts with a body mass index (BMI)

4585 Background: Patients (pts) with cisplatin-refractory or multiply relapsed germ cell tumors (GCT) have a poor prognosis and rarely achieve long term survival. Different combination chemotherapy...

e15029 Background: Resistance to chemotherapy is a potentially deadly and so far poorly understood event in germ cell tumors (GCT). To improve our knowledge on resistance to cisplatin (CDDP) in GCT, we compared the constitutive miRNA gene expression pattern in three CDDP resistant and corresponding paternal GCT cell lines. Methods: Three CDDP resistant cell lines were derived via long term culture under increasing concentrations of CDDP from well established GCT lines (NT2, NCCIT, 2102EP). CDDP resistance was determined using MTT assy. Total RNA was isolated from all 6 cell lines in three independent experiments. Aliquots of individual total RNA were reverse transcribed without preamplification using the so called “Megaplex pools without preamplification protocol for miRNA expression analysis protocol”. RTQ-PCR was performed using two different 384 well low density arrays covering about 800 known miRNA species of human origin. Statistical measures (e.g. coefficient of variation, CV, and 25-75% quantiles) ...

9085 Background: Elderly cancer patients (pts) are a heterogeneous population. Data from clinical trials in very old cancer pts is scarce, and little is known about treatment decisions and outcomes...

9549 Background: The internet based IN-GHO-Registry, a project of the Initiative for Geriatric Hematology and Oncology, supported by ORTHO BIOTECH, Division of Janssen-Cilag GmbH, Germany, prospectively collects data of elderly cancer pts. We report data from 1130 pts with solid tumors. Results of a CGA, physicians' and pts' judgement of fitness were studied in relation to mortality within 5–7 months. Methods: Demographic data, data on activities of daily living (ADL), instrumental activities of daily living (IADL), Karnofsky-Performance-Statuts (KPS), comorbidity (Charlson score), number of co-medication, mobility (Timed-Up&Go), cognition (MMSE), and depression (SCID screening) were collected. Age, results of CGA, physicians' rating (fit vs compromised vs frail) and pts' self rated fitness for treatment (Likert scale 1= very fit to 6 = very unfit) were analysed for association with death. Follow-up (FU) was scheduled 5–7 months after inclusion. Results: 1130 pts, mean age 76.3 years (SD 4.7; range 69–95), 44% male, were included. Main diagnoses were gastrointestinal tumors (38%), breast cancer (12%), and lung cancer (11%). Physicians rated 58% of pts fit, 37% compromised, and 5% frail. Pts' self rating of fitness: 1 = 19%, 2 = 30%, 3 = 28%, 4 = 17%, 5 = 5%, 6 = 1%. During FU (median 177 days), 272 pts (24%) died. Mean age (76.5 vs. 76.3; p=0.75) was not associated with death. However, sex (male vs female, p [Table: see text]

9551 Background: The PELICAN trial is a randomized phase III trial evaluating efficacy and safety of pegylated liposomal doxorubicin (PLD) versus capecitabine as first-line therapy in patients (pts) with metastatic breast cancer who are not considered candidates for combination chemotherapy. Randomisation was stratified according to age (cut-off 65 years) and anthracycline pretreatment. CGA results and adverse events (AEs), dose reductions, and number of applied cycles were evaluated. Methods: CGA comprised data on activities of daily living (ADL), instrumental activities of daily living (IADL), ECOG and Karnofsky performance score, comorbidity (CIRS-G), and number of comedication. According to CGA, pts were classified into groups 1–3 (Balducci 2000). Associations of CGA groups and single CGA items with outcome measures were assessed using Fisher's exact test, Wilcoxon test, and logistic regression. Results: Results of 210 pts, mean age 61 years (SD 11; range 22–85) were analysed. According to CGA, 102 pts were considered fit (group 1, 72%), 21 compromised (group 2, 15%), and 19 frail (group 3, 13%). Age was associated neither with overall AEs, nor with dose reductions or number of applied cycles. However, the following significant associations were observed: any AE and number of comedication (p=0.026); less hand foot syndrome (HFS) and CGA group 2/3 (p=0.032); and more constitutional symptomes and age >=65 as well as CGA group 2/3 (p=0.002 and 0.015, respectively). Comedication was the variable most frequently associated with single AEs (dermatology other than HFS, p=0.038; gastrointestinal, p=0.035; constitutional symptomes, p=0.002; pain, p=0.001; pulmonary, p [Table: see text]

20654 Background: The internet-based IN-GHO registry prospectively collects data of elderly cancer patients (pt) ≥70 y. In order to gain more insight into feasibility of cancer treatment, results o...

16012 Background: In patients (pts) treated for advanced germ cell tumors (GCT), radiologic procedures (RP), including computed tomography, and serum tumormarkers (TM) are performed as routine foll...

15567 Background: GCTs are the most common malignancy in young Caucasian males aged 20–40. With CDDP-based chemotherapy, highly effective treatment has been developed for the majority of patients with GCT. However, the molecular mechanisms underlying treatment response remain elusive. Therefore, we used a global proteomics approach to screen for changes induced in GCT cell lines by CDDP. Material and Methods: Global protein expression analysis of three GCT cell lines, NTERA2, 2102EP, and NCCIT, both untreated and treated with CDDP for 48 hours at the IC50, was performed using 2-dimensional gel electrophoresis. Differentially expressed proteins were analyzed using matrix-assisted laser desorption/ionization - time of flight (MALDI TOF) and Fourier transform ion cyclotron resonance (FTICR) mass spectrometry. Identification of proteins was performed using peptide mass-fingerprinting and NCBI/SwissProt and The Global Proteome Machine database search. Results: In total, 66 proteins were found to be differentially expressed comparing untreated and treated cells. Numbers of proteins showing upregulation under CDDP were 18 in NTERA2, 10 in NCCIT, and 9 in 2102EP, whereas 17 proteins were downregulated in NTERA2, 5 in NCCIT, and 7 in 2102EP. Identification of corresponding proteins was successful in 44 out of 66 spots (67%). Differential expression of 5 proteins was found in more than one cell line. One protein (Enolase 1) was consistently up-regulated in all three cell lines under CDDP. The identified proteins can be grouped into different categories: 15 are involved in DNA-binding/transcription, 8 in metabolism, 8 in protein folding/binding/chaperone, 6 in cell signaling/differentiation, 4 in apoptosis, and 3 in cytoskeleton/cell movement. Conclusion: The results of this screening approach identify factors differentially regulated during response to CDDP in GCTs, and reveal a specific response pattern for each cell line. This knowledge can help to identify therapeutic targets and factors potentially involved in treatment sensitivity and resistance. No significant financial relationships to disclose.

5084 Background: Treatment of pts with cisplatin-refractory GCT remains a therapeutic challenge. The present phase II study investigates the toxicity and activity of triple combination of gemcitabine/oxaliplatin/paclitaxel (GOP) in pts with multiple relapse or cisplatin- refractory GCT. Patients and Methods: Between 04/03 - 10/06, 41 patients (pts) refractory to cisplatin-based chemotherapy or with relapse after high-dose chemotherapy plus autologous peripheral blood stem cell transplantation (PBSCT) received 800 mg/m2gemcitabine, 80 mg/m2 paclitaxel, both on days 1+8, and oxaliplatin 130 mg/m2 on day 1 of a three week cycle for at least 2 cycles. Treatment was continued until tumor progression or a maximum of 8 cycles. Results: Pts were pretreated with a median number of 2 previous lines of platin-based chemotherapy (range, 1 - 3) and 78 % had relapsed after either first- (39%) or second-line high-dose chemotherapy (39%) with PBSCT. Responses: 5% of the pts achieved a complete response (CR), 47% a partial response and 20% a disease stabilisation. 20% of the pts underwent a secondary complete tumor resection resulting in a total NED-rate of 22%. After a median follow up time of 5 months (mos) (0 - 20+) 17% of the pts are continuously disease-free with a median response duration of 8 mos (1–17+). Median progression free and overall survival was 3 mos (1–17+) and 6 mos (1–19+), respectively. Toxicity was acceptable. 51% of 150 cycles could be applied without dose modifications or delay. Leucocytopenia grade 3/4 was observed in 15%, anemia in 7%, and thrombocytopenia in 49% of the patients. Grade 3/4 nonhematologic toxicities (diarrhoea, nausea and neurotoxicity) occurred in one patient each (2%). Conclusion: The triple combination chemotherapy with gemcitabine, oxaliplatin and paclitaxel is feasible and effective in patients with cisplatin-refractory or multiply relapsed germ cell tumors exhibiting an overall response rate of >50% and prolonged survival in about 20% of the pts. Despite a significant rate of grade 3/4 thrombocytopenia the overall toxicity profile is acceptable. No significant financial relationships to disclose.

14530 Background: Germ cell tumors (GCTs) are the most common malignancy in young Caucasian males aged 20–40. Even though highly effective treatment strategies have been developed for the majority of patients (pts) with GCT, long term survival rate in pts with primary mediastinal non-seminomas nowadays remains below 50%. The molecular mechanism underlying the increased resistance to treatment remains elusive. Therefore, a systematic approach of global proteome analysis was used as a screening tool to compare GCT cell lines of gonadal and extragonadal origin. Methods: Comparative protein expression analysis of two GCT cell lines, NTERA2 (of gonadal origin) and NCCIT (of mediastinal origin) was performed using 2-dimensional gelelectrophoresis. Differentially expressed proteins were analyzed using matrix-assisted laser desorption/ionization - time of flight mass spectrometry (MALDI-TOF). Identification of proteins was performed using peptide mass-fingerprinting and NCBI/SwissProt database search. Results: On average, 761 spots on each gel were detected. Of these, 66 proteins were found to be differentially expressed in an at least in 2-fold manner. Expression of 47 proteins was found to be higher in NCCIT than in NTERA2, 19 proteins showed lower expression in NCCIT than in NTERA2. All 66 spots were analyzed, and 35 (51%) proteins were positively identified by peptid mass fingerprinting and database search. These proteins could be grouped into different categories: 9 are involved in metabolism, 5 each in protein folding and DNA-binding/transcription, 4 in cytoskeleton/cell movement, 3 in cell signaling/differentiation, and 1 each in ion transport and membrane/vesicle trafficking. Of 7 proteins, the function is currently unknown. Conclusions: This analysis demonstrates that proteomic tools are valuable in studying similarities and differences between gonadal and primary extragonadal GCTs. Further characterization and functional analysis of selected proteins is ongoing, with special emphasis on factors potentially involved in treatment resistance and sensitivity. Furthermore, the acquired proteome map of untreated cells will help to identify cellular changes occuring under different experimental conditions in vitro like induction of differentiation and response to cytotoxic treatment. No significant financial relationships to disclose.