Traditional Prognostic Factors Research Articles

Análise do gene PTEN por hibridização in situ fluorescente no carcinoma de células renais

To evaluate the frequency of deletion of the PTEN gene in renal cell carcinoma (RCC) and its impact on the rates of overall and disease-free survival. We analyzed 110 patients with renal cell carcinoma who underwent radical or partial nephrectomy between 1980 and 2007. In 53 cases it was possible to analyse the PTEN gene by the method of fluorescent in situ hybridization using the technique of tissue microarray. For statistical analysis, patients were classified in two groups according to the presence or absence of the deletion. The mean follow-up time was 41.9 months. Hemizygous deletion was detected in 18 patients (33.9%), while the homozygous one was present in three (5.6%). Deletion was present in approximately 40% of the analyzed cases. Monosomy and trisomy were detected in nine (17%) and two patients (3.8%), respectively. In 21 patients (39.6%) the analysis of the PTEN gene by in situ hybridization was normal. There were no statistically significant differences in overall (p = 0.468) and disease-free (p = 0.344) survival rates between patients with or without deletion. Factors which were independent for overall survival: TNM clinical stage, symptoms at diagnosis, high Fuhrmann grade, performance status (ECoG) and tumor recurrence. Disease-free survival was influenced only by the clinical TNM stage. Deletion of the PTEN gene in RCC was detected with a frequency of approximately 40% and its presence was not determinant of lower survival rates, the traditional prognostic factors remaining as determinants of outcome.

Classification Of Acute Myelogenous Leukemia (AML) Based On Functionally Related Proteins Groups

BackgroundIn an era where the recognized heterogeneity of the pathophysiology of AML is increasing rapidly due to sequencing and other “omics” platforms, and where the number of available targeted therapeutics is also rapidly expanding, a means to individually match therapies to AML on a specific basis is needed. New targeted therapies modulate single pathways, so recognizing when a particular pathway is active is crucial, but mutations are rare within most of these pathways in AML. Furthermore pathway activation can arise by various means. Finally the interaction of multiple different activated pathways on the overall phenotype of a leukemic blast adds complexity. MethodsTo define the role of the pathway activation in AML we generated a custom made reverse phase protein array (RPPA) onto which were printed leukemia enriched (ficolled, CD3/CD19 depleted) cells from 511 newly diagnosed AML and 21 APL patients, including 49 with paired diagnosis and relapse samples. Both bone marrow (n=387) and peripheral blood (n=283) samples were used, with 140 cases having both. The RPPA was probed with 231 strictly validated antibodies (174 antibodies vs. total expression, 49 vs. phosphoproteins, 5 vs. cleaved forms, 3 vs. methylation sites). These proteins were divided on the basis of function into 21 Protein function groups (ProFnG) including: Apoptosis, Autophagy, Cell cycle, Creb, Cytoskeletal, Differentiation, Fli1, Hippo, Histone modifying, Hypoxia, Integrin & adhesion, MEK, P53, PI3K-AKT, PKC, SMAD, STAT, Transcription Ubiquitin and Wnt. ResultsEach ProFnGrp was analyzed to determine the optimal number of principal components (PrCp) and to identify noncontributory members, which were then discarded. Each PrCp (79 total) was then considered a variable and Hierarchical clustering was performed on continuous and binary signals (Sokal-Michener metric) (Figure 1). This defined 8 PrCp clusters and 8 patient groups(PatGp). Several categorical clinical features were unevenly associated with the PatGp including FAB (p=1.5e-12), WHO class (p=0.01), gender (p=0.03), Zubrod PS (p=0.005), antecedent hematological disorder (p=0.007), Cytogenetics (0.01), FLT3-ITD (p=0.003), Ras mutation (p=0.003). Likewise many continuous variables also correlated with PatGp: WBC (p=7e-14), %BM blast (p=2x10-16), % PB blast (p=2x10-16), Platelets (p=0.004), LDH (p=0.009), Albumin (p= 0.00001), Creatinine (p=0.007), CD13 (p=0.0002), CD33 (4.5xe-8), CD33 (1.1e-8). For all patients PatGp did not predict remission attainment. There was significant splay in overall survival and event free survival with median EFS durations ranging 52 , 100, 130 and 170 weeks ( p= 0.05) and in remission duration ranging from medians of 20- to 110 weeks. For intermediate cytogenetics Pat-Gps split into 4 with better outcomes (median OS 70-110 week, median RemDur 100-120 weeks) and 4 with worse outcomes (median OS 40-50 weeks, median RemDur 30-40 weeks). For unfavorable cytogenetics Pat-Gps also split into 4 with better outcomes (median OS 35-40 weeks, median RemDur 48-75 weeks) and 4 with worse outcomes (median OS 16-18 weeks, median RemDur 10-26 weeks). In a companion abstract we show the interaction between different protein functional groups. [Display omitted] ConclusionsAML could be classified based on protein functional groups. Although the classes were associated with cytogenetics the protein classifications gave prognostic information independent of cytogenetics and other traditional prognostic factors. Since this classification scheme is dependent on protein expression and functional activation states it may identify when particular pathways are being utilized and therefore be of use in triaging patients to targeted therapies. Disclosures:No relevant conflicts of interest to declare.

Resistance to Dasatinib in primary chronic lymphocytic leukemia lymphocytes involves AMPK-mediated energetic re-programming

Chronic lymphocytic leukemia (CLL) is the most common leukemia in adults in the western world. Although promising new therapies for this incurable disease are being tested in clinical trials, the therapeutic relevance of metabolic rewiring in chronic lymphocytic leukemia (CLL) is poorly understood. The aim of this study was to identify targetable metabolic differences in primary CLL lymphocytes by the use of Dasatinib. Dasatinib is a multi-tyrosine kinase inhibitor used to treat chronic myelogenous leukemia (CML) and is being tested in clinical trials for several cancers including CLL. This drug has been shown to be beneficial to CML patients suffering from diabetes by reducing their glucose plasma levels. In keeping with this previous observation, we report that Dasatinib induced glucose use while reducing lactate production, suggesting that this tyrosine kinase inhibitor decreases aerobic glycolysis and shifts glucose use in primary CLL lymphocytes. Our results suggest that primary CLL lymphocytes (independently of traditional prognostic factors) can be stratified in two subsets by their sensitivity to Dasatinib in vitro. Increased glucose use induced by Dasatinib or by inhibition of mitochondrial respiration was not sufficient to sustain survival and ATP levels in CLL samples sensitive to Dasatinib. The two subsets of primary CLL lymphocytes are characterized as well by a differential dependency on mitochondrial respiration and the use of anabolic or catabolic processes to cope with induced metabolic/energetic stress. Differential metabolic reprogramming between subsets is supported by the contrasting effect on the survival of Dasatinib treated CLL lymphocytes with pharmacological inhibition of two master metabolic regulators (mTorc1 and AMPK) as well as induced autophagy. Alternative metabolic organization between subsets is further supported by the differential basal expression (freshly purified lymphocytes) of active AMPK, regulators of glucose metabolism and modulators of AKT signaling. The contrasting metabolic features revealed by our strategy could be used to metabolically target CLL lymphocyte subsets creating new therapeutic windows for this disease for mTORC1 or AMPK inhibitors. Indeed, we report that Metformin, a drug used to treat diabetes was selectively cytotoxic to Dasatinib sensitive samples. Ultimately, we suggest that a similar strategy could be applied to other cancer types by using Dasatinib and/or relevant tyrosine kinase inhibitors.

CCAT2, a novel long non-coding RNA in breast cancer: expression study and clinical correlations

The clinical outcome of BC patients receiving the same treatment is known to vary considerably and thus, there is a compelling need to identify novel biomarkers that can select the patients that would benefit most from a given therapy and can predict the clinical outcome. The aim of this study was to determine the prognostic value of CCAT2, a novel long ncRNA recently characterized by our group and overlapping SNP rs6983267, in BC patients. We first evaluated by RT-qPCR and ISH the expression of CCAT2 in normal breast tissue and BC tissue and further analyzed CCAT2 expression in an independent set of 997 primary BC with regard to clinical, histological, pathological and other biological factors. Also, we explored the possibility of CCAT2 adding to the prognostic value of multivariate models that already included the traditional prognostic factors. Finally, we identified in in vitro models the impact of CCAT2 expression and SNP rs6983267 genotype on cell migration and chemoresistance. Our results revealed that although overexpressed in BCs in two out of three sets of patients, and having the highest expression in lymph node negative (LNN) disease, CCAT2 expression levels are informative solely for a subgroup of BC patients, namely for patients with LNP disease that have received adjuvant CMF chemotherapy. For this subgroup high levels of CCAT2 suggest the patients will not benefit from CMF containing adjuvant chemotherapy (shorter MFS and OS). Additionally, we found that CCAT2 upregulates cell migration and downregulates chemosensitivity to 5'FU in a rs6983267-independent manner.

Other Cancers in Patients With Chronic Lymphocytic Leukemia Requiring Therapy: Association With Prognostic Factors and Impact on Survival

Abstract 3896 Chronic lymphocytic leukemia (CLL) is associated with an increased incidence of other cancers (OC). Potential risk factors for OC in CLL include immune disturbances, shared risk factors, genetic predisposition and chemotherapy. While reports on OC in patients with CLL have been typically descriptive in nature, detailed information regarding the clinical features of OC, relationship with the time of diagnosis of CLL, patient characteristics, prognostic factors and survival have been missing. We report an analysis of OC in pts with CLL requiring therapy, with a post therapy observation time of 5 years or longer. We reviewed our database and selected pts with CLL seen at our center who underwent treatment and who have a follow-up of at least 5 years after therapy. We studied pts with an OC either before or after the diagnosis and/or treatment of CLL and compared them with patients without OC. Richter's transformation of CLL was not considered an OC in this analysis. A total of 1,364 pts with CLL referred between 1963 and 2007 with a median follow up time of 8.2 years (range 5–28) were studied. Among these, we identified 324 pts (24%) with OC. Sixty-eight pts were diagnosed with >1 OC [2 OC (62 pts), 3 OC (4 pts) or 4 OC (2 pts)], for a total of 400 individual OC. Of the 324 pts with OC, OC preceded the diagnosis of CLL in 117 pts (36%), with a median time from OC to CLL of 80 (0–455) months, OC occurred after the diagnosis of CLL but before treatment in 49 pts (15%), with median time from CLL to OC of 22 (0–131) months. OC occurred after treatment of CLL in 158 pts (49%), with a median time from treatment to OC of 94 (0–304) months. The type, frequency and timing of OC are summarized in [table 1][1]. Clinical characteristics and traditional prognostic factors were not significantly different between pts with and without OC, with the exception of age, as pts with OC are older than pts without OC at the time of referral (median age: 60 vs 55 years, p 5 years, the incidence of OC is 24%. Among pts with OC, there is a trend for higher number of pts with del11q. In pts with OC, OC itself is a major cause of death, while the vast majority of deaths among pts without OC are caused by CLL or CLL-related complications. | Site of OC | 1st case | Total cases | OC prior/at CLL | OC after CLL | |:---------------------------------------:| --------------- | ----------- | --------------- | ------------ | --- | | After diagnosis | After treatment | | NMSC[1][2] | 124 | 144 | 41 | 26 | 77 | | PROSTATE | 43 | 54 | 18 | 7 | 29 | | MELANOMA | 26 | 34 | 16 | 2 | 16 | | BREAST | 23 | 26 | 14 | 2 | 10 | | MDS/AML[2][3] | 16 | 18 | | | 18 | | LUNG | 14 | 21 | | 1 | 20 | | NEUROENDOCRINE | 3 | 6 | 1 | | 5 | | HEAD AND NECK | 12 | 16 | 3 | 2 | 11 | | UROTHELIAL CELL CANCERS | 11 | 12 | 7 | 2 | 3 | | GI[3][4] (including pancreas and liver) | 10 | 18 | 4 | 3 | 11 | | KIDNEY | 10 | 11 | 4 | 3 | 4 | | NHL[4][5] and MM[5][6] | 8 | 12 | | | 12 | | TESTIS/OVARY | 6 | 6 | 4 | | 2 | | THYROID | 6 | 7 | 5 | 1 | 1 | | SARCOMAS | 4 | 5 | 4 | | 1 | | UTERUS | 2 | 3 | 3 | | | | OTHERS[6][7] | 6 | 7 | 3 | | 4 | | TOTALS | 324 | 400 | 127 | 49 | 224 | * [↵][8]1 non-melanoma skin cancers; * [↵][9]2 myelodysplastic syndrome/acute myeloid leukemia; * [↵][10]3 gastrointestinal; * [↵][11]4 non-Hodgkin's lymphoma; * [↵][12]5 multiple myeloma; * [↵][13]6 includes: brain, nasopharyngeal, penis carcinoma, mesothelioma, unknown primary. Table 1. Distribution of OC (1364 pts). Disclosures: No relevant conflicts of interest to declare. [1]: #T1 [2]: #fn-1 [3]: #fn-2 [4]: #fn-3 [5]: #fn-4 [6]: #fn-5 [7]: #fn-6 [8]: #xref-fn-1-1 [9]: #xref-fn-2-1 [10]: #xref-fn-3-1 [11]: #xref-fn-4-1 [12]: #xref-fn-5-1 [13]: #xref-fn-6-1

The ‘Echo Heart Failure Score’: an echocardiographic risk prediction score of mortality in systolic heart failure

Although many transthoracic echocardiographic (TTE) measurements have been shown to predict outcome in heart failure (HF), whether incremental risk prediction is afforded by their combination is unknown. We developed a simple echocardiographic risk score of mortality in HF patients. We performed TTE in 747 systolic HF patients followed-up for 34 ± 23 months. The Cox hazard model was used to evaluate the association between 14 TTE parameters and death. The Echo Heart Failure Score (EHFS) was derived by assigning the value of 1 to each independent predictor when present, and 0 when it was absent, and then by summing the number. The 3-year risk prediction improvement was tested by adding the EHFS to a model containing clinical predictors, and by calculating the C index and net reclassification improvement (NRI). Five baseline TTE variables (end-systolic volume index, left atrial volume index, mitral E-wave deceleration time, tricuspid annular peak systolic excursion, and pulmonary artery systolic pressure) remained independent predictors of mortality. The mortality rate (per 100 patients/year) significantly increased with EHFS ranging from 0 to 5 (EHFS = 0, 2.7%; 1, 5.2%; 2, 10.1%; 3, 13.7%, 4, 29.7%; 5, 36.9%; P < 0.0001). Patients with EHFS ≥3 had a mortality hazard ratio of 3.58 (95% confidence interval 2.74-4.78) compared with EHFS <3. Adding EHFS to the base model improved the C index (from 0.74 to 0.81, P < 0.0001), yielding a continuous NRI of 0.63 (P < 0.0001). The EHFS, an easily obtainable echo score, improved risk prediction of death over traditional prognostic factors in systolic HF patients, and it may prove useful for risk stratification.

The Prognostic Significance of the Biomarker p16 in Oropharyngeal Squamous Cell Carcinoma

AimsThere is an increasing incidence of human papillomavirus (HPV)-positive oropharyngeal squamous cell cancers (OPSCC) mostly associated with favourable outcomes. p16 immunohistochemistry is a surrogate marker for HPV positivity in OPSCC. The prognostic strength of p16 over traditional prognostic factors is not fully characterised. In this study, we evaluated the clinical and demographic differences between p16-positive and -negative OPSCC and characterised its prognostic strength versus traditional prognostic factors. Materials and methodsFormalin-fixed, paraffin-embedded blocks and clinical information from 217 OPSCC patients, treated with radiotherapy (alone or in combination with other therapies) between 2000 and 2010 were collected retrospectively. Immunohistochemistry for p16 protein was carried out; cancer-specific survival (CSS), recurrence-free survival (RFS) and locoregional control (LRC) were calculated for both univariate and multivariate analyses. ResultsNinety-two per cent of the OPSCC originated from tonsil and tongue base sites, 61% were p16 positive. Patients with p16-positive OPSCC were younger (P < 0.0001), with lower alcohol (P = 0.0002) and tobacco (P = 0.0001) exposure. The tumours were less differentiated (P = 0.0069), had a lower T stage (P = 0.0027), higher nodal status (P = 0.014) and higher American Joint Committee on Cancer (AJCC) prognostic group (P = 0.0036). AJCC prognostic group was significant for RFS (P = 0.0096) and CSS (P = 0.018) in patients with p16-negative OPSCC, but not those with p16-positive tumours (P = 0.30 and 0.54). Other significant factors for CSS and RFS in univariate analysis were: pretreatment haemoglobin (P < 0.0001 and <0.0001), chemoradiotherapy (P = 0.005 and 0.03) and P16 status (P < 0.0001 and 0.0001). In multivariate analysis, p16 positivity was the strongest independent prognostic variable for both CSS, RFS and LRC (P < 0.0001, hazard ratio 4.15; 95% confidence interval 2.43–7.08), (P < 0.0001, hazard ratio 6.15; 95% confidence interval 3.57–10.61) and (P = 0.001, hazard ratio 3.74; confidence interval 1.76–7.95). ConclusionThis study shows that p16 is the single most important prognostic variable in OPSCC, surpassing traditional prognostic factors for both CSS and RFS. Furthermore, disease stage has no prognostic significance in p16-positive patients, highlighting the need for routine p16 assessment in OPSCC.

Correlation of contrast-enhanced ultrasound kinetics with prognostic factors in invasive breast cancer

To correlate contrast-enhanced ultrasound (CEUS) kinetic parameters with traditional and molecular prognostic factors in invasive breast cancer. Seventy-five invasive breast cancers were evaluated with contrast harmonic imaging after the injection of a bolus dose of 2.4 ml sulphur hexafluoride microbubble contrast agent. The lognormal function was used for quantitative analysis of kinetic data. These parameters correlated with traditional prognostic factors (tumour size, histological type, tumour grade, axillary lymph node status) and immunohistochemical biomarkers (ER, PR and HER2 status). Statistically significant correlation was found between time-to-peak and tumour grade (P value = 0.023), PR status (P value = 0.042) and axillary node status (P value = 0.025). Wash-out ratio, measured at 21 s was significantly associated with ER status (P value = 0.042) and PR status (P value = 0.026). Invasive breast carcinomas exhibiting earlier peak enhancement and faster elimination of microbubble contrast agent at CEUS are found to be associated with established predictors of poor prognosis.

The prospective MammaPrint MINT (Multi-Institutional Neo-adjuvant Therapy) study.

TPS11122 Background: Patients with locally advanced breast cancer (LABC) are often treated with neo-adjuvant chemotherapy to reduce the size of the tumor before definitive surgery. Complete pathologic Response (pCR) predicts better long term outcome. Genomics assays that measure specific gene expression patterns in a patient's primary tumor have become important prognostic and predictive tools for early breast cancer. This study is designed to test the ability of molecular profiling, as well as traditional pathologic and clinical prognostic factors to predict responsiveness to neo-adjuvant chemotherapy in patients with LABC. Methods: Women ≥ 18 yrs with histologically-proven invasive breast cancer T2(≥3.5cm)-T4,N0M0 or T2-T4N1M0, with measurable disease, adequate bone marrow reserves and normal renal and hepatic function who signed informed consent are enrolled. Axillary lymph nodes will be staged according to protocol. MammaPrint risk profile, BluePrint molecular subtyping profile, TargetPrint ER, PR and HER2 single gene readout, and the 56-gene TheraPrint Research Gene Panel will be analysed using the whole genome expression array. Patients will receive neo-adjuvant chemotherapy treatment according to protocol. Response will be measured by centrally assessed Residual Cancer Burden (RCB). Objectives are: (1) To determine the predictive power of MammaPrint and BluePrint for sensitivity to neo-adjuvant chemotherapy as measured by pCR. (2) To identify and/or validate predictive gene expression profiles of clinical response or resistance to neo-adjuvant chemotherapy. (3) To compare TargetPrint ER, PR and HER2 with local and centralized IHC and/or CISH/FISH assessment. (4) To identify correlations between TheraPrint and response to neo-adjuvant chemotherapy. (5) To compare BluePrint molecular subtype with IHC-based subtype classification. To achieve a difference of 20% in chemotherapy sensitivity for patients stratified by MammaPrint, a total of 226 samples is needed (significance level 0.05 and power of 0.90). So far 45 patients have been enrolled from multiple institutions. Clinical trial information: NCT01501487.

Multivariate analysis of prognostic factors in cancer of unknown primary (CUP) patients treated with site-specific therapy based on the 92-gene cancer classifier.

e15006 Background: Results from a single arm prospective study demonstrated overall survival of 12.5m in 194 patients that received first-line therapy directed by tumor classification using the 92-gene assay (JCO, 31:217-23). When the molecular assay predicted tumor types that were clinically more responsive, median survival was significantly improved when compared with predictions of more resistant tumors (13.4 v 7.6 m; p= 0.04).In this analysis, the survival benefit of assay-directed therapy was further evaluated after adjusting for pre-treatment prognostic factors. Methods: Cox proportional hazards regression was used to examine survival benefit of responsive tumor types (n=115) vs resistant tumor types (n=79) predicted by the molecular assay, after controlling for traditional prognostic factors. Results: Univariate Cox regression analysis (n=194) showed that ECOG status, responsive vs resistant tumor type, and assay probability were each significant prognostic factors. In multivariate analyses, the prognostic value of only ECOG status and the assay prediction of responsive vs resistant tumor type were maintained. Conclusions: These data demonstrate that the 92-gene assay results remained a significant independent variable affecting survival outcome in these patients after adjustment for prognostic heterogeneity, and provide further support of assay accuracy in molecular classification of CUP. Clinical trial information: NCT00737243. [Table: see text]

OP053: The role of sLea, sLex and CLA in oropharyngeal carcinoma: Preliminary report

Purpose Tumor-associated carbohydrate antigens (TACAs) result from the aberrant glycosylation that is seen as a frequent event in several human cancers cells. Sialyl Lewis (X) and Sialyl Lewis (A) are terminal carbohydrate structures belonging to the blood group Lewis. Their expression is strongly associated with prognosis in certain tumours, but their potential prognostic implications in oropharyngeal cancer patients are largely unexplored. Aim of the work was to define both the pathogenetic and the prognostic role of sLea, sLex and CLA, containing sLex antigenic determinants, in a large series of oropharyngeal squamous cell carcinomas (OPSCCs). Materials and methods 88 OPSCCs were analysed by immunohistochemistry for sLea, sLex and CLA. Statistical evaluations were carried out using Pearson’s Chi Square Test, one way analysis of variance (ANOVA) and the Tukey’s HSD test to compare distribution of clinical and pathological features among patients with the different immunohistochemical scores of CLA, sLea and sLex expression. Cox analysis was applied to determine the single contribution of covariates on survival rate. Only values of p Results In all neoplastic tissues we have observed higher levels of CLA, sLea and sLex than in normal mucosa ( p Discussion and conclusions Our work have underlined the key-role of TACAs in oropharyngeal carcinogenesis and in the prognostic stratification of patients. To our knowledge, the present work is the first largest study to highlight the existence of a statistical association between TACAs and traditional prognostic factors in oropharyngeal cancer.

Preoperative hydronephrosis and diabetes mellitus predict poor prognosis in upper urinary tract urothelial carcinoma

Introduction: We assess the impact of traditional prognostic factors, tumour location, degree of hydronephrosis and diabetes mellitus (DM) on the survival of patients treated for upper urinary tracturothelial carcinoma (UUTUC).Methods: From January 2004 to March 2010, we analyzed data from 114 patients with UUTUC who underwent nephroureterectomy with a bladder cuff excision. Median patient age was 71 years and median follow-up was 26.5 months. The influence of traditional prognostic factors, including DM, tumour stage, grade, location and degree of hydronephrosis, on recurrence-free survival (RFS) rates were analyzed using Kaplan-Meier analysis and Cox proportional hazards regression model.Results: Among 61 renal pelvis and 53 ureteral tumour cases, recurrence was identified in 71 cases (62.3%). Kaplan-Meier analysis showed that degree of hydronephrosis was associated with RFS (p = 0.001). DM and degree of hydronephrosis were independent factors for RFS in Cox proportional regression analysis (HR=1.8 CI:1.01-3.55, p = 0.04), (HR=3.7, CI: 2.0-6.5, p = 0.001). All patients with ureteral tumour had no worse prognosis than those with renal pelvis tumour, but the pT2 patients with ureteral tumour had a worse prognosis than those with renal pelvis tumour with a median RFS of 9 months (range: 2.6-15.3 months) and 29 months (range:8.0-13.2 months), respectively (p = 0.028).Conclusions: Tumour location is not a factor influencing RFS, except in the pT2 stage. However, severe hydronephrosis is associated with a higher recurrence in UUTUC. Also, DM is related to disease recurrence. Further prospective studies are needed to establish the prognostic significance of DM in large populations.

Prognostic value of proliferation markers expression in breast cancer

In this study, immunohistochemical expression of five proliferation markers: Ki-67, aurora-A kinase, survivin, B-Myb and cyclin B1, was analyzed. Consecutive 215 tumor samples from breast cancer patients operated from 2002 to 2003 were analyzed using the TMA ("tissue microarray") method. The median follow-up was 95 months (from 7.8 to 107 months). Statistically significant correlations between expression levels in five proliferation markers, and correlations between some of the proliferation markers and traditional prognostic factors were found. Statistically significant prognostic influence of aurora-A kinase, survivin and B-Myb expression levels on overall and disease-free survival was found, and cyclin B1 expression level on disease-free survival. A multivariate analysis confirmed survivin and B-Myb expression as independent prognostic factors of overall (p = 0.0195; p = 0.0004) and disease-free survival (p = 0.0107 and p = 0.0205) in breast cancer patients.

Prognostic Factors Including Ki-67 and p53 in Bacillus Calmette-Guérin-Treated Non-Muscle-Invasive Bladder Cancer: A Prospective Study

Objectives: To prospectively evaluate the prognostic utility of the traditional prognostic factors and molecular markers p53 and Ki-67 in a homogeneous series of patients with non-muscle-invasive bladder cancer (NMIBC) treated with Bacillus Calmette-Guérin (BCG). Patients and Methods: 192 intermediate- and high-risk NMIBC cases were enrolled. The variables in study were age, stage, grade, focality, tumour size, presence of associated carcinoma in situ, recurrence rate before BCG, maintenance for BCG, Ki-67 and p53. The endpoints considered were recurrence-free survival, progression-free survival, cancer-specific survival (CSS) and overall survival (OS). Results: T stage resulted in being associated with CSS, whereas age with OS. BCG maintenance was a significantly favourable independent predictor of OS, CSS, recurrence and progression. In univariate analysis, the labelling index of Ki-67 was significantly associated with OS, CSS and progression. Multivariate analysis, however, confirmed this association only for OS. On the contrary, the labelling index of p53 was a significant predictor of recurrence, both in uni- and multivariate analyses, but with a HR inferior to 1. Conclusions: Ki-67 was an independent predictor of survival. p53 overexpression showed a significant yet inverse correlation with recurrence, thus showing little clinical utility. Age, stage and maintenance were confirmed as independent predictors of BCG response.

The Prognostic Value of DNA Ploidy Determination in Endometrial Cancer

Aneuploidy, defined as an abnormal quantity of DNA in cells nuclei, is the most frequently observed genetic abnormality in cancer cells. Alterations in cell cycle control and chromosomal missegregation very often result in the accumulation of excess genetic material. These genetic reassortments simultaneously imbalance lots of structural and regulatory proteins. This chromosomal instability may be associated with mutations in tumor suppressor genes or loss of function of mismatch repair genes. These are common pathways of cancerogenesis in endometrial cancer. Endometrial cancer represents the most common female genital tract malignancy and is generally associated with favourable outcomes for affected patients. Despite this, recurrence rates and diseaserelated deaths are consistently reported in published series, also in so-defined low-risk groups of patients. Hence, is reasonable to believe that traditional prognostic factors–surgical stage, histologic type, tumor grading, myometrial deep of invasion, involvement of vascular spaces–do not definitely respond to the clinical needs of a comprehensive management. The determination of DNA ploidy in endometrial cancer has been widely investigated in the last decades and clear evidence is available of its direct correlation with prognosis. Despite this, poor acceptance and skepticism are common believes in the scientific community. Aim of the paper has been the attempt to underline, upon scientific strength, the importance and clinical potential usefulness of this determination in gynecological oncological practice.

Prognostic factors for patients with diffuse large B cell lymphoma and transformed indolent lymphoma undergoing autologous stem cell transplantation in the positron emission tomography era

In the positron emission tomography (PET) era, traditional prognostic factors may not apply for patients with relapsed/refractory (R/R) diffuse large B cell lymphoma (DLBCL) undergoing autologous stem cell transplantation (ASCT). Moreover, little is known about prognostic factors in patients transplanted for transformed indolent lymphoma (TIL). We conducted a retrospective study of 143 patients with R/R DLBCL and TIL who were transplanted in the last decade and had a post-salvage PET scan. We examined prognostic factors in both groups, and constructed a prognostic score for DLBCL patients. For patients with DLBCL, post-salvage PET response was an important prognostic factor. Advanced age and symptomatic relapse were also significantly associated with outcome. A simple score could stratify patients into three risk groups with 4-year post-ASCT overall survival of 84%, 59%, and 10%, and 4-year progression-free survival of 67%, 41% and 0% (P<0.0001 for both). However, none of those factors (including PET response to salvage) appeared relevant for patients with TIL, despite their comparable overall outcome. Our prognostic score for DLBCL patients undergoing ASCT may be useful for prognostication, for stratification in clinical trials, and to motivate the design of new strategies for patients in the high-risk group, who may not derive benefit from standard ASCT.

Abstract OT3-4-02: MINT I: Multi-Institutional Neo-adjuvant Therapy, MammaPrint Project I

Abstract Background: Women with locally advanced breast cancer (LABC) are often treated with neo-adjuvant chemotherapy to reduce the size of the tumor prior to surgery, to enable breast conserving surgery and to observe the clinical effect of therapy in real time. Studies have shown that the 25–27% of individuals who have a pathologic complete response (pCR) to neoadjuvant therapy have a survival advantage of 80% in 5 years, which is double the expected survival of the remaining patients without pCR. If patients who are likely to show a pCR could be identified prior to initiation of therapy, it would enable more informed treatment decisions – patients likely to respond would be served well by current neoadjuvant chemotherapy protocols, while those unlikely to respond may be better suited to innovative new strategies for drug discovery [von Minckwitz et al. JCO 2006]. Genomics assays, which are widely used to provide prognostic and predictive information in early breast cancer, have the potential to provide information on the likelihood of a patient with LABC responding to neo-adjuvant therapy [Glück et al. ASCO 2012]. Trial design: MINT I is a prospective study designed to test the ability of molecular profiling, as well as traditional pathologic and clinical prognostic factors, to predict responsiveness to neo-adjuvant chemotherapy in patients with LABC. MammaPrint risk profile, BluePrint molecular subtyping profile, TargetPrint estrogen receptor (ER), progesterone receptor (PR) and HER2 single gene readout, and the 56-gene TheraPrint Research Gene Panel will be analyzed on a fresh tumor specimen using the whole genome array. Patients will receive neo-adjuvant chemotherapy pre-specified in the protocol. Response will be measured centrally. pCR is defined as the absence of invasive carcinoma in both the breast and axilla at microscopic examination of the resection specimen, regardless of the presence of carcinoma in situ. Eligibility: The study will include women ≥18 years with histologically-proven invasive breast cancer T2 (≥3.5cm)-T4, N0M0 or T2-T4N1M0, adequate bone marrow reserves and normal renal and hepatic function who signed an IRB approved informed consent. Objectives: The objectives of the study are to: 1. Determine the predictive power of MammaPrint and BluePrint for sensitivity to neo-adjuvant chemotherapy as measured by pCR.2. Compare TargetPrint ER, PR and HER2 with local and centralized IHC and/or CISH/FISH assessment.3. Identify correlations between TheraPrint and response to neo-adjuvant chemotherapy.4. Identify and/or validate predictive gene expression profiles of clinical response or resistance to neo-adjuvant chemotherapy.5. Compare BluePrint with IHC-based subtype classification. Statistical methods: Standard statistical tests such as the Pearson Chi-square test will be used to characterize and evaluate the relationship between chemoresponsiveness and gene expression patterns. Accrual: A total of 226 eligible patients will be enrolled from multiple institutions. To date (June 06, 2012), 31 patients have been enrolled. Clinical trial registry number: NCT01501487. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr OT3-4-02.

External validation on a prospective basis of a nomogram for predicting the time to first treatment in patients with chronic lymphocytic leukemia

A nomogram that incorporates traditional and newer prognostic factors to identify patients with chronic lymphocytic leukemia (CLL) who are at high risk of receiving therapy was developed by investigators at The University of Texas M. D. Anderson Cancer Center (MDACC). Because the model required validation before its extensive use could be recommended, the authors sought to externally validate the nomogram in an independent, community-based cohort of patients with CLL. In total, 328 previously untreated patients with newly diagnosed, asymptomatic, Binet stage A CLL from different primary hematology centers who were registered on a prospective basis during 2006 to 2010 on an observational database of the Italian Lymphoma Study Group were considered suitable for external validation of the model. A total point score was calculated for each patient using a formula proposed by MDACC investigators, and the median score was 19.9 (range, 0-69.5). Furthermore, when the score was evaluated as continuous variable (ie, by measuring the risk of each point increase), the total point score was associated with the time to first treatment (hazard ratio [HR], 1.04; 95% confidence interval [CI], 1.02-1.05; P < .0001). Receiver operating characteristic analysis identified a point score of 25 (area under curve; 0.64; sensitivity, 61.5; specificity, 72.1; P < .0001) as the best threshold capable of separating patients who needed therapy from patients who did not (HR, 3.27; 95% CI, 2,07-5.18; P < .0001). The prognostic index category also remained a predictor of the time to first treatment when the analysis was limited to patients with Rai stage 0 disease (HR, 4.05; 95% CI, 2.25-7.52; P < .0001). Finally, a goodness-of-fit test demonstrated that the nomogram model had a significantly good fit at 2 years (correlation coefficient [r(2) ] = 0.966; P = .002). The current results confirmed the ability of a newly developed prognostic index to predict the time to first treatment among previously untreated patients with CLL who had early disease and extended the utility of the model to those with Rai stage 0 disease. In addition, the actual and predicted time to first treatment outcomes revealed good agreement, suggesting that, externally, the results provided by the model are well calibrated. Cancer 2013. © 2012 American Cancer Society.

Other Cancers in Patients with Chronic Lymphocytic Leukemia Requiring Therapy: Association with Prognostic Factors and Impact On Survival

AbstractAbstract 3896Chronic lymphocytic leukemia (CLL) is associated with an increased incidence of other cancers (OC). Potential risk factors for OC in CLL include immune disturbances, shared risk factors, genetic predisposition and chemotherapy. While reports on OC in patients with CLL have been typically descriptive in nature, detailed information regarding the clinical features of OC, relationship with the time of diagnosis of CLL, patient characteristics, prognostic factors and survival have been missing. We report an analysis of OC in pts with CLL requiring therapy, with a post therapy observation time of 5 years or longer. We reviewed our database and selected pts with CLL seen at our center who underwent treatment and who have a follow-up of at least 5 years after therapy. We studied pts with an OC either before or after the diagnosis and/or treatment of CLL and compared them with patients without OC. Richter’s transformation of CLL was not considered an OC in this analysis. A total of 1,364 pts with CLL referred between 1963 and 2007 with a median follow up time of 8.2 years (range 5–28) were studied. Among these, we identified 324 pts (24%) with OC. Sixty-eight pts were diagnosed with >1 OC [2 OC (62 pts), 3 OC (4 pts) or 4 OC (2 pts)], for a total of 400 individual OC. Of the 324 pts with OC, OC preceded the diagnosis of CLL in 117 pts (36%), with a median time from OC to CLL of 80 (0–455) months, OC occurred after the diagnosis of CLL but before treatment in 49 pts (15%), with median time from CLL to OC of 22 (0–131) months. OC occurred after treatment of CLL in 158 pts (49%), with a median time from treatment to OC of 94 (0–304) months. The type, frequency and timing of OC are summarized in table 1. Clinical characteristics and traditional prognostic factors were not significantly different between pts with and without OC, with the exception of age, as pts with OC are older than pts without OC at the time of referral (median age: 60 vs 55 years, p<.0001). We next analyzed the distribution of the newer prognostic parameters in these two groups. Fifty-three % of pts with OC had unmutated IGHV genes, 65% were ZAP70+, 22% CD38+, 8% had del17p, 27% del11q, 22% +12, 20% negative FISH, and 23% del13q. Sixty-one % of pts without OC had unmutated IGHV genes, 58% were ZAP70+, 26% CD38+, 6% had del17p, 17% del11q, 22% +12, 26% negative FISH, and 29% del13q. A trend toward significance (p.07) for a higher incidence of del11q was observed in pts with OC. At the time of this analysis 536 (39%) pts have died and 828 (61%) are alive. One hundred seventy six/324 (54%) pts with OC are alive, vs 652/1040 (63%) pts without OC (p<.007). Median overall survival (OS) has not been reached in either pt group at a median follow up of 95 (61–284) months for pts with OC and 98 (61–340) months for pts without OC. Causes of death were: progression of CLL (99), CLL-related complications (121), OC (43), complications of stem cell transplant (16), events unrelated to CLL (39) and unknown causes (218). In pts with OC, the OC itself was the cause of death in 37%. Other causes of death were CLL/CLL-related complications in 44%, complications of stem cell transplant in 8%, and events unrelated to CLL in 11%. In contrast, in pts without OC CLL/CLL-related complications accounted for 83% of deaths, complications of stem cell transplant for 4%, and events unrelated to CLL for 13%. In conclusion, in pts with CLL requiring therapy and with post therapy follow up of >5 years, the incidence of OC is 24%. Among pts with OC, there is a trend for higher number of pts with del11q. In pts with OC, OC itself is a major cause of death, while the vast majority of deaths among pts without OC are caused by CLL or CLL-related complications.Table 1Distribution of OC (1364 pts).Site of OC1st caseTotal casesOC prior/at CLLOC after CLLAfter diagnosisAfter treatmentNMSC1124144412677PROSTATE435418729MELANOMA263416216BREAST232614210MDS/AML216180018LUNG14210120NEUROENDOCRINE36105HEAD AND NECK12163211UROTHELIAL CELL CANCERS1112723GI3 (including pancreas and liver)10184311KIDNEY1011434NHL4 and MM58120012TESTIS/OVARY66402THYROID67511SARCOMAS45401UTERUS23300OTHERS667304TOTALS324400127492241non-melanoma skin cancers;2myelodysplastic syndrome/acute myeloid leukemia;3gastrointestinal;4non-Hodgkin's lymphoma;5multiple myeloma;6includes: brain, nasopharyngeal, penis carcinoma, mesothelioma, unknown primary. Disclosures:No relevant conflicts of interest to declare.

BOTH MD Anderson Cancer Center MODEL and German Score Work in Predicting Time to First Treatment in EARLY Chronic Lymphocytic Leukemia: Results of an External Validation Analysis

AbstractAbstract 3934In chronic lymphocytic leukemia (CLL) there has been a limited published work on the development of models predicting time to first treatment that incorporate traditional and novel prognostic factors. We wondered whether the nomogram recently proposed by M D Anderson Cancer Center (MDACC) investigators that includes clinical and newer prognostic variables (i.e., three or more involved lymph node sites, increased size of cervical lymph nodes, presence of 17p deletion or 11q deletion by FISH, increased serum LDH, and unmutated IGHV mutation status) could work when applied to a prospective community-based CLL patient cohort.We considered suitable for the purpose of this study 328 previously untreated patients with newly diagnosed asymptomatic Binet stage A CLL from different primary hematology centers registered on a prospective basis during the period 2006–2010 on an observational database of Gruppo Italiano Studio Linfomi (GISL).Using the formula proposed by Wierda et a total point score was calculated for each patient. A value ranging between 0 and 69.5 (median, 19.9) - therefore similar to total point scores reported by the MDACC group (median, 21.0; range, 0 – 87.4) – was found. As a continuous variable (i.e., measuring the risk of each point increase), total point score was associated with time to first treatment (HR 1.04; 95% CI 1.02–1.05; P < 0.0001). Receiver Operating Characteristic analysis identified a point score of 25 [area under curve (AUC) 0.64, sensitivity 61.5 specificity 72.1; P<0.0001)] as the best threshold capable of separating patients who needed therapy from patients who did not (HR, 3.27; 95% CI, 2,07–5.18; P<0.0001). The prognostic index category remained a predictor of time to first treatment also when analysis was limited to patients in Rai stage 0 (HR,4.05; 95% CI: 2.25–7.52; P<0.0001). Finally, the goodness of fit showed that the nomogram model had significant good fit at 2 years (r2=0.966; P=0.002).We next wondered whether the German score based on 7 prognostic variables (i.e., age, gender, b2-microglobulin, sTK, IGVH mutational status, ECOG performance status and hierachic type of FISH abnormalities) and originally designed for predicting overall survival could forecast time to first treatment in patients with early disease. Since sTK was not available a slightly modified version of German model based on 6 prognostic variables was used. The risk score calculated in each patient by summing single ratings of adverse factors, allowed the identification of two different risk categories scoring respectively 0–2 and 3–5 whose clinical outcomes were different with respect to time to first treatment (HR 2.19, 95% CI: 1.15–2.20; P=0.005).In conclusion, this analysis shows that both models proposed by MDACC investigators and German group provide interchangeable tool for assessing time to first treatment. Interestingly, in this validation analysis the pattern of distribution of patients into a low and high-risk category revealed a high degree of concordance between models (P<0.0001; Mann-Whitney test). Disclosures:Morabito:Celgene: Honoraria.