Independent Predictor Of Shorter Survival Research Articles

Vascular and tumor cell expression of VEGFR2 and molecular subtyping: An innovative biomarker approach in bladder cancer.

497 Background: Treatment approaches combining anti-angiogenic therapy with chemotherapy have shown promising clinical results in metastatic bladder cancer (BC). In order to interrogate the potential clinico-pathologic and molecular basis of these findings, we evaluated VEGFR2, vascular density and molecular subtyping in a series of BC patients. Methods: A custom-TMA with primary BC tissues from 117 patients (mean age 71 yrs; range 38-99 yrs; M:F = 83:34) treated at a single institution, was stained and scored (0-3) for CD34 (vascular density) and for VEGFR2 on tumor vessels and cells. CK5/6 and GATA3 IHC was scored by a pathologist to identify main molecular classes of BC. The association between clinico-pathologic variables, VEGFR2, CD34 and molecular subtypes was analyzed by Fisher’s exact test and ANOVA. Univariate and multivariate Cox proportional models were used for survival analysis. Results: Of 112 analyzable BC tissues, 41% were muscle-invasive (MIBC) vs. 56% non-muscle invasive (NMIBC) and 3% undetermined. Compared to NMIBC patients those with MIBC had shorter overall survival (p < 0.001). The main molecular subtypes included basal (11%), mixed baso-luminal (28%) and luminal (61%). Compared to luminal and baso-luminal subtypes, the majority of basal BCs were muscle invasive (p = 0.036). VEGFR2 expression was higher in tumor vessels but variable in tumor cells. 74% of BCs showed high-medium levels of VEGFR2 (scores 3-2) in tumor vessels and 88% had high-medium levels of tumor vascular density. Within basal, luminal and baso-luminal subtypes, 58%, 78% and 71% had high-medium levels of vascular VEGFR2 (p = 0.52), while 83%, 91% and 82% had high-medium levels of tumor vascular density (p = 0.08). Survival analyses showed increasing patient age, higher stage and lower VEGFR2 levels as independent predictors of shorter survival. Conclusions: Given the observed complexity of BC regarding VEGFR2 expression and vascular density across molecular subtypes, further investigation is warranted to understand how the frequent expression of VEGFR2 on tumor vasculature and variable expression in tumor cells relates to the efficacy of anti-angiogenic agents across these subtypes in bladder cancer.

MiR-483-5p promotes IGF-II transcription and is associated with poor prognosis of hepatocellular carcinoma.

The human insulin-like growth factor-II (IGF-II) gene transcribes four mRNAs (P1 mRNA-P4 mRNA), and P3 mRNA overexpression contributes to hepatocarcinogenesis. IGF-II-derived miR-483-5p is implicated in the development of cancers. Here, we investigated the involvement of miR-483-5p in P3 mRNA overexpression regulation and its role in hepatocellular carcinoma. Our results showed that miR-483-5p up-regulated P3 mRNA transcription by targeting the 5′-untranslated region (5′UTR) of P3 mRNA in hepatocellular carcinoma. The mechanism was involved in recruiting of an argonaute 1(Ago1)-argonaute 2 (Ago2) complex to the P3 mRNA 5′UTR and the P3 promoter of IGF-II gene by miR-483-5p, accompanied by increased enrichment of RNA polymerase II and activating histone marks histone 3 lysine 4 trimethylation (H3K4me3), histone 3 lysine 27 acetylation (H3K27ac), and histone 4 lysine 5/8/12/16 acetylation (H4Kac) at the P3 promoter. High miR-483-5p expression was an independent predictor for shorter survival of HCC patients. The findings suggest that miR-483-5p promotes P3 mRNA transcription by recruiting the Ago1-Ago2 complex to the P3 mRNA 5′UTR and is associated with poor prognosis of HCC. Our results display a potential new model for miRNAs to up-regulate gene expression.

Self-Expandable Metal Stent Use to Palliate Malignant Esophagorespiratory Fistulas in 88 Patients

PurposeTo identify predictors associated with clinical outcomes (initial clinical failure, stent patency, and survival) after self-expandable metal stent (SEMS) placement for malignant esophagorespiratory fistulas (ERFs). Materials and MethodsUsing logistic and Cox regression analyses, this study reviewed 88 patients (mean age 59.4 y ± 8.4; 84 men [95.5%] and 4 women [4.5%]) who underwent fluoroscopic SEMS placement for palliating malignant ERF from January 2000 to December 2016. ResultsTechnical success was achieved in all patients. Initial clinical success was achieved in 78.4% (69/88; 95% confidence interval [CI], 68.7%–85.7%). Among the 69 patients in whom initial clinical success was achieved, aspiration symptoms recurred in 37.7% (26/69; 95% CI, 27.2%–49.5%). Overall major complication rate was 25.0% (22/88; 95% CI, 17.1%–35.0%). Cumulative stent patency and cumulative survival rates at 1, 3, 6, and 12 months were 72.8%, 38.9%, 32.4%, and 21.6% and 81.4%, 51.9%, 30.5%, and 13.3%, respectively. Stricture of the upper esophagus was an independent predictor of initial clinical failure (odds ratio, 3.760; 95% CI, 1.207–11.811) and shorter stent patency (hazard ratio [HR], 2.036; 95% CI, 1.170–3.544). Initial clinical failure was an independent predictor of shorter survival (HR, 2.902; 95% CI, 1.587–5.305). ConclusionsSEMS placement offers sufficient short-term relief despite considerable major complications. Stricture of the upper esophagus is an independent predictor of initial clinical failure and shorter stent patency. Initial clinical failure is an independent predictor of shorter survival.

The Significance of Pretreatment Thrombocytosis and Its Association With Neutrophilia in Patients With Surgically Treated Endometrial Cancer.

The aim of this study was to investigate the prognostic significance of a pretreatment thrombocytosis and its association with neutrophilia in patients with surgically treated endometrial cancer. The baseline characteristics and outcome data of 508 patients with surgically treated endometrial cancer between January 2000 and December 2010 were collected and retrospectively reviewed. The patients were separated into 4 groups according to their platelet counts and the neutrophil counts, and the progression-free and overall survival rates of the 4 groups were compared. A Cox proportional hazards regression model was used to explore the independent prognostic factors. Pretreatment thrombocytosis was found to be associated with advanced stage (P = 0.0186), nonendometrioid histology (P = 0.0139), a deeper myometrial invasion (P = 0.0103), lymphovascular space involvement (P = 0.0404), cervical involvement (P = 0.004), positive peritoneal cytology (P = 0.0198), lymph node metastasis (P = 0.0301), and more frequent treatment failure (P = 0.0006). Multivariate analysis demonstrated that an older age (hazard ratio [HR], 2.54; 95% confidence interval [CI], 1.46-4.51; P = 0.0009), advanced clinical stage (HR, 5.27; 95% CI, 2.94-9.86; P < 0.0001), lymphovascular space involvement (HR, 3.37; 95% CI, 1.74-7.07; P = 0.0002), and pretreatment thrombocytosis (HR, 4.99; 95% CI, 2.47-9.39; P < 0.0001) were significant predictors of survival. When examined according to clinical stage, pretreatment thrombocytosis was prognostically significant only in patients with stage III-IV disease. The neutrophil count in patients who display thrombocytosis was significantly greater than that observed in patients without thrombocytosis (median, 6702 vs 4406/μL; P < 0.0001). Moreover, patients who displayed both thrombocytosis and neutrophilia had significantly shorter survival than that in those with either thrombocytosis or neutrophilia alone. Presence of thrombocytosis at the time of the initial diagnosis is an independent predictor of shorter survival in patients with advanced-stage (stages III-IV) endometrial cancer. Moreover, pretreatment thrombocytosis and concurrent neutrophilia are an independent predictor of shorter survival regardless of clinical stage.

SIX1 overexpression predicts poor prognosis and induces radioresistance through AKT signaling in esophageal squamous cell carcinoma.

The Sineoculis homeobox homolog 1 (SIX1) protein has been found to be overexpressed in several human cancers. However, its expression pattern and biological roles in esophageal squamous cell carcinoma (ESCC) remain unexplored. This study examined the clinical significance of SIX1 in 119 ESCC tissues. It was found that SIX1 protein was upregulated in 36.9% (44/119) cases. SIX1 overexpression was an independent predictor for short survival of ESCC patients. siRNA knockdown and plasmid transfection were carried out in ESCC cell lines. SIX1 depletion inhibited cell growth, invasion, and colony formation, whereas its overexpression facilitated in vivo and in vitro cell growth, invasion, and colony formation. The apoptosis rate induced by X-ray irradiation was substantially increased by SIX1 knockdown in Eca-109 cells. Ectopic overexpression of SIX1 in TE-1 cells dramatically enhanced resistance to irradiation. Western blot analysis showed that SIX1 depletion downregulated cyclin E, matrix metalloproteinase-2 (MMP-2), Bcl-2 expression and upregulated Bim expression. SIX1 overexpression exhibited the opposite effect on these proteins. In addition, it was found that SIX1 could positively regulate extracellular signal-regulated kinase (ERK) and AKT signaling pathway. ERK inhibitor abolished the effect of SIX1 on MMP-2 expression. AKT inhibitor treatment blocked the role of SIX1 on anti-apoptotic protein Bcl-2. In conclusion, this study demonstrates that SIX1 overexpression predicts poor survival in ESCC patients and confers radioresistance through activation of AKT signaling pathways.

MicroRNA-203 predicts human survival after resection of colorectal liver metastasis.

BackgroundResection of colorectal liver metastasis (CRLM) can be curative. Predicting which patients may benefit from resection, however, remains challenging. Some microRNAs (miRNAs) become deregulated in cancers and contribute to cancer progression. We hypothesized that miRNA expression can serve as a prognostic marker of survival after CRLM resection.ResultsMiR-203 was significantly overexpressed in tumors of short-term survivors compared to long-term survivors. R1/R2 margin status and high clinical risk score (CRS) were also significantly associated with short-term survival (both p = 0.001). After adjusting for these variables, higher miR-203 expression remained an independent predictor of shorter survival (p = 0.010). In the serum cohort, high CRS and KRAS mutation were significantly associated with short-term survival (p = 0.005 and p = 0.026, respectively). After adjusting for CRS and KRAS status, short-term survivors were found to have significantly higher miR-203 levels (p = 0.016 and p = 0.033, respectively).Materials and MethodsWe employed next-generation sequencing of small-RNAs to profile miRNAs in solid tumors obtained from 38 patients who underwent hepatectomy for CRLM. To validate, quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was performed on 91 tumor samples and 46 preoperative serum samples.ConclusionsAfter CRLM resection, short-term survivors exhibited significantly higher miR-203 levels relative to long-term survivors. MiR-203 may serve as a prognostic biomarker and its prognostic capacity warrants further investigation.

Early stridor onset and stridor treatment predict survival in 136 patients with MSA.

To evaluate the predictive value of stridor and its latency of onset and to investigate the role of stridor treatment in a cohort of patients with multiple system atrophy (MSA) referred to a tertiary center. We retrospectively identified patients diagnosed with MSA referred to our department beginning in 1991 and evaluated at least yearly during the disease course. Stridor was defined as present when confirmed by a whole night video-polysomnography and as early if presenting within 3 years of disease onset. Survival data, from disease onset to time of death, were calculated with Kaplan-Meier curves. Predictors were identified in univariate and multivariable Cox regression analyses. We included 136 patients with MSA; 113 were deceased at the time of study. Stridor was diagnosed in 42 patients, and 22 presented early stridor onset. Twelve of the 31 patients treated for stridor received tracheostomy, and 19 received continuous positive airway pressure. Overall survival did not differ between patients with and without stridor, while patients with early stridor onset had a worse prognosis than those developing this symptom later. In the stridor subgroup, early stridor onset was an unfavorable survival predictor. Stridor treatment was significantly associated with survival in our population. The Kaplan-Meier curve did not reveal significant differences in survival between the 2 treatments even though there was a trend toward longer disease duration in patients receiving tracheostomy. Our results demonstrated that early stridor onset is an independent predictor for shorter survival and that tracheostomy could control stridor, influencing disease duration.

High expression of TRF2, SOX10, and CD10 in circulating tumor microemboli detected in metastatic melanoma patients. A potential impact for the assessment of disease aggressiveness.

Circulating tumors cells (CTCs) can be detected in the blood of metastatic melanoma patients (MMPs) both as isolated circulating tumor cells (iCTCs) and circulating tumor microemboli (CTMs), but their clinical significance remains unknown. The aim of this work was to evaluate the prognostic impact in metastatic cutaneous melanoma of CTMs and iCTCs identified by a cytomorphological approach using the isolation by size of tumor cell (ISET) method. We characterized the phenotype of CTCs using anti‐PS100, anti‐SOX10, anti‐CD10, and anti‐TRF2 antibodies. 128 MMPs and 37 control healthy individuals with benign nevi were included in this study. Results were compared to the follow‐up of patients. 109/128 (85%) MMPs showed CTCs, 44/128 (34%) with 2 to 6 CTMs and 65/128 (51%) with 4 to 9 iCTCs. PS100 expression was homogeneous in iCTCs and heterogeneous in CTMs. SOX10, CD10, and TRF2 were mainly expressed in CTMs. None of the control subjects demonstrated circulating malignant tumor cells. Overall survival was significantly decreased in patients with CTMs, independently of the therapeutic strategies. In conclusion, the presence of CTMs is an independent predictor of shorter survival from the time of diagnosis of MMPs.

Body mass index and age are additional prognostic factors in patients with metastatic renal cell carcinoma treated with tyrosine kinase inhibitors

PurposeTyrosine kinase inhibitors (TKIs) are the preferred treatment drugs for metastatic renal cell carcinoma (mRCC). The aim of this study was to analyze prognostic factors for overall survival (OS) in patients with mRCC treated with TKIs. Materials and methodsClinical data on 155 patients enrolled in 5 clinical trials conducted at our hospital from 2006 to 2014 were reviewed retrospectively. All patients received first-line TKI therapy (sunitinib, sorafenib, pazopanib, or famitinib). Survival rates were determined by the Kaplan-Meier method. ResultsMedian OS (mOS) was 36.2 months. A total of 4 of the 5 adverse prognostic factors identified by the Memorial Sloan-Kettering Cancer Center (MSKCC) were found to be independent predictors of shorter survival, anemia, hypercalcemia, lactate dehydrogenase>1.5×upper limit of normal, and diagnosis to treatment time<1 year. In addition, we found that age≤45 years (P = 0.002), low or high body mass index ([BMI]<19 or>30kg/m2) (P = 0.003), and presence of≥3 metastatic sites (P = 0.000) were also independent adverse prognostic factors. According to the MSKCC model, the mOS time in the favorable-risk, intermediate-risk, and poor-risk groups were 46.6, 30.4 and 16.7 months, respectively (P = 0.005). When we integrated age and BMI into the MSKCC model to set up a 7-factor prognostic model, we found the mOS time for these 3 groups was 71.1, 41.6 and 15.3 months, respectively (P = 0.000). ConclusionAge and BMI are additional independent prognostic factors for patients with mRCC receiving vascular endothelial growth factor receptor-targeted TKI treatment, and the MSKCC prognostic model is also applicable to them. A 7-factor prognostic model might help to identify patients with the best prognosis. Further studies are needed to confirm these findings.

Low Vitamin D Levels Are Associated with Inferior Survival Following Azacitidine Treatment in Patients with Myelodysplastic Syndrome

▪Introduction: Azacitidine (AZA) therapy has become the recommended first-line treatment for patients with high-risk myelodysplastic syndromes (MDS) and oligoblastic acute myeloid leukemia (AML; bone marrow blasts <30%). However, only around 50% of patients achieve objective responses with AZA therapy and the vast majority of responding patients have disease progression within 2 years resulting in dismal survival rates.Vitamin D (VitD) is a central regulator of calcium and bone homeostasis and affects multiple signaling pathways controlling proliferation, apoptosis and differentiation. More than 30 years ago, VitD was demonstrated to induce in vitro differentiation of AML blast cells. Consequently, VitD alone or in combination with other differentiating agents has been used for treatment of MDS patients showing inconsistent and rather limited efficacy. We retrospectively tested the hypothesis that VitD levels prior to start of treatment are predictive of overall survival (OS) in newly diagnosed MDS and oligoblastic AML patients receiving upfront AZA therapy. In addition, the antiproliferative effects of AZA in combination with different VitD derivatives were investigated in vitro.Patients, materials and methods: A total of 58 patients treated at our center between 2006 and 2014 had serum samples available for assessment of VitD status. Serum samples were collected at a median of 18 days prior to start of AZA treatment and cryopreserved at -80°C. VitD status was assessed by measurement of circulating 25-hydroxyvitamin D levels applying a standard chemiluminescent immunoassay (DiaSorin Deutschland GmbH, Dietzenbach, Germany). Overall survival (OS) was estimated using the method of Kaplan and Meier. Survival times were measured from the date of AZA treatment start. Comparison of OS between the VitD groups was done using the logrank test and by cox regression adjusting for known confounders.For in vitro analyses, a tetrazolium based MTT assay was used for assessment of growth inhibition of two different AML cell lines (HL60 and MOLM13) after exposure to AZA alone or in combination with VitD (1alpha,25-dihydroxyvitamin D and 25-hydroxyvitamin D; both from Cayman Chemical, Ann Arbor, MI, USA) or the VitD antagonist TEI-9647 (kindly provided by Teijin Pharma Ltd., Tokyo, Japan). Drug interactions were analyzed using the median-effect method of Chou and Talalay and combination index (CI) values were calculated according to the classic isobologramm equation with CI<1, CI=1 or CI>1 corresponding to synergism, additivity or antagonism, respectively. In cases where no antiproliferative activity upon single-agent treatment was assessable (low-dose VitD and TEI-9647), an analysis in terms of sensitization (potentiation) or inhibition of AZA activity was performed instead.Results: Estimated median follow-up time was 14.2 months. Median serum VitD level prior to AZA treatment was 33 nM (range 11-102 nM). A total of 18 patients underwent allogeneic stem cell transplantation (alloSCT) following AZA therapy and follow-up was censored at the time of alloSCT. Patient, disease and treatment characteristics did not differ significantly between the low (≤33 nM; n=29) and high (>33 nM; n=29) VitD group. Estimated one-year survival was 72% (95% CI 54-90%) in the high VitD group, which was significantly longer as compared to the low VitD group (41%, 95% CI 20-62%, p<0.05; Figure 1). In multivariate analysis with OS as endpoint, VitD (per 10 nM decrease, HR 1.83 95% CI 1.06-3.16, p=0.03) and adverse cytogenetics (HR 2.58 95% CI 1.10-6.06, p=0.03) were independent predictors of shorter survival. In vitro treatment of HL60 and MOLM13 cells with AZA in combination with VitD produced synergistic and additive antiproliferative effects. Addition of nanomolar concentrations of VitD to AZA resulted in potentiation of AZA activity. Conversely, combination with TEI-9647 resulted in inhibition of AZA activity.Conclusions: Our study suggests that higher VitD levels were associated with a survival advantage following upfront AZA therapy. As compared to AZA monotherapy combination treatment with VitD resulted in enhanced cytotoxic effects in vitro. VitD repletion/supplementation during AZA treatment should therefore be explored. [Display omitted] DisclosuresLuft:Immundiagnostik AG: Research Funding.

Clinical Significance and Pathopysiological Function of the Tim-3/Galection-9 Pathway in Myelodysplastic Syndromes

Abstract Background: Galectin-9 (Gal-9), a member of the galectin family, plays a crucial role in inflammation and tumorigenesis involving angiogenesis, cell adhesion, immune escape, and cancer cell survival. In acute myeloid leukemia (AML), Gal-9 is highly expressed in some AML cell lines and binds with T-cell immunoglobulin- and mucin-domain-containing molecule-3 (Tim-3) on T cells and leukemic stem cells. The Tim-3/Gal-9 pathway may be associated with disease progression by inducing T-cell dysregulation and mediating autocrine proliferation of leukemic cells. However, the role of the pathway in myelodysplastic syndromes (MDS) remains unclear. In the current study, we investigated the prognostic impact of plasma Gal-9 levels and the pathophysiological roles of Gal-9 and Tim-3 in MDS. Methods and results: 1) To investigate the prognostic impact of Gal-9, plasma Gal-9 levels were measured by ELISA in 92 MDS patients: 30 with refractory anemia (RA); 3 with RA with ringed sideroblasts (RARS); 18 with RA with excess blasts (RAEB); 14 with RAEB in transformation (RAEB-t); and 29 with acute leukemia transformed from MDS (AL-MDS). The plasma Gal-9 level increased with disease progression (mean ± SD: control 3.4 ± 0.9; RA, 9.3 ± 5.7; RARS, 10.2 ± 6.8; RAEB, 9.7 ± 6.5; RAEB-t, 17.2 ± 9.6; AML-MDS, 20.1 ± 14.1 ng/ml). Furthermore, patients with AL-MDS in International Prognostic Scoring System (IPSS) high-risk categories (Intermediate-2/High) had significantly higher Gal-9 levels compared with IPSS low-risk (Low/Intermediate-1) patients (Low/Intermediate-1 vs Intermediate-2/High, p = 0.002; Low/Intermediate-1 vs AML-MDS, p = 0.004). The overall survival of MDS patients with plasma Gal-9 higher than the normal level (10 ng/mL) was significantly shorter than that of other patients (log-rank p &lt; 0.001) even in lower-risk (RA, RARS) cases (log-rank p = 0.0369). Moreover, multivariate analysis showed that a high level of Gal-9 was an independent predictor of shorter survival in MDS patients as well as IPSS. 2) Bone marrow mononuclear cells were isolated from some MDS patients, and Tim-3 expression on granulocytes, monocytes, lymphocytes, and blasts was analyzed using flow cytometry (FCM). Tim-3 expression on blasts tended to be higher in AL-MDS patients than in MDS patients. 3) To investigate whether MDS cells can produce Gal-9, we analyzed Gal-9 levels in supernatants of cell cultures of 3 human AL-MDS cell lines, F-36P, SKM-1, and MDS-L, and MDS blasts obtained from an AL-MDS patient by ELISA. Gal-9 was detected in all cell culture supernatants and its concentration was higher when cultured in medium containing 10% fetal bovine serum (FBS) than in 2.5% FBS medium in F-36P and SKM-1 cells. The concentration of Gal-9 produced from MDS blasts was increased time dependently in 2-6 day culture. Furthermore, recombinant human Gal-9 enhanced the cell proliferation of F-36P cells. 4) After analyzing Tim-3 expression in MDS cell lines using real-time quantitative PCR and FCM, all cell lines expressed relatively low levels of Tim-3 mRNA. Extracellular expression of Tim-3 was detected and induced by adding cell culture supernatants of the human stromal cell line HS-5 in F-36P cells. To elucidate the signaling pathway inducing Tim-3 expression, we investigated Tim-3 mRNA expression in F-36P cells treated with HS-5 supernatant with several signal transduction inhibitors (STAT3 inhibitor, U0126; MAPK/ERK; AG490; JAK2, LY294002; PI3K, PDTC; NF-κB inhibitor). Upregulation of Tim-3 gene expression in F-36P cells treated with HS-5 supernatant was inhibited by the MEK inhibitor U0126. These results suggest that the MAPK/ERK pathway is involved in Tim-3 expression induced by HS-5 supernatant. Conclusions: In MDS, the elevation of plasma Gal-9 levels is associated with disease progression, including leukemic transformation, and with shorter survival. Furthermore, its receptor Tim-3 is upregulated on blasts in AL-MDS patients. Our data suggest that the Gal-9/Tim-3 pathway plays a key role in MDS disease progression. Although further study is required to clarify the detailed functions of the interaction between Gal-9 and Tim-3, the current study could lead to the development of a new immunotherapeutic strategy via the blockade of this pathway in MDS. Disclosures No relevant conflicts of interest to declare.

Abstract C19: Use of liquid biopsies in clinical oncology: the UCSD Moores Cancer Center experience in 168 patients with diverse malignancies

Abstract There is a growing interest in using methods such as liquid biopsies to assess genomics of circulating tumor DNA (ctDNA) in patients with advanced cancer. Liquid biopsies are non-invasive, may provide access to shed DNA from multiple metastases, and the dynamic results of genomic evolution can be followed with repeat blood sampling. We report our initial experience using ctDNA testing in 168 consecutive patients with diverse cancers. Testing was performed using a panel of 54 cancer-related genes (digital sequencing), examining mutations (in all 54 genes) as well as amplifications in ERBB2, EGFR, and MET (Guardant Health, Inc. (Guardant360 test)). Fifty-eight percent of patients (98/168) had ≥ 1 alteration(s) identified in their plasma. In a multivariable analysis, gastrointestinal cancer (N = 13) was an independent predictor of a higher number of alterations (median = 2 alterations/patient; P = 0.001), while primary brain tumors (N = 56) correlated with fewer alterations (median = 0 alterations/patient; P = 0.019) detected in the plasma. Even so, 16 of 56 patients (28.6%) with primary brain tumors had discernible ctDNA alterations. Of the 98 patients with alterations, 71.4% had ≥ 1 anomaly potentially actionable by an FDA-approved drug. When comparing results of patients (N = 63) who had both a tissue and ctDNA test performed (and had aberrations in the tissue biopsy that were detectable by the ctDNA panel), 22 individuals (35%) had ≥ 1 alteration in common between the tissue and ctDNA test (median time interval between the tissue biopsy and the blood draw = 2.7 months for these 22 patients). In contrast, 41 patients had no alterations in common between tissue and ctDNA, and in these 41 individuals, the median time between tissue biopsy and blood draw was 14.4 months (P = 0.006). Overall concordance rates were 70.3% for TP53 and EGFR, 88.1% for PIK3CA, and 93.1% for ERBB2 alterations. In addition, we observed a strong correlation between cases with ≥ 1 alteration reported with ctDNA ≥ 5% and shorter overall survival (median = 4.0 months versus not reached; P&amp;lt;0.001 in multivariable analysis; median follow up time = 6.1 months). Finally, 5 of 12 patients (42%) matched to a therapy according to their ctDNA test results achieved stable disease ≥ 6 months (N = 2) or partial response (N = 3). Taken together, ctDNA testing via liquid (blood) biopsies demonstrated frequent potentially actionable aberrations, including in primary brain tumors. The longer the time elapsed between tissue and ctDNA procurement for testing, the less chance that genomic alterations would be consistent between the two specimens. Higher percentage of ctDNA was an independent predictor for shorter survival. Finally, preliminary results suggest that patients matched to cognate targeted treatments based on ctDNA can achieve clinical benefit. Overall, these observations indicate that ctDNA genomic analysis may be a valuable tool for evaluating patients with cancer. Citation Format: Maria C. Schwaederle, David E. Piccioni, Santosh Kesari, Hatim Husain, Sandip P. Patel, Razelle Kurzrock. Use of liquid biopsies in clinical oncology: the UCSD Moores Cancer Center experience in 168 patients with diverse malignancies. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr C19.

Paraneoplastic pemphigus in a patient with Chronic Lymphocytic Leukemia (CLL)

*test repeated due to value >100 units/mL serum ME-monkey esophagus, DSG-desmoglein, Abstracts patients <60 years (1.27; p1⁄40.027). Higher SIR was shown for secondary leukemia, melanoma and head and neck cancers, while a lower SIRwas found for lung, digestive tract, and bladder cancers. The development ofOCwas associatedwith shorter survival: 16.2 years for patients with OC vs. 22.9 years for patients without OC (p1⁄40.035). Factors that independently predicted development of OC were advanced age, male gender and lower platelet count. Conclusions: We confirmed in a large cohort of LTS patients with CLL that the overall incidence of OC is increased as compared to the general population. An increased risk of OC may persist indefinitely in patients with CLL, independently from their treatment status. In these patients the presence of OC represents an independent predictor of shorter survival.

Multiple primary cancers involving lung cancer at a single tertiary hospital: Clinical features and prognosis.

BackgroundThe development of other primary cancers in patients with lung cancer is unfortunate and uncommon, although the frequency is increasing. The aim of this study was to determine the clinical features and prognosis in patients with multiple primary cancers (MPC) involving lung cancer.MethodsAfter a retrospective review of 1644 patients who were newly diagnosed with primary lung cancer between 1998 and August 2012 at a tertiary hospital, 105 patients were included.ResultsThe median age at the time of lung cancer diagnosis was 67 years, and 68 patients were male. Synchronous primary cancers occurred in 47% of the study population (49/105). Among those with metachronous cancer (56/105), the median interval between the diagnosis of lung cancer and another malignancy was 47.1 months; 21 patients were diagnosed with lung cancer as the first primary tumor. The most frequent type of other malignancy was urogenital (30%), followed by gastrointestinal (30%) and thyroid malignancies (16%). Advanced stage of lung cancer (hazard ratio (HR), 3.2; 95% confidence interval (CI), 1.8–5.7; P < 0.001), supportive care only as treatment for lung cancer (HR, 2.8; 95% CI, 1.3–6.0; P = 0.006), and head and neck cancer as another malignancy (HR, 3.9; 95% CI, 1.4–10.8; P = 0.010) were independent predictors of shorter survival from the time of diagnosis of the second primary cancer.ConclusionAdvanced lung cancer stage, symptomatic supportive care only without antitumor therapy for lung cancer, and head and neck cancer as another primary malignancy were poor prognostic factors in patients with MPC involving primary lung cancer.

Other Cancers in Long-Term Survivors of Chronic Lymphocytic Leukemia: Incidence and Prognostic Relevance

Introduction. The clinical course of chronic lymphocytic leukemia (CLL) is mostly indolent. About one third of the patients are managed with lifelong watch-and-wait (WW) and those who receive therapy often achieve a durable remission. As a result, the majority of patients with CLL will live with their disease for long periods of time, and be exposed to several complications, including the occurrence of other cancers (OC). Patients with CLL may have an increased incidence in OC. Published reports indicate an incidence of 3-27%, mostly in treated patients, however, very little is known on OC in patients with CLL not requiring therapy. Furthermore, observation time in published studies is limited to <5 years, and the incidence of OC in patients followed for longer than 10 years is unknown. We, therefore, studied the incidence and prognostic impact of OC in treatment-naïve patients with CLL followed for ≥10 years.Methods. We reviewed our database and identified all patients with CLL untreated at the time of referral. We selected long-term survivors (LTS), defined as patients with a follow-up ≥10 years, and analyzed the incidence and prognostic impact of OC in this population. Non-melanoma skin cancers were excluded since these were diagnosed and treated promptly in virtually all cases and felt not to have prognostic impact. Standardized incidence ratios (SIR) were calculated for OC occurring after the diagnosis of CLL that were reportable to the Surveillance, Epidemiology and End Results program.The estimated overall survival (OS) according to the presence of OC was plotted considering OC as a time-dependent covariate.Results. We identified 797 LTS of CLL seen at our institution between 1957 and 2003. Median age was 56 years (24-88). 57% of patients were males. Median follow-up for the entire population is 154 months (120-485). We recorded 383 OC in 286 (36%) patients. 76/286 (26%) patients had >1 OC (62 had 2 OC, 10 had 3, 2 had 4, 1 had 5 and 1 had 6).The firstOC preceded or was diagnosed concomitantly with CLL in 100 patients (35%), while in the remaining 186 (65%) it occurred later during the course of the disease. 570 patients (71%) required treatment for CLL. Median time to treatment was 18 months (0-454). In treated patients, the cumulative frequency of OC was 205/570 (36%) and in WW patients 81/227 (36%). The SIR for all OC was 1.2 (p = .034). Males and patients younger than 60 years had a significantly higher incidence of OC (SIR 1.31 and 1.27, respectively). Among OC types, secondary leukemia, melanoma and head and neck cancers had the highest observed-to-expected ratio. Surprisingly, lung, digestive tract, and bladder cancer had a lower-than-expected incidence (table). 474 patients (59%) are alive. 222/570 (39%) treated patients and 101/227 (44%) WW patients have died. The median OS was longer in patients without OC (279 months) vs. those with OC (189 months). Independent predictors of shorter survival in multivariate analysis included higher creatinine, the presence of OC, and older age.Discussion. This is the first study to address the incidence of OC in LTS of CLL, including WW patients. In our population, the frequency of OC is similar in treated and WW patients. Although the incidence of OC in LTS of CLL is higher compared to matched general population, the incidence of lung, digestive and bladder cancer is lower than expected. Reasons of this finding remain to be identified.The occurrence of OC is an independent predictor of shorter survival, thus constituting a relevant competing risk of mortality in LTS of CLL.VariableObservedExpectedPerson-yearsSIR (O/E)95% CI for O/EP -valueOverall148123.34109561.201.01 – 1.400.034Male9673.458851.311.06 – 1.580.013Female5249.9350711.040.78 – 1.360.67Age ≥60 years6054.3334161.100.84 – 1.420.44Age <60 years8869.0275401.271.02 – 1.570.027OC typeProstate2825.92118091.080.72 – 1.560.64Lung2029.08119420.690.42 – 1.060.04Breast1918.60118551.020.62 – 1.590.96Melanoma164.23119263.782.16 – 6.140.00Leukemia154.27120093.511.96 – 5.790.00Non-Hodgkin lymphoma66.38119960.940.34 – 2.051.00Digestive1640.4119370.400.23 – 0.640.00Colon819.42119720.410.18 – 0.810.006Pancreas24.83120240.410.05 – 1.490.18Rectal38.69120110.340.07 – 1.000.05Bladder311.18119930.270.05 – 0.780.009Multiple Myeloma21.98120121.010.12 – 3.641.00Lip30.021201515031.00 – 438.50.00Salivary gland20.031202666.668.00 – 240.060.00 DisclosuresNo relevant conflicts of interest to declare.

826P - Contribution of Tumor Burden and Body Composition Parameters As Prognostic Factors of Metastatic Renal Cell Carcinoma (Mrcc) Treated By Targeted Therapy

ABSTRACT Aim: We previously showed that tumor burden (TB) and skeletal muscle density (SMD) at baseline are independent prognostic factors for overall survival (OS) in mRCC. We aimed to analyze the respective roles of baseline TB and Heng score, baseline and 12 week- body mass index (BMI) (weight /height2) and body composition (BC) on OS in mRCC included in trials evaluating targeted therapy. Methods: Baseline and 12 weeks- TB, BMI, adipose tissue (AT), muscular muscle (MM) and SMD were assessed using previously reported techniques. To withdraw causes of early death, OS was estimated from 12 weeks after therapy initiation, using Kaplan-Meier method and compared across strata using log-rank test. To identify prognostic factors, impact of baseline and 12 week- body components were assessed in a multivariable Cox proportional hazards models adjusted for Heng risk groups. Results: 116 pts, treated with sorafenib (n = 35), sunitinib (n = 33) or everolimus (n = 18) or placebo (n = 30) with full data for TB, BMI and BC, have been analyzed. BMI was low ( 25 kg2/m2) in 3 (3%), 47 (41%) and 66 (56%) pts respectively. Median OS was 18 months, 95% Confidence Interval (CI) = [16;21]. Median estimates and hazard ratios derivated from the multivariable Cox model were: Median OS (weeks) Hazard Ratio (95%CI) p-value Heng risk High 6 1 - 0.0002 Intermediate 45 0.2 0.1 – 0.6 Favorable 112 0.1 0.0 – 0.3 TB >= 19 cm 76 1 - 0.0002 9.5-19 cm 85 1.2 0.7 – 2.1 161 0.4 0.2 – 0.7 BMI Normal 112 1 - 0.007 Low 154 0.8 0.2-3.2 High 66 1.9 1.2 – 3.1 SMD and BC were not independently associated with OS. Conclusions: High/intermediate risk Heng score, large TB and high BMI are independent predictors of shorter survival in pts alive after 12 weeks of treatment. Mechanisms underlying the surprising deleterious effect of high BMI could not be determined from body composition impact analysis. After adjustment on TB, high muscle density is not protective any more. Disclosure: Y. Loriot: Advisory board Bayer; B. Escudier: Advisory board Bayer, Pfizer, Novartis Honoraria from Bayer, Roche, Pfizer, Genentech, Novartis, AVEO, GSK. All other authors have declared no conflicts of interest.

Comparison of outcomes between squamous cell carcinoma and adenocarcinoma in patients with surgically treated stage I-II cervical cancer.

To improve our understanding of cervical adenocarcinoma (AD) and evaluate the clinical and pathological variables affecting its prognosis, we retrospectively reviewed the medical records of 455 patients with cervical cancer [International Federation of Gynecology and Obstetrics stage I/II; 91 cases with AD and 364 with squamous cell carcinoma (SCC)] who underwent surgery at our hospital between January, 1995 and August, 2012 and compared the characteristics and prognoses between AD and SCC cases, including age, clinical stage, histological type, lymph node metastasis, lymphovascular space invasion (LVSI), cervical stromal invasion, parametrial invasion, vaginal invasion, corpus invasion, ovarian metastasis and tumor diameter. We used Cox regression analysis to determine independent prognostic factors. AD was found to have a significantly poorer prognosis in all the patients (P=0.001), stage I patients (P=0.001) and stage IB patients (P<0.05). The prognosis did not differ in patients who did not require postoperative treatment; however, patients who received postoperative treatment exhibited a significantly poorer prognosis (P<0.05). Patients with AD who received postoperative irradiation alone had a significantly poorer prognosis (P<0.05). The multivariate analysis identified LVSI (P=0.008), stromal invasion (P=0.024) and ovarian metastasis (P=0.032) as independent predictors of shorter survival. AD was associated with a worse prognosis compared to SCC in patients with stage IB disease, particularly in those who required postoperative treatment. Such patients may benefit from individualized postoperative treatments that differ from those applied for SCC.

Pretreatment neutrophil/lymphocyte ratio as a prognostic aid in colorectal cancer.

Colorectal cancers(CRC) are the third most common cancer in the western world, with surgery preferred for management of non-metastatic disease and post surgical treatment usually arranged according to the TNM staging system. However, there is still prognostic variation between patients who have the same stage. It is increasingly recognized that variations within disease course and clinical outcome in colorectal cancer patients are influenced by not only oncological characteristics of the tumor itself but also host response factors. Recent studies have shown correlation between the inflammatory response and clinical outcomes in various cancers. The neutrophil/lymphocyte ratio (NLR) has been described as a marker for immune response to various stimuli including cancer. Two hundred eighty-one CRC patients were included in our retrospective analysis, separated into two groups according to a cut-off value for the NLR. Patient data including age, gender, vertical penetration, anatomic location, and differentiation of the tumor, TNM stage, survival rate, and disease-free survival were analyzed for correlations with the NLR. Using ROC curve analysis, we determined a cut-off value of 2.2 for NLR to be best to discriminate between patient survival in the whole group. In univariate analysis, high pretreatment NLR (p=0.001, 95%CI 1.483-4.846), pathologic nodal stage (p<0.001, 95%CI 1.082- 3.289) and advanced pathologic TNM stage (p<0.001, 95%CI 1.462-4.213) were predictive of shorter survival. In multivariate analysis, advanced pathologic TNM stage (p=0.001, 95%CI 1.303-26.542) and high pretreatment NLR (p=0.005, 95%CI 1.713-6.378) remained independently associated with poor survival. High pre-treatment NLR is a significant independent predictor of shorter survival in patients with colorectal cancer. This parameter is a simple, easily accessible laboratory value for identifying patients with poorer prognosis.

Impact of NPM, TFF3 and TACC1 on the Prognosis of Patients with Primary Gastric Cancer

BackgroundNPM, TFF3 and TACC1 are molecular markers that play important roles in cell differentiation. Herein, we investigated their prognostic impact in patients with primary gastric cancer (GC) and determined whether they could be used as markers of more aggressive gastric carcinomas by detecting the extent of expression in human gastric carcinoma samples.Methodology/Principal FindingsTumor tissue specimens from 142 GC patients were retrospectively retrieved and immunohistochemically evaluated. Correlations between NPM, TFF3 and TACC1 over-expression and clincopathologic parameters, and their prognostic values were investigated with χ2, Kaplan-Meier method, and Cox uni- and multivariate survival models. NPM, TFF3 and TACC1 expression was significantly higher in GC patients with poorly differentiated histologic type than that in patients with well differentiated histologic type. NPM expression was significantly higher in patients with hepatic metastasis or recurrence than that in patients without metastasis. TFF3 expression was significantly higher in patients with positive lymph node metastasis than that in patients with negative lymph node metastasis. Age, lymph node metastasis, and TFF3 and TACC1 over-expression were significantly correlated with low survival (P<0.05, P<0.05, P = 0.005 and P = 0.009, respectively). Multivariate analysis showed that lymph node metastasis and TFF3 and TACC1 over-expression were independent prognostic factors.ConclusionsTFF3 and TACC1 over-expression in epithelial cells of surgically resected GC tissues was an independent predictor of short survival in GC patients. The prognosis was poorer in patients with positive expression of both TFF3 and TACC1 than that in patients with positive expression of TFF3 or TACC1 alone, or with negative expression of TFF3 and TACC1.

Genomic Complexity Versus Gene Mutations In Predicting Overall Survival In Adult Acute Myelogenous Leukemia

IntroductionGenomic complexity as measured through SNP array-based (SNP-A) genomic profiling is a strong negative predictor of survival outcome in adult acute myelogenous leukemia (AML). The recent discovery of multiple novel recurrently mutated genes in AML has led to the development of several prognostic models based on various combinations of genes along with the well-established risk factors of age and karyotype, but these models do not account for the strong effect of SNP-A-based genomic complexity. In this study, we seek to determine the relative importance of AML genomic complexity and gene mutation status on overall survival (OS) in AML. MethodsWe employed SNP-A genomic profiling of acquired copy number aberrations (aCNA) and copy neutral LOH (cnLOH) together with sequence analysis of 13 recurrently mutated genes to determine aCNA/cnLOH lesion load and gene mutational status for 156 consecutively collected previously untreated AML patients. AML cell samples were processed with a Ficoll gradient, negatively selected using Miltenyi microbead columns, and then further purified with flow cytometric cell sorting. Processed DNA isolated from highly purified AML blasts and paired buccal DNA was subsequently hybridized to Affymetrix SNP 6.0 arrays. aCNAs were visually identified using the dChip program in paired data displays and corroborated by algorithmic lesion scoring, and cnLOH was detected using internally developed software. Using the same DNA, we resequenced the recurrently mutated exons of NPM1, FLT3, CEBPA, IDH1, IDH2, NRAS, KRAS, and TP53, and all coding exons of RUNX1, ASXL1, TET2, DNMT3A, and BCORL1. Clinical data including age, overall survival, and cytogenetics were ascertained. Cytogenetic risk categories were assigned based on the SWOG criteria. Univariate analyses were performed using Kaplan-Meier estimates of survivor functions, and Cox proportional hazards models were used for multivariate analyses. ResultsAt the time of analysis, 119 (76%) patients had died. aCNA/cnLOH were common with ≥1, ≥2, and ≥3 lesions detected in 62%, 38%, and 26% of cases, respectively. Univariate overall survival analysis of all clinical and molecular variables demonstrated a significantly increased OS for mutations of NPM1 (p=0.01) and decreased OS for mutations of TP53 (p<0.001), aCNA/cnLOH load ≥2 vs. <2 (p=0.001), aCNA/cnLOH load ≥3 vs. <3 (p<0.001), age >60 (p<0.001), unfavorable vs. intermediate cytogenetics risk (p<0.001), and intermediate vs. favorable cytogenetic risk (p=0.01). Mutations in the other 11 genes analyzed were not prognostic. Multivariate analysis inclusive of either genomic complexity or gene mutations and always incorporating age- and cytogenetic-based risk categories demonstrated a statistically significant difference in the hazard of death for aCNA/cnLOH load ≥2 (HR 3.13, p=0.03), aCNA/cnLOH load ≥3 (HR 6.53, p=0.001), and TP53 mutations (HR 24.32, p<0.001).Within the total cohort, 103 patients (66%) were diagnosed with de novo AML. In this group of de novo AML, aCNA/cnLOH were again common with ≥1, ≥2, and ≥3 lesions in 56%, 29%, and 18% of cases, respectively. Statistically significant univariate variables adversely affecting OS included mutations of TET2 (p=0.023) and TP53 (p<0.001), aCNA/cnLOH load ≥2 vs. <2 (p=0.02), aCNA/cnLOH ≥3 vs. <3 (p<0.001), age >60 (p<0.001), unfavorable vs. intermediate cytogenetics risk (p=0.02), and intermediate vs. favorable cytogenetic risk (p=0.04). Similarly, multivariate analysis revealed a statistically significant difference in the hazard of death for aCNA/cnLOH load ≥3 (HR 11.3, p=0.002) and TP53 mutation (HR 74.8, p<0.001). ConclusionGenomic complexity is common in adult AML and constitutes a dominant independent predictor of short survival outcomes. Through use of multivariate analysis incorporating genomic complexity and the mutation status of 13 recurrently mutated genes and controlling for age- and cytogenetic-based risk, only genomic complexity and mutations of TP53 emerge as significant independent predictors of short overall survival. Disclosures:No relevant conflicts of interest to declare.