No Evidence Of Disease Patients Research Articles

The outcome of COVID-19 infection among Saudi oncology patients.

e23085 Background: Little data are available surrounding the outcomes and risk factors of COVID-19 in Saudi oncological patients. This study aims to elucidate the outcomes of COVID-19 infection among Saudi oncology patients and associated risk factors. Methods: We retrospectively reviewed the medical files of oncology patients who were diagnosed with COVID-19 at the King Faisal Specialist Hospital and Research Centre. The time from a positive PCR until death or last follow-up was calculated as COVID-specific survival. A p-value less than 0.05 was selected as the threshold for statistical significance. Results: 161 oncological patients were included in our analysis. The mean age was 53.83±15.81. 85 (52.8%) patients were female, 17 (10.6%) had a positive history of smoking, and 57 (35.4%) were obese. 70 (43.5%) patients had diabetes mellitus, 64 (39.8%) had hypertension, and 25 (15.5%) had dyslipidemia. The most common cancer in the group was breast cancer, which was present in 24 (14.9%), followed by non-Hodgkin lymphoma, colon cancer, and acute leukemia, respectively. 57 (35.4%) patients had no evidence of disease (NED) at presentation. Furthermore, 65 (40.4%) patients had received antineoplastic therapy within 30 days before infection. 25 (15.5%) patients had stage A COVID-19, 59 (36.6%) had stage B, 48 (29.8%) had stage C, and 29 (18%) had stage D. There was no difference between active and no NED patients in remission in regard to the stages of COVID-19 infection at presentation (p = 0.05). However, the stages of COVID were less severe in patients who received antineoplastic therapy (p = 0.02). 32 (19.9%) patients received tocilizumab during the acute phase of infection. Only 5 (3.1%) patients suffered COVID-specific death. Therefore, the median COVID-specific survival was not reached. Tocilizumab (p = 0.34), corticosteroids (p = 0.77), ACE inhibitors (p = 0.45) had no impact on COVID-specific death in our patient population. In patients who received steroid treatment, the duration of corticosteroid had no impact on survival (HR: 0.76, p = 0.39). Ventilation also had no impact on survival (p = 0.55). There was no difference between patients who received antineoplastic therapy within 30 days from infection and those who did not (p = 0.99). Finally, we noted no difference in survival between patients with active disease and NED patients (p = 0.17). Conclusions: COVID-related mortality is lower in Saudi oncology patients compared to other countries. Early management and admission of oncological patients may result in overall improved outcome. In line with some studies, antineoplastic therapy can reduce COVID-19 severity, possibly due to reduced ACE receptor expression.

Immunological signature of patients with thymic epithelial tumors and Good syndrome: Correlation with clinical outcome.

e20648 Background: Thymic epithelial tumors (TETs) are extremely rare and heterogenous neoplasms, associated with immune dysregulation. Recognized prognostic factors are TNM stage and the Masaoka-Koga classification. To the best of our knowledge, no study has been conducted to investigate the correlation between immunophenotype, level of cytokines, chemochines and clinical outcome in TETs patients. The aim of this study was to evaluate the correlation of immunological signature and long-term outcome in patients with TETs and Good Syndrome (GS). Methods: From May 2019 to October 2022, consecutive patients with TETs and GS, referred to the Rare Tumors Coordinating Center of Campania Region (CRCTR) of Federico II of Naples, were enrolled. The immunophenotype of monocytes, neutrophils, eosinophils, CD4+ T cells, CD8 + T cells, B-cells, NK cells and NKT-cells, T regulatory cells, and the evaluation of serum levels of cytokines, chemokines and growth factors were assessed using the 8-color immunophenotyping kit, Treg detection kit and pre-formed Kits by Bioplex multiplex, respectively. D’Agostino-Pearson normality test was used to evaluate whether the continuous data were normally distributed, and a two-tailed t-test for independent samples was used. Overall survival (OS) analysis was performed generating Kaplan-Meier curves and Cox Univariate models for the considered categorical stratifications. p-values < 0.05 were considered statistically significant. Results: A total of 24 consecutive TETs patients were included in the study: 13 (54.17%) patients were male and 11 (48.83%) were female, with a median age of 54.96 ± 9.6 years. Tumor histology included 20 thymoma, and 4 thymic carcinoma. Autoimmune diseases were found in 13 patients all of whom had thymoma. At the time of this analysis, 5 (20.83%) patients had no evidence of disease (NED) by imaging and were in follow-up, 17 (70.83%) patients were still alive. The analysis of leucocytes did not show statistically significant association with clinical outcome of TETs patients. On the contrary, a statistically significant correlation between serum concentration of IL-6 and OS was found with lower levels of IL-6 associated with poorer prognosis (HR: 9.654; p = 0.016). In addition, we found a statistically significant reduction of IL-2 (p = 0.04), IL-15 (p = 0.048), IL-5 (p = 0.03) and VEGF (p = 0.01) serum levels in patients with evidence of disease, as compared to NED patients. Conclusions: Overall, these findings suggest that measurement of IL-6 levels may be a useful parameter for identifying TETs patients with GS who have more aggressive diseases. Our preliminary data may be useful in the clinical management of this complex disease. Further studies are needed to better understand the pathophysiology and clinical implications of immunophenotype alterations in TETs patients.

549 An RNA-lipoplex (RNA-LPX) vaccine demonstrates strong immunogenicity and promising clinical activity in a Phase I trial in cutaneous melanoma patients with no evidence of disease at trial inclusion

BackgroundLipo-MERIT is an ongoing, first-in-human, open-label, dose-escalation Phase I trial investigating safety, tolerability and immunogenicity of BNT111 in patients with advanced melanoma. BNT111 is an RNA-LPX vaccine targeting the melanoma tumor-associated antigens (TAAs) New York esophageal squamous cell carcinoma 1 (NY-ESO-1), tyrosinase, melanoma-associated antigen 3 (MAGE-A3), and transmembrane phosphatase with tensin homology (TPTE). A previous exploratory interim analysis showed that BNT111, alone or combined with immune checkpoint inhibition (CPI), has a favorable adverse event (AE) profile, gives rise to antigen-specific T-cell responses and induces durable objective responses in CPI-experienced patients with unresectable melanoma.1 Here, we present preliminary data in patients with no evidence of disease (NED) at trial inclusion in the BNT111 monotherapy subgroup.MethodsPatients with stage IIIB/C and IV pre-treated cutaneous melanoma were intravenously administered with BNT111 using a prime/repeat boost protocol. Patients were treated in seven dose escalation cohorts (7.2 to 400 µg total RNA) and three expansion cohorts to further explore dose levels of 14.4, 50 and 100 μg. In this analysis, patients receiving BNT111 monotherapy were grouped as having evidence of disease (ED) or NED, and immunogenicity, efficacy and safety were evaluated. Vaccine-induced immune responses were analyzed using an interferon-γ enzyme-linked immune absorbent spot (ELISpot) assay directly ex vivo.ResultsAs of May 24, 2021, 115 patients have received BNT111 within the Lipo MERIT trial. Of 71 patients treated with BNT111 monotherapy, 38 patients had ED and 33 patients had NED after prior therapies. Baseline characteristics were similar between the two groups. ELISpot data revealed comparable BNT111-induced T-cell responses against at least one TAA in ED vs. NED patients (14/22 [64%] and 19/28 [68%] patients with available ELISpot-evaluable samples, respectively), suggesting that BNT111 has the ability to induce T-cell immunity irrespective of the presence of a detectable tumor. As previously reported for ED patients, vaccine-induced CD4+ as well as CD8+ T-cell responses were also observed in NED patients, with a substantial fraction of de novo induced responses undetectable prior to vaccination. In NED patients, clinical efficacy was promising; median disease-free survival was 34.8 months (95% confidence interval: 7.0–not reached). The safety profile was similar in ED vs. NED patients; 38/38 (100%) and 32/33 (97%) patients experienced related treatment-emergent AEs, respectively, of which the majority were mild-to-moderate flu-like symptoms.ConclusionsImmunogenicity and safety profiles of BNT111 monotherapy were comparable in ED and NED patients. Promising signs of clinical activity were observed in NED patients.AcknowledgementsThe authors would like to acknowledge Camilla West (BioNTech SE) for medical writing support.Trial RegistrationClinicaltrials.gov: NCT02410733; EudraCT No. 2013-001646-33.ReferencesSahin U, Oehm P, Derhovanessian E, et al. An RNA vaccine drives immunity in checkpoint-inhibitor-treated melanoma. Nature 2020;585(7823):107–112.Ethics ApprovalEthics & Institutional Review Board approval was obtained prior to initiation of the trial (2018-13393_21-AMG federführend).

Skindex-29 scores indicate poor quality of life in early stage mycosis fungoides

BackgroundQuestionnaire tools are increasingly being used to assess health related quality of life (HRQOL) in mycosis fungoides (MF) patients. However, a thorough understanding of the factors that lead to poor HRQoL in early stage disease and their distribution across patient subgroups is lacking. ObjectivesTo characterize factors affecting HRQoL as assessed by Skindex-29 in subgroups of patients with early stage MF seen at a multidisciplinary cutaneous lymphoma clinic. MethodsSkindex-29, a multidimensional survey that evaluates HRQoL (emotions, symptoms, function), was distributed to early stage MF patients. Overall and component scores were analyzed in three groups: no evidence of disease (NED), active disease with limited early stage (AD-T1), and active disease with more extensive early stage (AD-T2). Scores were also compared among patients receiving different treatment modalities. Results56 patients (9 NED, 36 AD-T1, and 11 AD-T2) were enrolled in the study. Overall Skindex-29 scores and scores for individual dimensions were comparable among the three sub-groups. Similarly, these scores did not significantly differ among treatment groups or after removal of patients with previous staging higher than IB. Analysis of individual questions revealed that NED patients reported higher scores for questions pertaining to anger and fatigue. ConclusionsEarly stage MF patients enrolled in this study had high overall Skindex-29 scores. Surprisingly, Skindex-29 scores of NED patients were comparable to those of patients with active disease, T1 and T2, mostly due to anger and fatigue. Even when skin involvement is minimal or absent, MF patients continue to report impaired HRQoL.

Abstract CT208: Prior vaccination with the autologous Tumor Lysate Particle Loaded Dendritic Cell (TLPLDC) Vaccine may impact clinical outcomes in melanoma patients treated with systemic therapies and re-vaccination

Abstract Background: Melanoma is an immunogenic cancer, yet only 50-60% of tumors respond to current immunotherapy. The autologous tumor lysate, particle loaded, dendritic cell (TLPLDC) vaccine may potentiate an immune response by stimulating T-cells. We are conducting a phase IIb double-blinded randomized trial of TLPLDC to prevent recurrence after resection. Patients who recurred were offered enrollment in an open-label TLPLDC vaccine trial along with standard therapy. Here, we describe characteristics and clinical outcomes of recurred patients that continued on open label TLPLDC vaccination. Methods: Patients with resected no evidence of disease (NED) stage III/IV melanoma were randomized to TLPLDC vs. empty yeast cell wall particle (YCWP) loaded DC in a 2:1 fashion. TLPDC is created by loading autologous tumor lysate into YCWP. This is then introduced ex vivo to the patient’s DC for phagocytosis. 1-1.5x106TLPLDCs are given via intradermal injection monthly x4 followed by boosters at 6 and 9 months. Patients who recurred after receiving TLPLDC (vaccine group, VG) or after empty YCWP (control group, CG) on the trial (primary endpoint) were offered open label TLPLDC vaccination with same dosing schedule along with standard therapy as determined by the patient’s treatment team. Disease status is measured by RECIST criteria. Results: To date, 23 (14 VG and 11 CG) patients have enrolled and been vaccinated, with median follow up (fu) of 7.86 months (mo). The patients received the following additional therapies: surgery, gamma-knife, check point inhibitor, radiation, TVEC, BRAF/MEK, and imantinib. No related toxicities > grade 2 were observed. Of 14 VG patients, 5 were NED when re-enrolled, 9 had measurable disease (MD). Of these, 11 have followed up data, 9 (82%) show disease control (5/5 NED patients remain NED, 4/6 with MD show SD), while 2 (18%) have progressed. Of 9 CG patients, 4 were NED and 5 had measurable disease at time of re-enrollment. Of these, 8 have followed up, 5 (62%) show disease control (2/4 NED remain NED, 1/4 with MD shows regression, 2/4 SD), while 3 (38%) have progressed (2/4 NED, 1/3 with MD). Conclusion: Open label administration of TLPLPDC vaccine after recurrence is demonstrated as a safe therapy and shows potential clinical benefit when combined SOC therapy. Further study is warranted to determine if prior exposure to TLPLDC may provide clinical benefit after the first episode of disease recurrence. Citation Format: Annelies T. Hickerson, Guy T. Clifton, Tommy A. Brown, Jessica L. Cindass, John W. Myers, Timothy J. Vreeland, Diane F. Hale, Kaitlin M. Peace, Doreen O. Jackson, Garth Herbert, Xianzhong Yu, Thomas E. Wagner, George E. Peoples. Prior vaccination with the autologous Tumor Lysate Particle Loaded Dendritic Cell (TLPLDC) Vaccine may impact clinical outcomes in melanoma patients treated with systemic therapies and re-vaccination [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr CT208.

Long-Term Survival of De Novo Stage IV Human Epidermal Growth Receptor 2 (HER2) Positive Breast Cancers Treated with HER2-Targeted Therapy.

An increasing proportion of human epidermal growth receptor 2 (HER2) positive (HER2+) metastatic breast cancer (MBC) is diagnosed as de novo stage IV disease. We hypothesize that a subset of these patients who achieve no evidence of disease (NED) status after multimodality HER2-targeted treatments may have prolonged progression-free survival (PFS) and overall survival (OS). Patients with de novo stage IV, HER2+ MBC (n = 483) diagnosed between 1998 and 2015 were identified at two institutions (Yale and MD Anderson Cancer Centers). Clinical variables, treatment details, and survival outcomes were compared between those who achieved NED and those who did not. All patients received trastuzumab, and 20% also received pertuzumab as first-line therapy. The median OS was 5.5 years (95% confidence interval [Cl]: 4.8-6.2). Sixty-three patients (13.0%) achieved NED; their PFS and OS rates were 100% and 98% (95% CI: 94.6%-100%), respectively, at 5 years and remained the same at 10 years. For patients with no NED (n = 420), the PFS and OS rates were 12% (95% CI: 4.5%-30.4%) and 45% (95% CI: 38.4%-52.0%) at 5 years and 0% and 4% (95% CI, 1.3%-13.2%) at 10 years, respectively. NED patients more frequently had solitary metastasis (79% vs. 51%, p = .005) and surgery to resect cancer (59% vs. 22%, p ≤ .001). In multivariate analysis, NED status (hazard ratio [HR]: 0.014, p = .0002) and estrogen receptor positive status (HR: 0.72; p = .04) were associated with prolonged OS. Among patients with de novo stage IV, HER2+ MBC, those who achieve NED status have a very high PFS and OS. Further randomized studies are required to fully understand the impact of systemic or locoregional therapy on achieving these excellent long-term outcomes. In this retrospective review at two institutions, it was demonstrated that 13% of patients with de novo stage IV, human epidermal growth receptor 2 positive metastatic breast cancer achieved no evidence of disease (NED) status with trastuzumab-based therapy plus/minus local therapies, and these patients had a very high progression-free survival (100%) and overall survival (98%) at both the 5- and 10-year time points. Achieving NED status may be an important therapeutic goal. However, further randomized studies are required to fully understand the impact of systemic or locoregional therapy on achieving these excellent long-term outcomes.

Abstract B079: Prospective study on the use of Raman molecular imaging to determine postoperative oncological outcomes in patients with prostate cancer: Analysis of a single center

Abstract Background: Prostate cancer (PCa) is the most common cancer in males and the second leading cause of cancer deaths. One of the most confounding problems faced by urologists is predicting the progression of PCa. Current prognostic tools for PCa rely solely on clinical and pathologic variables that cannot always accurately predict the progression of the disease. More accurate diagnostic tools to aid clinicians and patients to decide the course of treatment, even in the early stage of cancer, have long been sought. In a prospective study, we utilized Raman molecular imaging (RMI) to identify patients at risk for biochemical recurrence following a prostatectomy. RMI is a technology that combines the molecular chemical analysis capability of Raman spectroscopy with high-definition digital image visualization, enabling analysis of the molecular environment of prostate tissue by providing highly specific molecular imaging data. Objective: The aim of this study is to evaluate RMI as a methodology for predicting the outcome of PCa patients following radical prostatectomy. Methods: PCa patients who underwent prostatectomy were prospectively enrolled with informed consent. Surgeries were performed by two practiced surgeons in a single clinical center from 2009–2011, and 60 months’ follow-up time was recorded. Pathology data were reviewed by one genitourinary pathologist. Preoperative and postoperative data (preoperation prostate-specific antigen level [PSA], Nadir PSA, Gleason scores, pathologic stage, biochemical recurrence time) were recorded, and biochemical failure was defined as 2 consecutive increases in postoperative PSA (≥0.2 ng/mL). Patient identifiers were removed. Raman molecular images were collected from unstained prostatectomy tissue sections. Raman spectral data were extracted from epithelial and stromal regions, and multivariate statistical methodologies were applied to these spectra. Partial least squares discriminant analysis (PLS-DA) was employed in conjunction with leave-one-patient-out cross-validation to generate models based on the pathology data and RMI. Results: Initial data analysis was performed on a representative population of 38 PCa patients. In this population the mean (median) age at the time of diagnosis was 60 (61) years; the pathologic stage for 57.9% of the patients was T3 and for 42.1% of the patients T2. Nine patients (23.7%) progressed to biochemical failure in 60 months. Analysis of RMI performed on prostatectomy tissues from these patients indicates that the RMI data are distinguishable between those with biochemical failure (progressors) and those with no evidence of disease (NED). A PLS-DA model comprising 9 progressors and 29 NED patients differentiated between the two tissue classes with 89% sensitivity and 90% specificity, and with an area under the ROC curve of 0.87. In this model, classification accuracy was 90% and the p–value < 0.01. Analysis revealed most notable differentiation between progressors and NEDs when evaluating the epithelium and stroma as separate histologic elements. A PLS-DA model based on epithelial cells from 9 progressors and 10 NEDs discriminated between the two populations with 89% sensitivity and 100% specificity, with AUROC of 0.92 and a p-value < 0.01. A similar model differentiated between the stroma of progressors and NEDs with 100% specificity and 100% sensitivity, and a p-value well below 0.01. Conclusion: RMI is a novel technique that shows promise for identifying patients at risk of progression by visualizing molecular information not seen using other current methods. In Gleason 7 disease, RMI indicates distinctive chemical differences in patients who had biochemical failure postoperatively. This preliminary work lays the foundation for the further study of RMI for evaluating prostate tissue and developing an assay that may impact clinicians and patients with PCa. Citation Format: Arash Samiei, Ralph Miller, Jeffrey Cohen, Heather Gomer, Shona Stewart, Patrick Treado. Prospective study on the use of Raman molecular imaging to determine postoperative oncological outcomes in patients with prostate cancer: Analysis of a single center [abstract]. In: Proceedings of the AACR Special Conference: Prostate Cancer: Advances in Basic, Translational, and Clinical Research; 2017 Dec 2-5; Orlando, Florida. Philadelphia (PA): AACR; Cancer Res 2018;78(16 Suppl):Abstract nr B079.

Behavior of blood plasma glycan features in bladder cancer.

Despite systemic therapy and cystectomy, bladder cancer is characterized by a high recurrence rate. Serum glycomics represents a promising source of prognostic markers for monitoring patients. Our approach, which we refer to as “glycan node analysis”, constitutes the first example of molecularly “bottom-up” glycomics. It is based on a global glycan methylation analysis procedure that is applied to whole blood plasma/serum. The approach detects and quantifies partially methylated alditol acetates arising from unique glycan features such as α2–6 sialylation, β1–4 branching, and core fucosylation that have been pooled together from across all intact glycans within a sample into a single GC-MS chromatographic peak. We applied this method to 122 plasma samples from former and current bladder cancer patients (n = 72 former cancer patients with currently no evidence of disease (NED); n = 38 non-muscle invasive bladder cancer (NMIBC) patients; and n = 12 muscle invasive bladder cancer (MIBC) patients) along with plasma from 30 certifiably healthy living kidney donors. Markers for α2–6 sialylation, β1–4 branching, β1–6 branching, and outer-arm fucosylation were able to separate current and former (NED) cases from certifiably healthy controls (ROC curve c-statistics ~ 0.80); but NED, NMIBC, and MIBC were not distinguished from one another. Based on the unexpectedly high levels of these glycan nodes in the NED patients, we hypothesized that recurrence of this disease could be predicted by some of the elevated glycan features. Indeed, α2–6 sialylation and β1–6 branching were able to predict recurrence from the NED state using a Cox proportional hazards regression model adjusted for age, gender, and time from cancer. The levels of these two glycan features were correlated to C-reactive protein concentration, an inflammation marker and known prognostic indicator for bladder cancer, further strengthening the link between inflammation and abnormal plasma protein glycosylation.

Abstract 612: Nivolumab-induced changes associated with patient outcomes are disparate in metastatic melanoma patient Tregs versus conventional T cells

Abstract The PD1 blocking antibody nivolumab induces durable responses in metastatic melanoma patients. However, many patients do not respond and biomarkers differentiating patient response remain unidentified. To address this need, we investigated the effects of nivolumab on peripheral blood conventional CD4+ (Tcons) and regulatory T-cells (Tregs) of metastatic melanoma patients receiving nivolumab as adjuvant therapy or for metastatic disease. An increased ratio of PD1 expression on Tregs compared with Tcons was found at baseline patients with no evidence of disease (NED), while relapsing patients had a lower ratio (p<0.05). Intriguingly, NED patients had an increase in circulating Treg percentages post-nivolumab (p<0.05), which was not observed in relapsing patients. In vitro treatment of isolated T-cells with αPD1 resulted in increased proliferation (p<0.001). RNA-seq analysis of Tcons and Tregs showed distinct gene expression changes based on cell type and patient outcome. Pathway analysis demonstrated: upregulation of proliferation associated genes in NED Tregs, but not in relapse Tregs; downregulation of oxidative phosphorylation associated genes in NED patient Tregs and upregulation in NED Tcons; upregulation of heme metabolism associated genes in NED Tcons and downregulated in relapsing patient Tcons; and similar differences in MTORC1, heme metabolism and hypoxia pathways (all pathways q<0.01). Both Tcons and Tregs from non-relapsing patients had induction of phosphoSTAT3 (pSTAT3) expression post-nivolumab (p=0.012), which was absent in relapsing patients. This was also observed in a cohort of active disease patients (p=0.022), and the magnitude of change in pSTAT3 was positively correlated with patient survival (R2=0.49, p<0.05). Enhanced pSTAT3 expression was recapitulated in vitro in isolated T-cells (p<0.05). Post-nivolumab Tregs from patients with positive outcomes also had reduced suppressive function in mixed lymphocyte reactions (p<0.05), while patients with negative outcomes had no changes. In vitro treatment of Tregs with PD1 blocking antibodies resulted in a reduction in suppressive function (p<0.0001), while addition of a STAT3 inhibitor enhanced suppressive function (p<0.0001). These results suggest that nivolumab alters Treg signaling and metabolism pathways, hindering immune suppressive function. Collectively, our data demonstrate distinct effects of nivolumab on Tcons vs. Tregs, and highlight several potential biomarkers of patient benefit, warranting further exploration. Citation Format: David M. Woods. Nivolumab-induced changes associated with patient outcomes are disparate in metastatic melanoma patient Tregs versus conventional T cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 612.

Abstract IA08: Prostate cancer tumor dormancy and the development of metastases

Abstract Cancer dormancy refers to the prolonged clinical disease-free time between removal of the primary tumor and recurrence, which is common in prostate cancer (PCa). PCa disseminated tumor cells (DTC) can be detected in the bone marrow of patients with no evidence of disease (NED) and advanced disease (ADV). However, the molecular and cellular nature of DTC is unknown. Single cell transcriptomic analyses comparing DTC in NED versus ADV PCa patients identified a molecular signature associated with the p38 stress activated kinase pathway and dormancy. Understanding the mechanism by which dormant tumor cells are triggered to proliferate may provide new targets for therapy. We have demonstrated that LuCaP PCa xenografts, which appear dormant in vitro, can be induced to proliferate through contact with each other and bone marrow stroma. Constitutive activation of myosin light chain kinase (MLCK), a downstream effector of integrin-β1 and TGFβ2, in non-proliferating cells promoted cell proliferation. Additionally, gene expression analyses and in vitro data support a role for TGFβ2 in suppressing proliferation. Taken together, these data point to a potential mechanism by which PCa cells are released from dormancy to form lethal metastases. The early dissemination of cells from the primary to other sites most likely involves epithelial-mesenchymal transition (EMT)-like behavior. It has been proposed that further dissemination can occur where a metastasis can seed further metastases. TGFβ is a known driver of EMT which is associated with tumor aggressiveness and metastasis. We assessed the natural history of tumor dissemination and EMT-related markers TGFβ, Twist, Slug, and Zeb1 in bone and visceral metastases from castration resistant PCa (CRPC) patients. Importantly, nuclear Twist, Slug, and Zeb1 were only present in a subset of epithelial cells that had an EMT-like phenotype, suggesting only a subset of cells display an EMT-like behavior in any given CRPC metastasis. These studies highlight the biological relevance of dormancy, cell-contact and EMT-like behavior in the seeding and establishment of CRPC metastases. Citation Format: Colm Morrissey. Prostate cancer tumor dormancy and the development of metastases. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr IA08.

Highlights from Recent Cancer Literature

Highlights from Recent Cancer Literature

Carbon anhydrase IX specific immune responses in patients with metastatic renal cell carcinoma potentially cured by interleukin-2 based immunotherapy

The majority of clear-cell renal cell carcinomas (ccRCC) show high and homogeneous expression levels of the tumor associated antigen (TAA) carbonic anhydrase IX (CAIX), and treatment with interleukin-2 (IL-2) based immunotherapy can lead to cure in patients with metastatic renal cell carcinoma (mRCC). However, the involvement of CAIX specific CD8+ T cells and/or NK cells in the tumor eradication is unknown. We investigated T cell and antibody reactivity against overlapping 15-mer CAIX-peptides as well as HLA haplotype frequency and NK cell cytotoxicity in 11 patients with no evidence of disease (NED) following treatment with IL-2 based immunotherapy, and thus potentially cured. Immune reactivity in these patients was compared with samples from patients with dramatic tumor response obtained immediately at the cessation of therapy, samples from patients that experienced progressive disease during treatment and samples from healthy controls. We observed more focused but only weak and not consistent CAIX specific T-cells in the late observation and early observation response groups compared with the healthy control group. An increased frequency of the class II alleles HLA-DRB4 01:01, HLA-DPB 01:01 and HLA-DPB 03:01 was noted in the NED patients. In contrast, NK cytotoxicity was low even in the late observation response group as compared with controls. In particular, a HLA-B*40:01 restricted CD8+ T cell response recognizing the CAIX- derived peptide SEEEGSLKL was identified. This may have interest in future cancer vaccines, but more studies are needed to elucidate the immunological mechanisms of action in potentially cured patients treated with an immunotherapeutic agent.

1140 EVALUATION OF URINARY EXOSOMES IN BLADDER CANCER PATIENTS

You have accessJournal of UrologyBladder Cancer: Basic Research (III)1 Apr 20131140 EVALUATION OF URINARY EXOSOMES IN BLADDER CANCER PATIENTS Jennifer J. Ahn, Guillermo Salazar, Matthew Seaman, Anna Scott, Piruz Motamedinia, Crystal Castaneda, Wayne Comper, James M. McKiernan, and Leileata Russo Jennifer J. AhnJennifer J. Ahn New York, NY More articles by this author , Guillermo SalazarGuillermo Salazar New York, NY More articles by this author , Matthew SeamanMatthew Seaman New York, NY More articles by this author , Anna ScottAnna Scott New York, NY More articles by this author , Piruz MotamediniaPiruz Motamedinia New York, NY More articles by this author , Crystal CastanedaCrystal Castaneda New York, NY More articles by this author , Wayne ComperWayne Comper New York, NY More articles by this author , James M. McKiernanJames M. McKiernan New York, NY More articles by this author , and Leileata RussoLeileata Russo New York, NY More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.755AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Despite significant strides in understanding the molecular pathogenesis of bladder cancer (BCa), diagnosis and surveillance continue to rely upon cystoscopy and cytology. As a result, there has been substantial interest in the use of biomarkers for diagnosis and risk stratification. However, current assays rely on tumor cells to be shed into the urine, or examine proteins that may not be specific for cancer. We have previously shown that urinary exosomes and microvesicles (collectively referred to as exosomes) are a consistent and reproducible means of sampling high integrity genetic material, but this has yet to be investigated in the context of BCa. Here, we examined the feasibility of evaluating the urinary exosomes of bladder cancer patients. METHODS Following Columbia University Medical Center IRB approval, voided urine samples were collected from 12 patients. BCa status at time of collection, as determined by cystoscopy and biopsy, was high-grade (HG) tumor (n = 5), low-grade (LG) tumor (n = 2), or no evidence of disease (NED) (n = 5). Samples were stored at 4°C and 0.8 μm filtration was used to remove whole cells and debris. Urinary exosomal RNA (exoRNA) was isolated using an in-house technique (Exosome Diagnostics, NY) and analyzed using RT-qPCR for 18 genes previously described as being differentially expressed in BCa. To compare expression levels, genes were standardized to 18S and relative quantitation (RQ) was calculated using DataAssist software (v.3.0). RESULTS Of the 18 genes analyzed, ICAM1 showed significantly higher expression in patients with HG BCa, as compared to NED patients (RQ 37.0, p = 0.0084). A similar trend was seen when comparing patients with any BCa (HG or LG) to NED patients (RQ 14.2, p = 0.051). UPK2 expression levels were not significantly different between the groups HG versus NED (RQ 1.6, p = 0.958) or LG versus NED (RQ 1.0, p = 0.975). CONCLUSIONS We successfully isolated exosomal RNA from the voided urine of patients with concurrent bladder cancer, as well as those with no evidence of disease. Additionally, significant upregulation of ICAM1 was seen in patients with HG disease. Ultimately, this technique shows promise as a non-invasive method of assessing gene expression in bladder cancer, independent of examining urothelial cells. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e465-e466 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Jennifer J. Ahn New York, NY More articles by this author Guillermo Salazar New York, NY More articles by this author Matthew Seaman New York, NY More articles by this author Anna Scott New York, NY More articles by this author Piruz Motamedinia New York, NY More articles by this author Crystal Castaneda New York, NY More articles by this author Wayne Comper New York, NY More articles by this author James M. McKiernan New York, NY More articles by this author Leileata Russo New York, NY More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

The Clinical Effect of the Inhibitor of Apopotosis Protein Livin in Melanoma

Objective: The inhibitor of apoptosis protein (IAP) livin is frequently overexpressed in melanoma. Livin binds caspases and thereby inhibits apoptosis. We found that caspases cleave livin to produce a truncated form with a paradoxical proapoptotic activity. Methods: We assessed the correlation of livin expression with survival among 114 melanoma patients treated with an autologous melanoma vaccine. In 52 patients, resection resulted in no evidence of disease (NED) and 62 remained with active disease (WAD). Protein levels were assessed using Western blot. Results: We found livin protein expression in 44/114 samples (38.4%). Median overall survival was 1.4 years in NED patients with high levels of livin protein, 8.4 years in those with low-intermediate levels and not reached in patients who did not express livin (p = 0.025). The corresponding overall survival was 2.3 years among WAD patients with high levels of livin protein, 11.3 years in those with low-intermediate levels and, paradoxically, only 4.0 years in patients who did not express livin (p = 0.012). Conclusion: Livin protein expression may play a role in the progression of melanoma and correlates with survival. A high level of the protein is associated with a poor prognosis. However, in WAD patients low to intermediate level of livin, rather than absence of the protein, is associated with a favorable prognosis. This is probably due to the paradoxical proapoptotic activity of this important regulator of apoptosis.

Abstract PR4: Biochemical recurrence is not a definitive surrogate endpoint for development of clinically useful predictive models for post-prostatectomy patients

Abstract Introduction: Multivariable models that predict biochemical recurrence (BCR) can indicate patients at higher risk for developing clinical progression after prostatectomy. However, most patients that experience BCR do not metastasize or die from prostate cancer, making BCR an unreliable surrogate endpoint. In this study we compared the clinical and genomic characteristics of prostatectomy patients in order to better delineate the relationship between BCR and aggressive prostate cancer. Methods: Postoperative patients were retrospectively classified into three outcome groups: NED (no evidence of disease recurrence); BCR-only (patients that experienced BCR but didn't have progressive disease as evidenced by bone or CT scans); and CP (clinical progression with BCR plus positive CT or bone scans). Patient specimens were from the Mayo Clinic Radical Prostatectomy Registry, selected using a nested case-control design with 14 years median follow-up. RNA expression levels from FFPE tumor specimens were measured with 1.4 million feature whole-transcriptome oligonucleotide microarrays on 106 matching patients from each outcome group (total 318 patients). We analyzed the clinical covariates of the outcome groups in a pairwise manner (NED vs. BCR-only; NED vs. CP; CP vs. BCR-only), using a conditional logistic model in univariable and multivariable analyses. We performed feature reduction for the genomic data set using Kruskal-Wallis tests and regularized logistic regression. Principal Component Analysis (PCA) was used to quantify the variance of the genomic data with respect to the patient outcome groups. Results: Our initial hypothesis was that all three outcome groups would have different clinical and genomic characteristics. Univariable analysis of clinical variables failed to show statistically significant differences between NED and BCR-only groups. Multivariable analysis of the clinical variables also failed to show statistically significant differences between NED and BCR-only groups. Among the clinical variables only pathological Gleason score was statistically significant when comparing the BCR-only vs. CP and NED vs. CP groups. Genomic analysis found 363 coding and non-coding features that passed thresholds for differential expression between the three outcome groups. PCA established that the primary source of variance in the genomic data stemmed from the separation of CP from NED and BCR-only patients; whereas only a small amount of variance separates NED and BCR-only groups. The 363 features were further compared in the three outcome groups using Wilcoxon tests, adjusting for false-discovery using Benjamini-Hochberg. At a cutoff of p < 0.001 there were no features differentially expressed between NED and BCR-only, while 52 and 34 statistically significant features were found in the NED vs. CP and CP vs. BCR-only comparisons, respectively. Conclusion: Considering the clinical and genomic results together, we find that: 1) Patients that developed clinical progression were significantly different from those that did not, irrespective of whether they experienced BCR or not; 2) the NED and BCR-only groups were statistically very similar to each other. We therefore conclude that BCR may not be the most appropriate endpoint for predictive models that attempt to stratify patients at risk for progression. This abstract is also presented as Poster B44. Citation Format: Anamaria Crisan, Elai Davicion, R. Jeffery Karnes, Robert B. Jenkins, Mercedeh H. Ghadessi, Christine Buerki, Eric J. Bergstrahl, Nicholas Erho, Ismael A. Vergara, Karla V. Ballman, Peter C. Black, Timothy J. Triche. Biochemical recurrence is not a definitive surrogate endpoint for development of clinically useful predictive models for post-prostatectomy patients [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr PR4.

Role of RANK, RANKL, OPG, and CXCR4 Tissue Markers in Predicting Bone Metastases in Breast Cancer Patients

BackgroundThe RANK/RANKL/OPG system is active in primary cancers such as breast, prostate, and also in their bone metastases. CXCR4 chemokine receptor is highly expressed in human breast cancer cells and is believed to facilitate the homing of tumor cells to organs such as bone that express high levels of its ligand SDF1. Our study aimed to investigate whether the analysis of these markers with an inexpensive and simple test can help to predict bone metastases in breast cancer patients. Patients and MethodsMarker expression was evaluated by immunohistochemical staining in paraffin-embedded tissue sections of primary breast cancers from 40 individuals: 20 patients with bone metastases (BM), 10 with visceral metastases (VM; considered together as the relapsed group), and 10 with no evidence of disease (NED). ResultsRANKL was not detected in tumor cells. OPG- and RANK-positive tumors are found with similar frequency in NED (20%) and in relapsed patients (23% and 17%, respectively). However, in the latter subgroup, only RANK positivity was always associated with bone relapse. The frequency of CXCR4-positive tumors was three-fold higher in relapsed (30%) than in NED (10%) patients and positivity was always linked to bone metastases. Considering NED and VM patients together versus BM patients, we observed that CXCR4 expression, alone (P = .008) or in combination with RANK (P < .001), identified patients destined to relapse to bone. ConclusionOur results provide the first clinical evidence to support a pivotal role of combined CXCR4 and RANK expression in predicting bone relapse.

The Role of CXCR4 in the Prediction of Bone Metastases from Breast Cancer: A Pilot Study

Objective: The chemokine receptor CXCR4 is involved in tumor growth and homing of cancer cells to distant sites. The aim of our retrospective case-control study was to evaluate whether CXCR4 expression is more effective than conventional markers (estrogen receptor and HER-2) in predicting bone relapse in breast cancer. Methods: CXCR4 expression was evaluated by immunohistochemical staining in paraffin-embedded tissue sections of primary breast cancers from 20 patients with bone metastases (BM), 10 with visceral metastases (VM) and 10 with no evidence of disease (NED) at a median follow-up of 10.5 years (range 10.1–11.8). Results: Cytoplasmic CXCR4 expression was high in BM patients (45%, 95% CI 23–67), much lower in NED patients (10%, 95% CI 0–29) and negative in the VM group. CXCR4 coexpression in the nucleus and cytoplasm was observed in about half of the BM patients (45%) but never in NED or VM patients (p = 0.013). Conversely, estrogen receptor-positive and HER-2-negative status identified 80 and 95% of bone relapse patients, respectively, but did not discriminate between cases and controls. Conclusions: Our results suggest a pivotal role of CXCR4 expression as a predictor of BM in primary breast cancer. A larger study is ongoing to confirm these results.

685 EPITHELIAL-MESENCYHMAL TRANSITION IN PROSTATE CANCER DISSEMINATED TUMOR CELLS

You have accessJournal of UrologyProstate Cancer: Basic Research V1 Apr 2010685 EPITHELIAL-MESENCYHMAL TRANSITION IN PROSTATE CANCER DISSEMINATED TUMOR CELLS Melanie Ketchandji, Theodore Koreckij, Todd Morgan, Colm Morrissey, Paul Lange, Pete Nelson, Ilsa Coleman, Roger Coleman, Marty Kinnunen, Xiaotun Zhang, Bruce Montgomery, and Robert Vessella Melanie KetchandjiMelanie Ketchandji More articles by this author , Theodore KoreckijTheodore Koreckij More articles by this author , Todd MorganTodd Morgan More articles by this author , Colm MorrisseyColm Morrissey More articles by this author , Paul LangePaul Lange More articles by this author , Pete NelsonPete Nelson More articles by this author , Ilsa ColemanIlsa Coleman More articles by this author , Roger ColemanRoger Coleman More articles by this author , Marty KinnunenMarty Kinnunen More articles by this author , Xiaotun ZhangXiaotun Zhang More articles by this author , Bruce MontgomeryBruce Montgomery More articles by this author , and Robert VessellaRobert Vessella More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2010.02.1084AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES About 15-30% of patients diagnosed with prostate cancer (PCa) who undergo a radical prostatectomy (RP) later recur. PCa has a proclivity to metastasize to bone. Thus we characterized disseminated tumor cells (DTC) isolated from the bone marrow of PCa patients at various stages of disease progression to gain insight into the events associated with PCa metastases. We observed a loss of the differentiated epithelial phenotype with evidence of epithelial-mesenchymal transition (EMT) in early disease and subsequently mesenchymal-epithelial transition (MET) in advanced disease. METHODS Patients at various stages of disease progression underwent bone marrow aspirations either at the time of surgery or at follow-up. The aspirate was then enriched for epithelial cells using immunomagnetic beads conjugated to antibodies to CD45 and CD61 (negative selection) and to EpCam (positive selection). The enriched cells were stained with a FITC-BerEp4 antibody (anti EpCam) and pools of approximately 10-20 positive cells from 62 patients (33 at RP, 11 with no evidence of disease (NED), and 18 with advanced disease (ADV) were analyzed using cDNA microarrays with real-time PCR confirmation. Human tissue microarrays of fixed paraffin-embedded tissues from forty-four rapid autopsy patients were also used for immunohistochemical (IHC) analyses of EMT-related proteins of interest. RESULTS Gene expression analysis of DTC isolated from the 62 patients revealed a wide range of phenotypic heterogeneity. TWIST-1 (p=<0.001), S100A4 (p=0.002) and SNAIL1 (p=0.012), genes associated with EMT, were differentially expressed between the RP and ADV patients. Differences were also observed between the NED and ADV patients (TWIST-1 (p=0.002) and S100A4 (p=0.05)). Furthermore, IHC revealed that TWIST-1, S100A4 and SNAIL1 were present in PCa bone and soft tissue metastases. Ten of the forty-four RP and NED patients later developed biochemical recurrence. DTC from recurrent patients demonstrated increased expression of TWIST-1 (p=0.003) and decreased expression of E-cadherin (p=0.05) when compared to patients who did not recur. CONCLUSIONS The EMT related genes herein may have an important role in the development of PCa bone metastasis. The expression of E-cadherin and TWIST1 in DTC may predict recurrence in RP and NED patients. The molecular profiles of DTC suggest EMT is an early event in tumor cell dissemination followed by MET as the tumor cells become established within the bone marrow. Interestingly, some, EMT-related proteins continue to be expressed in clinically overt PCa bone metastases. Seattle, WA© 2010 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 183Issue 4SApril 2010Page: e267-e268 Advertisement Copyright & Permissions© 2010 by American Urological Association Education and Research, Inc.MetricsAuthor Information Melanie Ketchandji More articles by this author Theodore Koreckij More articles by this author Todd Morgan More articles by this author Colm Morrissey More articles by this author Paul Lange More articles by this author Pete Nelson More articles by this author Ilsa Coleman More articles by this author Roger Coleman More articles by this author Marty Kinnunen More articles by this author Xiaotun Zhang More articles by this author Bruce Montgomery More articles by this author Robert Vessella More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Disseminated tumor cells in prostate cancer patients after radical prostatectomy and without evidence of disease predicts biochemical recurrence.

Men with apparently localized prostate cancer often relapse years after radical prostatectomy. We sought to determine if epithelial-like cells identified from bone marrow in patients after radical prostatectomy, commonly called disseminated tumor cells (DTC), were associated with biochemical recurrence. We obtained bone marrow aspirates from 569 men prior to radical prostatectomy and from 34 healthy men with prostate-specific antigens <2.5 ng/mL to establish a comparison group. Additionally, an analytic cohort consisting of 98 patients with no evidence of disease (NED) after radical prostatectomy was established to evaluate the relationship between DTC and biochemical recurrence. Epithelial cells in the bone marrow were detected by magnetic bead enrichment with antibodies to CD45 and CD61 (negative selection) followed by antibodies to human epithelial antigen (positive selection) and confirmation with FITC-labeled anti-BerEP4 antibody. DTC were present in 72% (408 of 569) of patients prior to radical prostatectomy. There was no correlation with pathologic stage, Gleason grade, or preoperative prostate-specific antigens. Three of 34 controls (8.8%) had DTC present. In patients with NED after radical prostatectomy, DTC were present in 56 of 98 (57%). DTC were detected in 12 of 14 (86%) NED patients after radical prostatectomy who subsequently suffered biochemical recurrence. The presence of DTC in NED patients was an independent predictor of recurrence (hazard ratio 6.9; 95% confidence interval, 1.03-45.9). Approximately 70% of men undergoing radical prostatectomy had DTC detected in their bone marrow prior to surgery, suggesting that these cells escape early in the disease. Although preoperative DTC status does not correlate with pathologic risk factors, persistence of DTC after radical prostatectomy in NED patients was an independent predictor of recurrence.

Anti-Mesothelin Antibodies and Circulating Mesothelin Relate to the Clinical State in Ovarian Cancer Patients

Most human ovarian carcinomas express mesothelin, which is shed as a diagnostically useful biomarker. We applied an ELISA to measure antibodies to native mesothelin in serum from a series of patients with divergent clinical outcomes. The level of anti-mesothelin antibodies determined as OD(450 nm) and referred to as absorption units (AU) for 1:20 diluted serum was higher in patients who remained disease-free after therapy [no evidence of disease (NED); n = 14] than in patients whose disease recurred [clinical evidence of disease (CED); n = 21; P < 0.01]. Applying AU > or = 0.5 at a serum dilution of 1:20 as cutoff, 10 of 14 (71%) ovarian carcinoma patients with NED and 9 of 21 (43%) patients with CED had antibodies to mesothelin compared with 6 of 23 (26%) healthy women (P < 0.008) and 5 of 24 (21%) women with other benign gynecologic diseases (P < 0.003), whereas 7 of 9 (78%) of women with pelvic inflammatory disease were positive. Three of the 14 (21%) NED patients had circulating mesothelin detected as an AU > or = 0.2 at a serum dilution of 1:40 (P < 0.005) compared with 15 of 21 (71%) CED patients, and 9 of 14 (64%) NED patients (P < 0.0002) were positive for antibodies and negative for antigen compared with 1 of 21 (5%) CED patients. Although our data indicate that an antibody response to mesothelin is an important correlate of ovarian carcinoma, prospective studies are needed to show whether the measurement of such antibodies (alone or together with antigen) aids the diagnosis and monitoring of patients.