bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4955: Isolation and characterization of circulating tumor cells (CTCs) in breast and prostate cancer: Comparison of Harpoon CTC assay performance with the CellSearch CTC Test
Authors: Connelly Mark , Chianese David , Morano Carrie , Bui Thai , Powell Shemeeakah , Ngoubilly Noel , Sanders Renouard , Barber Tom , Kapur Ravi , Maheswaran Shyamala , Toner Mehmet , Haber Daniel 
Published: July 2016

Abstract
Circulating tumor cells (CTCs) can provide important information about a patient's prognosis. Currently, the CELLSEARCH® CTC Test (hereafter referred to as CS) is the only US Food and Drug Administration-cleared assay for isolating and enumerating CTCs. This technology uses a CTC antigen-dependent method to isolate CTCs that may limit its ability to detect CTCs with low or no epithelial cell adhesion molecule (EpCAM) expression. We evaluated the Harpoon CTC Isolator and Chip, which uses an antigen-independent, negative depletion method to enrich CTCs from patient blood samples. As a result of the negative depletion method, the final Harpoon Product (ie, CTC-enriched cell suspension) contains CTCs that express cytokeratin (CK) and EpCAM (canonical CTCs) as well as those with low or no expression of these antigens (non-canonical CTCs). After isolation, CTCs were spun onto microscope slides and imaged using a Vectra Multispectral Imaging System. We obtained performance data on an initial prototype Harpoon assay (Harpoon + Vectra [Hrp+Vec]) and compared CTC enumeration between Hrp+Vec and CS using blood samples from 36 breast and prostate cancer patients. From 7.5 mLs of blood, the Hrp+Vec assay recovered 79.0% to 93.5% of canonical EpCAM+/CK+ CTCs recovered by CS. The Hrp+Vec assay also successfully detected non-canonical CTCs (EpCAM-/CK+ or EpCAM+/CK-) in these samples. At the CS clinical cutoff for prognostic value, where ≥5 CTCs is associated with poor prognosis, the Hrp+Vec assay was 97% concordant with CS. Hrp+Vec also detected CTCs in 12.5% (2/16) more patient samples than CS. Finally, a consensus analysis of all Vectra scan results (including both canonical and non-canonical CTCs) demonstrated that the Hrp+Vec assay detected between 90.5% and 108.9% of the number of CTCs as identified by CS. These results indicate that, when compared with CS, the prototype Hrp+Vec assay detected CTCs with similar efficiency and identified more samples as CTC-positive in these breast and prostate cancer patients. Importantly, the Hrp+Vec assay did enrich non-canonical CTCs thought to be missed by CS, and ongoing efforts are aimed to further characterize these cell populations.
Citation Format: Mark Connelly, David Chianese, Carrie Morano, Thai Bui, Shemeeakah Powell, Noel Ngoubilly, Renouard Sanders, Tom Barber, Ravi Kapur, Shyamala Maheswaran, Mehmet Toner, Daniel Haber. Isolation and characterization of circulating tumor cells (CTCs) in breast and prostate cancer: Comparison of Harpoon CTC assay performance with the CellSearch CTC Test. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4955.
Abstract 4972: Detection of PSMA on circulating tumor cells from blood of prostate cancer patients using anti-PSMA Centyrin
Authors: Vaidya Bhavesh , Vulfson Peter , Sanders Renouard , Klein Donna , Goldberg Shalom , Jacobs Steve , Rao Chandra 
Published: July 2016

Abstract
Prostate specific membrane antigen (PSMA), a transmembrane glycoprotein, is a clinically validated marker of prostate cancer. PSMA is overexpressed in primary and metastatic prostate cancer when compared to normal prostate tissue which makes it an attractive target for immunotherapy. Several antibodies are being considered to target PSMA for the development of an anticancer drug. The expression levels of PSMA on tumor cells may play a role in response to treatment with anti PSMA drugs. Therefore, it is important to monitor PSMA expression on tumor cells before and during treatment. We used CellSearch® CTC assay to determine the presence of PSMA on circulating tumor cells (CTCs) using anti PSMA Centyrin as a biomarker. Centyrins are a class of alternative scaffold molecules which are proteins of small size (∼ 10 kDa). They are engineered to bind to target molecules with high specificity and sensitivity. The anti PSMA Centyrin was tested for specificity and sensitivity using tissue cultured tumor cell lines spiked into normal healthy blood and then compared to anti PSMA antibody. The anti PSMA Centyrin showed 100% positivity with the high PSMA expressing cell line (LNCap), 25% positivity with the low PSMA expressing cell line (22Rv1) and 0% positivity with the PSMA negative cell line (PC3-9). The anti PSMA Centyrin was further tested with prostate patient blood samples to determine the expression levels of PSMA on CTCs. The patient samples which contain CTCs are positive for anti PSMA Centyrin and the percentage of CTCs positive for PSMA ranged from 25-100%. Thus, PSMA expression on CTCs can be determined using CellSearch® system and anti PSMA Centyrin as a biomarker in a clinical trial.
Citation Format: Bhavesh Vaidya, Peter Vulfson, Renouard Sanders, Donna Klein, Shalom Goldberg, Steve Jacobs, Chandra Rao. Detection of PSMA on circulating tumor cells from blood of prostate cancer patients using anti-PSMA Centyrin. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4972.
Abstract 4974: Using serological proteomics analysis to identify anti-nucleophosmin 1 autoantibody in sera from prostate cancer patients as potential diagnositic and prognostic biomarker
Authors: li jitian , Dai Liping , Lei Ningjing , Xing Mengtao , Casiano Carlos A. , Zhang Jianying 
Published: July 2016

Abstract
Prostate specific antigen (PSA) level test in blood has been widely implemented for the early detection and prediction of prostate cancer (PCa). However, lack of specificity led to overdiagnosis, following by unnecessary biopsies and overtreatment. Therefore, novel serological biomarkers are urgently needed to complement PSA by enhancing its sensitivity and specificity in the diagnosis of PCa. Autoantibodies against intracellular tumor associated antigens (TAAs) are commonly found in various types of human cancers. Their utility in providing insights into tumor biology and as potential cancer biomarkers is well documented. Previous studies have demonstrated that sera from patients with PCa contain autoantibodies that react with TAAs. In this study, we identified and characterized nucleophosmin 1 (NPM1) with 33 kDa molecular weight as an autoantigen by using serological proteomics analysis (SERPA) approach. In the further validation study, it was observed that the level of anti-NPM1 in sera from patients with PCa was significantly higher than that from benign prostatic hyperplasia (BPH) and healthy individuals and receiver operating characteristic (ROC) curve analysis showed high diagnostic value for PCa (area under the curve (AUC):0.805). Interestingly, when considering anti-NPM1 combined with PSA level in sera from PCa patients at the early stage as well as BPH, 100% PCa patients could be identified correctly, while 69.2% patients with BPH who has elevated PSA level were found anti-NPM1 negative. The data suggested that anti-NPM1 can elicit humoral immune response in PCa and might be a potential biomarker for the immunodiagnosis of PCa, and it would be a supplementary of PSA examination to distinguish PCa from BPH.
Key words: prostate cancer; nucleophosmin 1 (NPM1); tumor-associated antigens (TAAs); autoantibodies; serological proteomics analysis (SERPA)
Citation Format: jitian li, Liping Dai, Ningjing Lei, Mengtao Xing, Carlos A. Casiano, Jianying Zhang. Using serological proteomics analysis to identify anti-nucleophosmin 1 autoantibody in sera from prostate cancer patients as potential diagnositic and prognostic biomarker. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4974.
Abstract 4975: Detection and characterization of circulating and disseminated tumor cells in patients with bladder and prostate cancer
Authors: Verdone James E. , van der Toom Emma E. , Pienta Kenneth J. , Gorin Michael A. 
Published: July 2016

Abstract
Introduction: Increasing evidence suggests that cancer cells display dynamic molecular changes in response to systemic therapy. Circulating tumor cells (CTCs) in the peripheral blood and disseminated tumor cells (DTCs) in bone marrow represent a readily available source of cancer cells with which to measure this dynamic process. To date, many strategies to isolate and characterize these cells have been described. These techniques, however, have been limited by the requirement for positive or negative selection thus resulting in cell losses. Herein we report a method to detect and characterize CTCs and DTCs in matched peripheral blood and bone marrow samples without need for a selection step.
Methods: Following blood or bone marrow aspirate collection, nucleated cells were extracted using a modified density-media centrifugation technique. Nucleated cells were spread on positively charged slides and stained with epithelial markers (Pan-cytokeratin and EpCAM) and a leukocyte marker CD45. High throughput scanning microscopy was performed followed by image processing and subsequent ranking for probability of being a cancer cell. Candidate cancer cells were defined as events which were positive for PanCK and/or EpCAM but negative for CD45. Candidate cells were independently confirmed with high resolution microscopy.
Results: Cytokeratin-positive, EpCAM-positive DTCs and CTCs were readily detectable in patients with clinically-localized bladder and prostate cancer without need for positive or negative selection.
Conclusions: Future work aims to incorporate disease specific makers to confirm that the observed cells truly represent cells of either bladder or prostate origin. This method offers the flexibility to preserve samples for downstream molecular assays including single cell isolation and sequencing. All told, this assay provides a novel means to detect and characterize CTCs and DTCs without positive or negative selection methods.
Citation Format: James E. Verdone, Emma E. van der Toom, Kenneth J. Pienta, Michael A. Gorin. Detection and characterization of circulating and disseminated tumor cells in patients with bladder and prostate cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4975.
Abstract 4982: BNC105 induces tumor micro-environment changes which enhance the efficacy of checkpoint inhibitor therapy in preclinical models
Authors: Inglis Daniel J. , Beaumont Donna M. , Leske Annabell F. , Scherer Michaela A. , Lavranos Tina C. 
Published: July 2016

Abstract
BNC105 is a Phase II stage drug that specifically targets tubulin causing rapid depolymerisation. The therapeutic window of BNC105 has been shown to be superior compared to similar compounds in its class with highly specific action on tumor vasculature and minimal off target activity. BNC105 shows evidence of strong anti-cancer efficacy in vitro and in animal models not only through destruction of tumour microvasculature but also through strong suppression of cancer cell proliferation. Complementary studies in models of chronic lymphocytic leukaemia have shown that BNC105 also activates pro-apoptotic proteins, which mediate cancer cell death. Recent studies have been conducted to assess changes in the tumour microenvironment that make BNC105 amenable to combination with immune checkpoint inhibitors.
Strong synergy with BNC105 has been seen with anti-PD-1 (97%TGI in combination compared to 40% with BNC105 and 74% with anti-PD-1 as monotherapies) and anti-CTLA-4 (70%TGI in combination compared to 27% with BNC105 and 14% with anti-CTLA-4 as monotherapies) using syngeneic MC38 and CT26 colorectal murine tumor models respectively. We hypothesised that the rapid BNC105 induced tumor microvasculature destruction and onset of tumor hypoxia and necrosis leads to an efflux of tumor antigens being presented to the immune system. This has the potential to elicit an immune response against tumors with a lower mutational load than those that would otherwise activate the immune system using checkpoint inhibitors alone. An increase in tumoral IFNgamma can be associated with the presence of antigen-stimulated lymphocytes within the tumor. IFNgamma levels, CD3+ and CD8+ cells were assessed in tumors treated with BNC105 to elucidate the changes in the recruitment of T cell populations within the tumor microenvironment. A more rapid rate of maturation of dendritic cells upon presentation of antigen is also a potential mechanism associated with changes in the tubulin content of the dendritic cell.
The elucidation of the mechanism whereby the disruption of micro-vasculature caused by BNC105 further validates the complementary functional role of BNC105 in immune activation with checkpoint inhibitors. These findings support the investigation of combining BNC105 with immune checkpoint inhibitors with the view of progressing such combinations to clinical evaluation.
Citation Format: Daniel J. Inglis, Donna M. Beaumont, Annabell F. Leske, Michaela A. Scherer, Tina C. Lavranos. BNC105 induces tumor micro-environment changes which enhance the efficacy of checkpoint inhibitor therapy in preclinical models. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4982.
Abstract 4984: Chemotherapy induces a coordinate autoantibody and T-cell response against tumor-associated antigens in pancreatic cancer patients
Authors: Novelli Francesco , Mandili Giorgia , Mazza Emanuela , Bulfamante Sonia Bulfamante , Giordano Daniele , Follia Laura , Cappello Paola 
Published: July 2016

Abstract
Pancreatic ductal adenocarcinoma (PDAC) is one of the most difficult cancer to treat, both for lack of effective screening method and for resistance phenomenon. At present surgical resection is the only potentially curative option, but the absence of early symptoms or clinical-pathological markers results in diagnosis at a late, inoperable stage in mostly of cases. Once diagnosed, a number of chemotherapy (CTX), radiation and combination therapy regimens are usually used to treat patients, but responses remain poor. However, chemotherapy is able to enhance tumor immunogenicity. Thus more immunogenic tumor associated antigens (TAA) can be induced by CTX and targeted by passive or active immunotherapy. We have previously identified a number of TAAs that are recognized by autoantibodies in PDAC patients. One of these, alpha-enolase (ENO1), was found to have good antigenic capability and immunization with DNA coding for ENO1 effectively enhanced the survival of genetically engineered mice that develop spontaneously PDAC. To discover TAAs that might be selected for immunotherapy, we have analyzed autoantibody response in 19 PDAC patients’ sera before and after CTX. The reactivity of PDAC patients’ sera obtained was analyzed on 2-dimensional gel electrophoresis proteome map of CF-PAC1 pancreatic cancer cell line and TAAs recognized by IgG autoantibodies were identified by mass spectrometry. The antibody response against 25 proteins was induced or up-regulated after CTX. Among CTX-induced TAA identified, only four, namely ENO1, FUBP1, CK8 and G6PDH were found able to ex novo induce or up-regulate IgG response in more than seven patients were selected and their ability to activate proliferation of autologous PBMC was evaluated following in vitro stimulation with the corresponding recombinant proteins. Notably, mostly of autologous T cells obtained after CXT displayed an increased proliferation in response to at least one or more of the four selected TAA after in vitro stimulation compared to autologous T cells obtained before CTX. Experiments are in progress to validate these CTX- induced TAA in a mouse model of PDAC and to evaluate the therapeutic value of the combination of CTX with the immunotherapy against these TAAs.
Citation Format: Francesco Novelli, Giorgia Mandili, Emanuela Mazza, Sonia Bulfamante Bulfamante, Daniele Giordano, Laura Follia, Paola Cappello. Chemotherapy induces a coordinate autoantibody and T-cell response against tumor-associated antigens in pancreatic cancer patients. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4984.
Abstract 4988: Reprogramming the immunologic microenvironment through radiation and Axl targeting
Authors: Aguilera Todd A. , Rafat Marjan R. , Castellini Laura , Kariolis Mihalis S. , vonEbyen Rie , Graves Edward E. , Giaccia Amato J. 
Published: July 2016

Abstract
There is increasing evidence that hypofractionated high dose ionizing radiation (RT) can enhance antitumor immune responses in many cancers. In some cases the combination of RT and checkpoint immunotherapy can enhance antitumor immune responses. Here, we developed a model to study the genetic, microenvironmental, and immunologic factors of immune mediated antitumor responses after radiation. Two different tumor clones from the same parental transgenic PyMT mammary carcinoma model reveal similar growth characteristics but different responses to RT primarily due to an antitumor immune response. The responsive Py117 tumors increase expression of PD-L1 after RT and the antitumor response is enhanced with immune checkpoint antibodies (Ab) targeting PD-1 and CTLA-4. However the unresponsive Py8119 tumors do not respond to radiation or combination immunotherapy. Key differences in the microenvironment include greater infiltration of suppressive macrophages, lower MHCI on tumor cells, and low T cell infiltrates. Upon profiling differences in the tumors we found that Axl a receptor tyrosine kinase of the TAM family associated with invasion and metastasis in a broad range of tumors was overexpressed on the surface of Py8119 cells but not Py117. We genetically knocked out Axl in Py8119 cells using CRISPR/Cas technology and found greater tumor latency and enhanced radiosensitivity in the mouse but not in culture. The effects are largely immune mediated as the RT response was diminished in nude mice. On analysis of the tumor cells we found that MHCI is significantly increased and secretion of a number of chemokines is diminished after Axl targeting. In the tumor microenvironment there was significant decrease in macrophages, increase in myeloid dendritic cells, and resulting influx of T cells by 10 days after RT. These data suggest that Axl may not only mediate invasion and metastasis but can influence immunoserveilance and response to therapy through suppression of antigen presentation and supporting a tumor promoting immune microenvironment through chemokine signaling.
Citation Format: Todd A. Aguilera, Marjan R. Rafat, Laura Castellini, Mihalis S. Kariolis, Rie vonEbyen, Edward E. Graves, Amato J. Giaccia. Reprogramming the immunologic microenvironment through radiation and Axl targeting. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4988.
Abstract 4999: Cellular mechanism of action of bispecific T-cell engager (BiTE®) antibody constructs targeting Folate Receptor 1
Authors: Bailis Julie M. , Orf Jessica N. , Sternjak Alexander , Belmontes Brian , Case Ryan , Grinberg Natalia , Chang Wesley , Hinkle Beth , Friedrich Matthias 
Published: July 2016

Abstract
Bispecific T-cell engaging (BiTE®) antibody constructs connect T cells to target-positive cells to induce cytotoxicity. Clinical proof of concept was achieved in B cell malignancies with the anti-CD19 BiTE® Blincyto®, and several BiTE® antibody constructs are currently in development for solid tumor indications. With the goal of developing a BiTE® for ovarian cancer, we focused on targeting the Folate Receptor (FOLR1). FOLR1 mRNA and protein are highly expressed in &gt;80% of ovarian tumor samples. The FOLR1 protein localizes to the cell membrane of ovarian tumor epithelial cells, whereas in normal tissues such as kidney tubules expression is restricted to the apical cell layer. We generated human FOLR1-BITE® antibody constructs that cross react to cynomolgus monkey and here present the preclinical characterization of these molecules. FOLR1-BiTE® antibody constructs had low nanomolar binding affinity for FOLR1 but did not bind to other FOLR family members. The FOLR1-BiTE® antibody constructs recognized diverse epitopes of the FOLR1 protein. In vitro, binding of FOLR1-BiTE® to CD3 on T cells and FOLR1 on tumor cells led to upregulation of CD25, CD69 and GITR on T cells, as well as release of cytokines including TNF, IFNγ, IL-6 and MCP-1. T cell activation by FOLR1-BiTE® was strictly target cell-dependent. Using time lapse imaging, we demonstrated that T cells were recruited to cancer cells expressing FOLR1 to form a cytolytic synapse and induce target cell lysis. Cytotoxicity mediated by FOLR1-BiTE® antibody constructs occurred even at very low target expression levels (1000 receptors per cell), and was not impacted by the presence of soluble receptor at levels found in ovarian cancer patients. FOLR1-BiTE® antibody constructs also mediated redirected lysis of cell lines that were resistant to standard of care chemotherapy. The in vivo activity of the FOLR1-BiTE® was explored in mouse xenograft studies where immunocompromised mice were implanted with mixtures of human T cells and human cancer cell lines. Daily intraperitoneal administration of the BiTE® inhibited tumor growth, even though the serum half-life of the BiTE® was just a few hours.
Citation Format: Julie M. Bailis, Jessica N. Orf, Alexander Sternjak, Brian Belmontes, Ryan Case, Natalia Grinberg, Wesley Chang, Beth Hinkle, Matthias Friedrich. Cellular mechanism of action of bispecific T-cell engager (BiTE®) antibody constructs targeting Folate Receptor 1. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4999.