bluuebunny/crossref_metadata_2025 huggingface dataset:
Multiplexed mass spectometic quantitation of HER1-3, cMET, and IGF1R in FFPE tumor samples: Implications for targeted therapy and resistance.
Authors: Hembrough Todd A. , Liao Wei-Li , Thyparambil Sheeno , Darfler Marlene , Krizman David , Burrows Jon
Published: March 2020

e21068

Background: The human EGF receptor family (HER’s) consists of two clinically validated drug targets (EGFR and HER2), and two receptors (HER3 and HER4) which are the subject of intensive preclinical and early clinical investigation. Although drugs inhibiting both EGFR and HER2 show significant antitumor activity in the clinic, the acquisition of resistance is a hallmark of these and other targeted therapies. In the case of both targets, one of the emerging resistance mechanisms is the co-expression of other receptor tyrosine kinases, including members of the EGFR superfamily, cMet and IGF1R. As an example, it was recently shown that HER2 co-expression mediates resistance in cetuximab treated head and neck cancer. Similarly, much attention has been paid to HER3 both as a bona fide drug target and a resistance mechanism. Methods: Using trypsin digestion mapping of recombinant proteins, we identified unique peptide sequences from each of these receptors, and built quantitative mass spectrometric (MS) assays which could be multiplexed into a single MS analysis of 1ug of tumor protein. Assays were preclinically validated on 10 formalin fixed cell lines, and FFPE human NSCLC primary tumor xenografts. Results: The validated multiplex assay was used to measure expression levels of HER1-3, cMET and IGF1R in two cohorts of clinical tumor tissue which had been treated with HER family antagonists. One, a set of gefitinib treated NSCLC tumors (N=15) , and a second, a cohort of advanced breast cancer tissues which had adjuvant treatment with trastuzumab(N=18). Here we present expression patterns for each of the RTKs studied, with the intent to begin to define the relationship between RTK expression and response to either gefitinib or trastuzumab treatment. Conclusions: It is important to not only understand primary mechanisms of tumor growth, but also mechanisms of resistance in patients undergoing targeted therapies. Our Liquid Tissue-SRM promises to be a platform which can deliver extremely high sensitivity, absolute specificity as well as multiplexing capabilities to assess critical oncology targets.
Panel of serum biomarkers to predict benefit from bevacizumab (BEV) in advanced NSCLC patients.
Authors: Braun Eduardo , Fidler Mary J. , Basu Sanjib , Gangaram Anjali , Walters Kelly Kaiser , Karmali Reem , Fhied Cristina , Lie Wen-rong , Borgia Jeffrey Allen , Bonomi Philip D.
Published: March 2020

e21069

Background: BEV has produced modest benefits in patients (PTS) with advanced NSCLC. Identification of positive predictors for BEV would have important implications for individual PTS and health care costs. Methods: We performed a prospective exploratory analysis to identify serum biomarkers as predictors of improved outcomes with BEV. Pre treatment sera were collected from 93 pts prior to initiation of first line treatment for advanced NSCLC. Treatment drugs, including BEV, were prescribed according to treating physician’s discretion. Seventy two serum biomarkers, relevant to angiogenesis and tumor progression, were recorded using Luminex immunobead platform. Serum levels were correlated with progression free survival (PFS) and overall survival (OS) and compared between patient treated with or without BEV containing regimens, BEV + and BEV- groups respectively. Log-rank and interaction p value tests were used to identify markers associated with longer PFS and OS in the BEV+ group but not in BEV- group. Results: Characteristics for each group were: BEV+ (n=43, median age 65 y/o, 72% smokers, 60% females, 100% non-squamous). BEV– (n=50, median age 64 y/o, 84% smokers, 50% female, 70 % non-squamous). The BEV+ group had longer PFS (5.8 vs. 3.0 mos, log-rank p= 0.039) and OS (13.1 vs. 8.5 mos, log-rank p =0.11) when compared to the BEV- group. High serum levels of these markers resulted in a differential decreased hazard in the BEV+ group: PDGF-AB/BB (interaction p <0.01 for PFS, p=0.04 for OS), FGF (interaction p=0.15 for PFS, p<0.04 for OS), tenascin-c (interaction p=0.18 for PFS, p=0.04 for OS), RANTES (interaction p=0.04 for PFS, p=0.6 for OS), epiregulin (interaction p=0.31 for PFS, p=0.04 for OS) and anti-HGF (interaction p=0.18 for PFS, p=0.03 for OS). In the BEV+ group higher levels of PDGF-AB/BB were associated with a better outcome (log-rank p=0.05 and p=0.01 for PFS and OS respectively). We did not find significant correlations between serum levels of VEGF, anti-VEGF or VEGFR and benefit from BEV. Conclusions: This exploratory analysis suggests that these biomarkers may have predictive value for BEV in NSCLC PTS and should be considered for further studies.
Performance of the high-definition circulating tumor cells (HD-CTC) assay in patients with non-small cell lung cancer (NSCLC).
Authors: Bazhenova Lyudmila , Nieva Jorge J. , Kolatkar Anand , Luttgen Madelyn , Marinucci Dena , Bethel Kelly , Kuhn Peter
Published: March 2020

e21074

Background: Technologies to isolate and study circulating tumor cells (CTCs) became available in recent years. To date, the most widely employed techniques to detect CTCs use EpCAM based enrichment protocols. However, those assays are only able to find CTCs in a minority of patients with NSCLC, thus limiting its potential applicability. We are reporting performance of a new enrichment-free HD-CTC assay in patients with NSCLC. Methods: This multi-institutional study recruited patients with all stages of NSCLC. Blood was collected at the time of the study entry, 3 weeks, 3m, 6m, 9m, and 12 m. CTCs were identified with enrichment free fluorescent labeling followed by automated digital microscopy. CTCs were physically and morphologically characterized by cytokeratin (CK) intensity, nuclear size, apoptotic changes, and aggregate formation. Cells were classified as HD-CTCs if they met strict morphologic and immunofluorescence characteristics. This assay has been previously validated in 15 normal controls. Results: 106 patients were recruited (I-10, II-3, III-34, IV-59), 78 were chemotherapy naïve. 85%, 79%, and 53% of patients with stages, I+II, III, and IV had detectable HD-CTCs. The chemotherapy naïve cohort had median HD-CTCs of 4.4/mL (range 0–515.6) and a mean of 44.1 (±95.2)/mL. No significant differences were detected in median HD-CTC count between tumor stages (p = 0.32). CK intensity, nuclear size, frequency of apoptosis and aggregate formation was also similar across stages. 28 patients with stage IV disease were analyzed with multiple draws over time. Mean CTC counts were averaged over multiple draws. In each individual patient there was a trend for increased CTC prevalence with advancement of the disease. Median survival was 244 days for patients with ≥5 CTC/mL vs. 304 days for patients with < 5 CTC/mL. HR 4.0, (p=0.0084). Conclusions: The HD-CTC assay can detect CTCs in a high percentage of patients with early and advanced NSCLC. Small subset analysis demonstrated the prognostic value for the enumeration of HD-CTCs. The high prevalence of HD-CTCs across stages could make this assay an ideal candidate for larger scale investigation into prognostic and predictive features of HD-CTCs.
Morphologic variation of circulating tumor cells in metastatic breast cancer patients visualized by immunofluorescent stain pattern.
Authors: Adams Daniel , Martin Stuart S , Charpentier Monica , Makarova Olga V , Zhu Peixuan , Li Shuhong , Amstutz Platte T , Tang Cha-Mei
Published: March 2020

e21079

Background: Isolation of circulating tumor cells (CTCs) using microfiltration is a growing utility in the field of CTC detection. The microfiltration approach can be used on peripheral blood as a non-invasive liquid biopsy for cancer detection and subtyping. We present a utilization of the CellSieve microfilter to subtype CTCs based on immunofluorescent staining pattern of cytokeratin filamentation and EpCAM surface marker expression. Our initial study on CTCs in patient blood indicates that disseminated CTC populations have high rates of phenotypic heterogeneity. Further detailed molecular analysis and patient tracking of these phenotypes may lead to individualized patient assessment based on CTC characterization. Methods: 7.5 mL of whole blood collected from metastatic breast cancer patients were diluted in a fixative solution. An 8 µm CellSieve precision microfilter was placed into a filter holder and the samples were passed through the filter (~ 90 seconds). The cells captured on the filter were fixed, permeabilized, and stained with DAPI, cytokeratin (FITC), EpCAM (PE), and CD45 (Cy5). Cells without CD45 staining were classified by their morphology, nuclear integrity and the presence of cytokeratin and EpCAM staining. Results: In our initial assessment, patient samples were found to have a number of phenotypic CTC subtypes. Cytokeratin filamentation was clearly seen on a number of CTCs while other cells presented with spotted patterns, implying CTCs in various stages of apoptosis. Later stage apoptosis, with segmented nuclear signature, was also seen in various samples. Cell clusters and cells in division were DAPI positive, while EpCAM positivity was negligible, 0-3 cells/sample, correlating with established data from the CellSearch CTC assay. Conclusions: In addition to enumeration, phenotypic variation of CTCs may be a valuable tool for the personalized care of cancer patients. We have shown that individual breast cancer patients have overlapping phenotypes of CTCs circulating in their peripheral blood. We have begun categorizing patients based on these phenotypes and plan to correlate them with overall prognosis.
Value of circulating epithelial tumor cells and circulating endothelial cells (CTCs/CECs) in patients with HER2-negative recurrent or metastatic breast cancer treated with bevacizumab (B) in combination with paclitaxel (P) and gemcitabine (G) as first-line therapy (AVALUZ trial).
Authors: Codes Manuel , Manso Luis , Ciruelos Eva M. , De la Haba- Rodriguez Juan , Galan Antonio , Baena Jose Manuel , Jaen Ana , Gil Miquel , Murias Adolfo , Blancas Isabel , Gonzalez Encarnacion , Perez Diego , Bayo Juan L. , Mel Jose Ramón , Garcia-Martinez Elena , Cubedo Ricardo , Salvador Javier
Published: March 2020

e21088

Background: CTCs in peripheral blood are an ideal source for the detection of tumor dissemination. Their prognosis significance has been demonstrated in metastasic breast carcinoma (Cristofanilli, NEJM 2004). Beacizumab in combination with CT, improves progression free survival (PFS) of first line treatments, may modify CTC and CEC levels. Aims of this study are the evaluation of the prevalence and kinetics of CTCs and CECs before and after treatment with B in pts with MBC Methods: Pts received B 10mg/kg/q2w combined with P 150mg/m2 and G 2000mg/m2 d1,15/q28d therapy until disease progression, unacceptable toxicity or withdrawal. CTC/CEC were measured in 7.5ml of blood at baseline and after the 1st cycle of treatment. Enumeration was performed by CellSearch System, Veridex Results: Data were available for 37 pts. Median of f-up was 16.2 m. The media of CTCs was 41 cells (min0-max845) in the 1st determination and 5 cells (min0-max99) in 2nd determination. Baseline CTCs>2 was associated with statistic lower PFS 10.8 m (95%CI:7.66-16.91) compared to those with CTCs<2, PFS 17.7m (95%CI:17.60-NA),p=0.046. Baseline CTCs>10 vs CTCs<10 was associated with statistic lower OS (14%vs45% of deaths), p=0.035. 92% (N=22) of pts that had stable disease/partial response, decreased or maintained CTCs value. CTC level was not correlated with CEC level, p=0.74. Media value of baseline CECs was 130 (min4-max1407) and 60.3 (min0-max349) in 2nd determination. Baseline CECs>200 was associated with lower PFS 8.2m (95%CI:0.6-10.8) compared to those with <200, PFS 16.9m (95%CI:8.78-NA),p=0.003. No difference was observed in OS. 74% of pts (N=14) that had stable disease/partial response, decreased or maintained CECs value. Baseline CTCs >5 was associated with a median PFS of 15.2 m (95%CI:7.6-16.9) Conclusions: Our study suggests significant correlations between levels of baseline CTCs and CECs and poor prognosis. Addiction of B to 1st-line CT was related with high reduction of CECs and CTCs count.
Molecular prognostic markers in colorectal cancer.
Authors: Ivanov Krasimir , Kolev Nikola , Tonev Anton , Tontchev Anton
Published: March 2020

e21100

Background: Patient 5-year survival rate from colorectal cancer in the last few decades remains at 40 to 45 percent. Nearly one-half of all patients who undergo a potentially curative resection will relapse because of undetected metastasis. Overall survival rate remains poor which strongly suggests that the dissemination of these cells occurs early in the disease process there is a need of diagnostic methods which evaluate tumor aggressiveness. The p-53, MUC-2, Ki-67, VEGF, Bax, Bcl-2, Stat, MMP2 are markers, which could provide useful individual information about biological potential and prognosis. Methods: We evaluate the expression of those markers in group of 72 patients with colorectal carcinoma. We observed the relations between: 1) type of operation; 2) histological type; 3) clinical stage; 4) individual risk index (IRI), based on evaluated expression of tumor markers; 5) impact on survival rate. Results: Achieving 100% successful rate in our study, we statistically analyzed the data received from marker’s expression. We calculated an IRI for every patient, based on the data from imm3unohystochemical expression by already mentioned molecular markers and divided the patients in 3 groups. A Group A, formed by 28 (38%) patients, which are with low clinical stage and worse molecular prognosis. A Group B, formed by 20 (27%) of patients had low tumor aggression and III or IV clinical stage and Group C, formed by the other 26 (35%) of the patients where is a correlation between the molecular and clinical stage prognosis. We found significant differences in survival rate in Groups A and B, according to clinical stage and our proposed IRI. Conclusions: Multivariate analysis revealed that the IRI is independent prognostic factor for the tumor outcome. There were many evidences for the need of alteration of the surgical behavior in different stages according to biological nature of the tumor. Analysis of a combination of immunohistochemical molecular markers with the conventional diagnostic methods for colorectal cancer allows better prediction of the patients’ prognosis and more accurate and individualized therapeutic strategy. That could decrease the incidence of recurrence-rate and low the mortality rate of this society-significant disease.
Prognostic value of stem cell quantification in stage II colon cancer.
Authors: Vaz M. Angeles , Martinez Juan Carlos , Devesa Manuel , Die Trill Javier , Abraira Victor , Riquelme Alejandro , Carrato Alfredo
Published: March 2020

e21104

Background: Cancer stem cells (CSCs) are a subset of tumor cells with capacity to self-renew and generate the diverse cells that comprise the tumor. They are considered cancer (Ca) initiating cells responsible for tumor recurrence and maintenance. These cells express pluripotency markers (CD133 and NANOG) and do not express markers of differentiation as cytokeratin 20 (CK20). The potential prognostic value of CSCs in colorectal cancer has been studied with conflicting results. However some of these series were based on heterogeneous situations, involving rectal and colon cancer (CC) and different tumor stages (I-IV). The aim of this study is to evaluate the prognostic value of CSCs in a highly homogeneous population of stage II CC. Methods: One hundred stage II CC patients (pts) treated by the same surgical team at Ramon y Cajal University Hospital between 1977 and 2005 were retrospectively analyzed. None of the pts received adjuvant chemotherapy. Inmunohistochemistry expression of CD133, NANOG and CK20 was scored on paraffin sections using four levels: <10%,11-25%, 26-50% and >50% positivity. Kaplan-Meier analysis and log rank test were used to compare survival. Results: Median age: 68 years (45-92). Median follow up: 5.8 years. Recurrent disease: 17 (17%). Expression of CD133 was shown in 60% of the tumors, in 95% for NANOG and 78% for CK20. No correlation was found among expression levels of CD133, NANOG or CK 20 and relapse-free survival (RFS) and overall survival (OS). However, a statistical significant correlation was found between established pathological prognostic factors and RFS and OS. Conclusions: Stem Cell quantification defined by CD133 and NANOG expression has no correlation with RFS or OS in this cohort of Stage II CC. However, a correlation with established prognostic pathological factors was found. Therefore, our results suggest that CSCs may not play a major role in early phases of CC.
Investigating cyclooxygenase-2 signaling in breast cancer-initiating cells.
Authors: Hall Carolyn S. , Walker Andrew , Laubacher Barbara A. , Singh Balraj , Lucci Anthony
Published: March 2020

e21107

Investigating cyclooxygenase-2 signaling in breast cancer-initiating cells Background: Only a small fraction of breast tumor cells, breast cancer-initiating cells (CSC), have the ability to initiate tumor growth and metastasis. Some characteristics of breast CSCs have been described in vitro by employing non-adherent CSC-enriching culture conditions, however, little is known regarding prostanoid signaling or prostaglandin E2 (PgE2) production in CSCs. In this study we measured cyclooxygenase-2 (COX-2) and prostaglandin E2 receptor expression in CSCs and assessed the effects of COX-2 and Ep4 inhibition on in vitro CSC spheroid formation and PgE2 production. Methods: CSCs were cultured in serum-free medium using ultra low attachment plates. Cells were treated with vehicle, a COX-2 inhibitor (Celecoxib), or an EP4 inhibitor (GW627368X) at concentrations of 0.1, 1, 5, 10, 50, or 100uM for 10 days. Spheroids were quantified using a Gelcount machine. Non-linear regression was used to calculate IC50 values. Western blots were performed using anti-COX-2, anti- EP1, 2, 3 and 4 receptor antibodies. Prostaglandin E2 production was measured using a PgE2 immunoassay kit. Results: COX-2 protein expression was observed for MDA-MB-231 and SUM149 CSCs. EP2 and EP4 protein was detected for MCF-7, MDA-MB-231, and SUM149 CSCs. The Celecoxib IC50 values for MCF-7 and SUM149 CSC spheroid formation were 0.5 and 2.2 uM, respectively; GW627386X IC50 values for MCF-7 and SUM149 CSC spheroid formation were 1.2 and 0.3uM, respectively. No significant differences in PGE2 production were observed for MCF-7 CSCs compared to adherent MCF-7 cells (11.5 ± 4.4 vs. 8.7 ± 2.1 pg/mL/106 cells; p=0.88). However, PgE2 production in MDA-MB-231 CSCs was significantly higher than adherent MDA-MB-231 cells (1507.0 ± 329.0 vs. 13.8 ± 5.7 pg/mL/106 cells; p≤0.001) and SUM149 CSC PgE2 production was significantly higher than SUM149 adherent cells (4932.9 ± 501.7 vs. 194.5 ± 31.0 pg/mL/106 cells; p≤0.001). 10uM Celecoxib treatment inhibited PGE2 production in MDA-MB-231 and MDA-MB-231 CSCs. Conclusions: MDA-MB-231 and SUM149 CSCs express COX-2, EP2 and Ep4 and produce high levels of PGE2. CSC spheroid formation is significantly decreased with COX-2 and EP4 inhibition.
Immunologic changes in patients with metastatic melanoma on bevacizumab and chemotherapy versus chemotherapy alone.
Authors: Mansfield Aaron Scott , Nevala Wendy Kay , Markovic Svetomir
Published: March 2020

e21112

Background: Metastatic malignant melanoma is difficult to treat and has a poor prognosis. Systemic therapy for metastatic melanoma has had limited success and novel drug combinations need to be tested. To that end, we have combined anti-angiogenic therapy (bevacizumab) with chemotherapy in clinical trials and have shown some clinical efficacy. Vascular endothelial growth factor (VEGF) is increasingly seen as an immunosuppressive factor that hampers dendritic cell maturation and reduces antigen presentation. We have recently shown that tumor-derived VEGF plays a role in Th-2 driven systemic chronic inflammation seen in advanced melanoma patients. Therefore, our data suggest that blocking tumor derived VEGF with the monoclonal antibody bevacizumab may impact immunity in such a way that augments anti-tumor immunity enhancing clinical efficacy. Methods: In order to understand the immune status in metastatic melanoma patients on chemotherapy (n=55) or chemotherapy plus bevacizumab (n=73), we measured levels of 42 cytokines, chemokines and growth factors in plasma by a mulit-plex bead assay. Matched pairs analysis was performed to determine if the average responses of the measured cytokines were different between the patients that received chemotherapy with bevacizumab or chemotherapy alone. The pre-treatment levels were compared with levels drawn four weeks after the first treatment. Results: There were significant differences in IL-1a, IL-1ra, IL-6, IL-8 and IP-10 between patients treated with chemotherapy alone relative to patients treated with chemotherapy plus bevacizumab. In all cases of patients treated with chemotherapy plus bevacizumab, the levels of these cytokines all decreased relative to pre-treatment samples. In patients treated with chemotherapy alone the levels of these cytokines increased relative to pre-treatment samples. Conclusions: Our data suggest that treating patients with the anti-angiogenic drug bevacizumab may affect the plasma cytokine milieu by reducing the level of many key inflammatory cytokines. Therefore, the use of bevacizumab in combination with chemotherapy may augment host immunity and elicit a more effective anti-tumor immune response.
Cellular responses to lactate in breast cancer.
Authors: Kennedy Kelly Marie , Dewhirst Mark W
Published: March 2020

e21119

Background: Clinical studies across different tumor types have found that high lactate levels correlate with increased metastases and poor patient prognosis, overall and metastasis-free survival; however, mechanisms are unknown. There is increasing evidence of cancer stem cells (CSC). Lactate inducing a stem-like phenotype may be one explanation for the increased metastases seen in high-lactate solid tumors. We hypothesized that lactate induces EMT in human breast cancer cells and induces/selects for a stem-like phenotype. Methods: We examined lactate levels in locally advanced breast cancer (LABC) biopsies using bioluminescence. Breast cancer cell lines (MCF7, T-47D, MDA-MB-231) were treated acutely (hours) or chronically (weeks) with lactate (0-40mM) and tested for cell viability (Annexin V, clonogenics), EMT markers (Snail-1), cellular detachment, and anchorage-independent cell growth (mammosphere assays). There was one “no treatment” control group. Experimental groups were treated with lactate in the context of complete media or glucose-free conditions (to mimic the typical tumor microenvironment). For the mammosphere assays, cells were treated both prior to (attached as a monolayer) and during sphere formation. Results: Lactate concentrations in the LABC biopsies were between 0-7.5mM. Lactate levels relevant to solid tumors (<40mM) were not toxic to the cell lines tested. Cellular detachment after lactate treatment was not statistically significant; however, the percentage of viable detached cells was significantly increased, and they retained colony-forming abilities. Preliminary data shows a slight increase in Snail-1 levels after chronic lactate treatment. Chronic lactate treatment had a slight but statistically significant effect on secondary mammosphere formation in MCF7s, indicating the important self-renewal properties characteristic of CSCs. Conclusions: The effect of lactate treatment alone on human breast cancer cell lines is subtle but significant. Phenotypic changes elicited by lactate are dependent upon lactate concentration and duration of treatment. Results thus far support the hypothesis that lactate affects expression of stem-like properties in breast cancer cell lines. Supported by BC083154 and R01 CA40355
Radiation-induced damage of the rat intestine after external beam irradiation of the liver.
Authors: Cameron Silke , Schwartz Antonia , Sultan Sadaf , Schaefer Inga-Marie , Rave-Fraenk Margret , Christiansen Hans , Ramadori Giuliano
Published: March 2020

e21128

Background: Radiotherapy alone or in combination with systemic therapies, is a major tool for local tumor control. Although organ-targeting techniques have greatly been improved over the past years, radiation enteropathy is a dose-limiting factor for effective radiotherapy. The out-of-field effects on the intestine, caused by radiation treatment of a parenchymatous organ, have not previously been studied. Methods: A single dose of 25 Gray was administered percutaneously to the liver of male Wistar rats after a planning CT-scan. Sham-irradiated animals served as controls. At 1, 6, 24, 96 hours, 1.5 and 3 months the duodenum, jejunum, ileum and distal colon were removed, washed and deep-frozen or prepared for paraffin staining. Results: All animals survived the treatment. Epithelial cell damage occurred in all small-intestinal segments. However, prolonged denudation of the villi together with destruction of the crypt lining was only observed in the ileum, which only received scattered radiation. Whilst the intestinal lining in the duodenum and jejunum was restored at the end of the study, in the ileum, tissue regeneration was deficient. In the colon, changes were minor. Immunhistochemical staining showed radiation mucositis with granulocyte (MP0+) infiltration from 1 to 24 hours in the duodenum and jejunum, when ED1+ macrophages, CD3+ T-lymphocytes, and CD34+ hematopoietic precursor cells were recruited, accompanied by an increase in the chemokines MCP-1, MIP-1α, MIP3α and Il-8. In the ileum, early granulocyte infiltration was delayed but continuous. Recruitment of macrophages and lymphocytes was deficient and induction of chemokines as of the adhesion molecules PECAM-1, ICAM-1 was lacking. Conclusions: Post-irradiation damage to the ileum was delayed and followed by an altered repair process with structural changes of the villi. The observed changes might result from a higher sensitivity to oxidative stress mechanisms with subsequent damage of the regenerative capacity of the crypt-villus axis, accompanied by a sustained “inflammatory response” and vascular damage with a lack of regeneratory cell recruitment.
Effects of radiotherapy and substance P on MIP-2 (macrophage inflammatory protein-2) levels in metastatic breast carcinoma cell lines.
Authors: Korcum Aylin Fidan , Kale Sule , Erin Nuray
Published: March 2020

e21130

Background: Breast cancer is ranked second among the most common cancers in the world. Radiotherapy (RT) is known to reduce the risk of the recurrence in cancer treatment. Substance P (SP), binds to the Neurokinin-1 (NK1) receptor which plays a role in inflammation, proliferation, angiogenesis and migration in cancer cells. MIP-2 (Macrophage inflammatory protein 2, IL-8 in human which has been currently understood to play a role in carcinogenesis is an inflammatory CXC chemokine. The aim of this study is to show whether MIP-2 levels alter with RT and whether SP has a role in this alteration. Methods: In this study, we investigated the effects of SP, NK1 receptor antagonist (NK1 RA) and RT on cell proliferation and MIP-2 secretion in 4T1 cells. 4T1 cells were derived from spontaneously formed breast tumors in Balb-c mice. We have cultured its subsets from metastatic lesions, and we named them as 4THM (heart metastasis), 4TLM (liver metastasis), 4TBM (brain metastasis). Cells were plated as 400-500 cells per well in a 96 well plate containing medium with 5% FBS. After 24-48 hours, cells were applied NK1RA and SP. After 3-4 hours of treatment, the group of RT was irradiated with 45 Gy. The proliferation of the cells was evaluated 48 hours after treatment. This proliferation was evaluated using WST cell proliferation reagent (WST-1, Roche). MIP-2 levels were measured with ELISA. Results: We observed that RT reduced the proliferation of cells in a ratio of 30,2 % in 4T1 cells, 43,9 % in 4THM cells, 44,7 % in 4TLM cells and 34 % in 4TBM cells. 4THM, 4TBM cells have the highest MIP-2 secretion, which was further increased after RT. Also, it was observed that SP, 10-8 M concentration decreased the RT-induced MIP-2 secretion. Conclusions: Increasing MIP-2 secretiondue to RT can precipitate cancer development and SP can be used as a therapeutic aim to prevent such an increase.
The efficacy of axillary ultrasound in the detection of nodal metastasis in breast cancer.
Authors: Lee Belinda , Lim Adrian K. , Krell Jonathan , Satchithananda Keshthra , Lewis Jacqueline S , Coombes R. Charles , Stebbing Justin
Published: March 2020

e21137

Background: Recent reports indicate a lack of survival benefit for axillary node dissection (ALND) versus sentinel lymph node biopsy in early breast cancer. To study this further we assessed the accuracy and effectiveness of ultrasound in detecting axillary nodal involvement in breast cancer patients, aimed at refining and evaluating our current clinical pathways as newly diagnosed invasive breast cancer patients routinely undergo pre-surgical axillary ultrasound. Methods: Ultrasound data were collected from consecutive breast cancer cases over 3 years. Images were reviewed by experienced radiologists and made the following assessments on size, morphology, fatty hilum and cortical thickness of the ipsilateral axillary nodes. The findings were correlated with histology outcomes following ALND. Results: 260 cases were included in the analysis, 113 (43.5%) had evidence of metastatic nodal involvement at final histology. Of these, 59/113 (52.2%) reported positive findings on ultrasonography. The overall positive predictive value of ultrasound for detecting metastatic nodal involvement measured 0.70. The negative predictive value was 0.61. The sensitivity was 52%, specificity measured 78% and the accuracy was 65%. The ultrasound morphological lymph node features with the greatest correlation with malignancy were absence of a fatty hilum (p=0.003) and increased cortical thickness (p=0.03). Cases with a metastatic nodal burden density of a least 20% were more likely to report an abnormal axillary ultrasound. (p=0.009). Conclusions: Axillary ultrasound has a low NPV and negative sonographic results do not exclude axillary node metastases with sufficient sensitivity, to justify its routine clinical use. Clinical pathways need to consider an evidence-based approach, focusing on the criteria by which we select breast cancer patients for axillary nodal dissection.
Expression of tumor endothelial markers (TEMs) in vitro in a canine hemangiosarcoma model of malignant angiogenesis.
Authors: Wypij Jackie M. , Fan Timothy M. , Pondenis Holly
Published: March 2020

e21145

Background: Expression of tumor endothelial markers (TEMs) in tumor-associated blood vessels presents opportunities for targeted therapy. TEM7 and TEM8 are selectively expressed and are associated with a worse prognosis in cancer patients. TEM expression is conserved across species, however species differences do exist as TEM7 is undetectable in murine tumor endothelium. Thus, further investigation of the role of TEMs in malignant angiogenesis is hindered by the lack of optimal animal tumor models. Canine hemangiosarcoma is relevant spontaneous model of malignant angiogenesis based on archetypal cell markers, endothelial functionality, growth factor/receptor expression, and angiogenic gene expression. The aims of this study were to characterize the in vitro expression of TEM7 and TEM8 in canine hemangiosarcoma as a novel model of malignant angiogenesis. Methods: Two canine hemangiosarcoma cell lines were assessed (Den and Fitz). Total RNA was isolated using standard technique, reverse transcribed to cDNA, and amplified by polymerase chain reaction (PCR) reaction using degenerate primers specific for human and murine TEM7 transcripts with forward and reverse primers. Protein expression of TEM7 and TEM8 in cell lysates was evaluated via Western blotting and cell surface expression was analyzed by flow cytometry. Polyclonal anti-TEM7 and anti-TEM8 antibodies (Sigma-Aldrich) were used with positive and negative controls. Results: Basal in vitro expression of TEM7 mRNA was confirmed in canine hemangiosarcoma, as well as in vitro protein expression of TEM7 and TEM8 via Western blotting and flow cytometry. Conclusions: The validation of TEM expression in canine hemangiosarcoma establishes TEM7 and TEM8 as promising targets for further evaluation in this novel model of malignant angiogenesis, and investigation of TEM-targeting is ongoing. This may represent a novel spontaneous animal tumor model for investigation of tumor vascular targeting.
Spatial and temporal variability of intratumoral 18F-FDG distribution in patients with cancer.
Authors: Bhatt Aashish D. , Li Xiao-Feng , Bhatt Geetika , Kloecker Goetz H. , Sharma Vivek R. , Civelek Ali Cahid
Published: March 2020

e21149

Background: We previously reported that in mice models, tumor microenvironment is complex and intratumoral 18F-FDG distribution is heterogeneous. There were apparent spatial and temporal intratumoral 18F-FDG distribution changes occurring even within 24-48 hour interval in human cancer cell xenografts grown in nude mice. We hypothesized that what was documented in mice models may also be applicable to human patients with cancer. The objective of this study was to investigate the intratumoral spatial and temporal distribution of 18F-FDG in cancer patients. Methods: Five patients who had repeat PET/CT scans within 1-28 days and with no interval therapy were identified. All patients had fasted overnight and had blood glucose level of less than 200 mg/dl. They were injected with 8.0-14.0mCi of 18F-FDG intravenously, and whole body PET/CT scans were performed 60-80 minutes later utilizing a Siemens PET/CT scanner. Each patient’s intratumoral FDG activity distributions were mapped and compared to each other utilizing magnified digital images of the tumors. Results: Intratumoral FDG activity is very heterogeneous. Within the same tumor, there is a wide range of high and low FDG uptake variations. Apparent change in intratumoral FDG distribution was found within as short as a 24 hour interval in the same patient. Although the global tumoral FDG activity and liver FDG activity measured as SUVmaximum and SUVaverage were not significantly different from one PET/CT to another, there were significant spatial changes and variations of the tumor FDG activity between the two PET/CT images. Conclusions: Intratumoral 18F-FDG uptake is very heterogeneous and manifests both temporal and spatial changes within as short as 24 hours even in untreated cancer patients. This should be taken into consideration during FDG PET based therapy decisions, for example in tumor and nodal staging or radiation therapy planning.
Using plasma proteomic analysis for venous thromboembolism risk stratification in patients with advanced gastrointestinal cancers.
Authors: Itakura Haruka , Holzer Alison K , Hofmann Lawrence V , Tsao Philip S
Published: March 2020

e21153

Background: Cancer patients represent 15-20% of the estimated 900,000 annual cases of venous thromboembolism (VTE) in the U.S., and thrombotic complications are the second most frequent cause of death in patients with cancer. Currently, no predictors or method exists by which patients with cancer can be accurately risk-stratified for thrombotic risk. Methods: Twenty-four patients with stage III/IV gastrointestinal (GI) cancers were followed for six months for incident VTE in a prospective cohort study. Twenty-nine biomarkers relevant to inflammation and thrombosis were tested for ability to discriminate between age- and sex- matched cancer patients with (n=10) and without incident VTE (n=14). Results: Expression levels of VCAM-1 and MMP-1 were statistically significantly elevated in VTE cases compared with controls in a series of univariate analyses. VCAM-1 remained statistically significant in a multivariate analysis. Conclusions: These findings suggest the role of VCAM-1 as a potential predictor of VTE in patients with GI cancers.
Assessment of feasibility and willingness of former heavy smokers to participate in chemoprevention trials to prevent lung cancer.
Authors: Kumar Nagi B. , Quinn Gwendolyn P. , Crocker Theresa , Alexandrow Mark , Gray Jhanelle Elaine , Estrella Timothy , Schell Michael J. , Sutton Steve , Haura Eric B.
Published: March 2020

e21155

Background: Over 50% of new lung cancers occur in former smokers, who often are seeking strategies to reduce their lung cancer risk. However, recruitment and retention of participants in chemoprevention trials continues to be costly and presents unique challenges. Evaluation of feasibility and knowledge of challenges are critical to inform design and ensure accrual in chemoprevention trials.The study assessed interest and willingness of former heavy smokers to participate in a chemoprevention clinical trial using a botanical agent to prevent lung cancer. Methods: An introductory letter and survey instrument that included the goal of the survey, epidemiological and smoking history, acceptability of trial procedures, perception of lung cancer risk and interest in participating in this trial were mailed to 500 consecutive, former heavy smokers with no cancer from a database of 826 subjects at the Moffitt Cancer Center. Results: 202 (40.4%) men and women returned completed surveys. 98% of respondents were over age 60 and 56% had an undergraduate education or higher. The average years smoked was 40.7 (SD 11.9) pack years. 76% believed there was a 50% chance or greater of developing lung cancer. In response to interest and motivation to participate, 92-96% reported interest in receiving free lung exams, health status monitoring and knowing their lung cancer risk. 88% were interested in being a part of a trial to evaluate a botanical agent for lung cancer prevention. Over 92% of subjects reported a willingness to comply with study requirements, multiple blood draws and trips to the Center, spiral CTs and chest x-rays. Subjects were relatively less enthusiastic (73-79%) about undergoing bronchoscopy, taking multiple study agents and possible assignment to a placebo arm. Conclusions: Our study strongly suggests feasibility, highlights potential challenges and the significant interest and willingness of former smokers to participate in chemoprevention trials.
Behavior of circulating tumor cells detected by EGF tagged SERS nanoparticles in head and neck cancer patients.
Authors: Beitler Jonathan Jay , Wang Xu , Qian Ximei , Wang Hong , Chen Zhuo Georgia , Khuri Fadlo Raja , Nie Shuming , Shin Dong M
Published: March 2020

e21161

Background: Quantification of circulating tumor cells(CTCs) is becoming central to the management of metastatic breast, colorectal and prostate cancers and the potential to improve management of head and neck squamous cell cancer (HNSCC) patients is very high. Our group has published our experience with nanotechnology, highly specific, surface-enhanced Raman spectroscopy(SERS) and Epidermal Growth Factor Receptor(EGFR) ligands to identify 7-720 CTCs/ml in 19 samples from HNSCC patients[1]. Our goal here is to understand the clinical behavior of CTCs in HNSCC for selected patients whom we believe our single channel assay has been accurate. Methods: Our methods for detection of CTCs using EGFR ligands have been previously described[1]. After comparing CTCs in 116 patient samples with and without gross disease, we examined 8 patients with decreasing sequential single channel assays of CTCs and no evidence of disease at last follow-up. Results: For 56 samples from patients with no gross disease, the mean CTC was 94/ml. For 61 samples from patients with gross disease the mean CTC was 193/ml (p=0.002). Among our 4 samples where we had initial disease and a non-zero, no evidence of disease, second sample, the absolute and percentage decrease per month in CTC was 121/ml/month(range 43-160) and 17%/month(range 11-25%). The decrease of 121 CTC/ml/month correlated well with 5 out of 6 other data sets. Conclusions: Targeting of EGFR for detection of CTCs in HNSCC patients appears very promising. Among +EGFR HNSCC patients who appear cured, the average decrease in CTC per ml was 121 per month and 17% per month. To further enhance the sensitivity and specificity of detection of CTCs, we are developing 3 SERS nanoparticles (multiplexed SERS) to target specific and distinct targets on the CTC surface while still maintaining a single tube assay system. 1. Wang, X., et al., Detection of circulating tumor cells in human peripheral blood using surface-enhanced Raman scattering nanoparticles. Cancer Research, 2011. 71(5): p. 1526-32.
Bevacizumab (Bev) with or without erlotinib as maintenance therapy, following induction first-line chemotherapy plus Bev, in patients with metastatic colorectal cancer (mCRC): Efficacy and safety results of the international GERCOR DREAM phase III trial.
Authors: Tournigand Christophe , Samson Benoit , Scheithauer Werner , Lledo Gérard , Viret Frédéric , Andre Thierry , Ramée Jean François , Tubiana-Mathieu Nicole , Dauba Jérôme , Dupuis Olivier , Rinaldi Yves , Mabro May , Aucoin Nathalie , Khalil Ahmed , Latreille Jean , Louvet Christophe , Brusquant David , Bonnetain Franck , Chibaudel Benoist , De Gramont Aimery ,   GERCOR
Published: March 2020

LBA3500^

The full, final text of this abstract will be available at abstract.asco.org at 12:01 AM (EDT) on Sunday, June 3, 2012, and in the Annual Meeting Proceedings online supplement to the June 20, 2012, issue of Journal of Clinical Oncology. Onsite at the Meeting, this abstract will be printed in the Sunday edition of ASCO Daily News.
Phase III randomized trial of definitive chemoradiotherapy (CRT) with FOLFOX or cisplatin and fluorouracil in esophageal cancer (EC): Final results of the PRODIGE 5/ACCORD 17 trial.
Authors: Conroy Thierry , Galais Marie-Pierre , Raoul Jean Luc , Bouche Olivier , Gourgou-Bourgade Sophie , Douillard Jean-Yves , Etienne Pierre-Luc , Boige Valérie , Martel-Lafay Isabelle , Michel Pierre , Llacer-Moscardo Carmen , Berille Jocelyne , Bedenne Laurent , Adenis Antoine ,   UNICANCER-GI/FFCD PRODIGE Intergroup
Published: March 2020

LBA4003

The full, final text of this abstract will be available at abstract.asco.org at 12:01 AM (EDT) on Saturday, June 2, 2012, and in the Annual Meeting Proceedings online supplement to the June 20, 2012, issue of Journal of Clinical Oncology. Onsite at the Meeting, this abstract will be printed in the Saturday edition of ASCO Daily News.
AURELIA: A randomized phase III trial evaluating bevacizumab (BEV) plus chemotherapy (CT) for platinum (PT)-resistant recurrent ovarian cancer (OC).
Authors: Pujade-Lauraine Eric , Hilpert Felix , Weber Béatrice , Reuss Alexander , Poveda Andres , Kristensen Gunnar , Sorio Roberto , Vergote Ignace B. , Witteveen Petronella , Bamias Aristotelis , Pereira Deolinda , Wimberger Pauline , Oaknin Ana , Mirza Mansoor Raza , Follana Philippe , Bollag David T. , Ray-Coquard Isabelle ,   AURELIA Investigators
Published: March 2020

LBA5002^

The full, final text of this abstract will be available at abstract.asco.org at 12:01 AM (EDT) on Saturday, June 2, 2012, and in the Annual Meeting Proceedings online supplement to the June 20, 2012, issue of Journal of Clinical Oncology. Onsite at the Meeting, this abstract will be printed in the Saturday edition of ASCO Daily News.
LUX-Lung 3: A randomized, open-label, phase III study of afatinib versus pemetrexed and cisplatin as first-line treatment for patients with advanced adenocarcinoma of the lung harboring EGFR-activating mutations.
Authors: Yang James Chih-Hsin , Schuler Martin H. , Yamamoto Nobuyuki , O'Byrne Kenneth John , Hirsh Vera , Mok Tony , Geater Sarayut Lucien , Orlov Sergey V , Tsai Chun-Ming , Boyer Michael J. , Su Wu-Chou , Bennouna Jaafar , Kato Terufumi , Gorbunova Vera , Lee Ki Hyeong , Shah Riyaz N.H. , Massey Dan , Lorence Robert M. , Shahidi Mehdi , Sequist Lecia V.
Published: March 2020

LBA7500

The full, final text of this abstract will be available at abstract.asco.org at 12:01 AM (EDT) on Monday, June 4, 2012, and in the Annual Meeting Proceedings online supplement to the June 20, 2012, issue of Journal of Clinical Oncology. Onsite at the Meeting, this abstract will be printed in the Monday edition of ASCO Daily News.
PARAMOUNT: Final overall survival (OS) results of the phase III study of maintenance pemetrexed (pem) plus best supportive care (BSC) versus placebo (plb) plus BSC immediately following induction treatment with pem plus cisplatin (cis) for advanced nonsquamous (NS) non-small cell lung cancer (NSCLC).
Authors: Paz-Ares Luis , De Marinis Filippo , Dediu Mircea , Thomas Michael , Pujol Jean-Louis , Bidoli Paolo , Molinier Oliver , Sahoo Tarini Prasad , Laack Eckart , Reck Martin , Corral Jaime Jesus , Melemed Symantha , John William J. , Chouaki Nadia , Zimmermann Annamaria , Visseren-Grul Carla , Gridelli Cesare
Published: March 2020

LBA7507

The full, final text of this abstract will be available at abstract.asco.org at 12:01 AM (EDT) on Monday, June 4, 2012, and in the Annual Meeting Proceedings online supplement to the June 20, 2012, issue of Journal of Clinical Oncology. Onsite at the Meeting, this abstract will be printed in the Monday edition of ASCO Daily News.
ABT-888 (veliparib) in combination with weekly carboplatin and paclitaxel in advanced solid tumors.
Authors: Rios-Perez Jorge Arturo , Beumer Jan H. , Appleman Leonard Joseph , Tawbi Hussein Abdul-Hassan , Chu Edward , Stoller Ronald G. , Belani Chandra Prakash , Jiang Yixing , Sobol Robert W. , Shepherd Stacie Peacock , Giranda Vincent L. , Chen Alice P. , Huggins-Puhalla Shannon Leigh
Published: March 2020

TPS1138

Background: The combination of paclitaxel and carboplatin is widely used for the treatment of patients with advanced solid tumors of diverse histologies. In breast cancer patients, a weekly regimen of paclitaxel has shown greater efficacy with comparable safety, when compared to every-three-weeks dosing (ECOG 1199). ABT-888 (veliparib) is an oral inhibitor of poly-ADP-ribose polymerase (PARP). Inhibition of PARP has been shown in preclinical studies to potentiate the effect of cytotoxic agents which induce DNA damage, such as platinum agents. The preclinical synergy of carboplatin with veliparib and the efficacy of the combination of paclitaxel with carboplatin supports exploration of this triplet regimen. Methods: This 3+3 phase I trial will seek to determine the maximum tolerated dose (MTD) of the combination of carboplatin (AUC 2), paclitaxel (80 mg/m2), and veliparib in patients with advanced solid tumors. Veliparib will be escalated beginning at 50 mg PO BID to a maximum of 200mg PO BID. Treatment will be given on a weekly basis over a 21-day cycle. There will be an expansion cohort of 6-12 patients with triple negative breast cancer at the maximum tolerated dose. This group of patients will undergo mandatory pre- and post-cycle 1 tumor biopsies. Secondary aims of the study include safety and toxicity of the combination, its pharmacokinetic and pharmacodynamic effects, documentation of any anti-tumor response, and assessment of the characteristics of the tumor specimens obtained in the expansion cohort that may contribute to efficacy. The latter will include whole genome microarray analysis to evaluate expression of genes involved in DNA repair pathways. Currently, the recommended phase II dose has not been determined and enrollment is ongoing on the last planned dose level (veliparib 200 mg BID).