bluuebunny/crossref_metadata_2025_split huggingface dataset:
Preclinical study of adoptive immunotherapy with natural killer cells in metastatic colorectal cancer.
Authors: Brugnatelli Silvia , Turin Ilaria , Manzoni Mariangela , Maestri Marcello , Montini Enrica , Caliogna Laura , Paulli Marco , Dionigi Paolo , Pietrabissa Andrea , Delfanti Sara , Rovati Bianca , Mariucci Sara , Maccario Rita , Pedrazzoli Paolo , Montagna Daniela 
Published: March 2020

e13103 Background: Several immune-based approaches for treatment of metastatic colorectal cancer (mCRC) are currently under investigation. Aims of our study were to grow and characterize tumor cells (TC) obtained from mCRC patients, and to investigate their susceptibility to autologous natural killer (NK) cell recognition and killing in vitro. Methods: Tumor samples and peripheral blood mononuclear cells from 10 mCRC patients undergoing surgery were collected. Establishment of TC lines that are representative of the original tumor and could be expanded in vitro for a significant number of passage was performed according with our previously described method (Turin I et al, Cytotherapy 2007). CD56+CD3- NK cells were incubated overnight with medium alone or activated with IL-2 (100U/ml) or IL-15 (20ng/ml) and tested against 51Cr-labeled primary TC in NK assay or in ADCC assay after pre-coating of TC with anti-EGFR monoclonal antibody cetuximab. Results: Eight stable mCRC lines have been obtained and characterized to confirm their neoplastic origin. Patient NK cells display low capacity to lyse autologous TC (median 12%, range 5%-17%). IL-2 and IL-15 incubation significantly potentiate NK cytotoxic capacity (median18%, range 6%-62% and median 35%, range 28%-65%, respectively). These results refer to an effector:target ratio of 25:1. ADCC assay has shown that cetuximab pre-coating of TC results on an increasing of their susceptibility to NK-mediated lysis. Phenotypical and molecular analysis of activating NK receptors and their ligands present on surface of TC are currently under evaluation. Conclusions: NK cells derived from mCRC patients display measurable cytotoxic activity against autologous primary TC. Susceptibility of NK-mediated lysis is strongly enhanced by pre-incubation of NK cell with cytokines and by pre-coating of TC with cetuximab. These findings, presented upon approval by the study’s data monitoring committee and undergoing confirmation in a larger series, may support a pilot study of NK cells pre-activated ex vivo for treatment of mCRC patients.
Effect of heparin and its derivatives on the progression of tumor growth in mouse Lewis lung carcinoma model.
Authors: Gray Angel , Hoppensteadt Debra , Hejna Matthew , Fareed Jawed 
Published: March 2020

e13115 Background: Preclinical evidence suggests that heparins have an effect on tumor progression independent of their anticoagulant activity. Heparins have also been shown to exhibit interactions with growth factors and other cellular receptors. This study was designed to investigate whether heparin and its derivatives are able to inhibit tumor growth. Methods: Female C57BL/6 mice were obtained at 6-8 weeks of age and were implanted with 5X105 LN7 tumor cells by dorsal subcutaneous injection of in the upper back. When tumors were first palpable, after 7-10 days of growth, mice were treated with subcutaneous injections of heparin, a low molecular weight heparin (LMWH), namely enoxaparin, an ultra LMWH, semuloparin or saline, daily for two weeks in a dose range of 1.0 – 0.25 mg/kg. After the treatment period, animals were sacrificed and the spleens and tumors were removed and their weight, volume, spleen weight and size were measured. Results: At the 1.0 and 0.5 mg/kg dosages, both enoxaparin (p<0.01) and semuloparin (p<0.01) showed a decrease in tumor volume compared to the saline control animals. At the 1.0 mg/kg dosage, the mortality was high in the heparin group due to bleeding. At 0.5 mg/kg heparin was not different from the saline control. In addition, at a dosage of 0.25 mg/kg, only semuloparin showed a difference compared to the saline control (p<0.01). Similar results were observed for the tumor weight. There were no significant differences noted in spleen weight or spleen size among these agents. The mortality rates between the mice treated with enoxaparin and semuloparin were comparable. Conclusions: These studies suggest that heparin and its derivatives are capable of inhibiting tumor growth in a dose dependent manner. Enoxaparin and semuloparin are more effective at reducing tumor growth compared to heparin. Clinical studies have shown that semuloparin is safe and effective for the prevention of venous thromboembolism in cancer patients and compares favorably to enoxaparin in terms of antithrombotic effect and safety profile. Therefore, semuloparin may be a better alternate for the prevention of cancer associated thrombosis.
A method for assessing tumor response to therapy and more precisely guiding treatment decisions so as to improve survival.
Authors: Wilkerson Julia , Amiri-Kordestani Laleh , Madan Ravi A. , Adesunloye Bamidele , Zhuang Sen Hong , Wells Samuel A. , Fojo Antonio Tito , Bates Susan Elaine , Stein Wilfred Donald 
Published: March 2020

e13122 Background: The response of tumors to chemotherapy is monitored using imaging data or tumor markers and this quantitative data provides a rich source for an objective response assessment and treatment decisions. Responses are usually assessed as categorical variables based on percentage increase or decrease in tumor size. Methods: We have developed mathematical equations that describe efficacy as a continuous variable, enabling the extraction of the appropriate rate constants for tumor growth and regression (decay), designated g and d, respectively. Both are used to describe the rates of tumor growth and regression for the fraction of tumor that is growing despite treatment and the fraction dying as a result of therapy, respectively. Results: Using data from randomized phase III trials in kidney and breast cancer, multiple myeloma, and medullary thyroid carcinoma; as well as phase II trials in prostate cancer we have shown that: (1) values of g but not those of d are strongly correlated (negatively) with patient survival; (2) g can be discerned early in treatment, before growth is demonstrated clinically, providing an early efficacy measure; (3) g typically does not change over time, even over years, suggesting resistance is intrinsic and predictable and does not worsen over time; (4) effective therapies both increase d, and reduce g; and (5) in every cancer studied, the evidence suggests tumor growth reverts to its pre-treatment rate when chemotherapy is discontinued. Conclusions: The observation that g remains stable allows one to predict the most likely outcome of continued therapy. The evidence indicates that the increase in g occurring after treatment discontinuation is due to a resumption of a pre-treatment growth rate and not a change in biology. Our hypothesis is that if a favorable growth rate that slows tumor growth can be identified, survival might be improved if therapies that achieve this favorable growth rate are continued despite crossing conventional disease progression boundaries. We plan a prospective test of this model to provide a more informed decision and better survival outcome by maximizing the benefit obtained from approved therapies.
Improved 5-year overall survival (OS) in breast cancer (CA) patients from 1994 to 2008.
Authors: Aljadir David Naji , Cummings Francis J. 
Published: March 2020

e13126 Background: A Canadian Study (Chia, ASCO 2003, Abstract 22) comparing OS across 4 time cohorts in patients with metastatic Breast CA demonstrated OS benefit at 1 and 2 years that appears related to the release of aromatase inhibitors (AI)/docetaxel and trastuzumab/capecitabine but not paclitaxel or vinorelbine. Methods: 5 yr overall survival (OS5) in breast CA patients (all stages) was separated into 3 time cohorts corresponding to dates of release of chemotherapy or hormonal treatments (cohort 1:1994-1997 (paclitaxel/vinorelbine), cohort 2:1998-2002 (AI /trastuzumab) and cohort 3:2003 (bevacizumab)) to determine if there was an impact on OS5 in all stages and if there was a relation to survival based on stage of disease. Data was extracted from the National Cancer Database (NCDB) for the US and Northeastern US (MA, ME, CT, RI, VT, NH). Results: The OS5 in the US was 80.0% [CI 80.6-80.9], 84.7% [CI 84.4-84.6] and 85% [CI 84.8-85.2] for cohorts 1, 2 and 3, respectively. The OS5 in the Northeast US was 82.3% [CI 81.8-82.8], 85.7% [CI 85.4-86.1] and 87.3% [CI 86.6-88.0] for cohorts 1, 2 and 3, respectively. Stage II 5 yr survival in the US was 78.9% [CI 78.7-79.1], 82.8% [CI 82.6-83.0] and 84.8% [CI 84.4-85.2] and in the Northeast US was 79.1% [CI 78.2-80], 82.4% [CI 81.7-83.1] and 83.7% [CI 82.2-85.3] for cohorts 1,2, and 3, respectively. 5 yr survival for stage III in the US was 53.8% [CI 53.2-54.3], 57.8% [CI 57.3-58.3] and 64.5% [CI 63.7-65.4] and in the Northeast US was 53.2% [CI 50.9-55.6], 58.8% [CI 56.7-60.8], and 70.7% [CI 67.3-74.1] for cohorts 1,2 and 3, respectively. Stage IV patients showed no change in 5 yr survival over the 3 cohorts, but showed improvement in 2 yr survival at the National Level (44.6%, 46.1%, and 46.7%) and in the Northeast US (46.7%,47.5% and 49.4%) for cohorts 1,2 and 3,respectively. Conclusions: 5 yr survival in stage II and stage III breast CA between cohorts 1 and 3 has improved, whereas stage IV survival at 5 yrs showed no improvement in any cohort, even though there was an improvement in stage IV 2 yr survival in the US and within the Northeast US. The improvement in OS5 is more pronounced between cohorts 1 and 2 than between cohorts 2 and 3, which appears to correspond to the addition of AI and trastuzumab.
Role of sphingosine kinase 1 (SphK1) on cetuximab resistance in colorectal cancer models.
Authors: Bianco Roberto , Rosa Roberta , Nappi Lucia , Formisano Luigi , Damiano Vincenzo , d'Amato Claudia , de Maio Ana Paula , Marciano Roberta , Malapelle Umberto , Troncone Giancarlo , Carlomagno Chiara , De Stefano Alfonso , Tortora Giampaolo , De Placido Sabino 
Published: March 2020

e13509 Background: Although EGFR inhibitors, such as the mAb cetuximab, represent an effective strategy in colorectal cancer (CRC), the clinical use of these agents is limited by intrinsic or acquired resistance. Alterations in the ‘sphingolipid rheostat’, or the balance between the proapoptotic molecule ceramide and the mitogenic factor sphingosine-1-phosphate (S1P), due to overactivation of sphingosine kinase 1 (SphK1), have been involved in the regulation of resistance to anticancer agents. Since some studies described cross-talks between SphK1 and EGFR-dependent signalling pathways, we investigated the contribution of SphK1 to cetuximab resistance in CRC models. Methods: We used CRC cell lines with both intrinsic or acquired resistance to cetuximab. In these models, we analyzed SphK1 expression/activation by using different tools, including the available drug fingolimod (FTY720), both in vitro and in vivo. We confirmed our data through a tissue microarray (TMA)-based analysis on CRC tissues. Results: SphK1 is overexpressed in CRC cells resistant to cetuximab. Higher doses of N,N-dimethylsphingosine (DMS), a potent competitive inhibitor of SphK1, are needed to achieve complete enzyme saturation and survival inhibition in resistant cells. Moreover, ceramide induces apoptosis less efficiently in resistant than in sensitive cells, consistently with the idea that increased SphK1 levels mediate S1P synthesis by ceramide in resistant cells. SphK1 contribution to resistance is supported by the demonstration that SphK1 inhibition by DMS or silencing via siRNA in resistant cells restores sensitivity to cetuximab, whereas exogenous SphK1 overexpression in wild-type cells confers resistance. Re-sensitization to cetuximab is observed after treatment with fingolimod, a S1P receptor inhibitor, both in vitro and in nude mice xenografted with CRC cells. Finally, a TMA-based analysis on CRC tissues revealed that SphK1 expression is related to K-Ras mutational status, a well-known determinant of cetuximab resistance. Conclusions: Our data could clarify the role of SphK1 in the onset of resistance to cetuximab, thus suggesting SphK1 inhibition as a part of novel targeting strategies for resistant cancer patients.
Relationship of alterations in the expression of nontargeted genes and suppression of bcl-2 by antisense oligonucleotides.
Authors: Rubenstein Marvin , Hollowell Courtney M.P. , Guinan Patrick 
Published: March 2020

e13513 Background: Antisense oligos have been employed against prostate cancer models targeting proteins associated with growth/apoptosis. However gene therapy may not be totally specific. Oligos against bcl-2 are in clinical trials and it was of interest: 1) To see whether therapy produces non-specific effects compromising efficacy; 2) To identify genes whose activity may have to be maintained/replaced. Methods: In LNCaP cells we evaluated monospecific (directed against bcl-2) and bispecific (directed against bcl-2 & epidermal growth factor receptor) oligos which comparably suppressed bcl-2. Cells were evaluated by PCR for expression of non-targeted genes associated with apoptosis (caspase-3, clusterin); androgen regulation (androgen receptor [AR], p300, creb binding protein [CBP]); cytokines (interleukins 2 [IL-2], 4 [IL-4], interferon [IFN]); antigen expression (prostate specific antigen [PSA], prostate membrane antigen [PSMA], prostatic acid phosphatase [PAP]); and autocrine growth (insulin like growth factor [IGF1]). Results: Most non-targeted genes were unaffected. However following suppression of bcl-2 activity by mono- and bispecific oligos, cells compensated by suppressing caspase-3 (apoptosis promoter) and enhancing AR and coactivator p300 expression. This suggests an ability to reverse bcl-2 suppression of apoptosis through decreased expression of a promoter and increased androgen sensitivity. It identifies caspase-3 as an essential gene whose expression must be either maintained/replaced when bcl-2 is therapeutically suppressed. Among the differentiation antigens, cell surface PSMA expression was increased by bispecific oligos; secretory PSA and PAP were not. Increased PSMA may to be due to induced IFN expression produced by bispecific oligos, attributed to a 2X intrastrand conformation not seen in the monospecific oligo. Increased PSMA expression could make prostate cancer cells more recognizable to cytotoxic T cells and potentiate prostate cancer vaccines. Conclusions: If gene therapy is to be effective, the mechanisms for compensatory changes which contribute to tumor resistance must be further identified/controlled.
Cellular response to the combined effect of trifluoperazine and ionizing radiation: A molecular study.
Authors: Gangopadhyay Sudeshna , Karmakar Parimal 
Published: March 2020

e13533 Background: It is known that some phenothiazines group of drugs have the potential to modulate ionizing radiation (IR) induced cellular damage. In our previous study, we have shown that Trifluoperazine (TFP), a phenothiazine derivative antipsychotic drug, modulate the effect of IR by inhibiting the IR induced DNA double strand break (DSB) repair, and raising the possibility of using TFP as an adjuvant to radiotherapy. In the present study we tried to explore the mechanism of IR induced DNA damage repair inhibition by TFP. Methods: Effect of TFP and IR on cell viability was determined by MTT assay. Study of possible signaling pathway leading to IR induced cell killing was checked using indirect immunolabeling and transfection. Results: It was observed that the use of TFP (2.5 µg/mL) reduces the LD50 from 13 Gy to 7.5 Gy. After transfecting the cells with GFP-tagged Ku80 plasmid, we have seen that in the gamma-irradiated cells, Ku80 was completely localized in the nucleus, indicating close association of Ku with the broken DNA ends. However, in the TFP (2.5 μg/mL) treated cells, Ku80 was retained in the cytoplasm after irradiation. Similarly, with indirect immunofluorescence experiments using antibody against DNA-PKcs, the catalytic component of DNA-PK, large bright dots of DNA-PKcs was present in the nucleus when cells were treated with IR alone. But, when the cells were irradiated in the presence of TFP, DNA-PKcs migrated to the nucleus partially, even after 4 h of irradiation. Conclusions: Thus, TFP prevents the trafficking of DNA-PK complex from cytoplasm to nucleus and thereby inhibits the DNA-PK mediated NHEJ. Taken together, our results suggest that TFP sequestered the DNA-PK complex, which is a major player in non homologous end joining (NHEJ) and enhance the effect of IR.
Inhibition of cardiomyocytes late INa with ranolazine blunts anthracyclines-cardiotoxicity in experimental models in vitro and in vivo.
Authors: Maurea Nicola , Coppola Carmela , Rea Domenica , Piscopo Giovanna , Riccio Gennaro , Barbieri Antonio , Palma Giuseppe , Gala Marianna , Luciano Antonio , Giudice Aldo , Capasso Immacolata , Scala Stefania , Iaffaioli Rosario Vincenz , De Lorenzo Claudia , Arra Claudio , Tocchetti Carlo G. 
Published: March 2020

e13539 Background: Anthracyclines produce a well-known cardiomyopathy through multiple mechanisms, which also include, among many, Ca2+ overload due to reduced SERCA2a activity and inappropriate opening of the RyR2, and impaired myocardial energetics. Anthracyclines generate Reactive Oxigen and Nitrogen Species (ROS and RNS), posing the heart at increased demand for oxygen, thus setting the stage for a metabolic ischemia that also activates late INa, the target of ranolazine (RAN). Here, we aim at assessing whether RAN, diminishing intracellular Ca2+ through its inhibition of late INa, and enhancing myocardial glucose utilization (and/or reverting impairment of glucose utilization caused by chemotherapy) blunts anthracyclines cardiotoxicity. Methods: To assess for toxicity in vitro, rat H9C2 cardiomyoblasts were pretreated with RAN (0.1-1mM) for 72 hours and then treated with doxorubicin (DOX, 0.1 mM) for additional 24 hours. Cells counts were assessed by Trypan exclusion test. To evaluate cardiac function in vivo, fractional shortening (FS) and ejection fraction (EF) were measured by echocardiography in C57BL6 mice, 2-4 mo old, pretreated with RAN (370mg/kg/day, a dose comparable to the one used in humans) per os for 3 days. RAN was then administered for additional 7 days, together with DOX (2.17mg/kg/day ip), according to our well established protocol. Results: After DOX, only 68% of the cells were viable. RAN alone did not affect cell survival, but blunted DOX toxicity, rescuing % cell survival to 87% (p=.01 vs DOX alone). In our in vivo studies, after 7 days with DOX, FS decreased to 50±2%, p=.002 vs 60±1% (sham), and EF to 81±2%, p=0.0001 vs 91±1% (sham). RAN alone did not change FS (59±2%) nor EF (89±1%). Interestingly, in mice treated with RAN and DOX, the reduction in cardiac function was milder: FS was 57±1%, EF was 89±1%, p=0.01 and 0.0009 respectively, vs DOX alone. Conclusions: RAN blunts DOX cardiotoxic effects in 2 different models, in vitro and in vivo. We plan to test RAN as a cardioprotective agent with other antineoplastic cardiotoxic drugs in our experimental models, and to better characterize the cardioprotective mechanisms of RAN in all these settings.
Screening of SirT1 activating compounds and their cytotoxicity in prostate cancer cell lines.
Authors: Fu Maofu , Pattabiraman Nagarajan , Wang Chenguang , Ju Xiaoming , Sauve Anthony , Pestell Richard G. 
Published: March 2020

e13545 Background: Our previous studies demonstrated that the human androgen receptor (AR) is an acetylated protein, and acetylation of the AR in the hinge region is essential for its ligand-dependent transactivation. The AR acetylation mimic mutant, ARK630Q, and a somatic mutation from the prostate cancer patient at the AR acetylating motif, ARK630T, showed enhanced receptor/coactivator binding and led to accelerated prostate cancer cell growth. SirT1, an NAD+-dependent class III HDAC, suppresses AR and inhibits prostate cell proliferation via deacetylation of AR. Deletion of the SirT1 gene in mice resulted in prostatic intraepithelial neoplasia (PIN) lesion formation associated with reduced autophagy. Our hypothesis is that small molecules with SirT1 activating properties could be potential novel agents for treatment of AR-expressing prostate cancer. Methods: We performed in-silico screening for small molecule antagonists of SirT1 based on computational model prediction of a known SirT1 activator, resveratrol, and identified potential activators for SirT1. The cytotoxicity of these compounds to the prostate cancer cell lines was tested by MTT assay. Results: Among the twelve compounds tested, three compounds, C5, C6 and C10, were found to inhibit AR expression in LNCaP cells and have cytotoxicity in several prostate cell lines including LNCaP, C4W2 and NeuT-transformed prostate epithelial cell line at concentration of 50 µM. Conclusions: Currently, we are testing the effects of these small molecules on the NAD+-dependent deacetylation activity of SirT1 and their effects on AR acetylation as its molecular mechanism of cell growth inhibition. The ultimate goal of this study is to develop early phase clinical trials in prostate cancer patients by blocking AR acetylation using these SirT1 activators.
A pharmacodynamic study of JI-101, an orally active inhibitor of VEGF-2, PDGF-β, and EphB4 receptors, in patients with refractory/recurrent ovarian cancer.
Authors: Werner Theresa Louise , Luebke Aaron Lynn , Wade Mark , Srinivas Nuggehally R , Abhyankar Dhiraj J. , Caligiuri Philip , Heilbrun Marta Elise , Sharma Sunil 
Published: March 2020

e13548 Background: JI-101 is an oral multi-kinase inhibitor, which targets VEGFR-2, PDGF-β, and EphB4. By targeting multiple angiogenesis signaling pathways, JI-101 has the potential advantage of inhibiting all vital stages of tumor angiogenesis, reducing tumor resistance, and enhancing anti-tumor efficacy. None of the currently approved angiogenesis inhibitors have any appreciable inhibition of EphB4, and therefore, this represents a novel mechanism of action. JI-101 has preclinical activity in various tumor models and has been well tolerated in phase 1 trials. Methods: Eleven women with refractory/recurrent ovarian cancer were enrolled with a median age of 54 yrs (range 52-76). All patients had adequate organ function and ECOG performance status ≤ 2. Each patient received single agent JI-101 at 200mg BID for 28 day treatment cycles. Toxicity and response were evaluated at two month intervals. Results: Eight of 11 patients were evaluable for response (3 were unevaluable due to inability to complete two cycles). A median of 3 cycles (range 2-6) was given. No grade 4 adverse events (AEs) were seen. The most common grade 2/3 hematological AEs were grade 2 anemia (1) and lymphopenia (1). The most common grade 2/3 non-hematological AEs were grade 3 hypertension (7), grade 3 bowel obstruction (2), grade 3 transaminitis (2), and abdominal pain (2). Six patients had stable disease (SD) at two months and two had progressive disease. Best response was SD at four months. Two-month progression free survival was 71% (90% CI 52-99%). Conclusions: JI-101 is well tolerated in refractory/recurrent ovarian cancer patients with the majority of patients with stable disease at two months. The most common toxicity was hypertension, which was easily controlled with anti-hypertensives. The novel mechanism of action of JI-101 is promising in ovarian cancer treatment and further prospective studies of this agent could be pursued in a less refractory patient population.