bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4857: Design and biological evaluation of substituted pyrrolo[3,2-d]pyrimidines as dual acting RTK and microtubule targeting agents
Authors: Gangjee Aleem , Shah Khushbu , Dybdal-Hargreaves Nicholas , Mooberry Susan , Bastian Anja , Ihnat Michael 
Published: July 2016

Abstract
Combination chemotherapy with antiangiogenic (cytostatic) and microtubule targeting (cytotoxic) agents (MTA) have shown significant promise and several studies with such combinations have progressed to the clinic. Single agents with both antiangiogenic activities as well as cytotoxicity would circumvent pharmacokinetic problems of multiple agents, might be effective at lower doses to alleviate toxicity, and delay or prevent tumor cell resistance. We have designed, synthesized and evaluated single agents that are novel pyrrolo[3,2-d]pyrimidine MTAs that also inhibit vascular endothelial growth factor receptor-2 (VEGFR-2), epidermal growth factor receptor (EGFR) and platelet-derived growth factor receptor â (PDGFRâ). Sixteen pyrrolo[3,2-d]pyrimidine analogs were synthesized with variations on the C-2 and the 4-anilino position. The importance of various substituents was evaluated based on their abilities to cause cellular microtubule depolymerization and inhibit proliferation of MDA-MB-435 cancer cells. Compounds with N5-Me were more potent than the N5-H analogs indicating that a conformational restriction induced by N5-Me could mimic the bioactive conformation. In addition, all C2-Cl compounds afforded 2 to 10-fold more potentIC50 values than their C2-Me counterparts as illustrated by C2-Cl analog AG336 (IC50 = 18.3 ± 5.0 nM) compared to its C2-Me analog AG85 (IC50 = 193 ± 39 nM). The most active antitubulin agents were evaluated against RTKs and demonstrated good inhibition for EGFR (IC50 = 29.5 - 91.5 nM), VEGFR-2 (182.3 - 543.6 nM) and PDGFRâ (IC50 = 204.4 - 450.2 nM), thus demonstrating dual-targeting ability. Preclinical assessment of AG165 at 75 mg/kg in mice in the NCI/ADR resistant tumor model demonstrated significant reduction of tumor volume compared to paclitaxel (10 mg/kg) treated mice [p = 0.0333], indicating antitumor activity in a multidrug resistant tumor model, in addition to excellent antitumor activity in MDA-MB-435 and MDA-MB-231 xenograft models. These results demonstrate the viability of these multitargeted single agents as potential clinically active antitumor agents.
Citation Format: Aleem Gangjee, Khushbu Shah, Nicholas Dybdal-Hargreaves, Susan Mooberry, Anja Bastian, Michael Ihnat. Design and biological evaluation of substituted pyrrolo[3,2-d]pyrimidines as dual acting RTK and microtubule targeting agents. [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 4857.
Abstract 4865: Metformin improves multiple cytokine producing ability of exhausted peripheral CD8+ T cells of cancer patients
Authors: Watanabe Mototsugu , Eikawa Shingo , Uehara Takenori , Kunisada Yuki , Toyooka Shinichi , Miyoshi Shinichiro , Udono Heiichiro 
Published: July 2016

Abstract
Background: We have recently shown that metformin, a prescribed drug for type 2 diabetes, can increase the number of CD8+ tumor infiltrating lymphocytes (CD8+ TILs) that can produce triple cytokines, thus, IFN-ã, IL-2 and TNF-á in mouse models. Increased numbers of such multifunctional CD8+ TILs have well correlated with tumor retardation, suggesting an underlying mechanism of metformin in tumor immunity. In the current study, we examined the effects of direct exposure of metformin in vitro to CD8+ T cells of cancer patients.
Methods: Peripheral blood mononuclear cells (PBMC) from cancer patients and healthy volunteers were obtained for the analysis. We cultured the PBMC with or without metformin overnight, followed by stimulation with phorbol 12-myristate 13-acetate (PMA) / ionomycin in vitro in various glucose concentrations for 6 hours. Expressions of markers for immune-check points (PD-1 and Tim-3) were examined together with accumulated intracellular cytokines, IFN-ã, IL-2 and TNF-á by flow cytometry analysis.
Results: PD-1 and Tim-3 expressions of cancer patients’ PBMC were apparently higher than those of healthy volunteers’. Increased production of multiple cytokines was observed in CD8+ T cells of almost all cancer patients by metformin pretreatment, whereas no changes were detected in the samples from healthy volunteers. Moreover, deprivation of glucose clearly limited the production of cytokines on CD8 + T cells in a cancer patient. These results suggested patient’ CD8 + T cells showed lower activity of glycolysis compared with a healthy volunteer and metformin might stimulate glycolysis followed by reversion of mitochondrial respiration in exhausted CD8 + T cells.
Conclusion: Metformin treatment might be useful in reversion of exhausted CD8+T cells in cancer bearing patients.
Citation Format: Mototsugu Watanabe, Shingo Eikawa, Takenori Uehara, Yuki Kunisada, Shinichi Toyooka, Shinichiro Miyoshi, Heiichiro Udono. Metformin improves multiple cytokine producing ability of exhausted peripheral CD8+ T cells of 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 4865.
Abstract 4893: Molecular detection of ALL in the peripheral blood is more sensitive than flow cytometric analysis of the bone marrow in patients with treatment-related hypocellularity
Authors: Smithers Hannah , Finney Olivia , Kirsch Ilan , Crossley Beryl , Jensen Michael , Gardner Rebecca 
Published: July 2016

Abstract
Measurable residual disease detection (MRD) is an important tool for early detection and monitoring relapse. Traditionally, bone marrow is collected and analyzed by flow cytometry (FC) to evaluate MRD status in acute lymphoblastic leukemia patients (ALL). More recently, highly sensitive molecular methods such as deep sequencing allow for higher sensitivity detection of MRD. Although the bone marrow is considered to be more representative of tumor burden than sampling the blood, bone marrow collection is more invasive, disruptive, and resource intensive than peripheral blood draws. ALL patients enrolled in the Seattle Children's Pediatric Leukemia Adoptive Therapy (PLAT-02) trial undergo MRD assessment pre and post CD19 CAR T cell infusion using FC analysis of a bone marrow sample. We have used high throughput sequencing (HTS) in paired blood and bone marrow samples from these patients to evaluate the complete B cell repertoire and assess MRD up to 63 days post-infusion. To-date, we have obtained 38 paired samples. Using HTS alone we have observed a significant correlation in both the frequency of total reads and the number of malignant cells per million in paired peripheral blood and bone marrow samples. 26/38 of the bone marrow samples were determined either aparticulate or hypocellular. Importantly, 8/14 of patients that were MRD negative in the bone marrow by FC were MRD positive in the blood by HTS. These data suggest, that, in cases where the bone marrow is hypocellular or aparticulate, peripheral blood samples evaluated by HTS are more sensitive than traditional flow cytometric analysis in the bone marrow for detection of MRD. The impact of B cell aplasia may be important in increasing the sensitivity of HTS in the detection of MRD. The ability to detect an ALL B cell clone in the blood through HTS has many clinical implications; blood can be sampled more often, is less of a burden to patients, and can provide earlier prediction of relapse.
Citation Format: Hannah Smithers, Olivia Finney, Ilan Kirsch, Beryl Crossley, Michael Jensen, Rebecca Gardner. Molecular detection of ALL in the peripheral blood is more sensitive than flow cytometric analysis of the bone marrow in patients with treatment-related hypocellularity. [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 4893.
Abstract 4896: A novel method for evaluating antibody-dependent cell-mediated cytotoxicity by flow cytometry using cryopreserved human peripheral blood mononuclear cells
Authors: Kitano Shigehisa , Yamashita Makiko , Aikawa Hiroaki , Kuchiba Aya , Hayashi Mitsuhiro , Yamamoto Noboru , Tamura Kenji , Hamada Akinobu 
Published: July 2016

Abstract
For predicting the clinical efficacy of antibody-dependent cellular cytotoxicity (ADCC) -dependent antibody therapy, analyzing the cytotoxic functions of effector cells such as NK cells against target cancer cells is thought to be necessary. The 51Cr release assay has long been the most widely used method for quantification of ADCC activity. However, the reproducibilities of these release assays are not adequate, and they do not allow evaluation of the lysis susceptibilities of distinct cell types within the target cell population. In this study, we established a novel method for evaluating cytotoxicity, which involves the detection and quantification of dead target cells using flowcytometry. CFSE (carboxyfluorescein succinimidyl ester) was used as a dye to specifically stain and thereby label the target cell population, allowing living and dead cells, as well as also target and effector cells, to be quantitatively distinguished. Furthermore, with our new approach, ADCC activity was more reproducibly, sensitively, and specifically detectable in not only freshly isolated but also in frozen human peripheral blood mononuclear cells (PBMCs) compared to the calcein-AM release assay. This assay, validated herein, is expected to be a standard assay to evaluate ADCC activity and finally contribute clinical development of ADCC dependent-antibody therapies.
Citation Format: Shigehisa Kitano, Makiko Yamashita, Hiroaki Aikawa, Aya Kuchiba, Mitsuhiro Hayashi, Noboru Yamamoto, Kenji Tamura, Akinobu Hamada. A novel method for evaluating antibody-dependent cell-mediated cytotoxicity by flow cytometry using cryopreserved human peripheral blood mononuclear cells. [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 4896.
Abstract 4909: The oncolytic peptide LTX-315 enhances tumor-specific immune responses and tumor regression in murine 4T1 breast cancer when combined with doxorubicin
Authors: Wang Meng Yu , Kristian Alexander , Camilio Ketil , Sveinbjornsson Baldur , Mælandsmo Gunhild Mari , Kvalheim Gunnar , Rekdal Øystein 
Published: July 2016

Abstract
The oncolytic effect of LTX-315 involves perturbation of the plasma membrane and distortion of the mitochondrial membrane with subsequent release of DAMPs (Damage-Associated Molecular Pattern molecules) such as ATP, cytochrome C and HMGB1 (1,2). Also multidomain proteins from the BCL-2 family seem to be partially involved in LTX-315 mediated killing (3). In addition, LTX-315 induces the hallmarks of immunogenic cell death (4). Preclinical studies have demonstrated that LTX-315 is able to induce complete tumour regression after intra-tumoral treatment (5,6). In this study the effect of combining intratumoral administration of LTX-315 and systemic treatment with Doxorubicin was investigated in the murine 4T1 breast cancer model. The combination treatment was investigated both in s.c. tumors and in tumors established in the mammary fat pad. The 4T1 cell were labeled with mcherry reporter gene in order to assess tumor growth by whole body imaging. LTX-315 (1 mg in 50 ìl) was injected intratumorally for three consecutive days. Doxorubicin (8mg/kg) was injected into the tail vein as a single dose together with the first injection of LTX-315. Tumor growth was assessed by caliper measurements and whole body luminescence. The combination of local administration of LTX-315 and systemic Doxorubicin, demonstrated significant synergy in both s.c. tumors and in tumors growing in the mammary fat pad. Furthermore, the combination induced a systemic tumour specific immune response since tumors did not develop in cured animals when re-challenged with tumor cells. The tolerable safety profile and novel mode of action of LTX-315 indicate a rationale for combining LTX-315 with other immunotherapies, targeted agents, and chemotherapy.
References:
1) Forveille et al., Cell Cycle (2015)
2) Eike et al., Oncotarget (2015)
3) Zhou et al., Oncotarget (2015)
4) Zhou et al. Cell Death & Differentiation (In rev)
5) Camilio et al., Cancer Immunol Immunother (2014)
6) Camilio et al., Oncoimmunology (2014)
Citation Format: Meng Yu Wang, Alexander Kristian, Ketil Camilio, Baldur Sveinbjornsson, Gunhild Mari Mælandsmo, Gunnar Kvalheim, Øystein Rekdal. The oncolytic peptide LTX-315 enhances tumor-specific immune responses and tumor regression in murine 4T1 breast cancer when combined with doxorubicin. [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 4909.
Abstract 4923: Expression of class III beta-tubulin (TUBB3) in 3580 colorectal cancers (CRCs) and correlation with clinico-pathological and molecular features
Authors: Xiu Joanne , Reddy Sandeep , El-Deiry Wafik S. 
Published: July 2016

Abstract
Class III beta-tubulin plays a crucial role in intracellular transport, meiosis and mitosis. High expression of TUBB3 has been shown to associate with poor prognosis and taxane resistance in various cancer types. CRC is known to be generally resistant to taxane therapy. We investigated expression of TUBB3 in 3580 CRCs and made correlations with clinicopathological and molecular parameters. 3580 CRC samples were evaluated by tumor profiling (Caris Life Sciences, Phoenix, AZ). Tests included Sanger or next generation sequencing (NGS), protein expression by immunohistochemistry (IHC) and gene amplification by in situ hybridization (ISH). TUBB3 expression was evaluated by immunohistochemistry (Ab: POLY, Covance) and expression higher than 2+, 30% was scored as positive. TUBB3 positive expression was observed in 37% (1320/3580) of the complete CRC cohort, specifically 27% in mucinous histology (164/617) and 17% in signet ring histology (29/171). While the expression was not associated with average patient age (59 years old) or gender (53% vs. 50% male), TUBB3 was more frequently expressed in tumors that originated from the left colon (370/1016 or 36%) than from the right colon (235/790 or 30%, p = 0.003). In the 1847 tumors taken from metastatic sites, 40% (746) overexpressed TUBB3 while in 1629 CRCs taken from primary tumors, only 34% (547) overexpressed TUBB3 (p&lt;0.0001). Interestingly, in tumors that overexpressed TUBB3, 65% also overexpressed cMET (828/1275) and 33% also overexpressed TLE3 (431/1299). This compares to 50% cMET expression (1096/2207, p&lt;0.0001) and 23% TLE3 expression (517/2225, p&lt;0.0001) in tumors that were negative for TUBB3. Microsatellite instability detected by fragment analysis was more prevalent in the TUBB3-negative cohort than the TUBB3-positive cohort (7.4% or 77/1046 vs. 2.9% or 16/558, p = 0.0002). Mutations in APC (62% vs. 56%, p = 0.0003) and KRAS (55% vs. 46%, p&lt;0.0001) were more frequent in TUBB3-positive tumors, while GNAS (2% vs. 5%, p&lt;0.0001) and SMAD4 (10.1% vs. 14.6%, p = 0.0005) mutations were significantly more frequent in tumors that were negative for TUBB3 expression. Thus, high expression of TUBB3 was found in 37% of CRCs, and was significantly associated with tumors that originated from the left colon and with tumors taken from metastatic sites. Distinct biomarker features detected by IHC and sequencing suggest that TUBB3 expression may carry theranostic importance which warrants further investigation in clinical trials.
Citation Format: Joanne Xiu, Sandeep Reddy, Wafik S. El-Deiry. Expression of class III beta-tubulin (TUBB3) in 3580 colorectal cancers (CRCs) and correlation with clinico-pathological and molecular features. [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 4923.
Abstract 4932: Stratification markers for the risk of recurrence after curative resection of stage II or III gastric cancer and potential clinical applications
Authors: Oshima Takashi , Kimura Yayoi , Sakamaki Kentaro , Miyagi Yohei , Yoshikawa Takaki , Rino Yasushi , Sentani Kazuhiro , Oue Naohide , Yasui Wataru , Imada Toshio , Munetaka Munetaka 
Published: July 2016

Abstract
Objective: Standard treatment for stage II or III gastric cancer is curative resection followed by adjuvant chemotherapy. Treatment outcomes are expected to be further improved by individualized therapy based on biomarker analysis. We extracted mRNA and protein from frozen specimens of gastric cancer and analyzed markers for stratifying the risk of recurrence after curative resection of stage II or III gastric cancer. We report our currently available findings.
Methods: Candidate markers for stratification of recurrence risk were identified by quantitative real-time polymerase chain reaction (Q-PCR) at the gene level and liquid chromatography-tandem mass spectrometry (LC-MS/MS) at the protein level. The study group for quantitative real-time PCR comprised 255 patients with stage II or III gastric cancer (training set, 145 patients; validation set, 110 patients). The relative expression levels of 127 genes were measured in gastric cancer tissue. Genes identified be independent predictors of poor outcomes in the training set were verified in the validation set. The study group for exploratory proteomics analysis LC-MS/MS comprised 24 patients with T4a Nx M0, Stage III gastric cancer (recurrence and death, 12 patients; relapse-free survival &gt;5 years, 12 patients). Proteins with max fold change &gt;2 and Anova (P) &lt;0.001 were searched.
Results: On Q-PCR, 5 genes; SPARC, HER2, CXCR4, INHBA, and VSNL1 were identified as candidate markers. On exploratory proteomics analysis by LC-MS-MS, 4 proteins were identified as candidate markers for recurrence risk stratification.
Conclusions: Promising candidate markers for stratification of the risk of recurrence after curative resection of stage II or III gastric cancer were identified at the gene and protein levels. These candidate markers are now being analyzed by tissue microarray, using specimens from about 500 patients each at two high-volume centers, with the goal of developing clinical applications.
Citation Format: Takashi Oshima, Yayoi Kimura, Kentaro Sakamaki, Yohei Miyagi, Takaki Yoshikawa, Yasushi Rino, Kazuhiro Sentani, Naohide Oue, Wataru Yasui, Toshio Imada, Munetaka Munetaka. Stratification markers for the risk of recurrence after curative resection of stage II or III gastric cancer and potential clinical applications. [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 4932.
Abstract 4934: Detection of HBV-host junction DNA sequences in urine of patients with hepatocellular carcinoma
Authors: Lin Selena , Trauger Evan R. , Song Benjamin P. , Lan Ling , Jongeneel Patrick M. , Thompson Emilie G. C. , Hoffman Malcolm C. , Jain Surbhi , Chang Ting-Tsung , Block Timothy M. , Song Wei , Su Ying-Hsiu 
Published: July 2016

Abstract
Human urine has been shown to contain DNA from circulation. In this study, we aim to provide unambiguous evidence that urine contains hepatocellular carcinoma (HCC)-derived DNA, and thus can be used for detecting HCC-associated DNA markers. We used novel hepatitis B virus (HBV)-host junction sequences (HBV-JSs), created by viral integration in the human genome, as unique markers to track the HBV-integrated DNA from tumor tissues to their corresponding urine samples. An HBV DR1-2 enriched Next-Generation sequencing (NGS) assay was developed to identify the major HBV-JSs in 15 HBV-HCC tissues. Upon validation of the major HBV-JSs by Sanger sequencing, a short-amplicon PCR assay tailed for each HBV-JS was developed to test the junctions’ existence in the corresponding urine DNA. 15 major HBV-JSs were identified by the HBV DR1-2 enriched NGS assay from 15 HBV related HCC (HBV-HCC) tissues, 13 of which were validated by Sanger sequencing. By using HBV-JS specific short-amplicon junction PCR assays, we detected and confirmed six of nine HBV-JSs for which there were matching urine samples. Urine contains detectable major HBV-JSs derived from HCC, and thus can be used for liquid biopsies to study not only the complexities of HBV-JS species during chronic HBV infection and carcinogenesis, but also other HCC-related DNA modifications for the early detection of HCC and disease management.
Citation Format: Selena Lin, Evan R. Trauger, Benjamin P. Song, Ling Lan, Patrick M. Jongeneel, Emilie G. C. Thompson, Malcolm C. Hoffman, Surbhi Jain, Ting-Tsung Chang, Timothy M. Block, Wei Song, Ying-Hsiu Su. Detection of HBV-host junction DNA sequences in urine of patients with hepatocellular carcinoma. [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 4934.