bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 1046: A breast cancer MET reporter cell line model for drug discovery and development
Authors: Enuameh Metewo S. , Shu Weiguo , Newman Robert , Kumari Sangeeta 
Published: August 2019

Abstract
World-wide, cancer metastasis continues to be the leading cause of death in cancer (DIZON et al. 2016). Although epithelial-to-mesenchymal transition (EMT) and mesenchymal-to-epithelial transition (MET) have been implicated in the incidence of cancer metastasis and drug resistance, their impact in cancer progression and patient survival is not fully understood (NIETO et al. 2016). This is partly due to the lack of suitable in vitro models. Thus, to facilitate the utility of the EMT concept in therapeutic development, we have utilized some of the basic biology of EMT/MET, to create a novel advanced in vitro model for use in both basic research and discovery of new anti-EMT drugs.
In breast cancer, vimentin (VIM) intermediate filament (IF) proteins are generally upregulated during EMT and down-regulated during MET (GILLES et al. 1999; THIERY and SLEEMAN 2006; RICHARDSON et al. 2012; LAMOUILLE et al. 2014). Here we employed CRISPR/Cas9 gene editing to generate a VIM RFP (red fluorescent protein) reporter in the MDA-MB-231 breast adenocarcinoma cell line. The VIM RFP C-terminal fusion gene at the endogenous VIM locus, enables end-point or real-time tracking of the MET status as cells transition from epithelial to mesenchymal phenotype under distinct conditions. We have validated the cell line at the nucleic acid (genomic and mRNA) and protein levels as well as in cell-based assays. Bio-functional evaluation of the MDA-MB-231 VIM RFP cell line shows sensitivity to metastatic breast cancer drugs axitinib (tyrosine kinase inhibitor) and U0126 (MEK1/2 inhibitor) via the inhibition of the inherent signaling pathways which impact EMT. These effects provide the basis for the use of this cell line in high throughput screening (HTS) applications including the discovery of new anti-EMT drugs for metastatic breast cancer. Furthermore, the MDA-MB-231 VIM RFP reporter cell line is also a convenient and sensitive model for studying the mechanisms of metastasis and for basic translational science research.
Citation Format: Metewo S. Enuameh, Weiguo Shu, Robert Newman, Sangeeta Kumari. A breast cancer MET reporter cell line model for drug discovery and development [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1046.
Abstract 1049: Targeting YAP/TAZ pathway inhibits Ewing sarcoma metastasis
Authors: Bierbaumer Lisa , Katschnig Anna M. , Radic-Sarikas Branka , Petro Jeffrey R. , Mühlbacher Karin , Aryee Dave N. , Pötsch Anna R. , Högler Sandra , Kenner Lukas , Uren Aykut , Kovar Heinrich 
Published: August 2019

Abstract
The term epithelial to mesenchymal transition (EMT) and its reverse (MET) describe the dynamic and reversible metamorphosis of cells from a highly organized to a loose migratory phenotype through cytoskeletal reorganization causing cellular plasticity. EMT/MET are inherent to normal embryonal development and wound healing, but cancer cells hijack the underlying mechanisms to enable oscillations between proliferation, invasion and migration that cause metastasis, the major killer of cancer patients. Recently, fluctuations of the oncoprotein EWS-FLI1 were identified to drive EMT/MET in Ewing sarcoma. We previously demonstrated that, in presence of EWS-FLI1, transcriptional co-activators MRTFB and TAZ are largely blocked from associating with TEAD and its target genes keeping tumor cells in a poorly migratory, highly proliferative state. In contrast, under EWS-FLI-low conditions, MRTFB and TAZ associate with YAP/TEAD complexes on chromatin resulting in cytoskeletal target gene activation and phenotypic transition to a highly migratory and low proliferative state. We therefore hypothesized that pharmacologic inhibition of YAP/TAZ/TEAD protein interaction should prevent Ewing sarcoma cells from EMT and consequently interfere with their metastatic potential. Verteporfin is a small molecule safely used in the treatment of age-related macular degeneration. Independent of its photosensitizing activity exploited in ophthalmology, it is a YAP/TAZ pathway -blocking compound. In vitro treatment of EWS-FLI1-low Ewing sarcoma cells with verteporfin resulted in decreased YAP/TAZ/TEAD complex formation in the nanomolar range, reversal of the de-repression of a EWS-FLI1 controlled EMT transcriptional signature, and inhibition of tumor cell migration in a Boyden chamber assay. Upon orthotopic implantation of TC71 Ewing sarcoma cells into the tibial crest of SCID beige mice, intra-peritoneal treatment of mice with 25mg verteporfin/kg/day before and after amputation of the affected limb led to a drastic decrease of lung metastases without affecting primary tumor growth and without obvious general toxicity. Therefore, YAP/TAZ pathway blockade holds promise as a potential metastasis-preventive strategy in the treatment of Ewing sarcoma patients with primary localized disease.
Citation Format: Lisa Bierbaumer, Anna M. Katschnig, Branka Radic-Sarikas, Jeffrey R. Petro, Karin Mühlbacher, Dave N. Aryee, Anna R. Pötsch, Sandra Högler, Lukas Kenner, Aykut Uren, Heinrich Kovar. Targeting YAP/TAZ pathway inhibits Ewing sarcoma metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1049.
Abstract 1050: Characterization of human cancer cell line xenografts in humanized mice
Authors: Stevens Christina , Maddage Christopher , Lasky Kerri , Rios-Doria Jonathan , Koblish Holly 
Published: August 2019

Abstract
Studying immune-oncology (IO) agents that target the human immune system in mice have posed challenges for oncology researchers. Since the commercial introduction of humanized mice, antitumor efficacy studies can now be performed with human cancer cells within mice bearing the components of a human immune system. However, development and characterization of these models is necessary to understand which model may be best for different agents. To this end, we characterized A375, A549, Caki-1, H1299, H1975, HCC827, HCT116, KU19-19, MDA-MB-231, and RKO human cancer cell lines in CD34+ humanized NSG mice for tumor growth rate and immune cell profiling in tumor and spleen compartments. We found that CD4+ T cell composition in both the spleen and tumor varied among the models, with A375, Caki-1, and RKO expressing higher levels of CD4+ T cells compared to the other models. Many of the models had moderate levels of CD8+ T cells in the tumor. Spleen analysis revealed higher NK cell populations in mice bearing Caki-1 tumors, while A549 tumor samples had a higher percentage of NK cells relative to the other models. Surprisingly, many of the tumor models showed an abundance of myeloid cells in the spleen and tumor compartments, although these were mostly comprised of B cells and dendritic cells and lower levels of macrophages and MDSCs. The expression level and percentage of cells expressing PD-1, and PD-L1 in the models were also determined. Furthermore, the antitumor response of select models to PD-1 and PD-L1 antibodies will be presented. In summary, these data demonstrate that there are tumor-intrinsic factors that influence the immune cell repertoire within tumors and spleen. These data may also aid in selection of tumor models to test antitumor activity of novel IO agents.
Citation Format: Christina Stevens, Christopher Maddage, Kerri Lasky, Jonathan Rios-Doria, Holly Koblish. Characterization of human cancer cell line xenografts in humanized mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1050.
Abstract 1051: Dianhydrogalactitol (VAL-083) reduces glioblastoma tumor growth upon bevacizumab-induced hypoxia, in vivo
Authors: Golebiewska Anna , Oudin Anais , Steino Anne , Niclou Simone P. , Bacha Jeffrey , Brown Dennis M. 
Published: August 2019

Abstract
Background: Standard-of-care for glioblastoma (GBM) includes surgery, radiation and temozolomide (TMZ). Nearly all tumors recur and 5-year survival is less than 3%. Unmethylated promoter status for O6-methylguanine-DNA-methyltransferase (MGMT) is a validated biomarker for TMZ-resistance. Second-line treatment with bevacizumab has not only failed to improve survival, but has also been shown to induce intratumor hypoxia, which is implicated in increased chemoresistance. VAL-083 is a bi-functional DNA-targeting agent that readily crosses the blood-brain barrier and accumulates in brain tumor tissue. VAL-083 induces DNA double-strand breaks at N7-guanine and cancer cell-death in GBM cancer stem cells (CSCs) and non-CSCs, independent of MGMT. We have previously shown that bevacizumab treatment upregulates expression of glucose transporters GLUT-1/GLUT-3 on GBM cells. We hypothesized that, based on its unique monosaccharide backbone structure, VAL-083 may benefit from bevacizumab-induced GLUT transporter upregulation leading to enhanced VAL-083 uptake and anti-tumor activity.
Methods: To investigate the in vivo anti-tumor effect of VAL-083+bevacizumab, we used a orthotopic patient-derived xenograft GBM model. All mice carried MGMT-unmethylated, temozolomide-resistant recurrent T16 GBM tumors as detected by MRI 35 days post-implantation. Mice were grouped into control, bevacizumab, VAL-083, and VAL-083+bevacizmab. Tumor progression was measured by MRI on days 49 and 56, and tumor growth rate was calculated for the entire study (day 35 vs. 56) and for the last 7 days (day 49 vs. 56).
Results: Tumors were significantly smaller in VAL-083-treated mice both compared to control (-83%, p&lt;0.001) and compared to bevacizumab-treated (-75%, p&lt;0.001) mice. Additionally, analysis of tumor growth in-time showed significantly reduced tumor growth rate for VAL-083+bevacizumab compared to VAL-083 alone (p&lt;0.01).
Conclusions: These results show strong in vivo anti-tumor efficacy of VAL-083 against MGMT-unmethylated, TMZ-resistant recurrent GBM. This effect was further augmented in combination with bevacizumab, providing rationale of clinical investigation of VAL-083 in combination with bevacizumab in the treatment of GBM.
Citation Format: Anna Golebiewska, Anais Oudin, Anne Steino, Simone P. Niclou, Jeffrey Bacha, Dennis M. Brown. Dianhydrogalactitol (VAL-083) reduces glioblastoma tumor growth upon bevacizumab-induced hypoxia, in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1051.
Abstract 1053: A patient-derived xenograft model demonstarates the effectiveness of combined oxaliplatin and 5-fluorouracil for advanced small bowel adenocarcinoma
Authors: Yamano Tomoki , Song Jihyung , Kimura Kei , Babaya Akihito , Hamanaka Michiko , Kataoka Kozo , Beppu Naohito , Noda Masafumi , Ikeda Masataka , Tomita Naohiro 
Published: August 2019

Abstract
Background: Combination oxaliplatin and capecitabine therapy is recommended to treat advanced small bowel adenocarcinoma (SBA). However, SBA is rare, and limited preclinical studies and randomized clinical trials have been conducted.
Methods: A patient-derived xenograft (PDX) was established from an advanced SBA patient with liver metastasis and peritoneal metastasis. Growth inhibition by oxaliplatin, irinotecan, 5-fluorouracil (5-FU), and 5-FU in combination with oxaliplatin, irinotecan, or paclitaxel was assessed using the PDX. Drug sensitivity against oxaliplatin, irinotecan, 5-FU, and paclitaxel was assessed using SBA cells from the PDX. The patient’s clinicopathological and genetic features were compared with the PDX to evaluate it as a preclinical model.
Results: 5-FU, but not oxaliplatin and irinotecan showed statistically significant growth inhibition compared to control group (P = 0.0024). Among the combination treatment with 5-FU, oxaliplatin showed the highest tumor growth inhibition rate of 0.84 or 1.1, which was 0.46 for irinotecan and 0.49 for paclitaxel, respectively. IC50 of SBA cells was significantly lower against oxaliplatin and higher against paclitaxel compared with colon cancer cell line HT29. These results corresponded to the clinical response to FOLFOX (folinic acid+5-FU+oxaliplatin) and FOLFIRI (folinic acid+5-FU+irinotecan). PDX pathological and genetic features resembled those of the patient.
Conclusions: We established a preclinical model comprising the PDX from SBA and SBA cells from the PDX. This model showed that the combination of oxaliplatin and 5-FU was a
reasonable standard therapy for advanced SBA. A preclinical model comprising PDX and tumor cells from PDX seems to be useful for rare tumors including SBA.
Citation Format: Tomoki Yamano, Jihyung Song, Kei Kimura, Akihito Babaya, Michiko Hamanaka, Kozo Kataoka, Naohito Beppu, Masafumi Noda, Masataka Ikeda, Naohiro Tomita. A patient-derived xenograft model demonstarates the effectiveness of combined oxaliplatin and 5-fluorouracil for advanced small bowel adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1053.
Abstract 1062: Development and characterization of patient-derived xenograft models for acute myeloid leukemia
Authors: Ulrich Michelle , Anderson Martha , Thurman Robert , Li Fu 
Published: August 2019

Abstract
Drug development for acute myeloid leukemia (AML) remains challenging, in part, due to the lack of preclinical models that reflect the biology of AML patients. For example, while the majority of AML patients express both CD33 and CD123, only a small fraction of AML cell lines express CD123. Similarly, most AML cell lines have a differentiated phenotype, which may bias the evaluation towards certain types of AML. To address these challenges, we developed a dozen patient derived xenograft (PDX) models to aid preclinical development of AML drugs. Bone marrow samples from 32 AML patients were implanted intravenously into NSG mice. Thirteen of them led to successful expansion of human CD45+ cells in the bone marrow, as demonstrated by flow cytometry. Twelve of these established disease in a secondary passage. Using multicolor flow cytometry analysis, we found that expression of CD33 and CD123 were preserved from AML patients and engrafted disease in NSG mice. Moreover, RNA-sequencing analysis demonstrated that gene expression largely remained consistent between the first two passages. Detailed genetic analysis demonstrated the preservation AML-relevant mutations, such as FLT3-ITD mutations and IDH2 mutations in the PDX model. In summary, we have successfully established AML PDX models that reflect the antigen expression and molecular genetics of AML patients. These models may facilitate the future development of antibody-based therapeutics.
Citation Format: Michelle Ulrich, Martha Anderson, Robert Thurman, Fu Li. Development and characterization of patient-derived xenograft models for acute myeloid leukemia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1062.
Abstract 1090: β-Hydroxy-β-Methylbutyrate supplementation preserves muscle mass and reduces tumor growth in obese mice
Authors: Coleman Michael F. , Liu Kristyn A. , Tang Xiaohu , Fabela Salvador , Lashinger Laura M. , Cui Zhengrong , Hursting Stephen D. 
Published: August 2019

Abstract
Background: Pancreatic ductal adenocarcinoma (PDAC) remains very challenging to treat with mean 5-year survival of approx. 6%, leading to PDAC’s status as the 4th most deadly cancer in the US. Diet induced obesity (DIO) has been shown to promote both increased incidence and growth of PDAC. Simultaneously DIO promotes muscle loss and the loss of immune surveillance in tumors. Cachexia, a chronic catabolic process in which muscle mass is lost, is a common feature of PDAC. We have previously shown that while leucine driven mTOR activation protects muscle mass in tumor bearing mice it also promotes tumor growth. Hence, we sought to test the potential of a leucine metabolite, β-hydroxy-β-methyl-butyrate, to protect against cachexia in a PDAC model while antagonizing DIO mediated tumor growth. We further sought to determine if HMB treatment would alter gemcitabine response in tumors from obese animals.
Methods: C57BL/6 mice consuming either control or high fat diet, bearing Panc02 tumors, were treated with HMB alone or in combination with gemcitabine. Muscle size was determined using Feret’s diameter measured from H&E stained cross sections. Immunohistochemistry staining was performed for Ki-67 and CD3. Tumor transcriptomic analysis was preformed using Affymetrics microarray. In vitro characterization HMB modulated signaling in C2C12 myotubes, Panc02 cells, and bone marrow derived macrophages treated with pro-inflammatory signaling molecules.
Results: a) HMB significantly increases muscle fiber size and mTOR signaling; b) HMB reduces DIO increased tumor volume; c) HMB significantly synergizes with gemcitabine to suppress PDAC growth in obese mice; and d) HMB reverts DIO mediate immune suppression and promotes tumor immune surveillance.
Discussion: Antagonism of DIO promoted PDAC tumor growth, by HMB, and promotion of immune surveillance may offer significant synergy with standard therapy or, immune targeted therapies in PDAC.
Citation Format: Michael F. Coleman, Kristyn A. Liu, Xiaohu Tang, Salvador Fabela, Laura M. Lashinger, Zhengrong Cui, Stephen D. Hursting. β-Hydroxy-β-Methylbutyrate supplementation preserves muscle mass and reduces tumor growth in obese mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1090.
Abstract 1096: Response of pancreatic ductal adenocarcinoma to gemcitabine using human organotypic slice cultures
Authors: Chang Jae Hyuck , Lim Chae Yoon , Lee Weon Sun , Lee Guk Jin 
Published: August 2019

Abstract
Background/Objective: The oragnotypic tumor slice culture model resembles the in vivo situation of tumor. It may serve as a proper ex vivo platform for response to anti-tumor therapies. We aimed to evaluate the response of pancreatic ductal adenocarcinoma (PDA) cells and helper, cytotoxic, and regulatory T cells to gemcitabine using human PDA slice culture model.
Methods: PDA tissues were obtained from patients who were operated on for pancreatic resection. The tumors were sliced by a vibratome, and the tumor slices were then cultured until 5 days with 0.1 μM, 1 μM, and 10 μM gemcitabine for 1 or 2 days. Tumor responses were evaluated by number of tumor cells and proportion of Ki-67 or caspase-3 positively expressed cells in the tumor. CD4+, CD8+, and FOXP3+ cells were analyzed by immunohistochemistry staining.
Results: Gemcitabine treatment to PDA slices exerted proportional cytotoxic effects in terms of tumor cell number, and Ki-67 and cleaved caspase-3 expression. Mean Ki-67 positive cells in PDA cells tended to decrease according to increase of gemcitabine concentration (case 1, 48 hr treatment; 65.7% in control vs. 27.9% in 0.1 μM, p=0.025). Mean caspase-3 positive cells in PDA cells tended to increase according to increase of gemcitabine concentration (case 1, 48 hr treatment; 8.6% in control vs. 18.6% in 0.1 μM, p=0.042). Mean PDA cells per x200 tended to decrease according to increase of gemcitabine concentration (case 2, 48 hr treatment; 78.3 in 1.0 μM vs. 27.3 in 10 μM, p=0.042). Mean CD4+, CD8+, and FOXP3+ cells/total cells tended to decrease according to increase of gemcitabine concentration; in case 3 (24hr treatment), mean CD4+ cells/total cells decreased as 2.6%, 1.6%, 1.5%, and 0.5%, mean CD8+ cells/total cells decreased as 2.6%, 1.5%, 1.4%, and 1.1%, and mean FOXP3+ cells/total cells were 0.2%, 0.3%, 0.2%, and 0.1% in control, 0.1 μM, 1 μM, and 10 μM gemcitabine, respectively. However, mean CD8+/FOXP3+ cell ratio did not significantly decrease, and even increase from 4.4 in 0.1 μM to 8.6 in 1 μM (p=0.039).
Conclusions: Tumoricidal effect of gemcitabine could be demonstrated using organotypic slice culture model. T cells in the tumor decreased according to increase of gemcitabine concentration; however CD8+/FOXP3+ cell ratio relatively maintained.
Citation Format: Jae Hyuck Chang, Chae Yoon Lim, Weon Sun Lee, Guk Jin Lee. Response of pancreatic ductal adenocarcinoma to gemcitabine using human organotypic slice cultures [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1096.
Abstract 1098: Enhanced AKT-dependent metabolic fitness and antitumor effect of T lymphocytes in lung cancer model
Authors: Kim Sung Eum , Pyo Kyoung-Ho , Lim Sang Bin , Yun Mi Ran , Kim Jae Hwan , Lee Ji Min , Jo Ha Ni , Xin Chun-Feng , Cho Jae Seok , Cho Byoung Chul 
Published: August 2019

Abstract
Introduction: PI3K signaling plays a pivotal role in balancing of immune responses. This study is to elucidate the role of PI3K-AKT signaling in regulating T cell metabolic fitness.
Methods: PI3K/AKT inhibitors including BKM120 were used to evaluate the contribution of PI3K-AKT signaling on the metabolic fitness of T cell. Glucose metabolism and T cell activity were measured by flow cytometry, IHC, RT-PCR. Oxygen consumption rate (OCR) and extra cellular acidification rate (ECAR) were examined by seahorse. In addition, anti-tumor effects of T cells were evaluated by adaptive transfer in LLC2 tumor bearing mouse.
Results: Compare to vehicle, BKM120-treated antigen-specific CD8+ T cells co-cultured with LLC2 cancer cells maintained cytotoxic effects under hypoglycemic and hypoxic conditions. Treatment of BKM120 significantly reduced tumor growth, compared to vehicle in syngeneic models (LLC2, KLN205), whereas it did not affect tumor growth in nude mice, which suggests role of activation of antitumor immunity, augmented by BKM120. Compared to vehicle, BKM120 significantly induced higher infiltration of CD8+ T cells and reduction of Foxp3+ regulatory T cell and CD11b+/Gr-1+ myeloid derived suppressor cells. Intriguingly, increased glucose uptake through overexpression of glucose transporter (GLUT1) was observed upon BKM120 treatment. Glut1 overexpression occurred through the release of feedback inhibition of mTORC2 followed by rebound AKT activation. In addition, glycolysis-related genes (Glut1, Hk, Pgam, Pdk and Pgm) and OCR/ECAR were significantly augmented in BKM120/selective PI3K-delta inhibitor (Idelalisib)-treated group. The enhanced glycolysis via PI3K signaling inhibition was significantly more prominent by Idelalisib, compared to BKM120 or selective PI3K-alpha inhibitor (BYL-719). The antigen-specific T cells pretreated with BKM120 ex vivo exhibited substantially more potent antitumor effects, compared to untreated T cells, in LLC2 tumor-bearing nude mice.
Conclusion: PI3K inhibitor, specifically selective PI3K-delta inhibitor, promotes AKT-dependent metabolic fitness of T cells which sustain anti-cancer effects under nutrient/oxygen-deprived tumor microenvironment. These data support a rationale for combination of selective PI3K-delta inhibitor with checkpoint inhibitors anti-PD1 for patient with Lung cancer.
Citation Format: Sung Eum Kim, Kyoung-Ho Pyo, Sang Bin Lim, Mi Ran Yun, Jae Hwan Kim, Ji Min Lee, Ha Ni Jo, Chun-Feng Xin, Jae Seok Cho, Byoung Chul Cho. Enhanced AKT-dependent metabolic fitness and antitumor effect of T lymphocytes in lung cancer model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1098.