bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2147: Generation of primary cell lines from advanced colon cancer: a tool for screening novel anticancer molecules.
Authors: Nair Rohini , Basu Aninda , Dhandapani Muthu , Majumder Biswanath , Ulaganathan Baraneedharan 
Published: August 2014

Abstract
For diversely heterogeneous diseases like cancer, a major challenge in the development of new drugs is using appropriate in vitro testing platform. Although high throughput screening of new compounds requires a large panel of genetically well characterized cell lines, their numbers are far less than requirement of biophrama. On top of this the translational potential of cell line panel is limited because most of the commercially available cell lines have been generated from metastatic site and highly transformed, therefore do not represent the true state of the disease. In this context, we have developed two early passage patient derived primary colorectal cancer (CRC) cell lines. Using this improved in vitro testing system we tested the efficacy of a number of anticancer agents against CRC.
Cell lines were generated by enzymatic disaggregation method. The tissues were obtained after informed consent from patient. The tumor tissues were sectioned into to small pieces (∼1mm) and were digested using 0.5% Collagenase for 2-4 hrs at 37°C. The single cell suspensions were cultured in DMEM supplemented with 10% fetal bovine serum. The epithelial cells were propagated up to passage 2 and were cryopreserved for further applications. Mutational (K-RAS, BRAF, PIK3CA, PTEN) and standard CRC specific marker (such as mucin) analyses were done in order to confirm the authenticity of these lines. The cell viability was evaluated using MTT assay following treatment with Standard of Care (SOC) agents (both single and combinations). Additionally for screening of selected FDA approved NSAIDs like Aspirin, Etoricoxib, Ibuprofen, Indomethacin, Mefenamic acid, Methotrexate and Nimesulide (repurposing) were performed at their respective CMAX concentrations after 48 hrs of treatment.
Our results showed that, the SOCs when tested as single agents did not show any significant cell death. However, when treated in combination with Irinotecan, Fluorouracil and Leucovorin in the presence and absence of Avastin a significant augmentation of cell death was observed in both the cell lines. Further, Mefenamic acid, Methotrexate, Ibuprofen and Aspirin exhibited a significant cytotoxic effect in both cell lines after 48 hrs of treatment. Together, these results demonstrate that COX-2 inhibitors could be developed as a as an effective anticancer therapeutics for colorectal cancer treatment and primary cell line model provides a rapid and more reliable screening platform.
Citation Format: Rohini Nair, Aninda Basu, Muthu Dhandapani, Biswanath Majumder, Baraneedharan Ulaganathan. Generation of primary cell lines from advanced colon cancer: a tool for screening novel anticancer molecules. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2147. doi:10.1158/1538-7445.AM2013-2147
Abstract 2152: The influence of didox on the efficacy and toxicity of doxorubicin.
Authors: Al-Abbasi Fahad A. , Assad Gihan F. , Al-Abd Ahmed M. , Abdel-Naim Ashraf B. 
Published: August 2014

Abstract
Didox, possess antioxidant propertieson the top of its DNA synthesis and repair inhibiting ability. Doxorubicin (DOX) is a major chemotherapy for liver cancer; however it evokes severe cardiotoxic effect which is attributed to DOX derived free radicals. Herein, we assessed the effect didox in enhancing DOX efficacy in liver cancer cells while protecting against its cardiotoxic effects. Combination with didox significantly reduced in the IC50 of DOX in Huh7 and HepG2 liver cancer cell lines. The combination index between DOX and didox indicated additive type interaction (CI-value of 0.81-0.9). Both DOX and didox significantly blocked the cell cycle in S-phase and their combination significantly aggravated that block. In addition, didox with DOX showed higher caspase-3 level compared to DOX alone. This enhancement effect might be attributed to didox induced PARP enzyme inhibition. On the other hand, didox (150 mg/kg daily) significantly protected the cardiomyocyte and decreased the intra cardiac reactive oxygen species induced by DOX treatment (15 mg/kg). This effect was coupled with reverting DOX induced cardiomegaly and cardio-pathological features. Didox significantly prolonged the median survival of DOX treated mice and reduced their mortality risk by 3.7 folds. In conclusion, didox significantly improved the cytotoxic effect of DOX in liver cancer cells and protected from its cardiotoxic side effects.
Citation Format: Fahad A. Al-Abbasi, Gihan F. Assad, Ahmed M. Al-Abd, Ashraf B. Abdel-Naim. The influence of didox on the efficacy and toxicity of doxorubicin. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2152. doi:10.1158/1538-7445.AM2013-2152
Abstract 2160: Development of selective MELK kinase inhibitors for cancer treatments.
Authors: Kowalczyk Piotr , Wegrzyn Paulina , Zawadzki Przemyslaw , Palacz Edyta , Trebacz Ewa , Wiklik Katarzyna , Milik Mariusz , Zarebski Adrian , Krawczynska Karolina , Brzózka Krzysztof 
Published: August 2014

Abstract
Maternal embryonic leucine zipper kinase (MELK) is an atypical member of the AMPK family of serine-threonine kinases that has been implicated in stem cell renewal, cell cycle progression, cytokinesis, mRNA splicing and apoptosis. MELK activity is correlated with its phosphorylation level, is cell cycle dependent and maximal during mitosis although direct upstream regulators of MELK kinase activity are unknown. This kinase is highly expressed in several types of solid cancers: colon, breast, ovary, lung, and brain and shows relatively low expression levels in normal tissues. MELK overexpression in patient-derived tumors strongly correlates with poor prognosis in glioblastoma and breast cancer. Moreover, siRNA mediated knockdown of MELK kinase significantly inhibits growth of tumor cell lines both in vitro and in vivo. Therefore, MELK kinase is an emerging and interesting target of significant potential for therapeutic intervention in cancer. In this study, we are reporting results for a series of new MELK kinase inhibitors that were developed at Selvita. Newly synthesized derivatives exert good selectivity and potency in MELK inhibitions with the low sub-micromolar range. Anticancer effects of these compounds were investigated in several cancer cell lines of solid tumor origin where the compounds were shown to induce cell death with low micromolar ED50 values. Compounds were also analyzed for their effects on cell death, proliferation, apoptosis, cell cycle parameters and ADME properties. Taken altogether, the presented data supports our rationale of using inhibitors of MELK kinases as a novel approach for the cancer therapy, especially for the treatment of glioblastoma and breast cancer.
Citation Format: Piotr Kowalczyk, Paulina Wegrzyn, Przemyslaw Zawadzki, Edyta Palacz, Ewa Trebacz, Katarzyna Wiklik, Mariusz Milik, Adrian Zarebski, Karolina Krawczynska, Krzysztof Brzózka. Development of selective MELK kinase inhibitors for cancer treatments. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2160. doi:10.1158/1538-7445.AM2013-2160
Abstract 218: R-Spondin (RSPO) signaling drives the growth of multiple human tumor types.
Authors: Gurney Austin , Chartier Cecile , Axelrod Fumiko , Cain Jennifer , Yen Wan-Ching , Murriel Christopher , Raval Janak , Fischer Marcus , Shah Jalpa , Ji May , Bond Christopher , Kapoun Ann , Lewicki John , Hoey Timothy 
Published: August 2014

Abstract
The R-spondin-LGR signaling axis is a recently appreciated pathway that promotes beta-catenin signaling in normal stem cell populations. Beta-catenin signaling had long been considered synonymous with Wnt signaling. We conducted a reporter-based screen for secreted beta-catenin signaling activities produced by a panel of human tumors. This exercise and further molecular characterization efforts discovered RSPO activity produced by numerous human tumors of multiple tumor types including ovarian, pancreatic, colon, breast and non-small cell lung cancer. Specific antibody antagonists of RSPO family members were developed. In minimally passaged human tumor xenograft models, anti-RSPO treatment markedly inhibited tumor growth in several tumor types. Moreover, RPSO blockade promoted tumor differentiation and reduced the frequency of tumor initiating cells. These data highlight the potential for therapeutic intervention with this newly appreciated signaling axis.
Citation Format: Austin Gurney, Cecile Chartier, Fumiko Axelrod, Jennifer Cain, Wan-Ching Yen, Christopher Murriel, Janak Raval, Marcus Fischer, Jalpa Shah, May Ji, Christopher Bond, Ann Kapoun, John Lewicki, Timothy Hoey. R-Spondin (RSPO) signaling drives the growth of multiple human tumor types. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 218. doi:10.1158/1538-7445.AM2013-218
Abstract 2180: Development of novel targeted therapies for non-small cell carcinoma by novel Midkine inhibitors.
Authors: Fukazawa Takuya , Maeda Yutaka , Hao Huifang , Yamatsuji Tomoki , Takaoka Munenori , Takigawa Nagio , Naomoto Yoshio 
Published: August 2014

Abstract
In the past few years a subset of driver mutations has been identified in non-small cell lung carcinoma (NSCLC), the most common type of lung adenocarcinoma. These driver mutations have differential sensitivities to various targeted therapies. The number of identified mutations in lung adenocarcinomas is limited, therefore identification of more molecular targets is needed in order to develop new treatments for NSCLCs. Efforts to find target mutations in lung adenocarcinoma with KRAS mutations are especially important because such adenocarcinomas account for 20% of lung adenocarcinoma, the most frequent type of lung adenocarcinomas.
Midkine is a heparin-binding cytokine that is induced by retinoic acid in embryonal carcinoma cells. It is involved in the development of cancer because of its mitogenic effect, promotion of angiogenesis, anti-apoptotic activity and transforming activity. Midkine expression is increased in various malignant tumors, including lung, esophageal, stomach, colon, hepatocellular, breast and pancreatic carcinomas, when compared with the level of expression in normal tissues. In a proof-of-principle effort to identify novel compounds that inhibit Midkine expression, we recently conducted cell-based high-throughput screening (HTS) of 44,000 compounds in the compound library using the Midkine promoter. We identified sixteen small molecules that inhibit Midkine promoter activity. Significantly, many of these inhibitors, which reduced Midkine protein, also inhibited proliferation in non-small cell lung cancer cells which express high levels of endogenous Midkine. We assigned the name iMDK1 to one of these compounds and assessed its ability to target non-small cell lung cancer cells. 48 hours after treatment, iMDK1 dose-dependently inhibited cell growth of Midkine expressing H441 pulmonary adenocarcinoma cells but did not impair the viability of normal proliferating human lung fibroblast cells. Hoechst 33342 staining and cell cycle analysis performed by flow cytometry demonstrated that the iMDK1 compound induced apoptosis in H441 cells. Furthermore, immunoblot analysis showed that iMDK1 inhibits the PI3 kinase pathway in H441 cells. These results indicate that iMDK1 might be a promising new therapeutic tool for the treatment of Midkine -expressing lung adenocarcinoma with KRAS mutations. Here we show a novel strategy to treat non-small cell lung cancer with Midkine inhibitors.
Citation Format: Takuya Fukazawa, Yutaka Maeda, Huifang Hao, Tomoki Yamatsuji, Munenori Takaoka, Nagio Takigawa, Yoshio Naomoto. Development of novel targeted therapies for non-small cell carcinoma by novel Midkine inhibitors. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2180. doi:10.1158/1538-7445.AM2013-2180
Abstract 2187: RNAi- and U1 small nuclear interference (U1i)-mediated gene knockdown reveals the functional relevance of Pim-1 kinase in colon carcinoma and glioblastoma.
Authors: Weirauch Ulrike , Thomas Maren , Grünweller Arnold , Hartmann Roland K. , Aigner Achim M. 
Published: August 2014

Abstract
Pim-1 is a constitutively active serine/threonine kinase overexpressed in various tumors. Its role as proto-oncogene is based on several Pim-1 target proteins involved in pivotal cellular processes, and Pim-1 overexpression has been linked to poor prognosis.
RNAi-based knockdown approaches were used to inhibit Pim-1 in colon carcinoma cells. We demonstrate anti-proliferative, pro-apoptotic and overall antitumor effects of Pim-1 inhibition. The analysis of the molecular effects of Pim-1 inhibition reveals a complex regulatory network, with therapeutic Pim-1 repression leading to major changes in oncogenic signal transduction with regard to p21Cip1/WAF1, STAT3, JNK, c-Myc and survivin, and in the levels of apoptosis-related proteins Puma, Bax and Bcl-xL. Furthermore, Pim-1 knockdown sensitizes tumor cells towards 5-FU treatment, thereby antagonizing a 5-FU-triggered Pim-1 upregulation. This effect is mediated through decreased miR-15b levels, and our studies identify miR-15b and miR-33a to regulate Pim-1.
There is first evidence of Pim-1 overexpression also in glioblastoma, but the functional relevance is so far unknown. U1 small nuclear interference (U1i) has recently been described as novel gene silencing mechanism. It employs short oligonucleotides, so-called U1 adaptors, for specific gene knockdown. We generated Pim-1 specific U1 adaptors and demonstrate their ability to induce Pim-1 knockdown in GBM cells. U1 adaptor transfection leads to anti-proliferative and pro-apoptotic effects.
To explore therapeutic in vivo applications, we complexed siRNAs or U1 adaptors with a low molecular weight polyethylenimine (PEI) that mediates protection and cellular internalization. Treatment of tumor xenograft-bearing mice with PEI/siRNA or PEI/adaptor nanoparticles exhibits antitumor effects based on the knockdown of Pim-1, thus offering novel therapeutic strategies.
Conclusions: We demonstrate that Pim-1 plays a pivotal role in several tumor-relevant signalling pathways, and establish the functional relevance of Pim-1 in colon carcinoma and in glioblastoma. Our results also substantiate the RNAi-mediated Pim-1 knockdown based on polymeric PEI/siRNA nanoparticles as a promising therapeutic approach, and we show that U1i represents an alternative to RNAi for the therapeutic knockdown of pathologically upregulated genes.
Citation Format: Ulrike Weirauch, Maren Thomas, Arnold Grünweller, Roland K. Hartmann, Achim M. Aigner. RNAi- and U1 small nuclear interference (U1i)-mediated gene knockdown reveals the functional relevance of Pim-1 kinase in colon carcinoma and glioblastoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2187. doi:10.1158/1538-7445.AM2013-2187
Abstract 2188: A screen to identify novel p53 family member activity-restoring compounds with a focus on p73.
Authors: van den Heuvel Antonius P. , Wang Wenge , Kopelovich Levy , El-Deiry Wafik S. 
Published: August 2014

Abstract
Tumor suppressor p53 family regulates gene transcription, and thereby cell cycle, apoptosis, metabolism and other processes. The p53 gene is found mutated in over 50% of all human cancers, and it has been of great interest to identify drugs that can restore p53 function. While p53 can display a wide variety of mutations, which potentially could complicate function restoration therapy, family member p73 gene mutation is rare. Therefore, we sought to identify novel compounds that are able to restore the activity of the p53 family, with a focus on p73 dependency in the presence of mutant p53. To this end, a functional cell-based screening assay was set up for cancer cell death-inducing small molecules that activate p53 pathway-dependent gene transcription. SW480 colon carcinoma cells, which carry R273H/R309S mutations in p53, and stably express a p53 family-responsive luciferase construct, were utilized, with parental cells and cells displaying stable knockdown of p73 being subjected to compounds from NCI Diversity Set 3. This set contains about 1600 compounds that are relatively well characterized and structurally rigid. After various treatment periods, p53 activity was assessed, as well as cell viability. Subsequently, a total of 26 compounds were selected for further investigation and validation. Follow up studies will include dose and time courses for p53 family activation and viability in a variety of cell lines that are wild-type, mutant or null for p53. Also mechanistic studies and cellular assays will be performed, with the hopes to identify novel drugs that are able to restore activity of the p53 family, and subsequent cancer cell death, and thus providing novel candidates for the treatment of human cancer.
Citation Format: Antonius P. van den Heuvel, Wenge Wang, Levy Kopelovich, Wafik S. El-Deiry. A screen to identify novel p53 family member activity-restoring compounds with a focus on p73. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2188. doi:10.1158/1538-7445.AM2013-2188
Abstract 2190: cIAP2 as a therapeutic target in malignancies.
Authors: Miura Koh , Karasawa Hideaki , Fujibuchi Wataru , Ohnuma Shinobu , Unno Michiaki 
Published: August 2014

Abstract
[Introduction] Inhibition of the apoptotic pathway is one of the factors that may be responsible for carcinogenesis and drug resistance, and the inhibitor of apoptosis protein (IAP) family is thought to prevent apoptosis through inhibition of caspases and pro-caspases and may affect anti-cancer drug sensitivity. [Purposes] The purpose of this study is to clarify the roles of cellular inhibitor of apoptosis 2 (cIAP2) gene, which is a member of IAP family, on apoptosis and 5-FU sensitivity in cancer cells. [Methods and Results] The expression profiles of human colon cancer cell line DLD-1, its 5-FU-resistant subclone DLD-1/FU and further 21 types of colon cancer cell lines were compared. 50% inhibitory concentration (IC50) of 5-FU in DLD-1/FU was over 100 times higher than that in DLD-1. With the analysis of expression profiles among 23 genes, we found that cIAP2 gene was up-regulated in DLD-1/FU compared with DLD-1 and other cell lines with low IC50. Next, the role of the cIAP2 gene was investigated for 5-FU resistance using RNA interference. The down-regulation of cIAP2 efficiently enhanced 5-FU sensitivity, the activation of caspase 3/7 and apoptosis under exposure to 5-FU. The immunohistochemistry of cIAP2 in cancer and corresponding normal tissues from colorectal cancer patients in stage III revealed that cIAP2 was more frequently expressed in cancer tissues than in normal tissues, and cIAP2-positive patients had a trend toward early recurrence after 5-FU-based chemotherapy. [Conclusion] IAPs are reported to be abnormally regulated and expressed in the majority of human malignancies at elevated levels. Among them, cIAP2 should be a possible therapeutic target in malignancies, especially in colorectal cancer.
Citation Format: Koh Miura, Hideaki Karasawa, Wataru Fujibuchi, Shinobu Ohnuma, Michiaki Unno. cIAP2 as a therapeutic target in malignancies. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2190. doi:10.1158/1538-7445.AM2013-2190
Abstract 2192: A novel mechanism of EML4-ALK rearrangement in a patient-derived cell line and its tumor growth inhibition by an ALK inhibitor CH5424802.
Authors: Kodama Tatsushi , Motoi Noriko , Ninomiya Hironori , Sakamoto Hiroshi , Kitada Kunio , Tsukaguchi Toshiyuki , Sato Yasuko , Ishii Nobuya , Terui Yasuhito , Hatake Kiyohiko , Ishikawa Yuichi 
Published: August 2014

Abstract
Purpose: EML4-ALK is a driver oncogene in non-small cell lung cancer (NSCLC) and has been developed into a promising molecular target for antitumor agents. Although EML4-ALK is reported to be formed by inversion of chromosome 2, other mechanisms of gene fusion remain unknown. Here we characterize the EML4-ALK fusion gene of a new cell line derived from a patient of NSCLC. In addition, the efficacy of ALK inhibitors against this new cell line was examined.
Experimental Design: An EML4-ALK-positive cell line, termed LC649, was established from pleomorphic carcinoma, a rare subtype of NSCLC. We investigated the chromosomal aberrations using fluorescence in situ hybridization (FISH) and comparative genomic hybridization (CGH). CH5424802, a selective ALK inhibitor, was evaluated in the antitumor activity against LC649 in vitro and in vivo xenograft model.
Results: Our initial characterization revealed that LC649 harbors variant 3 of EML4-ALK fusion with 2-fold copy number gain. Interestingly, CGH and metaphase-FISH analysis showed that LC649 contained, in addition to two normal chromosome 2, two aberrant chromosome 2 that were fragmented and scattered. These observations provided the first evidence that EML4-ALK fusion in LC649 cells was formed in chromosome 2 by a distinct mechanism of genomic rearrangement, termed “chromothripsis”. Additionally, CH5424802 inhibited phospho-ALK and suppressed tumor growth of the LC649 cells in vitro and in vivo in a xenograft model.
Conclusions: Our results suggested that chromothripsis may be a mechanism of oncogenic rearrangement of EML4-ALK. CH5424802 was effective against EML4-ALK positive tumors with ALK copy number gain mediated by chromothripsis.
Citation Format: Tatsushi Kodama, Noriko Motoi, Hironori Ninomiya, Hiroshi Sakamoto, Kunio Kitada, Toshiyuki Tsukaguchi, Yasuko Sato, Nobuya Ishii, Yasuhito Terui, Kiyohiko Hatake, Yuichi Ishikawa. A novel mechanism of EML4-ALK rearrangement in a patient-derived cell line and its tumor growth inhibition by an ALK inhibitor CH5424802. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2192. doi:10.1158/1538-7445.AM2013-2192