bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2564: The anticancer molecule TPEN induces DNA damage in human colon cancer cells
Authors: Gali-Muhtasib Hala , Rahal Omar , Fatfat Maamoun , Hankache Carla , Osman Bassam , Khalife Hala , Machaca Khaled 
Published: August 2015

Abstract
The maintenance of optimal metal levels is an essential aspect of cell homoeostasis. However, in many types of cancer these metal levels especially iron, zinc and copper diverge from normal levels. We have recently shown that the zinc chelator TPEN increases the generation of reactive oxygen species (ROS) which selectively kills colon cancer cells. We have also provided evidence that the redox cycling of copper is responsible for TPEN anticancer effects. In this study, we aimed to further decipher the mechanism of TPEN-induced cell death in colon cancer cells through studying its effect on DNA damage. HCT116 p53+/+ human colon cancer cells were seeded were treated with 5μM TPEN at 50% confluence The inhibition of cell growth was measured by MTT, while ROS production was measured by the DCFH Assay using flow cytometry. siRNAs against DNApk and Chk2 was used to investigate the involvement of these DNA damage sensors in TPEN activity. DNA damage was assessed by the comet assay. Phosphorylation of ATM, ATR, Chk1, Chk2 and H2AX by TPEN were detected immunocytochemically by multiparameter cytometry. Expression levels of Chk1/2, ATR and DNApK were determined by western blotting. We show that cell death by TPEN is associated with significant DNA damage, an effect that was dependent on ROS generation and on the redox cycling of copper, as evidenced by reversal of DNA damage in the presence of antioxidants (NAC, CAT) or the copper chelator neocuproine (Neo). DNA damage was associated with increased expression of p-H2AX and a significant activation of ATM/ATR signaling molecules, specifically p-ATM, p-ATR and p-Chk1. Interestingly, silencing DNApk and Chk2 reversed DNA damage caused by TPEN, suggesting the involvement of DNApk and ATM/ATR pathways in TPEN-mediated effects. This study shows for the first time the involvement of DNApk and Chk2 in TPEN-induced DNA damage and confirms our previous findings that the redox cycling of copper is the main mechanism by which TPEN induces cell death in human colon cancer cells.
Citation Format: Hala Gali-Muhtasib, Omar Rahal, Maamoun Fatfat, Carla Hankache, Bassam Osman, Hala Khalife, Khaled Machaca. The anticancer molecule TPEN induces DNA damage in human colon cancer cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2564. doi:10.1158/1538-7445.AM2015-2564
Abstract 2574: HIPEC for treatment of peritoneal carcinomatosis: 40°C is the critical threshold temperature for potentiating chemotherapy efficacy in vitro and in vivo
Authors: Schaaf Lea , Münch Marina , Steurer Wolfgang , Aulitzky Walter E. , Mürdter Thomas E. , van der kuip Heiko , Ulmer Christoph 
Published: August 2015

Abstract
Hyperthermic intraperitoneal chemotherapy (HIPEC) combines circulated chemotherapy with enhanced temperature in the abdominal cavity of peritoneal carcinosis (PC) patients. Unfortunately, a standardized HIPEC protocol is missing hindering systematic comparisons with regard to minimal effective temperatures. The purpose of the present study was to systematically analyse the temperature needed for potentiation of chemotherapy effects in vitro. We then asked if this temperature could be sustained in patients during HIPEC and evaluated possible effects on survival. We therefore established a cell line-based model to mimic HIPEC conditions used in clinic. We evaluated intracellular drug concentrations and long-term survival using different temperatures ranging from 38 to 42°C combined with Cisplatin and Doxorubicin. In parallel we evaluated the temperature reached in the clinical setting by measuring inflow and outflow, as well as in the peritoneal cavity in 36 PC patients. Finally we determined the influence of different temperatures on survival. In vitro, we found temperature-dependent effects on intracellular drug concentration and long term survival only at temperatures above 40°C. In patients, during HIPEC, constant temperatures were reached after 10 minutes in the peritoneal cavity. A temperature above 40°C for at least 45 minutes was achieved in 67% or 44% of patients with HIPEC in open or closed abdomen, respectively. Importantly, we observed a significantly enhanced overall survival as well as a trend towards longer recurrent free survival in those patients with HIPEC temperatures above 40°C.
In conclusion, hyperthermia significantly potentiated the chemotherapy effects only at temperatures above 40°C in vitro. Importantly, this temperature threshold was also critical for overall and recurrent-free survival of PC patients.
Citation Format: Lea Schaaf, Marina Münch, Wolfgang Steurer, Walter E. Aulitzky, Thomas E. Mürdter, Heiko van der kuip, Christoph Ulmer. HIPEC for treatment of peritoneal carcinomatosis: 40°C is the critical threshold temperature for potentiating chemotherapy efficacy in vitro and in vivo. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2574. doi:10.1158/1538-7445.AM2015-2574
Abstract 2580: Screening of a mixture-based synthetic combinatorial library identifies small molecules that inhibit the ability of GTP to displace mant-GDP from mutant G12D KRas
Authors: Kennedy Perry C. , Guilanotti Marc C. , LsVoi Travis , Sebti Said M. 
Published: August 2015

Abstract
Although mutant KRas significantly contributes to human oncogenesis and patient tumor resistance to therapy, there are no anticancer drugs directly targeting mt KRas available in clinic. A major effort in our laboratory is to identify novel compounds that bind mt KRas and inhibit potently and selectively KRas-driven malignant transformation. To this end, we have used a mixture-based synthetic combinatorial library (scaffold ranking library) containing over 6 million compounds in a mant-GDP assay to identify chemical scaffolds that inhibit the ability of GTP to displace mant-GDP from mt G12D KRas. Subsequent screening of the positional scanning libraries derived from the hit mixtures led to several individual compound hits with the most potent inhibiting the ability of GTP to displace mant-GDP from mt G12D KRas with IC50 of 61 μM. Current studies are focused on physical characterization of the hits to mt KRas and cellular activities of these hits in mt KRas-dependent and -independent human cancer cell lines.
Citation Format: Perry C. Kennedy, Marc C. Guilanotti, Travis LsVoi, Said M. Sebti. Screening of a mixture-based synthetic combinatorial library identifies small molecules that inhibit the ability of GTP to displace mant-GDP from mutant G12D KRas. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2580. doi:10.1158/1538-7445.AM2015-2580
Abstract 2585: Discovery of a potent covalent mutant-selective EGFR inhibitor - the journey from high throughput screening to EGF816
Authors: Lelais Gerald , Epple Robert , Michellys Pierre-Yves , Marsilje Thomas H. , Long Yun , McNeill Matthew , Chen Bei , Lu Wenshuo , Bursulaya Badry , DiDonato Michael , Jia Yong , Kasibhatla Shailaja , Li Chun , Matushansky Igor , Bender Steven 
Published: August 2015

Abstract
Epidermal growth factor receptor (EGFR) is a validated therapeutic target for lung cancer. First and second generation EGFR inhibitors (e.g., gefitinib, erlotinib and afatinib) have revolutionized treatment paradigms of non-small cell lung cancer (NSCLC) patients with oncogenic EGFR mutations. The use of EGFR tyrosine kinase inhibitors (TKI) provides superior efficacy compared to chemotherapy in patients with EGFR L858R or exon 19 deletion tumors. However, resistance inevitably develops after 8-12 months of treatment; most commonly via a secondary T790M point mutation at the gatekeeper residue of EGFR. Furthermore, responses are hindered due to treatment intolerance in the form of rash and diarrhea that are mediated by simultaneous inhibition of wild-type (WT) EGFR at doses required for mutant EGFR suppression. To overcome these limitations, we initiated a project to identify mutant-selective EGFR inhibitors that potently inhibit both activating and T790M resistance EGFR mutations while sparing WT EGFR.
In this presentation, we report our medicinal chemistry approach and optimization that led to the discovery of EGF816, a selective and potent covalent mutant-selective EGFR inhibitor with single digit nanomolar cellular target modulation on both activating and T790M resistance mutations. In addition, we will also report validated clinical efficacy data from the first patient treated with EGF816.
Citation Format: Gerald Lelais, Robert Epple, Pierre-Yves Michellys, Thomas H. Marsilje, Yun Long, Matthew McNeill, Bei Chen, Wenshuo Lu, Badry Bursulaya, Michael DiDonato, Yong Jia, Shailaja Kasibhatla, Chun Li, Igor Matushansky, Steven Bender. Discovery of a potent covalent mutant-selective EGFR inhibitor - the journey from high throughput screening to EGF816. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2585. doi:10.1158/1538-7445.AM2015-2585
Abstract 2588: Anti-tumor effect of afatinib, an irreversible EGFR/HER2 dual inhibitor, in lung cancers harboring HER2 oncogene
Authors: Suzawa Ken , Toyooka Shinichi , Sakaguchi Masakiyo , Ohtsuka Tomoaki , Watanabe Mototsugu , Hashida Shinsuke , Maki Yuho , Yamamoto Hiromasa , Soh Junichi , Asano Hiroaki , Tsukuda Kazunori , Miyoshi Shinichiro 
Published: August 2015

Abstract
HER2 is a member of the EGFR family of tyrosine kinase receptors and plays important roles in the pathogenesis of certain human cancers. In non-small cell lung cancer (NSCLC), HER2 amplification or mutations are reported. However, little is known about the benefit of HER2-targeted therapy for NSCLC harboring HER2 alteration. In this study, we investigated the anti-tumor effect of afatinib, an irreversible EGFR/HER2 dual inhibitor, in lung cancers harboring HER2 oncogene alteration, including the novel HER2 mutations in transmembrane domain which we recently identified. Ectopically overexpressing HER2 wild-type or mutant (G776V,Cins, A775insYVMA, G660D, and V659E) in normal bronchial epithelial cells, BEAS2B, showed constitutive autophosphorylation of HER2 and activation of downstream signaling. They were sensitive to afatinib, but insensitive to EGFR-TKI (gefitinib). Furthermore, we examined anti-tumor activity of afatinib and gefitinib in several NSCLC cell lines, and investigated the association between their genetic alteration and sensitivity to afatinib treatment. Afatinib down-regulated the phosphorylation of HER2, EGFR, and their downstream signalings, and induced anti-proliferative effect through a G1 arrest and apoptotic cell death in HER2 altered cells (H2170, Calu-3, and H1781). Whereas, HER2 non-dependent NSCLC cells (A549, H1299, and H1993) were insensitive to afatinib. Our results strongly suggest that afatinib is a therapeutic option for NSCLC patients with HER2 alteration.
Citation Format: Ken Suzawa, Shinichi Toyooka, Masakiyo Sakaguchi, Tomoaki Ohtsuka, Mototsugu Watanabe, Shinsuke Hashida, Yuho Maki, Hiromasa Yamamoto, Junichi Soh, Hiroaki Asano, Kazunori Tsukuda, Shinichiro Miyoshi. Anti-tumor effect of afatinib, an irreversible EGFR/HER2 dual inhibitor, in lung cancers harboring HER2 oncogene. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2588. doi:10.1158/1538-7445.AM2015-2588
Abstract 2589: Investigations of covalent binding efficiency for third-generation EGFR irreversible inhibitors AZD9291 and CO-1686
Authors: Qin Zhen , Sun Xuebing , Liu Ye , Du Yunguang , Wang Miao , Hu Nan , Zhang Jiye , Peng Hao , Wang Lai , Wei Min , Luo Lusong 
Published: August 2015

Abstract
Patients with non-small cell lung cancer with activating EGFR mutations initially respond to first-generation and second-generation EGFR inhibitors. However, clinical efficacy is limited by acquired resistance, frequently driven by the EGFR T790M mutation. In addition, patients oftentimes experience severe side effects including rash and diarrhea due to the inhibition profile against wild-type EGFR. AZD9291 and CO-1686 are the leading third-generation irreversible EGFR inhibitors in clinical development. They selectively inhibit T790M EGFR, while sparing wild-type EGFR. In this study, we investigated the covalent binding efficiency of these two irreversible EGFR inhibitors using biochemical and cellular assays. We discovered that these two inhibitors had dramatic difference in covalent binding efficiencies in WT and mutant EGFR binding at biochemical and cellular levels. In pEGFR cellular assay, AZD9291 displayed much prolonged inhibition of pEGFR than CO-1686. In CO-1686 treated cells, recovery of pEGFR had reached 50% of control levels only after 4 hours. In the meantime, pEGFR in AZD9291 treated cells had no recovery even after 24 hours. These data may suggest different covalent bond formation efficiency and reversibility between different types of irreversible EGFR inhibitors. This may be one of the important factors for consideration when develop third-generation EGFR inhibitors.
Citation Format: Zhen Qin, Xuebing Sun, Ye Liu, Yunguang Du, Miao Wang, Nan Hu, Jiye Zhang, Hao Peng, Lai Wang, Min Wei, Lusong Luo. Investigations of covalent binding efficiency for third-generation EGFR irreversible inhibitors AZD9291 and CO-1686. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2589. doi:10.1158/1538-7445.AM2015-2589
Abstract 2592: Anti-tumor activity of icotinib (BPI-2009H) and BPI-403B in the BxPC-3 pancreatic cancer cell line
Authors: Wilde Victoria Lynn , Zhang Don , Peng Jirong , Green Michael A. , Greco Michael N. , Costanzo Michael 
Published: August 2015

Abstract
Introduction: The purpose of this research was to investigate the anti-tumor effect of BPI-2009H and BPI-403B, an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) in the human pancreatic cancer cell lines BxPC-3 and Panc-1, in vitro and in vivo and determine the maximum tolerated dose (MTD) of the compound in mouse.
Experimental Design: In vitro, human pancreatic cancer cell lines, BxPC-3 and Panc-1 were exposed to varying concentrations of BPI-2009H and BPI-403B, the concentration of compound required to inhibit growth by fifty percent (IC50) was estimated. In vivo, tumor growth inhibition of low dose BPI-2009H was tested in xenografted nude mice with BxPC-3 pancreatic cancer tumors. The maximum tolerated dose (MTD) was established via oral administration of low, mid and high doses to Harlan mice of both BPI-2009H and BPI-403B. Upon determination of the MTD, the in vivo efficacy of high dose BPI-2009H was evaluated via tumor growth inhibition study on xenografted nude mice with BxPC-3 pancreatic cancer tumors.
Results: BPI-2009H, as a single agent, inhibited the growth of BxPC-3 cells in a dose dependent manner, with a relative IC50 value of 25.8 μM. BPI-403B, as a single agent, inhibited growth of BxPC-3 cells in a dose dependent manner as well, with a relative IC50 value of 2.7 μM. BPI-2009H and BPI-403B were tested up to 50μM in Panc-1 cells with no apparent effect at the upper limit of the study for either of the compounds. In vivo, the mice treated with BPI-403B demonstrated a statistically significant tumor growth delay, 14.3 days. 20% of the mice experienced tumor regression. The compound was well tolerated with minimal body weight loss. At low doses, BPI-2009H did not show any statistically significant tumor growth delay, but upon increasing the dose to the MTD, significant signs of anticancer activity was demonstrated, with an increase in tumor growth delay.
Conclusions: The in vitro and in vivo findings show that both BPI-2009H and BPI-403B have antiproliferative effects against BxPC-3 pancreatic cancer cells. These compounds are both well tolerated and may prove to be an alternative option of EGFR-TKI to erlotinib treatment in pancreatic cancer cases.
Citation Format: Victoria Lynn Wilde, Don Zhang, Jirong Peng, Michael A. Green, Michael N. Greco, Michael Costanzo. Anti-tumor activity of icotinib (BPI-2009H) and BPI-403B in the BxPC-3 pancreatic cancer cell line. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2592. doi:10.1158/1538-7445.AM2015-2592
Abstract 2593: Combination therapy with EGFR-TKI and cMET-TKIs in non-small cell lung cancer: the golden duo
Authors: Van Der Steen Nele , Deschoolmeester Vanessa , Wouters An , Lardon Filip , Rolfo Christian , Pauwels Patrick 
Published: August 2015

Abstract
Intro: The use of targeted therapies confers great responses in non-small cell lung cancer patients, but ultimately results in acquired resistance, e.g. against epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) due to cMET amplification. Therefore, combination therapy with EGFR-TKI and cMET TKIs in cell lines with a different EGFR and cMET background was investigated.
Materials & methods: All experiments were performed in 5 cell lines with a different EGFR and cMET background: the cMET amplified squamous lung cancer cell line EBC-1, the HGF-secreting myeloma cell line JJN3, the HCC827 and H1975 lung adenocarcinoma cell lines with an activating EGFR exon 19 deletion and an L858 as well as a T790M resistance mutation, respectively. Finally LUDLU-1 is a squamous lung cancer cell line, wild-type for both EGFR and cMET.
The cytotoxicity of combination therapy with erlotinib (EGFR-TKI) and crizotinib (cMET-ALK-ROS1-TKI) or PHA665752 (cMET-TKI) was determined by the sulforhodamine B assay after 72h of simultaneous/sequential treatment (Table 1).
Results: For all treatments, the JJN3, EBC-1 and HCC827 cell lines showed no synergistic effect. For the H1975, a decrease in resistance could be observed, albeit not significant. For LUDLU-1, a trend towards synergism was observed when erlotinib was administered simultaneously with crizotinib (p = 0.074) and PHA665752 (p = 0.055). Also a significantly synergistic effect was observed when treatment with erlotinib was followed by PHA665752 (p = 0.014).
Discussion: Synergism between erlotinib and crizotinib/PHA665752 depends on the genetic background of the cells and the treatment schedule. This was most pronounced in the EGFR and cMET wild-type cell line. These results open up the discussion to include EGFR and cMET wild-type patients into future clinical trials.
Cell lineErlotinib (μM)Crizotinib (μM)PHA665752 (μM)Erlo+CrizErlo = &gt;CrizCriz = &gt;ErloErlo+PHAErlo = &gt;PHAPHA = &gt;ErloEBC-10-100-0.025-0.050-0.1-0.150-0.025-0.050-0.1-0.20-0.3-0.40-0.1-0.2JJN30-100-2-30-2-30-2-30-5-70-5-70-5-6HCC8270-50-3-50-3-50-1-20-7-90-7-90-4-5H19750-100-2-30-2-30-1-20-6-70-6-70-2-3LUDLU-10-100-3-50-3-50-2-30-7-80-7-80-3.5-4
Table 1: Therapeutic concentrations per cell line & treatment schedule: control-IC20-IC40.
Citation Format: Nele Van Der Steen, Vanessa Deschoolmeester, An Wouters, Filip Lardon, Christian Rolfo, Patrick Pauwels. Combination therapy with EGFR-TKI and cMET-TKIs in non-small cell lung cancer: the golden duo. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2593. doi:10.1158/1538-7445.AM2015-2593
Abstract 2595: Efficacy of cetuximab and mutant selective EGFR inhibitor WZ4002 in EGFR T790M and non-T790M models of erlotinib resistant non-small cell lung cancer
Authors: Tricker Erin M. , Xu Chunxiao , Wong Kwok-Kin , Janne Pasi A. 
Published: August 2015

Abstract
Background: EGFR kinase inhibitors, including erlotinib, are effective treatments for patients with EGFR mutant lung cancer. However, resistance inevitably develops, most commonly (60%) mediated by an EGFR secondary mutation EGFR T790M. Mutant selective EGFR inhibitors (WZ4002, AZD9291, CO-1686) are effective in preclinical models and clinically (AZD9291 & CO-1686) in EGFR T790M patients (response rates (RR) &gt; 50%) but are less effective in patients with non-T790M mediated drug resistance (AZD9291 RR ∼10%). Combination of afatinib and cetuximab is effective in both T790M (32% RR) and non-T790M (25% RR) mediated resistance but is associated with significant toxicity. We evaluated whether cetuximab could add to the efficacy of a mutant selective EGFR inhibitor and whether the combination was also effective in a model of non-T790M mediated resistance.
Methods: Mice harboring xenografts from PC9GR (EGFR Del 19/T790M), H1975 (EGFR L858R/T790M), HCC827GR (EGFR Del 19/MET amplification) cells or genetically engineered mouse models (GEMMS) expressing EGFR L858R/T790M were treated with vehicle, WZ4002 alone, cetuximab alone or the combination. We compared the efficacy of the treatments and evaluated for tumor cures (defined as tumor disappearance and lack of regrowth following 6 weeks of drug withdrawal). In addition, we evaluated the impact of the treatments on EGFR signaling and apoptosis.
Results: In the EGFR T790M xenograft models (PC9GR and H1975), the combination of WZ4002 and cetuximab was more effective than either single agent. Only the combination led to tumor cures (5/15 in PC9GR and 3/18 in H1975) while none were observed with the single agents. In the HCC827GR xenografts, the combination of WZ4002/cetuximab was more effective than the single agents but did not lead to any cures. Evaluation of the tumors from treated mice revealed that the efficacy of the combination correlates with effective degradation of EGFR and MET (in HCC827GR). In the L858R/T790M GEMMs, the combination led to significantly greater tumor shrinkage compared to single agents. Evaluation of treated tumors revealed enhanced apoptosis in the combination treated mice.
Conclusions: The addition of cetuximab enhances the efficacy of the mutant selective EGFR inhibitor WZ4002 in vivo by increasing the single agent efficacy in both EGFR T790M and non-T790M models. Clinical trials of mutant selective EGFR inhibitors and cetuximab are warranted.
Citation Format: Erin M. Tricker, Chunxiao Xu, Kwok-Kin Wong, Pasi A. Janne. Efficacy of cetuximab and mutant selective EGFR inhibitor WZ4002 in EGFR T790M and non-T790M models of erlotinib resistant non-small cell lung cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2595. doi:10.1158/1538-7445.AM2015-2595