bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4448: Identification of a small molecule inhibitor of aldolase A for the targeting of hypoxic cancer cells
Authors: de Jong Petrus R. , Grandjean Geoffrey V. , Devkota Ashwini K. , Cho Eun Jeong , Dalby Kevin N. , Powis Garth
Published: August 2015

Cancer cells are critically dependent on glycolysis. Currently, there is no effective pharmacotherapy that exploits this metabolic vulnerability. We have uncovered a feed forward cycle of anaerobic glycolysis and hypoxia-inducible factor-1 (HIF-1). Glycolysis under hypoxic conditions normally maintains high ATP levels, which is driven by transcriptional activity of HIF-1. Glycolysis inhibition results in a cellular energy crisis, i.e. an increased AMP:ATP ratio, leading to AMPK-mediated phosphorylation of the HIF-1 co-activator p300/CBP. This prevents HIF-1 activity, although HIF-1 protein levels are unchanged, abrogating the feed forward cycle. We identified fructose-1,6-bisphosphate aldolase A (ALDOA) as a top glycolytic enzyme target for inhibiting hypoxic cancer cell glycolysis and HIF-1 activity. Our aim was to identify small molecule inhibitors for ALDOA that may yield compounds for further preclinical development. To this end, a primary high-throughput screen (HTS) was performed with 65,936 compounds using a fluorescence-based NADH oxidation aldolase (cell-free) assay. 640 compounds were further tested in a cherry-pick confirmation screen, from which 112 hits underwent concentration-response curve validation in cell-free assays. From these, 4 hits were further tested in cell-based assays by using colorectal, breast and pancreatic cancer cell lines. A lead compound that showed micromolar potency in inhibiting ALDOA, induced cell death under hypoxic conditions with IC50 values ranging between 2 - 8 μM in the cancer cell lines tested. It also inhibited extracellular flux in a real-time glycolysis assay, inhibited extracellular lactate production under hypoxic conditions as a measure of glycolysis, and blocked hypoxia responsive element (HRE) HIF-1 reporter activity but not HIF-1 protein levels. No effect of the lead compound was observed on mitochondrial respiration. Thus, this compound provides a valuable chemical probe for inhibiting glycolysis in cancer cells. Furthermore, the work provides proof-of-concept that targeting glycolysis with a small molecule inhibitor exerts potent antitumor effects in vitro and is currently being tested in preclinical models as a first-in-class oncological agent.

Citation Format: Petrus R. de Jong, Geoffrey V. Grandjean, Ashwini K. Devkota, Eun Jeong Cho, Kevin N. Dalby, Garth Powis. Identification of a small molecule inhibitor of aldolase A for the targeting of hypoxic 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 4448. doi:10.1158/1538-7445.AM2015-4448
Abstract 445: Tumor microenvironment influences survival of mantle cell lymphoma-initiating cells through FGF/FGFR1 signaling
Authors: Sehgal Lalit , Mathur Rohit , Berkova Zuzana , Khashab Tamer , Wang Xin , Romaguera Jorge E. , Rodriguez Alma E. , Neelapu Sattva , Samaniego Felipe
Published: August 2015

Introduction

Mantle cell lymphoma (MCL) represents an aggressive, incurable form of non-Hodgkin's lymphoma (NHL). The health complications associated with advanced age of MCL patients restrict treatment with intense chemotherapy. Translocation t(11;14), responsible for overexpression of cyclin-D1, is the hallmark of MCL. More detailed insight into MCL pathogenesis has been delayed until the recent development of a tissue culture system, using human mesenchymal stromal cells (hMSC), suitable for propagating primary MCL cells. We hypothesized that tumor-initiating cells are responsible for MCL relapse and chemoresistance and thus, identification of signals responsible for survival and maintenance of MCL-initiating cells (MCL-ICs) is essential for design of curative treatment strategies.

Methods

Isolates of primary MCL cells (n = 24) were co-cultured with human mesenchymal stem cells (hMSCs) and the content of MCL-ICs was analyzed by flow-cytometry based on marker expression profile; CD34-CD3-CD45+CD19-. Cytokine array was used to identify the soluble factors enriched in the co-cultures and the expression of these factors was confirmed by RT-PCR analysis. The signaling pathways employed by the newly-identified factors were blocked in 3 MCL cell lines (JVM2, Mino, Z138) to confirm their essential role in survival of MCL cells and, more importantly, for MCL-ICs.

Results

Co-cultures of primary MCL isolates with hMSCs supported the growth of MCL cells for over 4 weeks with continued presence of MCL-ICs (CD34-CD3-CD45+CD19-) representing about 1% of MCL cells. We found that IL-6 produced by hMSCs triggered an FGF/FGFR autocrine loop in MCL-ICs. The extent of FGFR expression correlated tightly with expression of SOX11, a pathology related negative prognostic marker in MCL. MCL cell survival and growth was regulated via the FGFR/mir101/ EZH2/ NF-κB/XIAP axis. Blocking of this signaling pathway with FGFR1 inhibitors consistently induced early reduction in XIAP levels and subsequently MCL cell death.

Conclusion

We established that propagation of primary MCL in co-cultures with hMSCs depends on an FGF/FGFR autocrine loop that enhances XIAP protein expression and thus, supports survival of MCL cells. We identified the factors essential for survival of MCL and MCL-ICs that present new targets for improved MCL treatment strategies.

Citation Format: Lalit Sehgal, Rohit Mathur, Zuzana Berkova, Tamer Khashab, Xin Wang, Jorge E. Romaguera, Alma E. Rodriguez, Sattva Neelapu, Felipe Samaniego. Tumor microenvironment influences survival of mantle cell lymphoma-initiating cells through FGF/FGFR1 signaling. [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 445. doi:10.1158/1538-7445.AM2015-445
Abstract 4462: The effects of NT-1044, a novel AMPK activator, on endometrial cancer cell proliferation and apoptosis
Authors: Roque Dario R. , Wysham Weiya W. , Zhou Chunxiao , Batchelor Ken , Brewster Wendy R. , Bae-Jump Victoria L.
Published: August 2015

Objectives: Obesity and diabetes are associated with increased risk and worse outcomes in endometrial cancer (EC). Anti-diabetic biguanide drugs such as metformin may have anti-tumorigenic effects by behaving as AMPK activators and mTOR inhibitors. Metformin requires organic cation transporters (OCTs) for entry into cells, and NT-1044 (NovaTarg Therapeutics) is an AMPK activator designed to have greater affinity for two of these transporters, OCT1 and 3. We sought to compare the effects of NT-1044 on cell proliferation and apoptosis in human EC cell lines.

Methods: Cell proliferation was assessed in two EC cell lines, ECC-1 and Ishikawa, by MTT assay after exposure to NT-1044. Apoptosis was analyzed by Annexin V-FITC assay. Cell cycle progression was evaluated by flow cytometry. Phosphorylated (phos)-S6, phos-AMPK, Cyclin D-1 and CDK-6 were evaluated by Western immunoblotting.

Results: NT-1044 significantly inhibited proliferation in a dose-dependent manner in both EC cell lines after 72 hours of exposure (IC50 218 uM for ECC-1; 87 uM for Ishikawa). Treatment with NT-1044 resulted in G1 cell cycle arrest and apoptosis in both cell lines. NT-1044 increased phosphorylation of AMPK and decreased phosphorylation of S6, a key downstream target of the mTOR pathway. Expression of Cyclin-D1 and CDK-6 also decreased in a dose dependent-fashion in both cell lines.

Conclusions: NT-1044 suppressed EC cell growth through G1 cell cycle arrest, inducing apoptosis, and inhibition of the mTOR pathway. More work is needed to determine if this novel biguanide will be beneficial in the treatment of women with EC, a disease strongly impacted by obesity and diabetes.

Citation Format: Dario R. Roque, Weiya W. Wysham, Chunxiao Zhou, Ken Batchelor, Wendy R. Brewster, Victoria L. Bae-Jump. The effects of NT-1044, a novel AMPK activator, on endometrial cancer cell proliferation and apoptosis. [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 4462. doi:10.1158/1538-7445.AM2015-4462
Abstract 4465: Sequential combination of docetaxel with a novel SHP-1 agonist enhanced anti-tumor effect in triple negative breast cancer cells
Authors: Tseng Ling-Min , Liu Chun-Yu , Yang Shiu-Ping , Wang Wan-Lun , Su Jung-Chen , Wu Chia-Yun , Shiau Chung-Wai , Chen Kuen-Feng
Published: August 2015

Background: Interfering the oncogenic STAT3 signaling is considered as promising anti-cancer strategy. STAT3 signaling can be negatively regulated by the protein tyrosine phosphatase SHP-1 through direct dephosphorylation of p-STAT3 (Tyr705). Our previous studies have confirmed SC-43 as a novel SHP-1 enhancer which down-regulate p-STAT3 and STAT3-related downstream effectors (Breast Cancer

Research 2013). Here, we tested the efficacy of SC-43 in combination with docetaxel in triple negative breast cancer cells.

Methods: TNBC cell lines (HCC-1937, MDA-MB-468, MDA-MB-231) were used for in vitro studies. Cell viability was examined by MTT assay. Combination index was determined using Calcusyn anaysis. Apoptosis was examined by flow cytometry and western blot. Signal transduction pathways in cells were assessed by western blot. In vivo efficacy of

SC-43 in combination with docetaxel was tested in nude mice with breast cancer xenografts.

Results: Sequential combination of docetaxel with SC-43 showed enhanced anti-proliferative activity in all tested TNBC cells. Synergistic effect was demonstrated when docetaxel combined with SC-43 at a dose ratio of 1 to 10, showing combination index below 1.0. Sub-G1 and western blot analysis showed that SC-43 enhanced docetaxel-induced apoptosis. In addition, combining SC-43 with docetaxel efficiently suppressed p-STAT3 and downstream effectors. Ectopic expression of STAT3 reduced the enhanced cytotoxicity by the combination therapy. Previous studies has shown SC-43 enhanced SHP-1 activity, here we showed that SC-43 in combination with docetaxel significantly enhanced SHP-1 activity. More importantly, intraperitoneal injection of SC-43 with tri-weekly docetaxel showed enhanced anti-tumor growth in xenografted tumor mice.

Conclusion: Our results suggest that the novel SHP-1 agonist SC-43 enhanced docetaxel-induced cytotoxic killing by SHP-1 dependent STAT3 inhibition in human triple negative breast cancer cells. (Supported by Yen Tjing-Ling Medical Foundation CI-104-07; NSC-102-2325-B-075-006 and MOST 103-2325-B-075-002; and MOHW103-TD-B-111-02)

Citation Format: Ling-Min Tseng, Chun-Yu Liu, Shiu-Ping Yang, Wan-Lun Wang, Jung-Chen Su, Chia-Yun Wu, Chung-Wai Shiau, Kuen-Feng Chen. Sequential combination of docetaxel with a novel SHP-1 agonist enhanced anti-tumor effect in triple negative breast 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 4465. doi:10.1158/1538-7445.AM2015-4465
Abstract 4469: A novel class of inhibitors of the immunosuppressive enzyme indoleamine 2,3-dioxygenase 1 (IDO1) with potential for the treatment of cancer
Authors: Ching Lai-Ming , Palmer Brian D. , Tomek Petr , Flanagan Jack U. , Henare Kimiora
Published: August 2015

A broad range of cancers express the tryptophan catabolising enzyme indoleamine 2,3-dioxygenase 1 (IDO1) as a mechanism of suppressing the endogenous anti-tumour immunity. High IDO1 expression by their tumours is associated with poor patient prognosis and has prompted the discovery of small molecule inhibitors of the enzyme for development as anti-cancer agents. Towards this end, we carried out a high throughput screen of 42,000 small molecules in the Walter and Eliza Hall Institute for Medical Research compound library and retrieved over 30 good hits with diverse structures that inhibited recombinant human IDO1 with potencies in the low μM range. Initial structure activity relationship studies of the best hits led us to focus on one chemical class, which, although less potent than some of the other hits on the list, exhibited a number of features that make it more favourable for development as a therapeutic agent. SN35837, the non-optimised hit from the high throughput screen is highly cell permeable and displays improved potency in inhibiting cytoplasmic hIDO1 in cell-based assays, compared to that for enzymatic assays. Complete inhibition of IDO1 function is seen at concentrations that has no measurable effects on cell viability. The imidazole lead from New Links Genetics, NLG919 has decreased activity in cells, whilst Incyte's hydroxyamidine inhibitor INCB024360 has similar potency in cellular and enzymatic assays. The maximum tolerated dose SN35837 in C57Bl/6 mice given as a single intraperitoneal bolus dose was 500 mg/kg. Mice tolerated daily intraperitoneal or subcutaneous administrations of SN35837 at 150 mg/kg. Statistically significant inhibition of growth of subcutaneously implanted GL261 gliomas and Lewis Lung carcinomas in syngeneic hosts were achieved after daily administration of 75 mg/kg SN35837. Preliminary studies indicate that the non-optimised hit also synergises when given in combination with anti-PD1 and anti-CTLA-4 antibodies. We conclude that derivatives of SN35837 may provide effective and potent inhibitors of IDO1 suitable for development as anti-cancer agents.

Citation Format: Lai-Ming Ching, Brian D. Palmer, Petr Tomek, Jack U. Flanagan, Kimiora Henare. A novel class of inhibitors of the immunosuppressive enzyme indoleamine 2,3-dioxygenase 1 (IDO1) with potential for the treatment of 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 4469. doi:10.1158/1538-7445.AM2015-4469
Abstract 4471: The antitumor activity of hyaluronic acid nimuselide-NH2 bioconjugate (CA102N) in human colorectal cancer (CRC)
Authors: Tchaparian Eskouhie
Published: August 2015

Carrier mediated drug delivery systems are promising next generation therapeutics for targeted delivery and improved efficacy and safety of cytotoxic drugs. Hyaluronic Acid conjugated drugs (HACD) are novel carrier-mediated drug delivery systems (Targeting Carrier™) characterized by CD44-mediated endocytosis of Hyaluronic Acid (HA) and intracellular drug release. The present study investigates the in vitro and in vivo antitumor activity of CA102N, an HA- Cyclooxygenase-2 inhibitor conjugate, HA-Nimesulide, currently in preclinical development. Cyclooxygenase-2 (COX-2) inhibitors are a class of compounds that are associated with anticancer properties. CA102N was able to selectively inhibit HCT116 and HT29, CRC cancer cell growth in vitro with an IC50 of 57.7 and 77.14 μM respectively. In in vivo studies, strong anti-tumor effects and rapid tumor shrinkage was observed within 3 days posttherapy in mice bearing HT-29 xenografts treated with either 1.5 or 4.5 mg/kg of relative amount of the conjugate. Further, full and sustained regression of tumors was also attained during the entire observational period (14 days) after the last dose. No treatment related toxicity was noticed in mice. HACD drug delivery systems may offer great advantages in the development of a new class of bioconjugated drugs for intracellular delivery and enhanced safety of potent cytotoxic drugs.

Citation Format: Eskouhie Tchaparian. The antitumor activity of hyaluronic acid nimuselide-NH2 bioconjugate (CA102N) in human colorectal cancer (CRC). [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 4471. doi:10.1158/1538-7445.AM2015-4471
Abstract 4472: Plitidepsin shows antitumor activity in patient-derived tumor xenografts and hematologic malignancies
Authors: Maier Armin , Kelter Gerhard , Vuaroquaux Vincent , Avilés Marin Pablo M. , Cuevas Carmen , Galmarini Carlos M. , Fiebig Heinz H.
Published: August 2015

Plitidepsin is a new marine-derived anti-tumor agent, originally isolated from the tunicate Aplidium albicans that is now fully obtained by total chemical synthesis. The compound is currently being evaluated in a pivotal phase III trial in patients with refractory/relapsed multiple myeloma. Previous in vitro studies in 26 hematological cell lines demonstrated that plitidepsin was more active in multiple myeloma and non-Hodgkin lymphoma compared to CML, AML and ALL derived cell lines as determined in a 4 day proliferation assay in suspension cultures. We have recently shown that plitidepsin interacts selectively with eEF1A2 (eukaryotic translation elongation factor 1 alpha 2) in tumor cells, which results in apoptotic cell death. To identify target tumor types for future clinical studies, the inhibition of colony formation of plitidepsin was assessed in vitro in 116 established patient derived xenografts tumor models from 23 different tumor types, including solid tumors and hematological malignancies such as myeloma, leukemia and lymphoma. The anti-tumor activity of plitidepsin varied depending on the concentration used, achieving a mean IC70 (inhibitory concentration of 70%) value of 3 nM (range between 0.03 nM and >1 μM), which can be also achieved in patients treated using the clinical recommended dose or less. 39 out of 116 tumors showed above average sensitivity (IC70 < 1nM) and IC70 values in these tumors were about 9-fold lower than the mean of all tumors as tested. Sensitive tumor types were seen among different tumor types, such as hematological malignancies, pleuramesothelioma, melanoma, small cell and non-small cell lung cancer (both adeno and large cell), as well as renal cancer. In conclusion plitidepsin showed a broad differential antitumor activity in vitro in hematological malignancies and in solid tumors. Bioinformatic analyses are currently underway to identify biomarkers and/or gene signatures that can predict sensitivity or resistance to plitidepsin for selecting patients being responsive to the drug.

Citation Format: Armin Maier, Gerhard Kelter, Vincent Vuaroquaux, Pablo M. Avilés Marin, Carmen Cuevas, Carlos M. Galmarini, Heinz H. Fiebig. Plitidepsin shows antitumor activity in patient-derived tumor xenografts and hematologic malignancies. [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 4472. doi:10.1158/1538-7445.AM2015-4472
Abstract 448: The role of the Nanog transcription factor in tumor immune escape
Authors: Robitschek Emily , Mao Chih-Ping , Peng Shiwen , Hung Chien-Fu , Wu TC
Published: August 2015

The emergence of cancer stem cells and escape from immune surveillance play major roles in the progression of many cancer types. We recently established a link between these two “hallmarks” of cancer by demonstrating that they are both driven by overexpression of Nanog, a pivotal transcription factor in the self-renewal of pluripotent stem cells. In particular, we found that tumor cells acquire Nanog expression in the course of immune selection, and that Nanog confers a stem-like and immune-resistant phenotype to these cells. However, the way in which Nanog causes this to occur remains largely unknown. We reasoned that Nanog-driven immune escape may be in part due to the effect of this transcription factor on the tumor microenvironment. Therefore, here we investigated the relationship between Nanog, immune selection, and the properties of the tumor microenvironment.

We found that mice bearing tumor cells that underwent immune selection, compared to those bearing parental tumor cells (without immune selection), had markedly elevated levels of the cytokine IL-6 as well as of myeloid-derived suppressor cells (MDSCs) in the tumor microenvironment; both IL-6 and MDSCs have been well-documented to promote cancer immune escape. We further investigated the role of Nanog in setting up this microenvironment. To this end, we have devised an in vivo system that enables us to examine the relationship between Nanog expression in the tumor microenvironment and tumor growth in real-time.

Taken together, we infer from these data that not only is Nanog induction a consequence of immune selection, but Nanog also plays an essential role in tumor immune escape. In fact, we found in multiple mouse and human tumor models that Nanog is necessary and sufficient for immune escape. Additionally, these results suggest that Nanog mediates immune escape by converting the tumor microenvironment into an immune-inhibitory site, through the induction of IL-6 and MDSCs. These studies authenticate the role of Nanog in cancer progression as a central instigator of tumor immune escape, and demonstrate its potential both as an important diagnostic/prognostic index and as a therapeutic target.

Citation Format: Emily Robitschek, Chih-Ping Mao, Shiwen Peng, Chien-Fu Hung, TC Wu. The role of the Nanog transcription factor in tumor immune escape. [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 448. doi:10.1158/1538-7445.AM2015-448
Abstract 4486: Antiangiogenic platinum through glycan targeting
Authors: Peterson Erica J. , Berners-Price Susan J. , Bezos Anna , Bohlman Lisa , Katner Samantha J. , Daniel A. Gerard , Chang Chih-Wei , von Itzstein Mark , Parish Christopher R. , Farrell Nicholas P.
Published: August 2015

Heparanase is an endo-β-D-glucuronidase that cleaves heparan sulfate glycosaminoglycans (HS-GAGs) in the extracellular matrix and basement membrane. Cancer cells that aberrantly express heparanase potentiate tumor progression, invasion and metastasis in two distinct ways; (1) cleavage of HS-GAGs releases growth-factors to directly activate growth receptors and (2) degradation of the heparan sulfate structural component of the extracellular matrix (ECM) allows metastatic spread of cancer cells. Recently, we determined that accumulation and cytotoxicity of polynuclear platinum compounds (PPCs), including the Phase II clinical trial compound, BBR3464 (+4), are dependent on the presence of cell-surface GAGs. Here, we demonstrate that high affinity PPC-GAG binding provides a new approach to glycan-based targeting by protection against enzymatic cleavage by heparanase. It was determined by NMR spectroscopy, that PPCs, especially the higher charged compound, TriplatinNC (+8), inhibit heparanase cleavage of the oligosaccharide, Fondaparineaux. Further, using the human umbilical primary cell line, HUVEC, it was determined that PPCs reduce heparanase cleavage of the GAG-bound growth factor, bFGF, from ECM. The end result of inhibition of heparanase cleavage is a reduction in tumor invasion and angiogenesis. Using the matrigel invasion assay, we show that sub-cytotoxic doses of PPCs, but not cisplatin, reduces serum-induced invasion of HCT116 cells through basement membrane. In an ex vivo rat aorta model, PPCs exhibit antiangiogenesis activity, measured by the inhibition of new blood vessel growth sprouting from the original aortic ring. Together, these encouraging results support the potential to combine anti-metastatic and cytotoxic activity in the development of dual-function platinum-based drugs.

Citation Format: Erica J. Peterson, Susan J. Berners-Price, Anna Bezos, Lisa Bohlman, Samantha J. Katner, A. Gerard Daniel, Chih-Wei Chang, Mark von Itzstein, Christopher R. Parish, Nicholas P. Farrell. Antiangiogenic platinum through glycan targeting. [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 4486. doi:10.1158/1538-7445.AM2015-4486
Abstract 4494: Physical and metabolic stability of SH7139, a new drug candidate for non-Hodgkin's lymphoma targeting HLA-DR10
Authors: Balhorn Rod , Hok Saphon , Balhorn Monique Cosman
Published: August 2015

Oncology drugs must remain sufficiently stable during storage, following formulation, after introduction into the bloodstream and during their entry into cells to ensure accurate dosing. After uptake by cells, the biologically active form of the drug also needs to remain stable long enough to achieve an optimal anti-tumor response. SH7139, our most advanced small molecule therapeutic for non-Hodgkin's lymphoma, was designed to target and bind to a unique site on HLA-DR10, a cell surface receptor found on B-cell lymphocytes and over-expressed on many B-cell derived malignancies. Previous studies conducted with lymphoma and other cell lines have shown that SH7139 binds selectively and is highly cytotoxic only to tumor cells expressing HLA-DR10. SH7139 has also been shown to be highly effective in treating human Burkitt's lymphoma (Raji) in a mouse xenograft model. All of the studies performed to date suggest SH7139 meets the necessary stability requirements. No degradation of the drug has been observed during storage at -20°C over a period of 30 months. SH7139 is also stable when dissolved in water or saline over a wide range of pH (pH 2 to 9). Instability is only observed at a highly alkaline pH. At pH 12, a slow auto-catalytic cleavage reaction releases a fragment of one of the recognition elements. The amide bonds linking the three recognition elements to the lysine and miniPEG scaffold of SH7139 remain stable in plasma. Hepatocytes metabolize SH7139 at a moderate rate and produce two primary metabolites. The same metabolites are produced by both canine and human hepatocytes, and the smaller product, 2-[3-chloro-5-(trifluoromethyl)pyridin-2-yl]oxyaniline, is rapidly glucuronidated to facilitate its excretion. Both the physical and metabolic stability of SH7139 are consistent with the drug's observed anti-lymphoma activity. This research was supported by the National Cancer Institute Phase II SBIR award R44CA159843-02 to SHAL Technologies Inc. Part of this work was performed under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344.

Citation Format: Rod Balhorn, Saphon Hok, Monique Cosman Balhorn. Physical and metabolic stability of SH7139, a new drug candidate for non-Hodgkin's lymphoma targeting HLA-DR10. [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 4494. doi:10.1158/1538-7445.AM2015-4494
Abstract 4515: A phase 1 randomized, open-label, fixed-sequence, 2-period study of the effect of multiple doses of rifampin on palbociclib (PD-0332991) pharmacokinetics in healthy volunteers
Authors: Hoffman Justin T. , Plotka Anna , O'Gorman Melissa , Chang Andrew , Kosa Maha , Loi Cho-Ming , Gallo-Stampino Corrado , Wang Diane D.
Published: August 2015

Background

Rifampin is a strong inducer of multiple Phase 1 and Phase 2 metabolic enzymes including cytochrome (CYP) P450 3A4 and sulfotransferases (SULT). In vitro, the metabolism of palbociclib occurs primarily through CYP3A4 and SULT2A1; thus, coadministration of rifampin and palbociclib could result in decreased exposure of palbociclib.

Methods

This open-label, 2-period, fixed-sequence study assessed the effect of multiple oral doses of rifampin on single-dose palbociclib pharmacokinetics (PK) in 15 healthy volunteers (ClinicalTrials.gov; NCT01953731). Each subject received the following treatments: Period 1, oral palbociclib 125 mg on day 1; Period 2, oral rifampin 600 mg for 12 days and oral palbociclib 125 mg on day 8 (washout of ≥12 days between the 2 palbociclib doses). Serial blood sampling for palbociclib PK was performed predose and up to 120 hours post-palbociclib dose in both periods. Plasma concentrations of palbociclib were measured using validated high-performance liquid chromatography tandem mass spectrometry methods. Palbociclib plasma PK parameters were estimated using standard non-compartmental methods.

Results

Median time to maximum plasma concentration (Tmax) and mean half-life (t1/2) were shorter with palbociclib plus rifampin relative to palbociclib alone. Adjusted geometric mean palbociclib AUCinf (area under the curve from time zero to infinity) and Cmax (maximal plasma concentration) following treatment with rifampin (242.5 ng•h/mL and 15.52 ng/mL, respectively) were considerably lower relative to palbociclib administered alone (1568 ng•h/mL and 51.42 ng/mL). Generally, study treatments were well tolerated with no reported serious adverse events; 1 subject discontinued due to liver function test abnormalities after receiving rifampin alone for 7 days in Period 2.

Conclusions

Treatment with multiple doses of rifampin with palbociclib decreased total (AUCinf) and peak (Cmax) palbociclib exposure by approximately 85% and 70%, respectively, versus palbociclib alone. Palbociclib was well tolerated when administered alone or in combination with rifampin. Based on these results, concurrent administration of palbociclib with strong CYP3A inducers should be avoided.

Citation Format: Justin T. Hoffman, Anna Plotka, Melissa O'Gorman, Andrew Chang, Maha Kosa, Cho-Ming Loi, Corrado Gallo-Stampino, Diane D. Wang. A phase 1 randomized, open-label, fixed-sequence, 2-period study of the effect of multiple doses of rifampin on palbociclib (PD-0332991) pharmacokinetics in healthy volunteers. [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 4515. doi:10.1158/1538-7445.AM2015-4515
Abstract 4529: Pharmacokinetics (PK) of LOXO-101 during the first-in-human Phase I study in patients with advanced solid tumors: Interim update
Authors: Burris Howard A. , Shaw Alice T. , Bauer Todd M. , Farago Anna F. , Doebele Robert C. , Smith Steven , Nanda Nisha , Cruickshank Scott , Low Jennifer A. , Brose Marcia S.
Published: August 2015

Background

LOXO-101 is the first highly selective and potent small-molecule inhibitor of TRKA, TRKB, and TRKC to be dosed clinically. It has approximately 100-fold selectivity over other kinase and non-kinase targets. We report here the interim results of the first-in-man Phase I study which evaluates the safety, maximum-tolerated dose (MTD), and PK of LOXO-101 in patients with advanced solid tumors.

Methods

This is an ongoing open-label, multicenter, 3+3 dose escalation Phase I study of LOXO-101. Patients with solid tumors refractory to standard therapy, with normal hematopoietic and major organ function were eligible for study. LOXO-101 was administered orally as a single dose, followed by QD or BID doses for continuous 28-day cycles. Receptor occupancy was determined with an Emax model based on unbound LOXO-101 plasma concentrations.

Results

To date, data are available for 8 patients including patients treated at each of the first two dose levels, 50 mg QD and 100 mg QD. LOXO-101 has been well tolerated; the MTD has not been reached and no adverse events have been reported more than once other than fatigue (two patients; related but not serious) and syncope (two patients; unrelated and serious).

Maximum plasma concentrations of LOXO-101 were reached 30-60 minutes following dosing and exposure increased in approximate proportion with dose. Mean values of Cl/F, V/F, and half-life were 0.5 L/h/kg, 2 L/kg, and 3.5 h, respectively, and were independent of dose. Exposure was similar following the single dose and on Day 1 and Day 8 of repeated dosing in all subjects. The unbound drug levels of LOXO-101 appear sufficient for approximately 98% inhibition of TRKA/B/C at peak concentrations with once-daily dosing of 50 or 100 mg.

Conclusions

LOXO-101 has been well tolerated and has sufficient systemic exposure for robust inhibition of the TRKA, TRKB, and TRKC kinases without inhibition of other kinases. With dose escalation continuing, additional data and follow up will be provided at time of presentation.

Citation Format: Howard A. Burris, Alice T. Shaw, Todd M. Bauer, Anna F. Farago, Robert C. Doebele, Steven Smith, Nisha Nanda, Scott Cruickshank, Jennifer A. Low, Marcia S. Brose. Pharmacokinetics (PK) of LOXO-101 during the first-in-human Phase I study in patients with advanced solid tumors: Interim update. [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 4529. doi:10.1158/1538-7445.AM2015-4529
Abstract 4556: Identification of novel agent for HCC treatment by GNMT-promoter orientated cell-based drug screening platform
Authors: Kant Rajni , Tung Lucas , Lu Chung-Kuang , Chen Yi-Ming , Yen Chia-Hung
Published: August 2015

Glycine N-methyltransferase (GNMT) a tumor suppressor gene for hepatocellular carcinoma (HCC) plays very important role in liver detoxification and tumorigenisis. GNMT expression is highly down-regulated in HCC. Recent studies have shown that overexpression of GNMT reduce tumor growth and sensitize HuH-7 cells to rapamycin treatment in vitro and in vivo. Therefore, enhancing GNMT expression is an attractive approach for HCC therapy. In present study we developed a GNMT-promoter driven luciferase reporter assay as a drug screen platform for the identification of novel drugs for prevention and treatment of HCC. We identified crude extract 317 as highest activator of GNMT promoter activity from a Chinese medicine library containing 472 herbal extracts. Partial purified fraction “F2-6” was isolated by bioassay-guided fractionation from crude extract 317 and validated for enhancement of endogenous GNMT expression at both mRNA and protein levels. Fraction F2-6 inhibited growth of liver cancer cell lines in vitro. Moreover, orally administrated F2-6 can delay and reduce growth of HCC xenograft tumor in vivo. Next, we identified the active component in fraction F2-6 named as compound P. Furthermore, we demonstrated that combination of sorafenib with compound P led to additive toxicity on HCC cells in vitro and in vivo. The cDNA microarray and real time PCR analysis showed that compound P altered cell cycle-associated genes such as CDK6, CCND1, CDKN1C, MYC and PLK1. Finally, Compound P treatment induced apoptosis in huh7 cells were confirmed by increased sub-G1 population and caspase 3/7 activity. Collectively, our results suggest that compound P has potential to be a novel agent for prevention and treatment of HCC.

Citation Format: Rajni Kant, Lucas Tung, Chung-Kuang Lu, Yi-Ming Chen, Chia-Hung Yen. Identification of novel agent for HCC treatment by GNMT-promoter orientated cell-based drug screening platform. [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 4556. doi:10.1158/1538-7445.AM2015-4556
Abstract 4568: Inhibition of autophagy in EBV-positive Burkitt's lymphoma cells enhances EBV lytic genes expression and replication
Authors: De Leo Alessandra , Colavita Francesca , Ciccosanti Fabiola , Fimia Gian Maria , Lieberman Paul , Mattia Elena
Published: August 2015

Autophagy, an important degradation system involved in maintaining cellular homeostasis, serves also to eliminate pathogens and process their fragments for presentation to the immune system. Several viruses have been shown to interact with the host autophagic machinery to suppress or make use of this cellular catabolic pathway to enhance their survival and replication. Epstein Barr virus (EBV) is a γ-herpes virus associated with a number of malignancies of epithelial and lymphoid origin in which establishes a predominantly latent infection. EBV lytic cycle characterized by the sequential expression of immediate early (IE), early and late antigens results in the production of infectious particles which allow the virus to spread. In this study we analyzed the relationship between EBV and autophagy after inducing the virus productive cycle in Burkitt's lymphoma (BL) cells. By monitoring autophagy markers and EBV lytic genes expression, we demonstrate that autophagy is enhanced in the early phases of EBV lytic activation but decreases thereafter concomitantly with increased levels of EBV lytic proteins. In a cell line defective for late antigens expression, we found an inverse correlation between EBV early antigens expression and autophagosomes formation indicating that early after activation, the virus is able to suppress autophagy. We report that inhibition of autophagy by Bafilomycin A1 or shRNA knockdown of beclin1 gene, enhance EBV lytic genes expression as well as intracellular viral DNA and viral progeny yield. Taken together, these findings indicate that viral replication induces an autophagic response which can inhibit the further expression of EBV early lytic products. Moreover, our findings open the possibility to utilize pharmacological modulators of autophagy to control EBV infection and treat EBV-related lymphomas.

Citation Format: Alessandra De Leo, Francesca Colavita, Fabiola Ciccosanti, Gian Maria Fimia, Paul Lieberman, Elena Mattia. Inhibition of autophagy in EBV-positive Burkitt's lymphoma cells enhances EBV lytic genes expression and replication. [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 4568. doi:10.1158/1538-7445.AM2015-4568
Abstract 4574: Genomic copy number variations characterize prognosis of oropharyngeal squamous cell carcinoma, including p16 positive subset, after surgery-based treatment
Authors: Jung Yuh-S. , Rhie Arang , Kim Jong-Il , Lee Jin Soo , Ryu Junsun , Park Weon Seo
Published: August 2015

Background High-risk human papillomavirus (HPV)-related (+) oropharyngeal squamous cell carcinoma (OSCC) exhibits unique clinical and molecular characteristics, compared to HPV-unrelated - OSCCs. As different patterns of genomic instability is expected in HPV-driven carcinogenesis, we questioned if patterns of genomic copy number variations (CNVs) differ depending on HPV association, and predict prognosis, after surgery-based treatment for OSCCs in Korean population. Methods Fifty-eight OSCCs patients, who underwent surgery-based treatment and followed for 42 months on median, were enrolled. HPV status was determined by p16 immunohistochemistry (IHC), and HPV DNA PCR. Genome-wide CNV analysis was performed using array-based comparative genomic hybridization of genomic DNA from paraffin-embedded specimen. Clinical association and survival analyses were followed. Results p16 predicted the overall survival (HR = 0.27, CI: 0.39-0.80, p = 0.0006), better than HPV DNA PCR (HR = 0.83, CI: 0.66-1.29, p = 0.64) or smoking history (HR = 2.18, CI: 0.92-5.18, p = 0.071). In overall, number of CNVs did not significantly differed between p16+/- groups. However, 30 locus showed different CNV pattern between these groups. PRDM2 was amplified only in p16+ group, while EGFR and 11q13.3 was more amplified in p16- patients. Loss of FGF18 lead to worse prognosis, and gain of CDK10 and RAD18 was linked to better prognosis in all patients. Amplification of HRAS and loss of KDR lead to better OS in p16+ group. Conclusions After surgery-based treatment, p16+ OSCCs exhibits different CNV patterns at some loci, compared with p16- counterpart. Specific patterns of CNVs do predict better survival, especially in p16+ OSCCs. In the future, clinical trials should stratify patients based on these patterns to apply escalated treatment to obtain better survival.

Citation Format: Yuh-S. Jung, Arang Rhie, Jong-Il Kim, Jin Soo Lee, Junsun Ryu, Weon Seo Park. Genomic copy number variations characterize prognosis of oropharyngeal squamous cell carcinoma, including p16 positive subset, after surgery-based treatment. [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 4574. doi:10.1158/1538-7445.AM2015-4574
Abstract 4586: Association of FUT2 gene variant, gut bacteria and colorectal adenomas
Authors: Okunola Winifred , McCoy Amber , Dulal Santosh , Keku Temitope O.
Published: August 2015

The fucosyltransferase 2 (FUT2) gene is responsible for the expression of the ABO blood group antigens. These antigens serve as anchors for a variety of microbes in the gut and may confer increased susceptibility to microbial infections and diseases. We assessed the association between a genetic variant in the FUT2 gene (rs516246) and mucosal-adherent gut bacteria in relation to colorectal adenomas. Genomic DNA from 200 consented screening colonoscopy subjects was genotyped for FUT2 rs516246 using a TaqMan genotyping assay. qPCR was performed to target the 16S rRNA gene for specific bacteria. Statistical analysis included t-test, logistic regression Chi-squared tests and Spearman's correlation test. Results show that, except for age and waist-hip ratio, patient characteristics were similar for cases and controls. Higher abundance of E. coli and Eubacteria was associated with adenoma status. There was a null association between FUT2 genotype and adenomas. Similarly, gut bacteria and adenoma status was not strongly influenced by FUT2 genotype. In conclusion: the FUT2 (rs516246) gene variant does not appear to influence the relationship between gut bacteria and colorectal adenomas.

Citation Format: Winifred Okunola, Amber McCoy, Santosh Dulal, Temitope O. Keku. Association of FUT2 gene variant, gut bacteria and colorectal adenomas. [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 4586. doi:10.1158/1538-7445.AM2015-4586
Abstract 4588: Whole genome hypo- and hyper-methylation sequencing among Africa American patients with colorectal cancer
Authors: Ashktorab Hassan , Shakoori Afnan , Zarnogi Shatha , Sun Xueguang , Varma Sudhir , Lee Edward , Shokrani Babak , Laiyemo Adeinko O. , Washington Kareem , Brim Hassan
Published: August 2015

Background: Colorectal cancer (CRC) involves epigenetic changes including DNA hypomethylation, a paradigm shift in the role of DNA methylation alterations in colorectal carcinogenesis.

Aim: Here we performed Reduced Representation Bisulfite Sequencing (RRBS) on a normal, and a tumor patient's tissue DNA to elucidate hyper- and hypomethylaed target genes in colorectal cancer progression in this population.

Methods: Genomic DNA was isolated from fresh frozen surgical tissues from a patient with normal colon, and from a carcinoma patient. RRBS was performed on these DNA samples for hyper- and hypomethylation targets identification. Alignment, mapping and CpG methylation analyses were performed. Preferential hypomethylated pathways were determined using Ingenuity Pathway Analysis (IPA).

Results: We identified the hyper (6360 CpG sites) and hypo-methylation (4845 CpG sites) status of top genes in the CpG Island within promoter regions (19 and 18 genes respectively). Top hyper- and hypomethylated CpG Island outside promoter regions (16 and 18 genes, respectively) were also identified. Among these top genes, CDH4 and SOX21 were reported to be hypermethylated in CRC and SCUBE2 (112 methylated CpG) mapped by IPA to the Hedgehog, TGF-β, and VEGF Signaling pathways was reported to play a role in breast cancer. Among the top hypomethylated genes, ACOT9 (97 CpGI sites) and RIN2 (40 CpGI sites) might play a significant role in CRC in AAs, since IPA mapped these markers to the Wnt/β-catenin, VEGF and PTEN signaling pathways. ACOT9 was found to play a role in breast cancer.

Conclusion: This work provides insight into differential CpG island hyper- and hypo-methylation profiles in CRC and provides a window into the more complex epigenetic events associated with CRC, including the hypomethylation of known and novel genes. Investigations into the possible roles of the novel gene targets in the context of early and prognostic methylation biomarkers are underway.

Citation Format: Hassan Ashktorab, Afnan Shakoori, Shatha Zarnogi, Xueguang Sun, Sudhir Varma, Edward Lee, Babak Shokrani, Adeinko O. Laiyemo, Kareem Washington, Hassan Brim. Whole genome hypo- and hyper-methylation sequencing among Africa American patients with colorectal 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 4588. doi:10.1158/1538-7445.AM2015-4588
Abstract 4613: MDM4 SNP 34091 (rs4245739) effect on risk of breast, colon, lung, prostate, endometrial and ovarian cancer
Authors: Gansmo Liv B. , Bjørnslett Merete , Dørum Anne , Salvesen Helga , Romundstad Pål , Hveem Kristian , Vatten Lars , Lønning Per Eystein , Knappskog Stian
Published: August 2015

Background

MDM4 enhances MDM2's E3 ligase activity causing ubiquitin-proteasome-dependent degradation of p53. Thus, elevated levels of both MDM4 and MDM2 may result in p53 inactivation and elevated cancer risk. A single nucleotide polymorphism in the MDM4 3′ UTR (SNP34091A>C; rs4245739) has been found to exert biological function as the SNP34091C allele creates a putative target site for hsa-miR-191 leading to specific hsa-miR-191 down-regulation of MDM4. Further, the SNP34091AA genotype is associated with increased risk for both recurrence and tumor related death in estrogen negative ovarian cancer patients. In the present study we assessed the potential effect of MDM4 SNP34091A>C on cancer risk in six major cancer forms.

Materials and methods

We analyzed 1717 breast-, 1331 lung-, 1531 colon-, 2501 prostate cancer cases and 3747 healthy controls form the same population based study (CONOR) as well as 1404 endometrial- and 1699 ovarian cancer cases from hospital based studies.

All samples were genotyped for MDM4 SNP34091A>C using custom Light-SNiP assay on a LightCycler 480 II instrument. Potential associations between MDM4 SNP34091 and cancer risk were estimated by calculating Odds Ratio (OR) with 95% confidence intervals (CI). All statistical analyses were performed using the IBM SPSS 19 software.

Results

We observed no significant association between MDM4 SNP34091A>C status and cancer risk in any of the analyzed cancer forms. Interestingly, stratifying the ovarian cancer samples according to grade and histology, we observed a reduced risk of high grade serous ovarian cancer in patients harboring the MDM4 SNP34091AA genotype (OR = 0.80; 95% CI = 0.65 - 0.98). Stratifying according to MDM2 SNP309 status we found the MDM4 SNP34091A allele to be associated with increased risk for breast cancer (OR = 2.10; 95% CI = 1.08 - 4.10) in patients carrying the SNP309 GG genotype.

Conclusions

The data presented here indicate that the effect of MDM4 SNP34091 on cancer risk may be tissue specific and that there may be cooperative effects with MDM2 SNP309.

Citation Format: Liv B. Gansmo, Merete Bjørnslett, Anne Dørum, Helga Salvesen, Pål Romundstad, Kristian Hveem, Lars Vatten, Per Eystein Lønning, Stian Knappskog. MDM4 SNP 34091 (rs4245739) effect on risk of breast, colon, lung, prostate, endometrial and ovarian 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 4613. doi:10.1158/1538-7445.AM2015-4613