bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract PR01: Understanding the role of BCL11A in triple-negative breast cancer
Authors: Lazarus Kyren , Caldas Carlos , Liu Pentao , Khaled Walid T.
Published: May 2016

Triple-negative breast cancer (TNBC) has poor prognostic outcome compared to other subtypes of breast cancer. The molecular and cellular mechanisms underlying TNBC pathology are not fully understood. We analyzed both METABRIC and TCGA data found the transcription factor BCL11A is overexpressed in TNBC including basal-like breast cancer (BLBC) and that its genomic locus is amplified in up to 38% of BLBC tumors. Moreover, analysis of the TCGA data revealed that the BCL11A locus is hypo-methylated almost exclusively in BLBC tumors but not other subtypes. We show that exogenous BCL11A overexpression promotes tumor formation, whereas its knockdown in TNBC cell lines suppresses their tumorigenic potential in xenograft models. In the mouse DMBA-induced tumor model, Bcl11a deletion substantially decreases tumor formation, even in p53-null cells and inactivation of Bcl11a in established tumors causes their regression. At the cellular level, Bcl11a deletion causes a reduction in the number of mammary epithelial stem and progenitor cells. Mechanistically, proteomics studies have revealed that BCL11A interacts with a specific set of chromatin modifiers in tumor cells but not in normal cells - the significance of which is currently under investigation. This study highlights the potential for targeting BCL11A as an approach for TNBC-targeted therapy. To this end we are currently using the Crispr/Cas9 system to generate endogenous reporter cell lines for high-throughput drug screens to identify potential inhibitors of BCL11A.

Citation Format: Kyren Lazarus, Carlos Caldas, Pentao Liu, Walid T. Khaled. Understanding the role of BCL11A in triple-negative breast cancer. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research; Oct 17-20, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(2_Suppl):Abstract nr PR01.
Abstract A11: N-Myc and STAT interactor (NMI): A regulator of developmental signaling and EMT
Authors: Samant Rajeev S. , Pruitt Hawley C. , Metge Brandon J. , Shevde Lalita A.
Published: May 2016

The NMI protein associates with known key transcription factors such as Myc and STATs and serves as a critical regulatory co-factor that dictates specialized functions. While a physiological function for NMI is yet unknown, it has potential roles in pathologies ranging from viral infection to cancer. We will present pioneering work from our laboratory showing that NMI impacts two major developmental signaling pathways - TGFβ and Wnt pathways.

EMT is one of the key phenomena underlying normal embryonic and organ development, with a vital role in metastatic progression. We discovered that the expression of NMI is significantly downregulated in metastatic breast tumors. We recapitulated this by stably silencing the expression of NMI in breast cancer cells. Abrogating NMI expression from epithelial-like breast cancer cells enabled cells to assume a mesenchymal-like phenotype. Detailed molecular and functional investigations revealed that this mesenchymal transition was facilitated by decreased STAT5 signaling that caused concomitant reduction in SMAD7 expression and manifestation of a TGFβ-driven EMT program.

In contrast, breast cancer cells restored for NMI expression showed autophagic vacuoles and LC3 processing. We found that NMI expression restricts Wnt/β-catenin signaling by upregulation of the secreted Wnt inhibitor, DKK1. Thus NMI prompts activation of GSK3-β, a key kinase upstream of the TSC1/TSC2 complex. Inhibition of GSK3-ββin NMI expressing cells activated mTOR signaling and decreased the cells’ autophagic response. Autophagy is a determinant of cellular survival through dormancy and cytotoxic drug insult. Rapid progression of autophagy leads to cell death. In agreement with our molecular analyses, we determined that abrogation of NMI expression rendered cells resistant to cisplatin and doxorubicin.

Our findings elicit interesting possibilities about the role of NMI in regulating EMT and autophagy. Evidence from our investigations strongly suggests that loss of NMI leading to aberrant activation of multiple developmental signaling pathways, may be a vital event in the progression of breast cancer characterized by reduced autophagy and stabilization of transcription factors that function as major drivers of EMT.

Citation Format: Rajeev S. Samant, Hawley C. Pruitt, Brandon J. Metge, Lalita A. Shevde. N-Myc and STAT interactor (NMI): A regulator of developmental signaling and EMT. [abstract]. In: Proceedings of the AACR Special Conference: Developmental Biology and Cancer; Nov 30-Dec 3, 2015; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(4_Suppl):Abstract nr A11.
Abstract B04: IKBKE integrates Gli and mTOR-AKT activation during Kras-induced pancreatic tumorigenesis
Authors: Rajurkar Mihir , Fernandez-Zapico Martin , Lewis Brian , Mao Junhao
Published: May 2016

Pancreatic Ductal Adenocarcinoma (PDAC) is one of the deadliest malignancies lacking effective therapeutic strategies. Here we show that the non-canonical IκB-related kinase, IKBKE, acts downstream of the Gli transcription factors and engages in reciprocal regulation of Gli signaling in pancreatic cancer cells. We find that loss of IKBKE inhibits the initiation and progression of pancreatic tumors in mice carrying pancreatic specific KRAS activation. Mechanistically, IKBKE does not play a major role in NF-κB activation in pancreatic cancer; but is critical for AKT activation in vivo. Moreover, we find that IKBKE promotes reactivation of AKT post-inhibition of mTOR in PDAC cells, and that combined inhibition of IKBKE and mTOR synergistically blocks pancreatic tumor growth. Together, our findings highlight the functional interaction of IKBKE with Hh/Gli and mTOR/AKT signaling in pancreatic cancer, and suggest IKBKE inhibition as a strategy to improve efficacy of mTOR inhibitors in the clinic.

Citation Format: Mihir Rajurkar, Martin Fernandez-Zapico, Brian Lewis, Junhao Mao. IKBKE integrates Gli and mTOR-AKT activation during Kras-induced pancreatic tumorigenesis. [abstract]. In: Proceedings of the AACR Special Conference: Developmental Biology and Cancer; Nov 30-Dec 3, 2015; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(4_Suppl):Abstract nr B04.
Abstract B21: DNp63 promotes stem cell activity in mammary gland development and basal-like breast cancer by enhancing Fzd7 expression and Wnt signaling
Authors: Chakrabarti Rumela , Sinha Satrajit , Kang Yibin
Published: May 2016

Emerging evidence suggests that cancer is populated and maintained by tumor initiating cells (TICs) with stem-like properties similar to that of adult tissue stem cells. Despite recent advances, the molecular regulatory mechanisms that may be shared between normal and malignant stem cells remain poorly understood. Here we show that the DNp63 isoform of the Trp63 transcription factor promotes normal mammary stem cell (MaSC) activity by increasing the expression of the Wnt receptor Fzd7 and thereby enhancing Wnt signaling. Importantly, Fzd7-dependent enhancement of Wnt signaling by DNp63 also governs tumor initiating activity of the basal subtype of breast cancer. These findings establish DNp63 as a key regulator of stem cells in both normal and malignant mammary tissues and provide direct evidence that breast cancer TICs and normal MaSCs share common regulatory mechanisms.

Citation Format: Rumela Chakrabarti, Satrajit Sinha, Yibin Kang. DNp63 promotes stem cell activity in mammary gland development and basal-like breast cancer by enhancing Fzd7 expression and Wnt signaling. [abstract]. In: Proceedings of the AACR Special Conference: Developmental Biology and Cancer; Nov 30-Dec 3, 2015; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(4_Suppl):Abstract nr B21.
Abstract IA08: Oxidative stress inhibits distant metastasis by human melanoma cells
Authors: Piskounova Elena , Agathocleous Michalis , DeBerardinis Ralph J. , Morrison Sean J.
Published: May 2016

Solid cancer cells commonly enter the blood and disseminate systemically but are highly inefficient at forming distant metastases for poorly understood reasons. We studied human melanomas that differed in their metastasis histories in patients and in their capacity to metastasize in NSG mice. All melanomas had high frequencies of cells that formed subcutaneous tumors, but much lower percentages of cells that formed tumors after intravenous or intrasplenic transplantation, particularly among inefficient metastasizers. Melanoma cells in the blood and visceral organs experienced oxidative stress not observed in established subcutaneous tumors. Successfully metastasizing melanomas underwent reversible metabolic changes during metastasis that increased their capacity to withstand oxidative stress, including increased dependence upon NADPH-generating enzymes in the folate pathway. Anti-oxidants promoted distant metastasis in NSG mice. Folate pathway inhibition using low-dose methotrexate, ALDH1L2 knockdown, or MTHFD1 knockdown inhibited distant metastasis without significantly affecting the growth of subcutaneous tumors in the same mice. Oxidative stress thus limits distant metastasis by melanoma cells in vivo.

Citation Format: Elena Piskounova, Michalis Agathocleous, Ralph J. DeBerardinis, Sean J. Morrison. Oxidative stress inhibits distant metastasis by human melanoma cells. [abstract]. In: Proceedings of the AACR Special Conference: Developmental Biology and Cancer; Nov 30-Dec 3, 2015; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(4_Suppl):Abstract nr IA08.
Abstract IA11: C. elegans GLP-1/Notch signaling: Direct targets and visualization in stem cells
Authors: Sorensen Erika B. , Lee ChangHwan Lee , Sorokin Elena , Groth Amy C. , Kimble Judith
Published: May 2016

My laboratory focuses on the in vivo regulation of stem cell self-renewal and differentiation in the germline tissue of the small nematode C. elegans. In this system, Notch signaling from a single-celled mesenchymal niche promotes maintenance of germline stem cells (GSCs)(1). My talk will focus on two broadly important questions. First, what key downstream effectors of Notch signaling are transcriptionally activated in stem cells to maintain their totipotent state? Second, how does Notch-dependent transcriptional activation maintain a pool of stem cells? We have known for some time that an RNA regulatory network acts downstream of Notch signaling to drives the choice between self-renewal and differentiation. We recently identified two Notch-dependent direct target genes, lst-1 and sygl-1, that fully account for the role of GLP-1/Notch signaling in GSC control (2). My talk will report recent efforts to understand lst-1 and sygl-1, to identify additional direct Notch targets in GSCs and to visualize Notch signaling within the stem cell pool.

References:

(1) Austin and Kimble (1987) Cell 51:589-599

(2) Kershner et al. (2014) PNAS,111(10):3739-3744.

Citation Format: Erika B. Sorensen, ChangHwan Lee Lee, Elena Sorokin, Amy C. Groth, Judith Kimble. C. elegans GLP-1/Notch signaling: Direct targets and visualization in stem cells. [abstract]. In: Proceedings of the AACR Special Conference: Developmental Biology and Cancer; Nov 30-Dec 3, 2015; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(4_Suppl):Abstract nr IA11.
Abstract IA21: YAP/TAZ biology in development and cancer: Hippo signaling and mechanotransduction
Authors: Piccolo Stefano
Published: May 2016

We study how cells sense their environment and use this information to build and maintain tissues with specific form, size and function, and how these systems are corrupted in diseases. At the centerpiece of these events is the activity of the transcriptional coactivators YAP and TAZ. Enhanced YAP/TAZ activity is emerging as a hallmark of multiple human tumors. I will discuss the cell and tissue-level mechanisms that lead to unrestrained YAP/TAZ activity, in turn essential for tumor formation and for tissue regeneration upon injury. I will also present new evidence on the function of YAP/TAZ in regulating the biology of normal somatic stem cell explanted from adult tissues.

Citation Format: Stefano Piccolo. YAP/TAZ biology in development and cancer: Hippo signaling and mechanotransduction. [abstract]. In: Proceedings of the AACR Special Conference: Developmental Biology and Cancer; Nov 30-Dec 3, 2015; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(4_Suppl):Abstract nr IA21.
Abstract IA22: Mechanisms of evasive resistance in cancer
Authors: Dazert Eva , Heim Markus , Beerenwinkel Niko , Hall Michael N.
Published: May 2016

Cancer is a major health problem due to the failure of current therapies to effectively eradicate the disease. Extensive research over decades has led to the development of therapies that target cancer-specific signaling pathways. However, tumors escape such therapies by activating compensatory signaling pathways, a process referred to as ‘evasive resistance’. The identities of the alternative signaling pathways and the functional interconnections that underlie evasive resistance remain widely unknown. Elucidating mechanisms of evasive resistance is currently a major challenge in cancer research. We integrate clinical, molecular, and computational sciences to understand the signaling defects that enable tumors to evade therapy. Within the framework of rigorously designed clinical studies, hepatocellular carcinoma (HCC) tissue is isolated before therapy, during treatment, or at the time of tumor progression. HCC was chosen as the focal cancer based on medical importance, accessibility to repeated sampling, and ethical considerations. The tumor tissue is obtained by needle biopsy and immediately snap frozen to preserve in vivo tumor properties. High- and low-throughput experimental and computational methods are then applied to determine, the underlying signaling defects. This endeavor will elucidate mechanisms of evasive resistance and will ultimately improve cancer diagnosis, treatment and clinical outcome. Recent progress in this ambitious project will be described.

Citation Format: Eva Dazert, Markus Heim, Niko Beerenwinkel, Michael N. Hall. Mechanisms of evasive resistance in cancer. [abstract]. In: Proceedings of the AACR Special Conference: Developmental Biology and Cancer; Nov 30-Dec 3, 2015; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(4_Suppl):Abstract nr IA22.
Abstract IA30: BAF complex structure and function in development and disease
Authors: Kadoch Cigall
Published: May 2016

Recent exon-wide sequencing studies in human cancer have revealed a striking frequency of mutations in the genes encoding subunits of the mSWI/SNF (BAF) ATP-dependent chromatin remodeling complexes we recently determined these mutations to be broadly recurrent in over 20% of all human cancers. To investigate the underlying mechanism, our group has studied rare, genomically well-defined cancer types, namely human synovial sarcoma (SS) in which 100% of tumors have a precise translocation involving a specific subunit, SS18, and BAF47-deficient malignant rhabdoid tumors. The utility of these disease settings has provided us with a powerful foundation upon which to understand the oncogenic mechanisms directed by altered structure and function of chromatin remodeling complexes. Here, we present studies focused on BAF complex pathway-of-assembly, complex subunit and associated protein factor composition of oncogenic BAF complexes, and mechanisms underlying BAF and polycomb opposition.

Citation Format: Cigall Kadoch. BAF complex structure and function in development and disease. [abstract]. In: Proceedings of the AACR Special Conference: Developmental Biology and Cancer; Nov 30-Dec 3, 2015; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(4_Suppl):Abstract nr IA30.
Abstract PR02: The SWI/SNF component Arid1a regulates regenerative capacity and carcinogenesis in a dose-dependent fashion
Authors: Sun Xuxu , Wang Sam , Maples Thomas , Li Lin , Nguyen Liem , Chuang Jen-Chieh , Sorouri Mahsa , Zhang Shuyuan , Wu Linwei , Celen Cemre , Zhu Hao
Published: May 2016

It is unclear if regenerative capacity can be dissociated from cancer risk, a question that is highly relevant for liver carcinogenesis. Frequent SWI/SNF and Arid1a loss-of-function mutations suggest tumor suppressive roles, but the functional impact of these mutations in tissue homeostasis and cancer is unclear. Mice without Arid1a possessed improved healing abilities after an array of injuries. After liver resection, chemical hepatocyte injury, and toxic bile duct injury, Arid1a deficient livers exhibited increased regeneration, reduced tissue damage, and improved organ function. Moreover, global homozygous Arid1a loss potentiated soft tissue healing after ear hole punch and hematopoietic recovery after irradiation. The chromatin state as reprogrammed by Arid1a loss restricted access to promoters by lineage-specific transcription factors that ordinarily suppress cell cycle reentry, thus increasing regeneration after injury. Intriguingly, the homozygous Arid1a deficient state protected mice from chemical injury induced hepatocellular carcinoma (HCC) and extended survival in a MYC-driven hepatoblastoma model. Since Arid1a mutations are most frequently heterozygous in human HCC genome studies, we examined the haploinsufficient state in these mouse models and observed accelerated carcinogenesis and metastasis. Transcriptome analysis identified dysregulated differentiation, proliferation, and metastasis programs. These models show that full deletion of Arid1a can enhance mammalian regeneration without increasing cancer risk, but that Arid1a’s tumor suppressor functions are highly sensitive to dose.

This abstract is also presented as Poster A16.

Citation Format: Xuxu Sun, Sam Wang, Thomas Maples, Lin Li, Liem Nguyen, Jen-Chieh Chuang, Mahsa Sorouri, Shuyuan Zhang, Linwei Wu, Cemre Celen, Hao Zhu. The SWI/SNF component Arid1a regulates regenerative capacity and carcinogenesis in a dose-dependent fashion. [abstract]. In: Proceedings of the AACR Special Conference: Developmental Biology and Cancer; Nov 30-Dec 3, 2015; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(4_Suppl):Abstract nr PR02.
Abstract A16: Restoration of gefitinib sensitivity and upregulation of BIM by simvastatin in T790M mutated non-small cell lung cancer cells
Authors: Young Mo Jin , Uk Lim Jeong , Young Lee Hwa , Kyoung Kim In , Haak Lee Sang
Published: May 2016

Objectives: Epidermal growth factor receptor (EGFR) gene mutation sensitize non-small cell lung cancer (NSCLC) cells to EGFR inhibitors. As the first line treatment on NSCLC, the tyrosine kinase inhibitors (TKIs) are effective, however T790M acquired resistant mutation reduces the therapeutic effect. Statins, 3-hydroxy-3-methyl-gluteryl-CoA (HMG-CoA) reductase inhibitors have pleotrophic effects to induce cell cycle arrest and death in certain type of cancer cells by inhibition of cholesterol synthesis and protein prenylation. The antitumor activities of statin in NSCLC cells inhibiting EGFR signaling proteins including PI3K/Akt and MAPK had been reported, but those in gefitinib resistant lung cancer cells harboring T790M mutation were not known. Recently, BH3-only polypeptide BIM (BCL2 like 11) was reported as one of the molecular mechanisms of T790M resistance in NSCLC cells. This study clarified the roles of simvastatin in the regulation of EGFR signaling and BH3-only protein BIM.

Methods: The cytotoxic effects of simvastatin on the gefitinib sensitive (HCC827, E716-A750del) and resistant (H1975, T790M + L858R) NSCLC cells were compared. Cell proliferation, intrinsic apoptosis and the expression of EGFR signaling proteins were evaluated. Expression of BIM was determined with western blot analysis and RT-PCR in H1975 cells after the time-dependent treatment of simvastatin or gefitinib.

Results: After the treatment of 2μM simvastatin for 24hrs and 48hrs, H1975 cells showed significantly decreased viability compared to HCC827 cells, (50% vs. 98% and 40% vs. 95%). Expression of Bax, cleaved caspase 3 and p-PARP proteins were increased in H1975 cells which showed the intrinsic apoptosis from simvastatin. Activity of EGFR signaling proteins, phosphorylation of EGFR, Akt and Erk were not affected with 2μM simvastatin for 12, 24 and 48 hours in H1975 cells. With gefitinib (1μM) treatment, BIM was suppressed in H1975 cells compared to HCC827 cells. Notably, BIM was induced after the exposure of simvastatin (2μM) for 12, 24 and 48 hours in H1975 cells.

Conclusions: T790M mutation, major mechanism of acquired resistance to EGFR-TKI, was associated with decreased BIM proteins. Simvastatin restored the expression of BIM to induce mitochondrial apoptosis in cells harboring T790M mutation and not in gefitinib sensitive mutation. Our study suggests the possibility of simvastatin to overcome gefitinib resistance with T790M mutation.

Citation Format: Mo Jin Young, Lim Jeong Uk, Lee Hwa Young, Kim In Kyoung, Lee Sang Haak. Restoration of gefitinib sensitivity and upregulation of BIM by simvastatin in T790M mutated non-small cell lung cancer cells. [abstract]. In: Proceedings of the AACR Special Conference: Metabolism and Cancer; Jun 7-10, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(1_Suppl):Abstract nr A16.
Abstract A28: Bioenergetic analysis of primary human mammary epithelial cells (hMECs)
Authors: Henchey Elizabeth M. , Schneider Sallie S. , Jerry D. Joseph , Yadava Nagendra
Published: May 2016

We have demonstrated reversible suppression of the tumor suppressor protein p53 by respiratory chain deficiency. Further, a recent study from a different group also suggests genetic inactivation of p53 by the impairments of oxidative metabolism, and mutations in mtDNA and TP53 gene coexist in cancers. Therefore, we predict that variation in oxidative metabolism can result in differences in p53 response among individuals. Conditions that suppress oxidative metabolism can impair p53, and thereby promote cancer development. Toward testing this hypothesis, our approach is to quantify the variation in respiratory activity of normal human mammary epithelial cells (hMECs), and then determine the correlation between respiratory activity and p53 response. Thus, we have performed in situ respirometry using 24-well Seahorse Bioscience's Extracellular Flux (XF) Analyzer on hMECs (at passage 2) isolated from breast cancer patients (n=23). The hMECs were isolated from cancer affected (AB) and non-affected (NAB) breasts and differences in parameters affecting mitochondrial bioenergetics under normal and treated (IGF1, TNFα;) conditions were assessed. Our data show 6-9 fold variation in hMECs basal respiratory activity (n=23). Further, 48%, 30% and 22% of patients showed either no difference in mitochondrial respiratory capacity upon addition of pyruvate (-2±5%), or showed at least 10% increase (26±18%) or decrease (-15±3%), respectively. Treatments with selected cytokines such as IGF1 and TNFα; for 24 hours altered the mitochondrial bioenergetics of hMECs. Overall they reduced respiratory activity in cells; and altered response to pyruvate addition in some individuals. Interestingly, matched normal hMECs from affected (AB) and not affected (NAB) breasts showed differences in the respiratory activity even at passage 2 suggesting that local environments play a critical role in the bioenergetics of normal cells and these changes do not revert ex vivo immediately when cells are removed from their environment.

Acknowledgements: This work was supported by funds from Rays of Hope and Incubator grants from the Baystate Health Foundation, CEAR at the PVLSI supported by an award (A00000000004448) from Massachusetts Technology Collaborative as administrator of the John Adams Innovation Institute.

Citation Format: Elizabeth M. Henchey, Sallie S. Schneider, D. Joseph Jerry, Nagendra Yadava. Bioenergetic analysis of primary human mammary epithelial cells (hMECs). [abstract]. In: Proceedings of the AACR Special Conference: Metabolism and Cancer; Jun 7-10, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(1_Suppl):Abstract nr A28.
Abstract A32: Structural and molecular mechanisms of human hexokinase 2 in cancer metabolism and apoptosis
Authors: Rabeh Wael
Published: May 2016

Cancer utilizes glucose at elevated levels to support its growth and proliferation, historically known as “ Warburg effect”. Targeting glucose metabolism in cancer cells to limit its growth will enhance patients' survival rate. Hexokinase catalyzes glucose phosphorylation, and is a major step in regulation of glycolysis. Four isozymes are present in human with Hexokinase 2 (HK2) being the most active and specifically expressed in verity of different cancers. In addition, gene expression profiling experiments of different types of cancer showed high expression levels of HK2, and various biological studies highlighted the importance of HK2 in tumor metastasis making it an ideal target to characterize cancer metabolism and for the development of new class of cancer therapeutics.

We solved the crystal structure of human HK2 in complex with glucose and glucose-6-phosphate (PDB code: 2NZT), where it is a homodimer with catalytically active N- and C-terminal domains linked by a seven-turn helix. Through biochemical and biophysical characterization of HK2, we found that the N-terminal domain not only catalyzes a reaction but it thermodynamically stabilities the entire enzyme, where deletion of the N-terminal helix altered the stability and catalytic activity of the full-length enzyme. Also, conformation of the N-terminal active site but not C-terminal is important in stabilizing the enzyme. Understanding the structural and molecular mechanisms of human HK2 in cancer metabolism and apoptosis will accelerate the design and development of new class of cancer therapeutics.

Citation Format: Wael Rabeh. Structural and molecular mechanisms of human hexokinase 2 in cancer metabolism and apoptosis. [abstract]. In: Proceedings of the AACR Special Conference: Metabolism and Cancer; Jun 7-10, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(1_Suppl):Abstract nr A32.
Abstract A55: Novel nilotinib derivatives inhibit cell proliferation by targeting 5' AMP-activated protein kinase in hepatocellular carcinoma
Authors: Su Jung-Chen , Hsueh Ting-ting , Huang Jui-Wen , Shiau Chung-Wai
Published: May 2016

5' AMP-activated protein kinase constitutes a network for energy homeostasis, cellular metabolic and proliferation, thus providing a potential therapeutic target for metabolic diseases including diabetes and cancers. In response to the increasing cellular ratio of AMP /ADP to ATP, the binding of AMPK and AMP or ADP promote or maintain the activation of AMPK through the phosphorylation of threonine 172, leading to phosphorylate and activate downstream substrates for cellular processes. Our previous study has shown that nilotinib, a BCR-ABL kinase inhibitor for the treatment of chronic myeloid leukemia, induces AMPK activation in carcinoma cells. We further apply the scaffold of nilotinib to design and synthesize a series of new derivatives which show the higher potency in activating AMPK and inhibiting cell proliferation in hepatocellular carcinoma. These new agents pave a road for the development of hepatocellular carcinoma and other metabolic diseases.

Citation Format: Jung-Chen Su, Ting-ting Hsueh, Jui-Wen Huang, Chung-Wai Shiau. Novel nilotinib derivatives inhibit cell proliferation by targeting 5' AMP-activated protein kinase in hepatocellular carcinoma. [abstract]. In: Proceedings of the AACR Special Conference: Metabolism and Cancer; Jun 7-10, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(1_Suppl):Abstract nr A55.
Abstract A58: Temporal dynamics underpin metabolism-driven cancer therapy cross-resistance
Authors: Goldman Aaron , Dhawan Andrew , Medhi Ragini , Kohandel Mohammad , Sengupta Shiladitya
Published: May 2016

The acquisition of resistance to chemotherapy is the underlying cause of relapse leading to metastasis and mortality. It is clear that the metabolic states of cancer cells contribute to cancer therapy failure. However, the inherent plasticity of cancer cells and the complexity of intratumoral heterogeneity have obscured a clear understanding for the role of metabolism in the development of resistance. Here, we used a computational biology approach in-tandem with functional assays at the single cell level to build a mechanistic understanding of an adaptive, metabolic cell-state-switch which leads to cross-therapy resistance of chemotherapies in cancer. We show that cytotoxic chemotherapies induce a temporally-dependent cell behavior via 1. induction of cell surface scaffold-kinase interactions 2. mitochondrial-induced reactive oxygen species (ROS) which are then requisite to drive 3. redox stress-mediated glucose uptake. Interestingly, we identified that the early-enhanced mitochondrial ROS promotes a delayed glucose shunt towards the pentose phosphate pathway (PPP) which was mediating cross-drug resistance. Using pharmacologic inhibitors of proteins in the PPP as well as inhibitors of upstream glycolysis intermediates, we were able to restore cancer cell sensitivity to combination therapies producing robust tumor responses in otherwise resistant cancer cells. These findings unveil a systems analysis of metabolic plasticity leading to therapy failure and provide novel strategies for treatment.

Citation Format: Aaron Goldman, Andrew Dhawan, Ragini Medhi, Mohammad Kohandel, Shiladitya Sengupta. Temporal dynamics underpin metabolism-driven cancer therapy cross-resistance. [abstract]. In: Proceedings of the AACR Special Conference: Metabolism and Cancer; Jun 7-10, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(1_Suppl):Abstract nr A58.
Abstract A74: Preclinical evaluation of a fatty acid synthase inhibitor in pancreatic cancer
Authors: Ahmed Roshan A.
Published: May 2016

Fatty Acid Synthase (FASN) is a key enzyme in lipid biosynthesis and is overexpressed in many cancers, including pancreatic ductal adenocarcinoma (PDA). Upregulation of FASN in pancreatic cancer is correlated to poor prognosis, poor differentiation, and advanced tumor grade. In contrast, most normal cells have very low rates of de novo fatty acid synthesis and negligible FASN expression, making FASN inhibition an attractive therapeutic option.

Current strategies for treating pancreatic cancer have severe side effects and low response rates. An alternative strategy is to target the changes in metabolic state that arise during tumor development, including an increase in anabolic processes such as de novo fatty acid synthesis. Both the de novo fatty acid synthesis pathway and FASN have been targeted previously. However past agents suffered from issues of poor bioavailability, metabolic stability, and potency. In addition several of the drugs induced significant side effects in mice, including weight loss and decreased appetite.

IPI-9119 is an irreversible inhibitor of FASN with excellent pharmacological properties. This agent is highly potent and has been shown to inhibit FASN and have anti-proliferative effects in pancreatic cancer cell lines and in vivo. We utilize a clinically relevant genetically engineered mouse model of PDA and an advanced preclinical infrastructure to evaluate IPI-9119 as a therapeutic for PDA. Preliminary in vivo studies demonstrated effective delivery of IPI-9119 to pancreatic tumor tissues, resulting in near-complete inhibition of FASN function. Short-term studies found elevated levels of apoptosis and reduced proliferation in IPI-9119 treated tumors. Combination with the genotoxic agent gemcitabine did not alter drug delivery or biochemical efficacy. These results will serve as a foundation for a comprehensive preclinical evaluation of IPI-9119 in which we will explore pharmacology, tolerability, and mechanisms of IPI-9119 action.

Citation Format: Roshan A. Ahmed. Preclinical evaluation of a fatty acid synthase inhibitor in pancreatic cancer. [abstract]. In: Proceedings of the AACR Special Conference: Metabolism and Cancer; Jun 7-10, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(1_Suppl):Abstract nr A74.
Abstract B05: Inhibition of ER/Stat3 signaling pathways drive the evolution of ER+ metastatic breast cancer through the generation of ER-independent OXPHOShi self-renewing cells
Authors: Sansone Pasquale , Berishaj Marjan , Chang Qing , Strillacci Antonio , Giacco Ferdinando , Bromberg Jacqueline
Published: May 2016

A better understanding of the molecular features of the metastatic switch might reveal new cancer cell liabilities to develop novel therapeutics for metastatic cancer patients. Here we demonstrated that the generation of dormant and tumor-initiating cells takes place in the tumor microenvironment through a metabolic reprogramming in luminal (ER+) breast cancer. The expression of the surface markers interleukin 6 receptor (IL6R) and CD44 identify distinct tumor initiating and dormant cells. We demonstrated that IL6Rlo/CD44hi cells originate in hypoxic niches and self-renew following re-oxygenation via estrogen receptor alpha (ER)-dependent up-regulation of oxidative phosphorylation (OXPHOS). Conversely, IL6Rhi/Stat3hi cells are dormant populations arising in an environment defined by recurrent episodes of hypoxia/normoxia. These cells were characterized by decreased OXPHOS and ER activities. Reducing Stat3 expression resulted in cells resistant to hypoxia/re-oxygenation-mediated dormancy. Molecularly, Stat3lo cells had reduced ER signaling, high OXPHOS and increased metastatic capacity. Thus we propose that inhibition of ER/Stat3 signaling pathways drive the evolution of ER+ metastatic breast cancer through the generation of ER-independent-OXPHOShi self-renewing cancer cells.

Citation Format: Pasquale Sansone, Marjan Berishaj, Qing Chang, Antonio Strillacci, Ferdinando Giacco, Jacqueline Bromberg. Inhibition of ER/Stat3 signaling pathways drive the evolution of ER+ metastatic breast cancer through the generation of ER-independent OXPHOShi self-renewing cells. [abstract]. In: Proceedings of the AACR Special Conference: Metabolism and Cancer; Jun 7-10, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(1_Suppl):Abstract nr B05.
Abstract B09: Mitochondrial respiration controlled by survivin directs mitochondrial dynamics and tumor cell invasion
Authors: Rivadeneira Dayana B. , Caino M. Cecilia , Seo Jae Ho , Angelin Alessia , Douglas Wallace C. , Languino Lucia R. , Altieri Dario C.
Published: May 2016

Recent studies have focused on unraveling the molecular processes by which cancer cells undergo metabolic reprogramming. The switch from oxidative phosphorylation to glycolysis allows cancer cells to adapt to harsh conditions, evade cell death and perpetuate cell growth and cell invasion. Survivin, a member of the Inhibitor of Apoptosis (IAP) gene family, is multifunctional protein. Survivin plays a critical role in promoting cell proliferation by regulating cell division, and protecting cancer cells from apoptosis. Most importantly, survivin has been shown to be highly expressed in cancers, associated with poor disease outcome and metastatic dissemination. In addition, previous studies have identified a novel pool of survivin that localizes to mitochondria selectively in tumors. This pool of survivin prevents cancer cell apoptosis and contributes to tumor progression, but the underlying mechanism is uncertain. In this study, with the use of sirRNA and pharmacological approaches against survivin, as well as protein reconstitution experiments, we dissect the role of mitochondrial survivin in modulating tumor bioenergetics. Biochemically, this study identifies mitochondrial survivin as part of a multi-protein complex with mitochondrial chaperone TRAP-1 and oxidative phosphorylation Complex II subunits. Survivin recruitment to this complex ensures protein folding and oxidative phosphorylation activity. Interference with this pathway inhibits cellular respiration, ATP production and induces cellular starvation. Functionally, this multiprotein complex suppresses tumor cell migration, invasion and affects metastatic progression in vivo. Overall, this study identifies mitochondrial bioenergetics, regulated by mitochondrial survivin, as a modulator of tumor cell invasion and metastasis.

Citation Format: Dayana B. Rivadeneira, M. Cecilia Caino, Jae Ho Seo, Alessia Angelin, Wallace C. Douglas, Lucia R. Languino, Dario C. Altieri. Mitochondrial respiration controlled by survivin directs mitochondrial dynamics and tumor cell invasion. [abstract]. In: Proceedings of the AACR Special Conference: Metabolism and Cancer; Jun 7-10, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(1_Suppl):Abstract nr B09.
Abstract B35: Association between acute leukemias and levels of leptin in blood: A meta-analysis
Authors: Dendi Vijaya SenaReddy , Aloor Sivakanth , Gadde Rahul , Kunkalla Kranthi , Reddy Roopesh Reddy Sadashiva , Gryn Jeffrey
Published: May 2016

Background: Leptin, a secretory protein of adipocytes, shown to induce differentiation, proliferation, anti-apoptotic and activation of hematopoietic cells through paracrine interaction in the bone marrow microenvironment. Existing literature provides conflicting evidence as to the association between levels of serum leptin and acute leukemia's.

Purpose: The aim of this study is to conduct a meta-analysis to investigate the association between serum leptin levels and presence of acute leukemia's.

Methods: We searched MEDLINE, CIINHAL and COCHRANE databases for studies reporting serum leptin levels in the peripheral blood and bone marrow with and without acute leukemia's [Acute lymphoblastic leukemia (ALL)/Acute myleogenous leukemia (AML)]. We included case controls, cohort and cross-sectional studies. We calculated the weighted standardized mean difference (SMD) in serum leptin levels between the acute leukemia's patients and control groups.

Results: Our search strategy yielded 70 articles and we included 10 studies enrolling 354 participants with 176 patients and 178 controls of matched age, sex and BMI (body mass index). We included 108 ALL patients compared with 126 controls and136 AML patients compared with 104 controls. The median age of the ALL and AML group compared to controls was 8.12 yrs. (IQR 7.17 - 26.25) and 42 yrs. (IQR 28.8 – 52.88). The median percentage of females of ALL and AML compared to controls was 45 (IQR 27 – 66) and 52.5 (IQR 47 – 60). The unweighted median serum leptin levels in the ALL and AML groups were 21.2 ng/ml (13.4 – 27.4) and 29.9 ng/ml (5.55 – 55.83) compared to 9.43 ng/ml (6.6 – 17.8) and 31.5 ng/ml (6.43 – 89.77) in the control groups.

The SMD's of having abnormal serum leptin levels of ALL and AML compared to controls was 1.27 (95% CI 0.53 - 2.00) P = 0.001 and – 0.97 (95% CI -1.39 - -0.55) P <0.001.

Conclusion: Increased and decreased serum leptin levels are significantly associated with ALL and AML respectively. Further investigation into the molecular mechanism underlying leptin's role in acute leukemia's and its use as an adjunctive screening tool are warranted.

Citation Format: Vijaya SenaReddy Dendi, Sivakanth Aloor, Rahul Gadde, Kranthi Kunkalla, Roopesh Reddy Sadashiva Reddy, Jeffrey Gryn. Association between acute leukemias and levels of leptin in blood: A meta-analysis. [abstract]. In: Proceedings of the AACR Special Conference: Metabolism and Cancer; Jun 7-10, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(1_Suppl):Abstract nr B35.
Abstract B39: PTP1B deficiency potentiate prostate cancer invasiveness by sensitizing Pten-null tumors to high-fat diet
Authors: Labbé David P. , Uetani Noriko , Vinette Valérie , Aubry Isabelle , Migon Eva , Sirois Jacinthe , Haigh Jody J. , Lessard Laurent , Bégin Louis R. , Trotman Lloyd C. , Paquet Marilène , Tremblay Michel L.
Published: May 2016

Specific diets can affect the risk and progression of prostate cancer (PCa). However, the interplay between diet and genetic alterations remains ill defined. Here we show that progression of PCa that is driven by Pten loss is mostly unresponsive to a high fat diet; however, in the absence of protein tyrosine phosphatase Ptpn1 (which encodes PTP1B) mice that are fed a high fat diet develop a highly invasive disease that is characterized by increased cell proliferation and Akt activation. Together with the finding that prostate-specific PTP1B overexpression does not initiate PCa by itself, we conclude that PTP1B act as an environment-dependent tumor suppressor in the context of Pten-null prostate tumors. PTP1B is a validated therapeutic target at the crossroad of metabolism (diabetes, obesity) and cancer (breast), and is currently being investigated in clinical trials. Due to PTP1B's nutrient sensing capabilities, we suggest that a careful monitoring of the balance between improving metabolic syndrome and promoting oncogenic effects under particular diets be pursued when using PTP1B-targeted therapeutics.

Citation Format: David P. Labbé, Noriko Uetani, Valérie Vinette, Isabelle Aubry, Eva Migon, Jacinthe Sirois, Jody J. Haigh, Laurent Lessard, Louis R. Bégin, Lloyd C. Trotman, Marilène Paquet, Michel L. Tremblay. PTP1B deficiency potentiate prostate cancer invasiveness by sensitizing Pten-null tumors to high-fat diet. [abstract]. In: Proceedings of the AACR Special Conference: Metabolism and Cancer; Jun 7-10, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(1_Suppl):Abstract nr B39.
Abstract B47: Unraveling tumor metabolism with in silico IHC multiplexing supported by automated imaging analysis
Authors: Harring Suzana Vega , Canamero Marta , Korski Konstanty , Marchal Georges , Grimm Oliver , Ferreira Claudia , Bredno Joerg , Chefd`hotel Christophe , Gaire Fabien
Published: May 2016

The aim of our study is to understand and characterize tumor biology and its metabolism by evaluating some of the most relevant hallmarks of cancer like hypoxia, apoptosis, tumor proliferation, angiogenesis and immune status.

We selected from our tissue bank FFPE blocks from 3 main tumor indications (CRC, GC, RCC) analyzed with 9 different markers: CA9, CC3, Ki67, CD34, aSMA, Podoplanin, CD3, CD8 and FOXP3 in consecutive sections. We applied 2 triplex, one duplex and one single IHC assays. After quality check made by board certified pathologist, all slides were scanned with the iScan-HT scanner (VENTANA) at 20x magnification and the whole slide sections were analyzed with help of automated imaging algorithms developed in house.

Tumors harbor different metabolic states. Our cutting-edge approach enables comprehensive understanding of the interactions among different tumor components with respect to tumor metabolism and immune infiltration. Our automated image analysis tool is able to reconstruct and understand the architectural patterns of different tumor metabolic states and to show distribution of hypoxia (CA9), proliferation (Ki67), cell death (CC3), angiogenesis (CD34/aSMA/Podoplanin) and immune cells (CD8/CD3, FOXP3).

Deep understanding of tumor biology and metabolism is crucial for guiding drug development and overcoming challenges in clinical translation.

Citation Format: Suzana Vega Harring, Marta Canamero, Konstanty Korski, Georges Marchal, Oliver Grimm, Claudia Ferreira, Joerg Bredno, Christophe Chefd`hotel, Fabien Gaire. Unraveling tumor metabolism with in silico IHC multiplexing supported by automated imaging analysis. [abstract]. In: Proceedings of the AACR Special Conference: Metabolism and Cancer; Jun 7-10, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(1_Suppl):Abstract nr B47.
Abstract B53: The role of RPL39 induced autophagy in resistance of breast cancer stem cells
Authors: Dave Bhuvanesh , Choi Dong Soon , Wong Helen , Liu Yi , Gunamalai Prem Siddharth , Chang Jenny C.
Published: May 2016

Introduction: Autophagy is an evolutionarily conserved lysosomal pathway for degradation of cytoplasmic proteins, macromolecules, as well as organelles, and is a survival pathway under nutrient and environmental stress. Tumor initiating cells (TICs) survive standard radiation, and chemotherapy that ultimately cause breast tumor recurrence and metastasis. Our previous studies have identified autophagy as the mechanism of survival in treatment-resistant populations in patient samples. Recent evidence from our group has defined for the first time two previously unidentified cancer genes, ribosomal protein L39 (RPL39) and myeloid leukemia factor 2 (MLF2) that play a crucial role in tumor initiation and metastasis. Additionally, RPL39 and MLF2 genes in TIC self-renewal utilize hypoxia and reactive nitrogen species for their action. In this study, we aim to determine the mechanism by which RPL39 and MLF2 modulate TICs and their relationship to tumor microenvironment.

Experimental Procedures: Analysis of clinical samples was performed using low density array analysis. This data was then confirmed using real time PCR analysis. The mechanism of action of RPL39 as related to autophagy was identified utilizing western analysis.

Results and Discussion: We confirmed our in silico bioinformatics based findings by using a real time PCR low density array of 84 genes related to autophagy in 9 paired pre- vs. post-treatment clinical samples, where 60 out of 84 genes demonstrated a statistically significant change (p<0.05) in this pathway. Using western analysis in three cell lines (SUM159, BT549, HS578T); we have demonstrated the activation of mTOR dependent translational initiation complex in TIC survival. Our results demonstrate that RPL39 binds to EIF4G and 4EBP-1 as an immunoprecipitated complex. Downstream analysis describes a RPL39-dependent modulation in 4EBP-1 and EIF4G which can be altered by over- or under- expression of the gene. In conclusion, TICs survive therapy by activating the autophagy pathway using RPL39-dependent inducible nitric oxide synthase process, through 4EBP-1 and EIF4G. Understanding this process may allow us to target TICs and breast tumor metastasis.

Citation Format: Bhuvanesh Dave, Dong Soon Choi, Helen Wong, Yi Liu, Prem Siddharth Gunamalai, Jenny C. Chang. The role of RPL39 induced autophagy in resistance of breast cancer stem cells. [abstract]. In: Proceedings of the AACR Special Conference: Metabolism and Cancer; Jun 7-10, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(1_Suppl):Abstract nr B53.