bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4292: Indole-3-carbinol (I3C) enhances efficacy of Gemcitabine in leiomyosarcoma
Authors: Suwal Sujit , Moran Alexandra , Paz-Mejia Ana , Flor Lorena , Picado Omar , Azab Basem , Trent Jonathan , Pendola Fiorella , Livingstone Alan S. , Yakoub Danny
Published: July 2017

Introduction: Adjuvant Gemcitabine have recently been shown to improve progression free survival in some types of leiomyosarcoma. Adequate cellular uptake of the drug remains a challenge. We aimed to evaluate the role of nucleoside transporter activator (I3C) in increasing the in vitro efficacy of gemcitabine in the treatment of leimyosarcoma.

Methods: Leiomyosarcoma cells (SKLMS) were incubated with DMEM+10% FBS + 1% Antibiotics. Dose response curves were generated for gemcitabine and I3C to identify the IC-50 values. Triplicates of cells were then plated in 96 well plates (3000 cells/well) and allowed to adhere. The adherent cells were further treated with gemcitabine in the presence or absence of I3C and compared with controls (DMSO treatment). MTS cell viability assays were done and absorbance was monitored in a micro-plate reader. Flow cytometry was performed with Propidium Iodide (PI) staining. Scratch assays were done to assess cell migration.

Results: Viability assay showed significant increase in the cytotoxicity of gemcitabine in presence of I3C compared to controls. Flow cytometry data showed gemcitabine alone treatment has arrested the cell cycle at G0/G1 phase without increase of cell death. On the contrary, use of gemcitabine in combination with I3C increased the cell death by at least three folds compared to control as well as gemcitabine alone treated cell population. Scratch assay over a 16 hour period showed that the rate of cellular migration significantly dropped by 1.5 folds upon treatment with gemcitabine combined with I3C compared to gemcitabine alone. Western blots showed increased expression of membrane transporter hENT-1 with I3C treatment.

Conclusion: The use of I3C enhanced cellular uptake, transport and cytotoxicity of gemcitabine in SKLMS cells. Further studies are warranted and underway to elaborate on intracellular mechanisms of action to optimize the use of I3C as a potential adjunct in preclinical and clinical leiomyosarcoma treatment.

Note: This abstract was not presented at the meeting.

Citation Format: Sujit Suwal, Alexandra Moran, Ana Paz-Mejia, Lorena Flor, Omar Picado, Basem Azab, Jonathan Trent, Fiorella Pendola, Alan S. Livingstone, Danny Yakoub. Indole-3-carbinol (I3C) enhances efficacy of Gemcitabine in leiomyosarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4292. doi:10.1158/1538-7445.AM2017-4292
Abstract 4297: ADCs inhibiting the all-powerful osmotic transducer
Authors: Prudent James R. , Marshall David , Malavasi Fabio , Chillemi Antonella
Published: July 2017

Purpose

Extracellular drug conjugates (or EDCs) are a new type of ADC that selectively inhibit the all-powerful Na,K-ATPase, an extracellular membrane ion pump required for life . To determine if EDCs may be valuable clinical candidates, we further assessed their safety, efficacy and mode of action.

Experimental Design

Cynomolgus monkeys were infused with EDCs at various levels and monitored by heart telemetry, blood cell levels and serum examination. Additionally, cultured cells were subjected to EDCs and monitored in real time and by high resolution dual fluorescent labeling to better understand the mechanism of cell death.

Results

We tested the efficacy of the EDCs in a number of xenografts models and observed strong activity (regression and cures). Little to no tumor reduction was observed for the free antibodies or with control-EDCs where the antibody target was not expressed.

To test for safety, each cynomolgus monkey were infused with one of three EDCs to better understand the MTD of each EDC. In all cases, MTD was determined to be between 5 and 10mgs/kg where EDC serum levels reached between 100 and 200ugs/ml. At the maximum safe dose tested (5mgs/kg), heart beat and other observed parameters were found to be normal with the exception of the blood cells expressing the EDC targets (EDC targeting CD20 at this high level was found to drop all B-cells to a nondetectable level which rebounded to normal on day 58).

To further understand the EDC mechanism, cells exposed to the EDCs and monitored visually in real-time or by dual fluorescence microscopies. All cells exposed to the EDCs showed membrane swelling and extracellular localization of the EDC.

Conclusion

These results show that targeted inhibitors of the Na,K-ATPase ion pump have promise as future anticancer medicines. First, EDCs appear safe in nonhuman primates at efficacious doses in rodent xenografts. In addition, an identified mechanism of extracellular necrotic death is novel in the field of oncology. Our hope is that cancers that up-regulate MDR and/or anti-apoptotic mechanisms will be sensitive to EDCs. Continuing efforts to identify the full death pathway of this new type of ADC should lead to identifying patients most likely to respond to EDC based therapies.

Citation Format: James R. Prudent, David Marshall, Fabio Malavasi, Antonella Chillemi. ADCs inhibiting the all-powerful osmotic transducer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4297. doi:10.1158/1538-7445.AM2017-4297
Abstract 4298: FTY720 induces necroptosis in lung cancer by modulating ceramide signaling at the plasma membrane
Authors: Nganga Rose
Published: July 2017

Sphingolipids are important signaling molecules in cells and have recently been explored as cancer therapy targets. FTY720 (Fingolimod, Gilenya) is an FDA approved sphingosine analogue drug used for the treatment of multiple sclerosis (MS). FTY720 is phosphorylated by sphingosine kinase 2 (SK2), to generate P- FTY720 to exert its immunosuppressive properties through binding to sphingosine -1 phosphate receptors (S1PRs). FTY720 also exhibits anti-cancer properties. Our previous studies indicated that one of the mechanism by which FTY720 induces cell death is through necroptosis. FTY720 directly binds to I2PP2A/SET (Inhibitor 2 of PP2A), consequently activating the tumor suppressor protein phosphatase 2A (PP2A). The activated PP2A then induces cell death by stimulating the activity of Receptor-Interacting Protein kinase-1 (RIPK1), involved in necroptosis signaling. Previous studies have shown that FTY720 can modulate sphingolipids metabolism in cells. However, little is known about the roles of FTY720 in ceramide signaling and regulation of necroptosis in lung cancer. We hereby seek to investigate the mechanisms of FTY720 in inducing necroptosis with regard to ceramide signaling. Preliminary data indicate that inhibitors of ceramide generation partially protect cells against FTY720-induced cell death. Interestingly, FTY720 and non-phosphorylated FTY720 analogues do not affect ceramide generation, but lead to the formation of cancer specific ceramide-multi-protein complexes at the plasma membrane. Our studies show a role of these complexes in inducing plasma membrane rupture, a key phenomenon in necroptosis. Further elucidation of these novel structures will be helpful in improving cancer therapeutics that further promote the formation these structures hence more cell death.

Citation Format: Rose Nganga. FTY720 induces necroptosis in lung cancer by modulating ceramide signaling at the plasma membrane [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4298. doi:10.1158/1538-7445.AM2017-4298
Abstract 4306: Combined effects of chemotherapy to reduce metastasis caused by insufficient hyperthermia
Authors: Lee Tae Hee , Bu Jiyoon , Kim Byoung Hyuck , Kim Young Jun , Kang Yoon-Tae , Moon Jung Eun , Cho Young-Ho
Published: July 2017

Hyperthermia therapy is one of the most widely studied non-surgical methods for breast tumors, which eliminates tumor by inducing acute stress on tumor cells. However, it has been extensively known that the viability of the tumor cells is highly influenced by the applied temperature. Insufficient thermal stress during hyperthermia treatments may alter tumor microenvironment by promoting epithelial to mesenchymal-like transition (EMT) and as a result, enhancing the outgrowth of residual tumor cells. Therefore, cells that have survived from sublethal thermal stress and experienced EMT may cause substantial clinical problems. In this case, hyperthermia requires additional therapy in order to promote cell death of more invasive tumor cells that have resisted to the thermal stimulus. In this study, we confirmed that the co-treatment of chemotherapy with hyperthermia may overcome the phenotypical transition caused by insufficient heat treatment. After exposing breast cancer cells (MCF-7) into two different temperature conditions (42°C and 47°C) for an hour, we have verified that 10.51 ± 1.71% and 18.27 ± 10.66% of cells experienced apoptosis or necrosis when the cells were exposed to 42°C and 47°C, respectively. At the same time, cancer cells showed higher invasiveness, spear-like morphology, and enhanced migratory behaviors as the exposed temperature increases, which is mainly shown among the cells that have experienced EMT. Further western blot assay and quantitative real-time polymerase chain reaction (qRT-PCR) using mesenchymal marker (vimentin) and epithelial marker (E-cadherin) also support that the mesenchymal-like phenotype has been highly increased on the cells that have resisted to the thermal stress. However, when chemotherapy was conducted after the heat treatment, cell viability was highly reduced. Paclitaxel (11.7 nM), cisplatin (3.3 μM), and combination of two anticancer drugs were treated for 24 hours on the cells that have been exposed to different temperatures, respectively. As a result, death rate of tumor cells has increased from 11.31 to 66.69%. Especially, when paclitaxel and cisplatin were co-treated, the death rate was up to 73.75 ± 4.37% after cells were exposed to 47°C. In conclusion, cancer cells that have survived from insufficient hyperthermia showed high potential to promote metastasis or recurrence but additional chemotherapy can successfully reduce the side effects induced by insufficient hyperthermia treatment.

Citation Format: Tae Hee Lee, Jiyoon Bu, Byoung Hyuck Kim, Young Jun Kim, Yoon-Tae Kang, Jung Eun Moon, Young-Ho Cho. Combined effects of chemotherapy to reduce metastasis caused by insufficient hyperthermia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4306. doi:10.1158/1538-7445.AM2017-4306
Abstract 431: IDH1-mutated gliomas rely on anaplerosis of glutamate and lactate whereas IDH1 wild-type gliomas rely on glycolysis and acetate anaplerosis
Authors: Khurshed Mohammed , Lenting Krissie , Molenaar Remco J. , Leenders William P. , Noorden Cornelis J. van
Published: July 2017

Hotspot mutations in isocitrate dehydrogenase 1 (IDH1MT) initiate low grade glioma (LGG) and secondary glioblastoma and induce neomorphic activity that converts α-ketoglutarate (α-KG) to the oncometabolite D-2-hydroxyglutarate (D-2-HG). This causes metabolic rewiring that is not fully understood and in vitro studies have shown that IDH1MT cancer cells rely on glutaminolysis, providing cells with α-KG via activities of glutaminase (GLS) and glutamate dehydrogenase (GLUD). Here, we first show by in silico analysis of 269 IDH1 wild-type (IDH1WT) and 408 IDH1MT gliomas, obtained from the The Cancer Genome Atlas (TCGA) database, that IDH1WT gliomas have high expression levels of genes encoding for enzymes that are involved in glycolysis and acetate anaplerosis. On the other hand, the tricarboxylic acid (TCA) cycle, rather than glycolytic lactate production, is the predominant metabolic pathway in IDH1MT gliomas and is driven by lactate and glutamate anaplerosis to facilitate production of α-KG, and ultimately D-2-HG. IDH1WT- and IDH1MT-related differences in expression were found in both LGG and glioblastoma. Furthermore, via in situ enzymatic activity mapping, we show in human gliomas and in xenocraft models that GLUD activity is increased and GLS activity is decreased in IDH1MT glioma, indicating that IDH1MT gliomas depend on glutamatolysis, rather than glutaminolysis. We show that transcript levels in our xenocraft models are in good agreement with our in silico analysis of the TCGA database. Finally, we confirmed the glutamate dependency of IDH1MT gliomas by MRS-flux analysis, whereas IDH1WT gliomas show high lactate production. Taken together, we show that IDH1WT gliomas have a typical Warburg phenotype and rely on acetate anaplerosis whereas IDH1MT gliomas are glutamate and lactate dependent. This metabolic rewiring in IDH1MT glioma, enables targeting of glutaminolysis rather than direct inhibition of IDH1MT for therapy. It diminishes the supply of glutamate-derived α-KG and directly inhibits the production of D-2-HG and simultaneously worsen the redox status of the glioma cells by inhibiting NAD(P)H production by GLUD. A candidate drug to inhibit GLUD is epigallocatechin-3-gallate gallate, a component of green tea that is currently receiving high interest as anti-cancer agent.

Note: This abstract was not presented at the meeting.

Citation Format: Mohammed Khurshed, Krissie Lenting, Remco J. Molenaar, William P. Leenders, Cornelis J. van Noorden. IDH1-mutated gliomas rely on anaplerosis of glutamate and lactate whereas IDH1 wild-type gliomas rely on glycolysis and acetate anaplerosis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 431. doi:10.1158/1538-7445.AM2017-431
Abstract 4310: SH003 induces apoptosis of DU145 prostate cancer cells by inhibiting ERK-involved pathway
Authors: Choi Yu-Jeong , Kim Myeong-Sun , Kang Soo-Yeon , Lee Kangwook , Ku Jin Mo , Hong Se Hyang , Kim Hyo In , Cheon Chunhoo , Ko Youme , Wen Huang Ching , Sasaki Yui , Kang Sohyeon , Kim Tai Young , Kim Ji Hye , Shin Yong Cheol , Ko Seong-Gyu
Published: July 2017

Herbal medicines have been used in cancer treatment, with many exhibiting favorable side effect and toxicity profiles compared with conventional chemotherapeutic agents. SH003 is a novel extract from Astragalus membranaceus, Angelica gigas, and Trichosanthes Kirilowii Maximowicz combined at a 1:1:1 ratio that impairs the growth of breast cancer cells. Our data demonstrate that SH003 induced apoptosis in DU145 prostate cancer cells by inhibiting ERK signaling. SH003 induced apoptosis of prostate cancer cells in dose-dependent manner, which was independent of androgen dependency. SH003 also increased intracellular ROS levels but this is not associated with its pro-apoptotic effects. SH003 inhibited phosphorylation of Ras/Raf1/MEK/ERK/p90RSK in androgen-independent DU145 cells, but not androgen-dependent LNCaP and PC-3 cells. Moreover, ERK2 overexpression rescued SH003-induced apoptosis in DU145 cells. Thus, our data conclude that SH003 induces apoptotic cell death of DU145 prostate cancer cells by inhibiting ERK-mediated pathway.

Citation Format: Yu-Jeong Choi, Myeong-Sun Kim, Soo-Yeon Kang, Kangwook Lee, Jin Mo Ku, Se Hyang Hong, Hyo In Kim, Chunhoo Cheon, Youme Ko, Huang Ching Wen, Yui Sasaki, Sohyeon Kang, Tai Young Kim, Ji Hye Kim, Yong Cheol Shin, Seong-Gyu Ko. SH003 induces apoptosis of DU145 prostate cancer cells by inhibiting ERK-involved pathway [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4310. doi:10.1158/1538-7445.AM2017-4310
Abstract 4318: Role of Bcl-2 in hematological cancer formation
Authors: Fortier Abigail V. , Koh Ally , Stanovski Lisa , Kepner Alisa , Jain Supriya , Ul-Islam Misha , Walter Ellie , Boise Lawrence
Published: July 2017

B cell leukemia/lymphoma 2 (BCL-2) is a 26 kilodalton, amphipathic, anti-apoptotic protein located on the outer mitochondrial membrane. BCL-2 interacts with other anti-apoptotic and pro-apoptotic factors in order to regulate apoptosis. If a structural change occurs on the BCL-2 gene, overexpression of the BCL-2 protein can result. This causes a change in the ratio of BCL-2 to its BCL-2 Homologous 3 (BH3) ligands, preventing the release of the apoptotic signal. The primary change associated with BCL-2 overexpression in lymphomas is a t(14;18) chromosomal translocation, which causes an increase in BCL-2 transcription. The upregulation of the oncogene allows for the survival of the mutated B cells by raising the apoptotic threshold, which then dislodges regular lymphocytes in the bone marrow or lymph nodes. Lack of cell death due to overexpression of BCL-2 has been linked to hematological malignancies, specifically lymphomas and myelomas. Both B-lymphocytic cancers can result from alterations of the immunoglobulin heavy chain locus on chromosome 14. While BCL2 translocation can cause Follicular Lymphoma, BCL2 over-expression is observed in many other tumor types and likely contributes to cancer progression and drug resistance in these tumors. Treatments to inhibit BCL-2 include attempts at activation and overexpression of the pro apoptotic BH3 proteins, BCL-2 Associated X Protein (BAX) and BCL-2 Associated K protein (BAK), and creation of BH3 mimetics. Indeed, the FDA approved the BCL2 inhibitor Venetoclax (ABT-199) earlier this year for the treatment of Chronic Lymphocytic Leukemia. These treatments could help cause cancer cell death by binding to targeted anti-apoptotic proteins and causing the signal for apoptosis to be released. This could neutralize the cancerous effects of the BCL-2 oncogene upregulation. Walton High School SMART (Students Modeling A Research Topic) Team used the computer program JMOL to create a 3D printed model of BCL-2 and composed a review poster of our secondary research to investigate and demonstrate the relationship between the structure and function of this molecule.

Citation Format: Abigail V. Fortier, Ally Koh, Lisa Stanovski, Alisa Kepner, Supriya Jain, Misha Ul-Islam, Ellie Walter, Lawrence Boise. Role of Bcl-2 in hematological cancer formation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4318. doi:10.1158/1538-7445.AM2017-4318
Abstract 4323: Metabolic reprogramming in acute myeloid leukemia cells by mesenchymal stromal cell-derived exosomes induces chemoresistance
Authors: Zhao Hongyun , Achreja Abhinav , Zhu Ziwen , Rawi Ahmed N. , Konopleva Marina , Andreeff Michael , Nagrath Deepak
Published: July 2017

Mesenchymal stromal cells play an important role in acute myeloid leukemia (AML) development. Altered cellular metabolism supports AML cells' survival in multiple aspects, such as drug resistance. Here, we demonstrate the role of MSC-derived exosomes in metabolic regulation of AML cells, and put forward a combinatorial strategy to sensitize AML cells to chemodrugs. Exosomes secreted by MSCs can reprogram the metabolic machinery following their internalization by AML cells. Through 13C tracing experiments and flux analysis, we elucidate that MSC-derived exosomes enhance oxidative phosphorylation and glutamine's entry into TCA cycle, which replenish the pool of carbon sources in mitochondria. Further, our work shows that inhibiting the interactions between MSCs and AML cells by targeting the metabolic regulation exerted by MSC-derived exosomes sensitizes AML cells to chemodrugs. Taken together, our work reveals a novel role of the TME in regulating the metabolic adaptation in AML cells and uncovers the improved strategy for AML therapy.

Citation Format: Hongyun Zhao, Abhinav Achreja, Ziwen Zhu, Ahmed N. Rawi, Marina Konopleva, Michael Andreeff, Deepak Nagrath. Metabolic reprogramming in acute myeloid leukemia cells by mesenchymal stromal cell-derived exosomes induces chemoresistance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4323. doi:10.1158/1538-7445.AM2017-4323
Abstract 4324: Estrogen promotes proliferation and apoptosis of endometrial carcinoma cell lines Ishikawa and HEC1A via endometrial stromal cells
Authors: Tao Xiang , Wu Jian , Yu Yinhua
Published: July 2017

Endometrial carcinoma is the leading malignancies of women in Western countries and the second leading cancer of female in China. As endometrial stromal cells (ESCs) are sharply decreased in endometrial carcinoma (EC) compared to normal endometrium, it is interested to explore the roll of ESCs in the tumorigenesis of endometrium. Unopposed estrogen is considered as the etiology of endometrioid carcinoma, but the underlying mechanism is needed to further explore. This study is to determine the biological effects of primary cultured ESCs on two endometrial cancer cell lines-Ishikawa and HEC1A. ESCs from normal, atypical hyperplastic and malignant endometrial specimens were collected, validated by immunohistochemistry and cultured in vitro, which were maintained in a medium containing 10 nM estradiol (E2) for three months to simulate the unopposed estrogen stimulation of EC patients. Microarray were performed to detect the alteration of gene expression, which were then confirmed by real time RT-PCR. Co-culture of ESC and EC cell lines were performed by Transwell plate. Cell proliferation was analyzed by CCK8 assay. Apoptosis was analyzed by flow cytometry based on PI/Anexin V staining. Our results showed E2 promoted the proliferation of three types of ESCs. E2 has no direct effect on cancer cell lines Ishikawa and HEC1A, but when they co-cultured with ESCs, which were stimulated by E2, the proliferation of cancer cells was promoted by all three type of ESCs. Moor interestedly, we found that the much obvious promoted effects were occurred by the ESCs that extracted from malignant samples. Estrogen could also reduce the apoptosis of both cancer cell lines when co-culturing with ESCs from normal specimen. Gene microarray and real time RT-PCR indicated that collagen 9 were up-regulated 2.3 times when ESCs cells treated by E2. In conclusion, Estrogen altered the microenvironment of stromal cells, which could lead to the proliferation of endometrial carcinoma cells via paracrine function. The alteration of collagen synthesis by ESCs might contribute to this communication between the epithelial and mesenchymal cells.

Citation Format: Xiang Tao, Jian Wu, Yinhua Yu. Estrogen promotes proliferation and apoptosis of endometrial carcinoma cell lines Ishikawa and HEC1A via endometrial stromal cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4324. doi:10.1158/1538-7445.AM2017-4324
Abstract 4339: Single cell RNA sequencing dissects cellular growth factor dependencies and oncogenic driver effects in an organoid model of gastric cancer
Authors: Chen Jiamin , Andor Noemi , Grimes Susan M. , Lau Billy , Ji Hanlee P.
Published: July 2017

Gastric cancer is a lethal malignancy with few therapeutic options. Gastric tumors rely on complex intercellular signaling “crosstalk” that enables tumor development, metastasis, and therapeutic resistance. To recapitulate the intercellular communications among various cell populations that exist in vivo, we are using a three-dimensional culture system to grow and manipulate mouse gastric tissue in vitro, otherwise referred to as organoids. Importantly, this organoid model contains epithelium with its endogenous mesenchymal niche and does not require exogenous Wnt stimulation. To systematically analyze the distinct cellular lineages and their interactions, we applied a massively-scaled single cell RNA-Seq platform to sequence thousands of individual cells from organoid cultures. With PCA and t-SNE analysis of the high-dimensional data generated from single cell RNA-seq, we characterized two major cell types, i.e. epithelial and mesenchymal cells. Leverage the information from single cell transcriptome profiles, we identified specific niche factors of the Wnt signaling pathways that are activated in different stomach cell lineages. These results suggest that the mesenchymal cell populations provide a potential source of the R-spondin, a Wnt agonist, that sustains the growth of epithelium. Furthermore, we compared cell populations from Cdh1-/-/Trp53-/- and Trp53-/-organoids, and characterized changes on the transcriptome profiles due to the loss of Cdh1, an early oncogenic event in diffuse gastric cancer development. Overall, using organoid model and high-throughput single cell RNA-Seq provides a novel approach to study early tumor transformation and critical cancer-stroma interactions.

Citation Format: Jiamin Chen, Noemi Andor, Susan M. Grimes, Billy Lau, Hanlee P. Ji. Single cell RNA sequencing dissects cellular growth factor dependencies and oncogenic driver effects in an organoid model of gastric cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4339. doi:10.1158/1538-7445.AM2017-4339
Abstract 434: Characterizing the role of serine metabolism in pediatric sarcomas
Authors: Issaq Sameer , Kidner Ria , Rohde Jason , Boxer Matthew , Helman Lee
Published: July 2017

Sarcomas represent a diverse group of malignancies with unique molecular and pathological characteristics. In order to improve sarcoma treatment, a better understanding of the alterations associated with specific sarcoma subtypes is critically important. Renewed interest in the altered metabolic properties of cancer cells has led to an exploration of targeting metabolic dependencies as a novel therapeutic strategy. Metabolism of the amino acid serine is frequently altered in cancer, supporting a number of critical biological processes, including protein, lipid, and nucleotide synthesis, and redox balance. The first, rate-limiting step in the serine synthesis pathway is catalyzed by the enzyme 3-phosphoglycerate dehydrogenase (PHGDH), which is overexpressed in several cancers. Previous work has shown that PHGDH loss or inhibition is selectively toxic to cancer cells with high PHGDH expression or increased flux through the serine synthesis pathway. In this study, we have characterized the dependency of pediatric sarcomas on serine metabolism by examining expression of PHGDH in Ewing sarcoma and rhabdomyosarcoma cell lines, and evaluating the effects of PHGDH inhibition and serine deprivation on cellular proliferation and bioenergetic properties. We show that PHGDH is highly expressed in pediatric sarcoma cell lines, and that PHGDH knockdown resulted in decreased proliferation, especially under conditions of serine limitation. Moreover, pharmacological inhibition of PHGDH resulted in a dose-dependent decrease in proliferation and mitochondrial bioenergetic function. Furthermore, individual sarcoma cell lines were differentially sensitive to serine deprivation, indicating that some sarcoma cells may depend on extracellular serine in addition to de novo serine synthesis. Our findings suggest that the dependency of pediatric sarcomas on serine metabolism should be further investigated in order to identify vulnerabilities that could be targeted for potential therapeutic benefit.

Citation Format: Sameer Issaq, Ria Kidner, Jason Rohde, Matthew Boxer, Lee Helman. Characterizing the role of serine metabolism in pediatric sarcomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 434. doi:10.1158/1538-7445.AM2017-434
Abstract 4349: Adverse & anticancer activities of 5-azaCdR & DNA methyltransferase (DNMT) isoform specific inhibitors: therapeutic implications
Authors: Cheishvili David , Szyf Moshe
Published: July 2017

Activation of methylated and silenced promoters of genes that suppress tumorigenesis has been the rationale behind the clinical use of DNMT family inhibitor 5aza for treating cancer. However, the adverse effects of general DNA methylation inhibition on tumors through activation of tumor promoting and prometastatic genes as well adverse effects on normal cells have not been comprehensively assessed, nor is it known which particular DNMT is responsible for regulating these adverse effects. A related critical question is whether such potential toxicity could be reduced by targeting a specific isoform of DNMT. We, therefore, performed a comprehensive molecular characterization of the effects of the pan DNMT inhibitor 5aza and isoform-specific inhibition of DNMT1, DNMT3a and DNMT3b by shRNA on cell growth and invasiveness, the methylome and transcriptome of the breast, liver and lung cancer cells as well as their normal primary cell counterparts. Our results suggest that 5aza causes general DNA methylation inhibition both as far as genomic features as well as the genes that are affected. This results in activation of genes involved in promoting cancer and metastasis in both cancer and normal cells. 5aza also induces rampant activation of retroviral elements and ectopic transcription initiation in gene bodies and intergenic regions. Second, isoform-specific exhibit a more limited and specific profile of activity on promoter methylation and gene expression. Third, the adverse profile of 5aza corresponds to the effects of specific inhibition of DNMT3a suggesting that the adverse effects of 5aza are partly mediated through inhibition of DNMT3a. Fourth, specific inhibition of DNMT1 results in an enriched inhibition of DNA methylation in promoters and inhibition of growth without triggering activation of tumor suppressor genes and a more favorable molecular footprint on cancer cells. Our data provide a comprehensive assessment of the impact of DNA methylation inhibition on normal and cancer cells and points to the potential adverse effects of such an approach. However, since the different DNMTs have a different molecular footprint, these adverse effects might be inhibited using isoform-specific DNMT inhibitors. Our data needs to be considered for further clinical development of DNMT inhibitors.

Note: This abstract was not presented at the meeting.

Citation Format: David Cheishvili, Moshe Szyf. Adverse & anticancer activities of 5-azaCdR & DNA methyltransferase (DNMT) isoform specific inhibitors: therapeutic implications [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4349. doi:10.1158/1538-7445.AM2017-4349
Abstract 4357: ESRP1 overexpression and its role in ovarian cancer
Authors: Kwon Mi Jeong
Published: July 2017

Epithelial splicing regulatory protein 1 (ESRP1) and 2 (ESRP2) are RNA binding proteins which are recently known to be involved in tumor progression via regulation of alternative splicing of cancer-associated genes. However, their expression and clinical significance in ovarian cancer remain unclear. Here, we found that ESRP1 and ESRP2 gene expression is upregulated in ovarian cancer cells compared with immortalized ovarian surface epithelial (IOSE) cells and their overexpression at protein level was also confirmed in ovarian cancer tissues. Consistent with TCGA data of ESRP1 gene amplification, higher ESRP1 gene copy number was observed in ovarian cancer cells compared with IOSE cells, whereas there was little correlation between gene copy number and gene expression in ovarian cancer cells. Importantly, their gene expression was inversely correlated with DNA methylation in ovarian cancer cells and ESRP2 overexpression in ovarian cancer was significantly associated with DNA hypomethylation (P= 0.036). Of note, TCGA data analysis showed that high ESRP1 gene expression is significantly associated with shorter 5-year survival of patients with ovarian cancer. Ectopic expression of ESRP1 in ESRP1 low-expressing mesenchymal type ovarian cancer cells increased cell proliferation, whereas it suppresses cell migration. Furthermore, our results revealed that the effect of ESRP1 overexpression on cell proliferation and migration is related to activation of the Akt/CyclinD pathway and increased of E-cadherin expression. In conclusion, our findings suggest that ESRP1 overexpression is associated with poor outcome of patients and ESRP1 may play a cancer-promoting role in ovarian cancer.

Citation Format: Mi Jeong Kwon. ESRP1 overexpression and its role in ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4357. doi:10.1158/1538-7445.AM2017-4357
Abstract 4361: An integrated genome-wide methylation analysis with gene expression in normal breast tissues identifies differentially methylated CpG loci associated with obesity
Authors: Weng Daniel Y. , Song Min-Ae , Brasky Theodore M. , Marian Catalin , Lan Renny S. , Llanos Adana A. , Spear Scott L. , Kallakury Bhaskar V. , Freudenheim Jo L. , Shields Peter G.
Published: July 2017

Introduction: Obesity is a risk factor for breast cancer, but the underlying mechanisms for this are only partially understood, and there has been only limited study of breast tissues molecularly before cancer develops.

Methods: Breast tissues from healthy women undergoing reduction mammoplasty and epidemiologic interviews (n=121) were profiled for the whole genome transcriptome (Affymetrix Human Transcriptome Arrays) and genome-wide DNA methylation (Infinium HumanMethylation 450 BeadChip array).

Results: After adjusting for confounding by age and race, 12,210 CpG dinucleotides with altered methylation levels correlated with body mass index (BMI) (10,808 positive correlations and1,402 negative correlations, FDR<0.05). Among them 4,170 BMI-associated hypermethylated CpG dinucleotides (-1.5 kb from transcription start site, 1st Exon, and 5’UTR) and 443 BMI-associated hypomethylated CpG dinucleotides were located in promoter regions. By integrating DNA methylation and mRNA expression data, we identified 310 methylated genes that correlated with gene expression (FDR<0.05). Of these, 242 genes had higher methylation status showing concurrent down-regulation in obese women, and 68 genes had lower methylation status showing concurrent up-regulation in obese women. Among the affected genes involved in diseases and disorders for inflammatory response, hereditary disorder, and immunological disease.

Conclusions: This study provides evidence that obesity epigenetically deregulates genes potentially involved in breast cancer that have functional relationships to gene expression.

Citation Format: Daniel Y. Weng, Min-Ae Song, Theodore M. Brasky, Catalin Marian, Renny S. Lan, Adana A. Llanos, Scott L. Spear, Bhaskar V. Kallakury, Jo L. Freudenheim, Peter G. Shields. An integrated genome-wide methylation analysis with gene expression in normal breast tissues identifies differentially methylated CpG loci associated with obesity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4361. doi:10.1158/1538-7445.AM2017-4361
Abstract 4368: The clinicopathological significance of glutathione S-transferase P1 hypermethylation in breast cancer, a meta-analysis and literature review
Authors: Liu Mingli
Published: July 2017

Background: Glutathione S-transferase P1 (GSTP1) has been reported to function as a tumor suppressor gene in various types of human cancers including breast cancer (BC). However, the association and clinicopathological significance between GSTP1 hypermethylation and BC remains unclear. The purpose of this study is to conduct a meta-analysis and literature review to investigate clinicopathological significance of GSTP1 methylation in BC.

Method: A detailed literature search was made in PubMed, EMBASE, Web of Science and Google Scholar databases. The data were extracted and assessed by two reviewers independently. Odds Ratios (OR) with 95% corresponding confidence intervals (CIs) were calculated. A total of twenty four relevant articles were available for meta-analysis.

Results: The frequency of GSTP1 hypermethylation was significantly increased in BC than in benign breast disease, the pooled OR was 7.09, p<0.00001. The rate of GSTP1 hypermethylation was significantly different between early stage I/II and later stage III/IV, OR was 2.05, p<0.0001. In addition, the rate of GSTP1 hypermethylation was also significantly different regarding the node involvement status. The rate of GSTP1 hypermethylation in node positive BC was higher than that in node negative BC, OR was 1.77, p=0.006. However, GSTP1 hypermethylation was not significantly associated with ER, PR and HER2 status.

Conclusion: In summary, our meta-analysis indicated that the frequency of GSTP1 hypermethylation was significantly increased in BC than in benign breast disease. The rate of GSTP1 hypermethylation in late stages of BC was higher than that in early stages. Our data suggested that GSTP1 hypermethylation could be a risk factor and a diagnostic biomarker of BC carcinogenesis. GSTP1 hypermethylation may have a role in the progression of BC. GSTP1 is a potential drug target for development of demethylation treatment for patients with BC.

Note: This abstract was not presented at the meeting.

Citation Format: Mingli Liu. The clinicopathological significance of glutathione S-transferase P1 hypermethylation in breast cancer, a meta-analysis and literature review [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4368. doi:10.1158/1538-7445.AM2017-4368
Abstract 4383: Novel fusion transcripts in colorectal cancer patients revealed by next-generation RNA sequencing
Authors: Choi Yuri , Kwon Chae Hwa , Park Hye Ji , Won Yeo-Jin , Park Do Youn
Published: July 2017

Although colorectal cancer (CRC) is the third most common type of cancer in the world, there is an insufficient research on molecular targets for prevention and therapies of CRC. Gene fusions are associated with tumorigenesis and tumor activation, and could be a standard to distinguish tumor heterogeneity for personalized therapy. In the present study, we performed next-generation RNA sequencing (150 tumor and 50 matched normal tissues) in order to discover novel fusion transcripts in CRC. A total 25 transcript candidates were detected by GFP, deFuse, and FusionMap algorithms. Among the candidates, 6 fusion transcripts were selected (B4GALT1-BAG1, GTF3A-CDK8, NAGLU-IKZF3, RNF121-FOLR2, STRN-ALK, and TAF4-NDRG3) after analyzing expression and function of the acceptor genes. All of them were validated by sanger-sequencing and were not found in another cohort. Overexpression of these fusion genes, except B4GALT1-BAG1, increased proliferation of CRC cells. In addition, overexpression of NAGLU-IKZF3 enhanced migration of CRC cells. These results reveal novel fusion transcripts and their tumorigenic effects in CRC. Ultimately, it could illuminate a genomic-driven strategy to develop personalized medicine.

Key words: fusion transcript, colorectal cancer, RNA-seq

Citation Format: Yuri Choi, Chae Hwa Kwon, Hye Ji Park, Yeo-Jin Won, Do Youn Park. Novel fusion transcripts in colorectal cancer patients revealed by next-generation RNA sequencing [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4383. doi:10.1158/1538-7445.AM2017-4383
Abstract 4384: Targeted sequencing reveals distinct and rare pathogenic variants in Caucasians with colorectal cancer
Authors: Mokarram Pooneh , Varma Sudhir , Azimi Hamed , Olumi Hasti , Safarpour Ali reza , Nickerson Michael , Brim Hassan , Ashktorab Hassan
Published: July 2017

PURPOSE: Next-generation sequencing (NGS) is currently used to establish mutational profiles in many multigene diseases such as colorectal cancer (CRC) which is on the rise in many parts of the developing World including in the Middle East. Little is known about its genetic hallmarks in these populations. AIM: To identify variants in 15 CRC-associated genes in patients of Iranian descent.

METHODS: CRC specimens from 63 patients were used to establish the variants’ profile on an Ion Torrent platform by targeted exome sequencing. To rule out technical artifacts, the variants were validated in 13 of these samples using an Illumina NGS platform. Validated variants were annotated and compared to variants from publically available databases. An in-silico functional analysis was performed. MSI status of the analyzed samples was established. RESULTS: There were 51 validated variants distributed on 12 genes: 22% MSH3 (n=11/51), 10% MSH6 (n=5/51), 8% AMER1 (n=4/51), 20% APC (n=10/51), 2% BRAF (n=1/51), 2% KRAS (n=1/51), 12% PIK3CA (n=6/51), 8% TGFβR2A (n=4/51), 2% SMAD4 (n=1/51), 4% SOX9 (n=2/51), 6% TCF7L2 (n=3/51), and 6% TP53 (n=3/51). Most known and distinct variants were in mismatch repair genes (MMR, 32%) and APC (20%). Among oncogenes, PIK3CA was the top target (12%). MSH3 variants were more frequent and predominantly homozygous in the analyzed population.

CONCLUSION: These results illustrate for the first time CRC mutational profile in Iranian patients. MSH3, MSH6, APC and PIK3CA genes seem to play a bigger role in the path to cancer in this population. This is especially true for MSH3 variants that were very frequent and predominantly homozygous as these will associate with the EMAST phenotype that has prognostic implications. These findings will potentially lead to informed genetic diagnosis protocol and targeted therapeutic strategies.

Citation Format: Pooneh Mokarram, Sudhir Varma, Hamed Azimi, Hasti Olumi, Ali reza Safarpour, Michael Nickerson, Hassan Brim, Hassan Ashktorab. Targeted sequencing reveals distinct and rare pathogenic variants in Caucasians with colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4384. doi:10.1158/1538-7445.AM2017-4384
Abstract 439: Metabolic flux analysis reveals targets to sensitize chemoresistance in acute myeloid leukemia induced by mesenchymal stromal cell-derived exosomes
Authors: Achreja Abhinav , Zhao Hongyun , Zhu Ziwen , Gerszberg Jonathan , Konopleva Marina Y. , Andreeff Michael , Nagrath Deepak
Published: July 2017

The tumor microenvironment has a pleiotropic role in supporting cancer cell growth, metastasis and drug resistance. Exosomes from mesenchymal stromal cells (MSC) were found to regulate metabolism of acute myeloid leukemia (AML) cells that led to a chemoresistant phenotype. Exosomes carry a host of proteins, nucleotides and metabolites that can induce metabolic reprogramming recipient cells via direct supply of metabolite cargo or through signaling. We utilized 13C tracer techniques and two metabolic flux analysis techniques to reveal the mechanism of metabolic reprogramming induced by MSC-derived exosomes. First, 13C metabolic flux analysis was used to quantify intracellular fluxes of central carbon, amino acids and fatty acid metabolism in AML cells cultured with and without exosomes in media with 13C substrates. Second, we employed a novel technique, exosome-mediated metabolic flux analysis (Exo-MFA), to estimate direct support provided by metabolite supply from exosome cargo by introducing 13C-labeled exosomes. The combined analysis dissected the metabolite supply from exosome cargo from the overall reprogramming of metabolic pathways due to exosomes. Our results revealed key metabolic pathways that could be targeted to inhibit exosome-induced reprogramming to reverse the chemoresistant phenotype.

Citation Format: Abhinav Achreja, Hongyun Zhao, Ziwen Zhu, Jonathan Gerszberg, Marina Y. Konopleva, Michael Andreeff, Deepak Nagrath. Metabolic flux analysis reveals targets to sensitize chemoresistance in acute myeloid leukemia induced by mesenchymal stromal cell-derived exosomes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 439. doi:10.1158/1538-7445.AM2017-439
Abstract 4394: Whole-genome, virome, and immuno-genome of liver cancer indicate its genomic feature associated with virus infection and inflammation
Authors: Nakagawa Hidewaki
Published: July 2017

Liver cancer is prevalent worldwide, mainly associated with virus infection and chronic inflammation, but its underlying molecular mechanism is quite heterogeneous. We performed virome and immuno-genomic analyses, in addition to whole-genome landscape of somatic alterations and transcriptome in 300 liver cancers. Our whole-genome analysis elucidated point mutations in non-coding regions, structural variations, and virus integrations (HBV and AAV), in addition to coding mutations. We discovered novel recurrently mutated coding and non-coding regions, such as NEAT1/MALAT1 lincRNAs, promoters, CTCF-binding sites, and novel regulatory regions, and known cancer genes (CDKN2A, APC, TERT) and novel cancer genes (ASH1L, NCOR1, MACROD2) that were recurrently affected by SVs. For virome analysis, we performed ultra-deep sequencing after capturing HBV sequences from DNAs of 100 liver cancers or adjacent HBV-infected liver tissues, which demonstrated genome-wide HBV-integrations (1794 sites) in liver cancers and infected liver tissues and eliminated pathological mechanism of viral integration. We also generated immuno-genomic profiles of these liver cancers, including HLA genotypes and mutations, neo-antigen presentation, immune signatures, and TCR repertories, which suggested some distinct immunological features and underlying molecular pathways of liver cancer associated with inflammation. These trans-omics data demonstrated comprehensive and distinct immuno-genomic feature of virus-related liver cancer, as well as its distinct whole-genome profiles, and can help to understand the molecular and microenvironment pathways underlying carcinogenesis and treatment response of liver cancer.

Citation Format: Hidewaki Nakagawa. Whole-genome, virome, and immuno-genome of liver cancer indicate its genomic feature associated with virus infection and inflammation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4394. doi:10.1158/1538-7445.AM2017-4394