bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2775: A novel immunostimulatory OX40/PD-L1 bivalent bispecific antibody (MEDI1109) for the treatment of patients with cancer
Authors: Oberst Michael D. , Kasturirangan Srinath , Sachs Clifford , Auge Catherine , Moynihan James , Rothstein Raymond , Hair James , Neal Francis , Mamidi Srinivas , Hanabuchi Shino , Watkins Amanda , Zheng Yanan , Rosenthal Kim , Freeman Daniel J. , Hammond Scott
Published: August 2018

Targeted immunotherapy utilizing agonist antibodies (Ab) to OX40 shows great promise in preclinical mouse models alone or in combination with various therapies including PD1/PD-L1 antagonist Abs. Ab-mediated signaling through OX40 requires higher-order clustering by Fcγ receptors to co-stimulate antitumor-specific T cells and attenuate regulatory T cell (Treg) immunosuppression, leading to antitumor activity. To date, OX40 agonists are well-tolerated but show limited activity alone or in combination with checkpoint blockade in clinical trials. We hypothesize that the antitumor activity of OX40 agonists in patients may be limited by insufficient intratumoral exposure and an inadequate amount of clustering by Fcγ receptors within the tumor microenvironment. To overcome these limitations, we engineered MEDI1109, a bivalent bispecific Ab composed of an OX40 agonist Ab molecularly fused to two single-chain variable fragments derived from a PD-L1 antagonist. The resulting bispecific Ab retained the functional properties of each parent Ab that included binding and agonizing OX40, binding and full blockage of the PD-L1/PD-1 pathway, depletion of OX40+ Tregs and overcoming Treg suppression. Emerging data suggest that the bispecific Ab also drives novel biology including (1) increased OX40 agonism through clustering by either PD-L1 and Fcγ receptors, (2) improved intratumoral exposure through PD-L1 targeting, (3) elimination of PD-L1 positive tumor cells by NK cell mediated antibody-dependent cellular cytotoxicity and (4) enhanced antitumor immunity through concurrent PD-L1/PD-1 blockade on tumor cells and OX40 co-stimulation of antitumor T cells. Intravenous administration of OX40/PD-L1 bispecific Abs to cynomolgus monkeys resulted in the proliferation of peripheral blood CD4+ and CD8+ total memory T-cell populations, and in the reduction of soluble PD-L1 levels in the plasma. These results demonstrate that MEDI1109 is a unique and potent immune modulating agent and may be utilized to enhance antitumor immunity in patients with cancer.

Citation Format: Michael D. Oberst, Srinath Kasturirangan, Clifford Sachs, Catherine Auge, James Moynihan, Raymond Rothstein, James Hair, Francis Neal, Srinivas Mamidi, Shino Hanabuchi, Amanda Watkins, Yanan Zheng, Kim Rosenthal, Daniel J. Freeman, Scott Hammond. A novel immunostimulatory OX40/PD-L1 bivalent bispecific antibody (MEDI1109) for the treatment of patients with cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2775.
Abstract 2815: Ganoderma lucidum chemosensitizes triple negative breast cancer to carboplatin mediated by DNA damage response
Authors: Suarez-Arroyo Ivette J. , Martinez-Montemayor Michelle M.
Published: August 2018

Management of triple negative breast cancer (TNBC) is challenging because of a lack of targeted therapy, its aggressive behavior and poor prognosis. Interest in using platinum agents, such as carboplatin as a line of treatment emerged from data suggesting frequent DNA repair defects in TNBC. Carboplatin resistance may occur due to multiple factors including increased activation of DNA repair mechanisms. Studies show that Ganoderma lucidum extract (GLE) sensitizes cancer cells to different anti-cancer therapies. Thus, we aim to investigate the effects of GLE in combination with carboplatin in the DDR signaling and in tumorigenesis. To study whether GLE sensitizes TNBC cells to carboplatin therapy, we treated SUM-149 and MDA-MB-231 cells with several concentrations of carboplatin, GLE, or both treatments simultaneously for 72h. Our results demonstrate that when carboplatin and GLE are used in combination the IC50s decreased in both breast cancer cell lines in comparison with treatments alone. Immunobloting assays revealed that GLE and/or the combination decrease the expression of 53BP1, p-CHK2, CHK2, p-CHK1, CHK1, p-ATR, p-ATM and p-BRCA1 in SUM-149. Accordingly, immunofluorescence studies to monitor the recruitment of DNA repair proteins 53BP1 and phosphorylated γH2AX at the lesion sites after treatments suggest that the GLE and/or carboplatin combination impair DDR. To determine tumor progression, severe combined immune deficient mice were injected with SUM-149 cells and then treated with carboplatin/GLE or carboplatin for 10 weeks. The combination significantly decrease tumor volume in comparison with the vehicle. Taken together, our results provide evidence of GLE's potential in chemosensitizing TNBC cells to carboplatin therapy affecting the DDR. These results highlight GLE's anti-breast cancer therapeutic potential.

Citation Format: Ivette J. Suarez-Arroyo, Michelle M. Martinez-Montemayor. Ganoderma lucidum chemosensitizes triple negative breast cancer to carboplatin mediated by DNA damage response [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2815.
Abstract 2871: Diosgenin inhibits the proliferation of MCF-7 breast cancer cells through the demethylation of miR-145 gene
Authors: Shi Fengqin , Lu Lingeng , Li Ya , Hou Li , Chen Xinyi , Rui Han , Wang Chong , Yue Ya
Published: August 2018

Background: Altered DNA methylation in microRNA (miRNA) host genes has been frequently associated with abnormal miRNA expression during the pathogenesis of cancer. miR-145 was identified as a tumor-suppressive miRNA that was down-regulated in several cancer types. Accumulating evidence has clearly demonstrated that a low expression of miR-145 in breast cancer was correlated with an aberrant hypermethylation in its host gene. Diosgenin is a steriod saponin, which is extracted from the tubers of Dioscorea wild yam. The anti-tumor mechanisms of diosgenin have been demonstrated via the modulation of multiple cell signaling pathways, which are associated with growth, differentiation, apoptosis and oncogenesis. However, there is little information regarding the epigenetic-based effects of Diosgenin in breast cancer. In our study, we hypothesize that Diosgenin has anti-proliferative effects on breast cancer cells through the demethylation of miR-145.

Material and Methods: MCF-10A breast epithelial cells and MCF-7 breast cancer cells were used to investigate the effects of Diosgenin on the expression and methylation of miR-145 using real time-PCR (RT-PCR) and methylation-specific PCR (MSP), respectively. An MTS proliferation assay was performed to assess the effect of Diosgenin on the proliferation of MCF-7 breast cancer cell.

Result: In comparison with MCF-10A breast epithelial cells, MCF-7 breast cancer cells showed the hypermethylation of miR-145, and significantly down-regulated miR-145 expression. Diosgenin treatment led to proliferation inhibition of MCF-7 breast cancer cells. In MCF-7 breast cancer cells, the Diosgenin treatment led to significantly up-regulated miR-145, whereas the methylation level of miR-145 significantly decreased.

Conclusion: Diosgenin, a natural compound, exhibits an anti-proliferative effect on breast cancer cells, and affects the methylation and expression of miR-145. These findings indicate that the demethylation of miR-145 may be a molecular mechanism underlying the anti-tumor of Diosgenin in breast cancer.

Citation Format: Fengqin Shi, Lingeng Lu, Ya Li, Li Hou, Xinyi Chen, Han Rui, Chong Wang, Ya Yue. Diosgenin inhibits the proliferation of MCF-7 breast cancer cells through the demethylation of miR-145 gene [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2871.
Abstract 289: Probing the non-enhancing component of glioblastoma: Targeting what is left behind
Authors: Peng Sen , Halperin Rebecca , Dhruv Harshil , Byron Sara , Legendre Christophe , Phillips Joanna , Prados Michael , Berens Michael , Tran Nhan
Published: August 2018

While genomic profiling and therapeutic selection support individualized GBM treatment, such therapeutic decision-making is usually made with reference to tumor obtained from the enhancing core region. GBM is known to be heterogeneous and exhibits a high resistance to standard therapies. To address whether non-enhancing tumor (representing the majority of tumor left behind after surgery) shows distinct genomic characteristics and therapeutic targets compared to the enhancing tumor core, we performed genome-wide exome-sequencing and RNA-sequencing for 12 patients with matched enhancing region and at least one non-enhancing region. Non-enhancing biopsies show a surprisingly high level of tumor content, with a median of 28% tumor cells and 6 of the 22 samples having greater than 50% tumor cells. Cognate non-enhancing and enhancing specimens demonstrated overall concordance in therapeutically actionable alterations (single nucleotide variants) and copy number alterations. However, non-enhancing regions were not genetically identical and did reveal additional and distinct variants compared to enhancing cores. For example, the non-enhancing region of patient 1 showed two nonsense NF1 mutations (R1534X; R2517X) while the enhancing region showed an NF1 frameshift mutation (F1247fs). Clonality analysis by LumosVar also indicated that 7 out of 12 patients harbored dissimilar cellular prevalence patterns between enhancing and non-enhancing regions. In addition, comparison of alternative polyadenylation between enhancing and non-enhancing regions uncovered distinct 3' UTR usage: e.g. SGMS2 and TOB1 tended to have longer 3' UTR in enhancing regions whereas longer 3' UTR of SYNPO and NOS1AP were

prevalent in non-enhancing regions. We posit that the enhancing component of glioblastoma probably underrepresents the genomic alterations in patients' tumors. Given non-enhancing tumor is left behind after surgical debulking, genomic profiling of this region would potentially reveal more accurate tumor vulnerabilities and lead to more effective therapy.

Supported by a grant from the Ben & Catherine Ivy Foundation.

Citation Format: Sen Peng, Rebecca Halperin, Harshil Dhruv, Sara Byron, Christophe Legendre, Joanna Phillips, Michael Prados, Michael Berens, Nhan Tran. Probing the non-enhancing component of glioblastoma: Targeting what is left behind [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 289.
Abstract 2916: Targeting lysophospholipid metabolism inhibits pancreatic cancer cell proliferation under nutrient-limiting conditions
Authors: Jong Petrus R. de , Maruggi Marco , Campos Alejandro D. , Brand Morgan A. , Lemos Robert , Scott David A. , Litherland Sally A. , Arnoletti J. Pablo , James Brian P. , Powis Garth
Published: August 2018

Patients with pancreatic ductal adenocarcinoma (PDAC) have a poor prognosis, and more effective systemic treatments for patients with local progression or metastasis (85% of cases) are needed. The pancreatic tumor microenvironment provides a rich source for novel drug targets. We aimed to identify and validate novel metabolic drug targets that are unique to hypoxic PDAC cells. Using bulk RNA sequencing in combination with metabolomics analyses in vitro, we previously found that PDAC cells negate the loss of intracellular unsaturated fatty acids in hypoxia by orchestrating the release of lysophospholipids (lyso-PLs) by cancer-associated fibroblasts, which are then taken up and stored in intracellular lipid droplets in hypoxic cancer cells. To confirm the relevance of these findings in vivo, we performed 3' droplet based single-cell RNA sequencing (scRNA-seq) combined with metabolomics analyses of intracellular and extracellular (tumor interstitial fluid) metabolites of MIAPaCa2 and patient-derived xenografts (PDX). Identification of cell lineages and subpopulations with hypoxic gene signatures was performed to correlate changes in metabolite levels with metabolic gene expression in vivo. This approach confirmed differential expression of lipid droplet-associated enzymes in hypoxic areas of the tumor, including lyso-PL acyl transferases (LPCAT1, LPCAT3), and phospholipases (LYPLA1, PLA2G15). We found that resistance of PDAC cell lines to pharmacologic treatment with inhibitors of fatty acid desaturases (FADS), reminiscent of hypoxia and nutrient starvation in vivo, was mediated by uptake of lyso-PLs from the medium. Importantly, genetic knockdown of LPCAT and LYPLA isoforms reversed the resistance to FADS inhibitors in culture in vitro and in vivo. Clinical relevance was demonstrated by mRNA expression analysis of PDAC patients from The Cancer Genome Atlas (TCGA) database, which showed that the expression of lyso-PL metabolizing genes is correlated with a significant worse prognosis (log-rank test, P=0.008). We are currently developing pharmacologic approaches to target LPCAT and LYPLA enzymes in hypoxic cancer cells as a novel approach for PDAC patients with unresectable disease.

Citation Format: Petrus R. de Jong, Marco Maruggi, Alejandro D. Campos, Morgan A. Brand, Robert Lemos, David A. Scott, Sally A. Litherland, J. Pablo Arnoletti, Brian P. James, Garth Powis. Targeting lysophospholipid metabolism inhibits pancreatic cancer cell proliferation under nutrient-limiting conditions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2916.
Abstract 2931: MTX-211, a dual and selective inhibitor of EGFR and PI3 kinase, shows promising activity in combination with MEK inhibition in preclinical models of pancreatic cancer
Authors: Frankowski-McGregor Christy , Maust Joel , Ziemke Elizabeth , Mumby Rachel , Delaney Amy , Rehemtulla Alnawaz , Whitehead Christopher , Sebolt-Leopold Judith S.
Published: August 2018

Pancreatic cancer is one of the deadliest forms of cancer, with median 5-year survival rates less than 10%. This disease is recalcitrant to chemotherapeutic approaches and recently approved therapies afford only modest improvements in survival. KRAS is the most commonly mutated gene in pancreatic tumors with an incidence rate exceeding 90%. Despite intensive efforts, KRAS has remained undruggable. Kinases acting both upstream and downstream of RAS continue to be exploited for the development of novel agents to attenuate signaling through this critical oncogene. While EGFR and MEK inhibitor monotherapies have been evaluated in the clinic for the treatment of pancreatic cancer, efficacy has been modest despite their relevance to MAP kinase (MAPK) pathway-altered tumors. Novel therapies are urgently needed to address adaptive signaling mechanisms that diminish the effectiveness of kinase-targeted approaches. MTX-211 is a first-in-class dual inhibitor of PI3K and EGFR kinase with a promising pharmaceutical profile and proven ability to potentiate the effectiveness of MEK inhibitor therapy in KRAS mutant colorectal tumors. The present study was undertaken to extend our evaluation of MTX-211 to include pancreatic cancer model systems. We have found that MTX-211 exhibits low micromolar potency against an extensive panel of primary models of pancreatic cancer and is highly synergistic with the MEK inhibitor trametinib. Preliminary data suggest that this synergy is driven by the ability of MTX-211 to target compensatory transcriptional activation of HER3 that occurs in response to MEK inhibition. A genetically engineered KRAS and p53 mutant (KPC) mouse model was transduced with a lentiviral construct encoding a caspase reporter, which was used to show that the combination of MTX-211 and trametinib elicited a significant increase in apoptosis over single agent controls. Results from an animal study conducted with the KPC model further showed that the combination of MTX-211 and trametinib significantly slowed growth of these aggressive tumors. Employing both KPC and patient-derived xenograft models of pancreatic cancer, efforts are underway to optimize therapeutic outcome in response to MTX-211-based combination treatment regimens.

Citation Format: Christy Frankowski-McGregor, Joel Maust, Elizabeth Ziemke, Rachel Mumby, Amy Delaney, Alnawaz Rehemtulla, Christopher Whitehead, Judith S. Sebolt-Leopold. MTX-211, a dual and selective inhibitor of EGFR and PI3 kinase, shows promising activity in combination with MEK inhibition in preclinical models of pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2931.
Abstract 3064: Oral cancer stem cells modulate Fusobacterium nucleatum to acquire the capability to induce tumor stemness switch
Authors: Pal Bidisha , Bhuyan Seema , Baishya Debabrat , Das Bikul
Published: August 2018

Background: Fusobacterium nucleatum is an oral bacteria that contributes to oral carcinogenesis. Its potential role in oral cancer is not known. Fusobacterium nucleatum (henceforth FN) is a gram-negative anaerobe associated with periodontal diseases and also colon cancer. The bacteria modulates the host cell signaling mechanism including the activation of beta-catenin signaling and p38 pathway. The Fap2 is the outer membrane proteins of FN that inhibit lymphocytes and NK cells. The bacteria also directly binds to NKp46 receptor of NK cells. These properties of FN may be exploited by oral CSCs to defend their own niche as a part of niche-defense mechanism against immune system. Hence, it is not surprising that oral CSCs may hijack this specific oral bacterium to enhance tumor stemness. We speculate that oral CSCs may hijack specific bacteria type present in oral mucosa, and enhance their stemness-inducing mechanisms to switch non-CSCs to CSCs.

Methods: We have established an in vitro model of oral cancer cells/FN host/pathogen interaction model. Using this model, we wanted to study potential role of the bacteria in cancer stemness. We also used the saliva of subjects with oral cancer to infect oral cancer stem cells and non-stem cancer cells.

Results: When the SCC-25 oral cancer cell line was treated with the saliva of subjects with oral cancer, the EpCAM+/ABCG2+ oral CSCs population was increased and showed the presence of FN. The FN recovered from CSCs exhibited high expression of FadA and Fap2, two surface proteins present in FN that can modulate signaling pathways of host cells. Importantly, FN recovered from the infected CSCs induced stemness in the non-stem cancer cell population of SCC-25, suggesting that CSCs may modulate the pathogen. Indeed, our preliminary observation indicates that FN recovered from the FN strain ATCC 25586 infected SCC-25 ABCG2+ cells showed increased expression of FADA and Fap2 as compared to the parental strain ATCC 25586.

Conclusion: Our data suggests that CSCs have the capability to reprogram F. nucleatum genome in order to make the bacteria a potent inducer of tumor stemness in non-stem cancer cells.

Citation Format: Bidisha Pal, Seema Bhuyan, Debabrat Baishya, Bikul Das. Oral cancer stem cells modulate Fusobacterium nucleatum to acquire the capability to induce tumor stemness switch [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3064.
Abstract 3208: Optimal array layouts for tumor treating fields therapy in glioblastoma: Oblique array layouts are superior to standard LR and AP positions
Authors: Mikic Nikola , Korshøj Anders
Published: August 2018

Introduction: Tumor treating fields (TTFields) is a new and effective treatment for glioblastoma. Two pairs of transducer arrays are placed on the scalp of the patient to deliver low intensity, alternating electric fields, which inhibit mitosis and cancer growth. In the present study, we investigated systematic variations in electrode array positions to identify optimal layouts, which induce the highest electric fields in the tumor tissue.

Method: The electric field distribution was calculated using finite element methods applied to a realistic computational head model. Array positions were rotated systematically in fifteen-degree intervals around a central cranio-caudal axis of the head to obtain thirteen different layouts in the same plane. Tumors were introduced in the computational mesh at nineteen different fronto-parietal positions in the same plane as defined by the electrode positions.

Results: Electrode array layout greatly affected the field distribution. For the vast majority of tumors, two optimal and oblique array orientations were identified. At these positions, field intensities were approximately 30-40% higher compared to the standard anterior/posterior and left/right positions. In addition, the two optimal layouts were oriented at ninety-degree interval to each other. Correspondingly, a single optimum layout was identified when analyzing the combination of two pairs of arrays. Each tumor position was associated with only one optimum (combined) layout, which was generally oblique relative to the sagittal plane (15-45 degrees). An oblique layout at forty-five degrees to the sagittal plane was effective for most tumor positions and in all cases it was superior to the default layout combination of anterior-posterior and left-right positions.

Conclusion: Our results provide guiding principles for optimal electrode placement, which will be helpful, especially in situations where the placement must be changed due to concurring clinical conditions such as shunts, scars, tissue rash etc. Patient specific models should be for more accurate TTF distributions. Additional research is needed to confirm these current results.

Citation Format: Nikola Mikic, Anders Korshøj. Optimal array layouts for tumor treating fields therapy in glioblastoma: Oblique array layouts are superior to standard LR and AP positions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3208.
Abstract 33: Calsequestrin 2 regulates proliferation, migration, and invasion in triple-negative breast cancer cells
Authors: Kim Ju Hee , Hong Bok sil , Heo Woohang , Han Jong Min , Han Wonsik , Noh Dong-Young , Moon Hyeong-Gon
Published: August 2018

Triple-negative breast cancer (TNBC) is the most heterogeneous and aggressive breast tumor subtype defined by absence of receptor for estrogen, progesterone, or HER2. However, the biologic mechanism for TNBC phenotype is still unclear. Here, we show that expression of Calsequestrin 2 (CASQ2), a Ca2+-binding protein, correlates with increase of proliferation, migration, and invasion, suggesting that intracellular Ca2+ may contribute to tumor growth and metastatic phenotype. CASQ2 is the main Ca2+-binding protein inside the sarcoplasmic reticulum of cardiomyocytes. CASQ2 forms a complex with ryodine receptor 2 (RyR2) luminal calcium release channel in cardiac muscle. Ca2+ is a sequestor and regulator of diverse cellular processes, and specific Ca2+ channels play important roles in cell proliferation and invasiveness of cancers. To know the role of CASQ2 TNBC cells, we established CASQ2-overexpressing stable cells in Hs578T (Hs578T-CASQ2) using retrovirus. Stimulation with caffeine triggered the remarkable increase of intracellular calcium in Hs578T-CASQ2 cells; in addition, the basal level of calcium in cells had much higher amount of Hs578T-CASQ2 than Hs578T. By contrast, calcium chelator BAPTM/AM blocked CASQ2-induced calcium release. Hs578T-CASQ2 cells showed higher level of proliferation, migration and invasion rate compared to Hs578T, which indicated that overexpression of CASQ2 related with cellular functions. We also found that CASQ2 overexpression elevates extracellular signal-related kinase (ERK) expression. In epidermal growth factor (EGF)-treated cells, Hs578T-CASQ2 cells had higher phosphorylated ERK compared to Hs578T, leading to the expression of epithelial-mesenchymal transition (EMT) marker, vimentin. These results indicate that CASQ2 overexpression increases the level of calcium and induces cell proliferation, migration and invasion through ERK signaling. Our findings from this study show a possible cause of migration and invasiveness in breast cancer cells. Taken together, these findings demonstrate that CASQ2 could be a new therapeutic target for breast cancer.

Citation Format: Ju Hee Kim, Bok sil Hong, Woohang Heo, Jong Min Han, Wonsik Han, Dong-Young Noh, Hyeong-Gon Moon. Calsequestrin 2 regulates proliferation, migration, and invasion in triple-negative breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 33.
Abstract 3469: Dual inhibition of the EGFR and Aurora A pathways in Kras mutated non-small cell lung cancers
Authors: Bagnyukova Tetyana , Egleston Brian , Kramer Mackenzie , Astsaturov Igor , Golemis Erica , Borghaei Hossein
Published: August 2018

Lung cancer is the leading cause of cancer related death in the US. Patients with advanced disease generally have a poor prognosis with a median survival of around 10 to 12 months with standard chemotherapy. In patients with non-squamous non-small cell lung cancers (NSCLC) incidence of KRAS mutations reach ~22%. Specific treatments for Kras mutated NSCLC have not been developed although recent reports suggest an advantage for treatment with immunotherapy over chemotherapy in the second line setting. Published reports using a synthetic lethal screening method show that the combined inhibition of the EGFR and Aurora A kinase pathways could have synergistic effect on viability in head and neck and colon cancer cell lines suggesting a potential benefit of such combination therapy in these patients (Astsaturov and colleagues, 2010). Our study aims to examine the effects of combination of erlotinib (E), an EGFR small-molecule inhibitor, and Alisertib (MLN8237, A), an inhibitor of Aurora A kinase, in lung cancer models with different KRAS status. Human NSCLC cell lines with both wild type and mutated KRAS showed weak or moderate sensitivity to both drugs in vitro. Treatment with both E and A had synergistic activity in all examined cell lines, wild type KRAS (H2228, H1299) or mutated KRAS (A549, H1299, H358, H460, and H62). Western blot analysis confirmed that combination treatment enhanced apoptosis compared to single drug use as evidenced by cleaved PARP. In xenograft models, CB17 SCID mice were treated with single drug or E+A for three weeks. H322M xenograft tumors bearing wild type KRAS were moderately sensitive to growth inhibition by E (10 mg/kg/day) and A (20 mg/kg/twice daily) alone and the combination had an additive effect. A549 and H358 xenografts were insensitive to E because of the mutated KRAS and moderately sensitive to A. However, the combination of E+A had a synergistic effect and significantly inhibited tumor growth. Western blot analysis of signaling pathways downstream of the EGFR and Aurora A in tumor xenografts shows synergistic inhibition after two-drug treatment. Our results suggest that combined inhibition of both pathways might be an effective treatment strategy for lung cancers harboring KRAS mutations.

Citation Format: Tetyana Bagnyukova, Brian Egleston, Mackenzie Kramer, Igor Astsaturov, Erica Golemis, Hossein Borghaei. Dual inhibition of the EGFR and Aurora A pathways in Kras mutated non-small cell lung cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3469.
Abstract 3508: Separate and combined effects of caloric restriction mimetics and autophagy inhibition on KRAS-driven pancreatic adenocarcinoma
Authors: Chen Xuewen , O'Flanagan Ciara H. , Coleman Michael , Der Channing J. , Hursting Stephen D.
Published: August 2018

Calorie restriction (CR) is one of the most effective dietary interventions for reducing cancer risk and progression. However, maintaining a dramatically reduced calorie intake over an extended time period is very challenging for most individuals. Thus a major effort is under way to identify mechanism-based pharmacologic approaches that mimic the anticancer effects of CR without the requirement of reduced calorie intake. We have established that CR exerts antitumor effects at least in part through altered metabolism, reduced circulating growth factors (such as insulin and IGF-1) and reduced signaling downstream of the insulin receptor and IGF-1R along the phosphoinositide 3-kinase/mTOR axis. We have previously shown CR reduced growth of KRAS-driven tumors in vivo, and the antitumorigenic effect was enhanced when combined with autophagy inhibition. Here we evaluated the effects of in vitro CR via glucose and serum modulation, as well as separate and combined effects of CR mimetics and autophagy inhibition on the KRAS-driven murine pancreatic adenocarcinoma (PDAC) cell line, FC1242. Glucose modulation alone via glucose restriction, metformin or AICAR treatment showed no effect on clonogenic growth of FC1242. In contrast, both cell viability and clonogenicity were significantly reduced by serum starvation, rapamycin treatment, IGF-1R/insulin receptor inhibition by BMS-754807, and sirtuin inhibition with nicotinamide, suggesting these cells are dependent on mTOR/IGF-1 signaling. The growth-suppressive effects of these CR mimetics were enhanced when combined with autophagy inhibition via chloroquine or verteporfin. Together, the study indicated the targeting of mTOR or IGF-1 signaling may most closely mimic the growth-suppressive effects of CR, and that co-treatment of these CR mimetics with autophagy inhibition may be an effective strategy for PDAC anticancer therapy.

Citation Format: Xuewen Chen, Ciara H. O'Flanagan, Michael Coleman, Channing J. Der, Stephen D. Hursting. Separate and combined effects of caloric restriction mimetics and autophagy inhibition on KRAS-driven pancreatic adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3508.
Abstract 3532: Bitter melon extract suppresses triple-negative breast cancer growth by inhibiting ACAT1
Authors: Shim So Hee , Steele Robert , Ray Ratna
Published: August 2018

Breast cancer is the most common malignant cancer diagnosed in women worldwide each year. Triple-negative breast cancers (TNBC), defined by the lack of expression of estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2), account for approximately 15% of breast cancers. Moreover, TNBC is usually more aggressive and metastatic with a worse prognosis. Bitter melon (Momordica charantia) is well known as a dietary supplement. Recently, we have shown an anti-cancer activity of bitter melon extract (BME) in breast cancer mouse models, although the mechanism is poorly understood. Furthermore, we became interested to understand the role of BME on cholesterol and lipid metabolism in TNBC. Our mass spectrometry data suggested that stored as a cholesteryl esters (CE) in Lipid droplets (LD) were much lower in BME treated TNBC cells. We found that not only CE but also LD was reduced in BME treated TNBC cells. The acyl-CoA:cholesterol acyl transferase-1 (ACAT-1) catalyzes the formation of CE from free cholesterols together with long-chain fatty acyl coenzyme A. Therefore, inhibition of ACAT-1 resulted in downregulation of CE, and upregulation free cholesterols. The attenuated CE is known to reduce cancer proliferation. Subsequent study demonstrated that BME treatment highly inhibited ACAT-1 in TNBC cells. The upregulated free cholesterol negatively regulates sterol regulatory element-binding proteins (SREBPs), which regulates lipogenesis protein such as fatty acid synthase (FASN) associated with cancer cell proliferation. We also observed that SREBP-1 and -2 were barely detectable, and FASN and low-density lipoprotein receptor (LDLR) were reduced in BME treated TNBC cells as compared to control cells. To our knowledge, this is the first report demonstrating that BME suppresses TNBC cell growth through ACAT-1 inhibition. BME supplement may potentially be an additional regimen in human breast cancer treatment.

Citation Format: So Hee Shim, Robert Steele, Ratna Ray. Bitter melon extract suppresses triple-negative breast cancer growth by inhibiting ACAT1 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3532.
Abstract 3641: Use of adjuvant interferon alfa-2b or ipilimumab 10 mg/kg for high-risk patients with melanoma, and associated adverse events and duration of therapy
Authors: Tarhini Ahmad , Gupte-Singh Komal , You Min , Gupta Amit , Kim Le Trong , Rao Sumati
Published: August 2018

OBJECTIVE: Adjuvant systemic therapy with interferon alfa-2b (IFN) or ipilimumab 10 mg/kg (IPI10) has been the standard for high-risk patients with melanoma who have undergone surgery; however, a high proportion of patients remain untreated. This analysis examines the adverse events (AEs) and duration of therapy (DoT) associated with IFN or IPI10. METHODS: A retrospective cohort study was conducted among each of the four US healthcare databases: OptumInsight, IMS PharMetrics, Truven Commercial, and Truven Medicare Supplemental. Patients receiving melanoma-related surgery (lymph node dissection) from July 1, 2006, to September 30, 2016, were identified. The study estimated the incidence rate (IR) per patient per year of AEs that commonly occur with IFN or IPI10 across different DoT levels among patients with melanoma following surgery. RESULTS: Of 46,401 eligible patients with melanoma surgery claims, 2423 (5%) were treated: 2316 with IFN and 107 with IPI10. Mean age was 51.7 years (60.5% male), and mean (standard deviation) Charlson comorbidity index score was 6.6 (2.8) and 7.5 (2.5) for patients prescribed IFN or IPI10, respectively. The DoT (median, mean (SD)) for IFN was 142, 182 (162) days and for IPI10 was 84, 73 (26) days. Similarly, the number of prescriptions (median, mean (SD)) for IFN were 20, 26 (28) and for IPI10 were 4, 3 (1). The overall AE IR per year was 4.4 in patients receiving IFN and 4.9 for those receiving IPI10. Among patients receiving IFN, the most common AEs were hepatitis, nausea, and fatigue (IRs: 0.92, 0.67, and 0.66, respectively). The most common AEs among IPI10-treated patients were fatigue, rash, nausea, and diarrhea (IRs of 0.95, 0.71, 0.66, and 0.63, respectively). AE rates were greater in patients with low DoT compared with those with a high DoT for most AEs in both IFN and IPI10 cohorts. CONCLUSION: Only 5% of high-risk patients with melanoma following lymph node dissection surgery were treated with adjuvant IPI10 or IFN, while the remaining were untreated. Patients treated with IPI10 or IFN had high AE rates impacting their ability to remain on therapy. Newer adjuvant treatments with more favorable AEs and DoT profiles are urgently needed for high-risk patients with melanoma.

Disclosure: This research was funded by Bristol-Myers Squibb.

Citation Format: Ahmad Tarhini, Komal Gupte-Singh, Min You, Amit Gupta, Le Trong Kim, Sumati Rao. Use of adjuvant interferon alfa-2b or ipilimumab 10 mg/kg for high-risk patients with melanoma, and associated adverse events and duration of therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3641.
Abstract 3659: Detection and validation of cancer immunotherapy biomarkers in blood
Authors: Xie Feng , Yu Jianjun , Jia Shidong
Published: August 2018

Immunotherapy response varies widely, making it difficult for physicians to know whether immunotherapy will be effective for a given patient. Indeed, ~80% or more patients with cancer fail to respond to checkpoint inhibitor immunotherapy. In addition to PD-L1 IHC staining, recent studies reported that patients with deleterious mutations in mismatch repair (MMR) genes, high tumor mutation burden (TMB) or microsatellite instability (MSI) are also associated with better clinical response. As tissue biopsy represents a practical challenge due to its insufficient quantity or lack of access, non-invasive molecular has emerged as an efficient complementary test and attracted increasing attention in clinical development of cancer immunotherapy.

With Predicine's gene RARDAR technology, we developed a blood-based PrediSeq NGS panel to capture genomic alterations in 180 cancer genes including tumor mutation burden (TMB) and microsatellite instability (MSI) and a Predi-CI (Cancer Immunotherapy) panel to measure dynamic expression change of 212 genes related to immune cells or immune responses.

Technical validation was performed to evaluate assay sensitivity, specificity and accuracy using reference samples with known genetic profiling. The panels have been further tested using tissue biopsy and plasma samples from cancer patients. The development of PrediSeq Panel and Predi-CI RNA gene expression tests offer a comprehensive solution to stratify and monitor cancer patients who may benefit from cancer immunotherapy.

Citation Format: Feng Xie, Jianjun Yu, Shidong Jia. Detection and validation of cancer immunotherapy biomarkers in blood [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3659.
Abstract 3672: The three-dimensional (3D) spatial lamin A/C organization in normal lymphocytes, and in Hodgkin and Reed-Sternberg cells
Authors: Contu Fabio , Szczurek Aleksander , Vanni Roberta , Knecht Hans , Mai Sabine
Published: August 2018

Classical Hodgkin Lymphoma (HL) is a B-Cell lymphoma comprising mononucleated Hodgkin cells (H) and diagnostic bi- to multi-nucleated Reed Sternberg cells (RS). Genomic instability in HL is characterized by telomere dysfunction caused by critically short telomeres and telomere uncapping. Super resolution microscopy showed significant increases in the amount of DNA-poor nuclear spaces in HL compared to normal primary lymphocytes. A direct 3D interaction between telomeres and shelterin protein TRF2 is progressively disrupted from H to RS cells by either massive upregulation or downregulation of TRF2. TRF2 interacts with nuclear matrix protein lamin A/C in the maintenance of the 3D genome organization, which is disturbed in H and RS cells. Therefore we hypothesized that the TRF2- telomere-lamin A/C interaction is also disturbed in H and RS cells and leads to organizational alterations of the genome in HL. To this end, we analyzed lamin A/C expression levels of lymphocytes, H and RS cells and found that lamin A/C is upregulated in cancer cells compared to both activated and resting lymphocytes. The regular homogeneous and round shaped pattern found in lymphocytes was replaced in H and RS cells by a more irregular one, characterized by the presence of lamin A/C invaginations and septa that subdivide the nuclei into multiple smaller compartments. Our data show no co-localization between telomeres and lamin A/C in the DNA-poor spaces. Lamin A/C bordered the DNA-poor spaces but was never found inside them or in association with inner telomere fragments. 3D TRF-2 staining showed upregulation of the protein compared to normal lymphocytes, in correlation with the observation made in EBV-negative patients with clinically aggressive disease. The analysis of pre-treatment Hodgkin's patients' samples will allow to determine whether the TRF2-telomere-lamin A/C binding is different in patients with recurrent versus non-recurrent disease.

Citation Format: Fabio Contu, Aleksander Szczurek, Roberta Vanni, Hans Knecht, Sabine Mai. The three-dimensional (3D) spatial lamin A/C organization in normal lymphocytes, and in Hodgkin and Reed-Sternberg cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3672.
Abstract 3705: Targeted anticancer drug delivery to Ewing's sarcoma using human anti-CD99 targeted hybrid polymerization liposomal nanoparticles
Authors: Kang Hyunggyoo , Nagy Jon , Triche Timothy
Published: August 2018

Most anticancer therapies are unspecific and therefore inevitably lead to off-target toxicity. To overcome these shortcomings, targetable hybrid polymerization liposomal nanoparticles (HPLNs) have been invented. Multiple innovative and proprietary technologies have been developed to enable creation of this unique form of nanoparticle therapy, including the use of cross linked lipids to improve both stability and bioavailability, modified polyethylene glycol (PEG) for both ‘stealthiness' (to prevent immune response to the nanoparticle) and for covalent attachment of targeting molecules such as tumor specific antibodies or peptides. Our present study showed the superior efficacy of human anti-CD99 targeted HPLNs in the treatment of Ewing sarcoma. Human monoclonal anti-CD99 antibody-targeted HPLNs encapsulating irinotecan (CD99-HPLN/Ir) can efficiently reach implanted Ewing sarcoma tumors in xenograft mice and dramatically reduce and/or eliminate the tumor burden. Complete tumor ablation has been observed at doses as low as 1 mg irinotecan/kg, treated twice per week. In other cases, animals who failed untargeted (HPLN/Ir) treatment showed complete tumor ablation after ‘salvage' treatment with CD99-HPLN/Ir. Our CD99 targeted HPLN/irinotecan formulation, termed NV103 showed much better efficacy than the commercial untargeted liposomal irinotecan (Onivyde™) and doxorubicin (Doxil™). Toxic side effects, normally associated with systemic administration of free irinotecan, were minimized or undetectable. Our nanoparticles showed excellent bioavailability. Even the untargeted HPLN/Ir improve drug bioavailability six-fold in a comparison experiment with the drug in liposome form (Onivyde™), with no discernable systemic toxicity. Antibody targeting of the HPLN irinotecan formulation (NV103) improves the efficacy 12-fold, over the untargeted liposome form. In conclusion, this formulation may be effective for treatment of other CD99-expressing tumors like certain leukemias, lymphomas and glioma. Ultimately, after development of other tumor-specific targeting agents and varied encapsulated therapeutics, this technology can be usefully adapted for the treatment of other cancers as well.

Citation Format: Hyunggyoo Kang, Jon Nagy, Timothy Triche. Targeted anticancer drug delivery to Ewing's sarcoma using human anti-CD99 targeted hybrid polymerization liposomal nanoparticles [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3705.
Abstract 3770: Characterization of novel dual A2A/A2B adenosine receptor antagonists for cancer immunotherapy
Authors: Galezowski Michal , Wegrzyn Paulina , Bobowska Aneta , Commandeur Claude , Dziedzic Katarzyna , Nowogrodzki Marcin , Obara Alicja , Szeremeta-Spisak Joanna , Dzielak Anna , Lozinska Iwona , Dudek Marcelina , Janiga Anita , Reus Jacek , Wronowski Marek , Swirski Mateusz , Radzimierski Adam , Ziembik Magdalena , Mierzwicka Joanna , Wojcik-Jaszczynska Katarzyna , Gocek Elzbieta , Grycuk Karolina , Golas Aniela , Pierzchala Olga , Zachmann Julian , Nowak Mateusz
Published: August 2018

Adenosine is the key immunometabolite responsible for immune tolerance in tumors. It is present in normal tissue in low concentrations, having various physiological functions. In the tumor, its concentration increases rapidly, as a result of overexpression of enzymes producing adenosine, additionally enhanced by hypoxia and inflammation. Adenosine inhibits the biological functions of T lymphocytes infiltrating the cancer tissue by binding to the A2A receptor. The affinity to A2B receptor is believed to attenuate the response of dendritic cells and other parts of innate system. Thus blocking simultaneously the effects mediated by both receptor subtypes with dual inhibitor seems to be a viable approach to a single agent cancer immunotherapy or a combination with current immunotherapeutic agents.

We have discovered a novel series of potent and selective A2A/A2B inhibitors. Best compounds present subnanomolar activity in both in vitro recombinant cellular model and in primary human cells. Our antagonists dose-dependently restore the adenosine agonist-impaired functional activity of CD4+ and CD8+ human T-lymphocytes (cytokine release assays). We observe also the relieve of adenosine-related suppression in NK cells cytotoxicity. Most importantly tested compounds present improved primary pharmacological profile in comparison to A2A inhibitors currently tested in clinical trials. Further validation in in vivo models showed efficacy standalone and in combination with checkpoint inhibitors.

Citation Format: Michal Galezowski, Paulina Wegrzyn, Aneta Bobowska, Claude Commandeur, Katarzyna Dziedzic, Marcin Nowogrodzki, Alicja Obara, Joanna Szeremeta-Spisak, Anna Dzielak, Iwona Lozinska, Marcelina Dudek, Anita Janiga, Jacek Reus, Marek Wronowski, Mateusz Swirski, Adam Radzimierski, Magdalena Ziembik, Joanna Mierzwicka, Katarzyna Wojcik-Jaszczynska, Elzbieta Gocek, Karolina Grycuk, Aniela Golas, Olga Pierzchala, Julian Zachmann, Mateusz Nowak. Characterization of novel dual A2A/A2B adenosine receptor antagonists for cancer immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3770.