bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4872: ONC201 induction of apoptosis in hPheo1 cell line supports use of this oral agent in ongoing phase 2 clinical trial against neuroendocrine tumors (NCT03034200)
Authors: Proudfit Austin M. , Anderson Peter M. , Gupta Neetu
Published: August 2018

ONC201 is an imipridone that induces TRAIL and causes apoptosis in cancer cell lines, but not normal cells. Recent data shows the dopamine-like DRD2 receptor binds ONC201 and pheochromocytoma-paragangliomas (PC-PG) have >10x more DRD2 than any other human cancer (TCGA data). We present pre-clinical data on dose response and time course of ONC201 against hPheo1, a PC-PG cell line, that support a recently opened phase 2 clinical trial of weekly oral ONC201 in neuroendocrine tumors (NETs) including PC-PG. The human pheochromocytoma cell line, hPheo1, was treated with ONC201 and studied for in vitro anti-cancer activity. Effects on cell viability/proliferation were measured via cell counting with trypan blue exclusion. Post-treatment target-protein levels were assessed via western blotting. Additionally, cells were stained with Annexin V and analyzed using flow cytometry for markers of apoptosis. Upon treatment with ONC201, hPheo1 cells experienced a dose-dependent decrease in cell viability and proliferation with peak effect at 72 h post-treatment (GI50=3.2 μM). Substantial inhibition of pro-survival kinases, Akt and ERK, were confirmed at 72 hours to further support the drug's efficacy and elucidate on a corresponding mechanism of action. The anti-proliferative effects of the drug were gradual over a 96 h time period, rather than an abrupt, intense response. In addition, apoptosis was confirmed at 24 and 96 h, with Annexin V+ cells detectable by FACS. Pre-clinical data show excellent anti-cancer efficacy of ONC201 against PC-PG cells at concentrations achievable in vivo. These findings suggest both anti-proliferative effects and induction of apoptosis lead to anti-cancer activity. The gradual influence of ONC201 on PC-PG cells, rather than rapid cell destruction argues against a scenario where immediate catecholamine release could result in severe hypertension. These pre-clinical data support safe use of ONC201 against PC-PG in vivo and other NETs in our ongoing Phase 2 clinical trial (NCT03034200; investigator-initiated IND132665).

Citation Format: Austin M. Proudfit, Peter M. Anderson, Neetu Gupta. ONC201 induction of apoptosis in hPheo1 cell line supports use of this oral agent in ongoing phase 2 clinical trial against neuroendocrine tumors (NCT03034200) [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 4872.
Abstract 4879: Mutation selectivity of the allosteric SHP2 inhibitor SHP099
Authors: Sun Xiaojun , Ren Yuan , Gunawan Steven , Teng Peng , Chen Zhengming , Lawrence Harshani R. , Cai Jianfeng , Lawrence Nicholas J. , Wu Jie
Published: August 2018

SHP2 (PTPN11) mediates oncogenic signaling and gain-of-function (GOF) SHP2 mutations are associated with leukemias and other types of cancer. Recently, SHP099 was identified as a novel allosteric SHP2 inhibitor that displayed high potency and selectivity to the wildtype SHP2 in vitro and inhibited protein tyrosine kinase oncogene-driven cancer cells. However, it was unclear whether SHP099 inhibits SHP2 mutants. Using SHP2 mutant-dependent TF-1 leukemia cells, we determined sensitivities of four common oncogenic SHP2 mutants to SHP099. SHP2D61Y-, SHP2A72V-, and SHP2E76K-expressing TF-1 cells were resistant, whereas SHP2E69K-expressing TF-1 cells were sensitive to SHP099 with a potency similar to that of cytokine-stimulated TF-1 cells expressing the wildtype SHP2. Consistently, SHP099 reduced active ERK1/2 and Bcl-XL and induced apoptosis of SHP2E69K-dependent TF-1 cells, but had no effects on the other SHP2 mutant cell lines. SHP2 knockout cells were unresponsive to SHP099, demonstrating specificity. Like the wildtype SHP2, SHP2E69K PTP was inhibited by SHP099 in vitro with a sub-micromolar IC50, whereas micromolar IC50s of SHP099 were observed against SHP2D61Y, SHP2A72V, and SHP2E76K. These results indicate that SHP2D61Y, SHP2A72V, and SHP2E76K mutants are resistant to SHP099 and that SHP2E69K is a SHP099-sensitive oncogenic mutant.

Citation Format: Xiaojun Sun, Yuan Ren, Steven Gunawan, Peng Teng, Zhengming Chen, Harshani R. Lawrence, Jianfeng Cai, Nicholas J. Lawrence, Jie Wu. Mutation selectivity of the allosteric SHP2 inhibitor SHP099 [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 4879.
Abstract 4880: Novel nano-molar small-molecule STAT3 inhibitor series with antitumor activities against human breast cancer
Authors: Yue Peibin , Lopez-Tapia Francisco , Brotherton-Pleiss Christine , Kubota Casie , Chen Jasmine , Tius Marcus , Turkson James
Published: August 2018

Constitutively-active Signal transducer and activator of transcription (STAT) 3 has been well validated as a significant mechanistic underpinning of many human cancers and a target for anticancer therapy. To date, no small molecule STAT3 inhibitor has been successfully developed for clinical application as a therapeutic agent, despite the discovery of many molecular entities that inhibit the protein. To advance the therapeutic development of small molecule STAT3 inhibitors, we focused on the existing lead compounds, BP-1-102, SH4-54, and SH5-07 (all three showing IC50 of 4-7 μM). All three leads are based on an N-methylglycinamide scaffold, with its two amine groups condensed with three different functionalities. Intensive medicinal chemistry campaign was directed towards understanding and optimizing the molecular determinants critical for enhanced STAT3-inhibitory potency and strong antitumor effects. We report the development of nanomolar potent inhibitors of STAT3 DNA-binding activity. Most notably, the analogs H127, H145, H171, H174, H181, and H182 preferentially inhibit STAT3 activity in DNA-binding assay in vitro, with IC50 of 300 - 800 nM, compared to little or no observed effect on STAT1 and STAT5 DNA-binding activity at concentrations up to 20 μM. Treatment of human breast cancer MDA-MB-231 and MDA-MB-468 cells with H171, H174 or H182 inhibited constitutive STAT3 DNA-binding activity and phosphorylation at Tyr705 in both time- and dose-dependent manner. H171, H174 or H182 further blocked the growth and viability, colony formation, and oncogenic transformation of the human breast cancer cells that harbor persistently active STAT3, with IC50 of 1.0 - 1.9 μM, compared to the IC50 of 3.8 - 8.1 μM against the viability of the immortalized breast epithelial cells that do not harbor persistently active STAT3. These compounds represent some of the first nanomolar potent small molecules observed to directly inhibit STAT3 activity, with potent antitumor cell effects.

Citation Format: Peibin Yue, Francisco Lopez-Tapia, Christine Brotherton-Pleiss, Casie Kubota, Jasmine Chen, Marcus Tius, James Turkson. Novel nano-molar small-molecule STAT3 inhibitor series with antitumor activities against human breast 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 4880.
Abstract 4883: Suppression of breast cancer by reprogramming glucose metabolism and energy production with HJC0152
Authors: Kim Hyejin , Dong Jiabin , Xu Jimin , Li Dengfeng , Zheng Zhi , Ye Na , Zhang Ziwei , Chen Haiying , Zhou Jia , Shen Qiang
Published: August 2018

Most mammalian cells use glucose as the primary fuel source. Glucose is metabolized via glycolysis to pyruvate, which is further transferred into mitochondria for generation of ATP through the Krebs cycle in normal condition. However, metabolism is characteristically reprogrammed and cancer cells or highly proliferative cells preferably generate ATP through lactate production by lactate dehydrogenase (LDH/LDHA), referred to as the Warburg effect or metabolic reprogramming toward anaerobic glycolysis. Efficient control of energy metabolism is the key to maintaining metabolic homeostasis, and disturbance in energy balance provokes diseases such as obesity, diabetes and cancer. However, the mechanisms underlying efficient energy metabolic homeostasis and breast cancer development are poorly understood. HJC0152, a novel small-molecule glucose metabolism modulator, was developed using structure-/fragment-based drug design strategies and molecular modeling techniques. Aggressively growing and metastatic breast cancer cells of triple-negative subtype (MDA-MB-231) treated with HJC0152 showed inhibited cell growth. Intriguingly, HJC0152 decreased activity and protein level of LDHA, which resulted in a decrease of lactate production in breast cancer cells. In addition, these cells also exhibited a decrease of glucose uptake by reducing Glut1 and HK2 protein level and modulation of the expression of glycolytic enzymes. Furthermore, HJC0152 treatment causes decreased mitochondrial membrane energy potential, resulting in significantly reduced level of intracellular ATP in MDA-MB-231 cells. Our findings suggest that HJC0152 is capable of reprogramming caner metabolism by modulating glucose metabolism and ATP production. These results may provide a rationale to develop HJC0152 as an effective therapeutics for cancer and other metabolic diseases with aberrant glucose metabolism. In addition, HJC0152 can serve as a molecular probing tool for elucidating the key factors responsible for developing breast cancer and other metabolic diseases.

Citation Format: Hyejin Kim, Jiabin Dong, Jimin Xu, Dengfeng Li, Zhi Zheng, Na Ye, Ziwei Zhang, Haiying Chen, Jia Zhou, Qiang Shen. Suppression of breast cancer by reprogramming glucose metabolism and energy production with HJC0152 [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 4883.
Abstract 4891: Co-inhibition of AXL and FN14 by pharmacologic inhibitors or genetic inhibition in NSCLC mice models
Authors: Suresh Dhananjay , Mukherjee Soumavo , Zambre Ajit , Ghoshdastidar Shreya , Yadavilli Sairam , Upendran Anandhi , Kannan Raghuraman
Published: August 2018

Non-small Cell Lung Cancer (NSCLC) patients who express EGFR activating mutations are treated with tyrosine kinase inhibitors (TKI) and have a median overall survival time of 22 months. TKI therapy showed only moderate success with poor (37%) survival rate. On the other hand, TKI therapy is ineffective in patients with active KRAS mutation. Acquired drug resistance is one of the major reasons for the failure of the TKI therapy. Studies have shown that overexpression of AXL is one of the possible reasons for the drug resistance. In our previous studies, we have shown that AXL inhibition in NSCLC drug-resistant cells resulted in FN14 survival activation. Subsequently, we have demonstrated that co-inhibition of both AXL and FN14 resulted in increased therapeutic efficacy. Based on these studies, we recognize that both AXL and FN14 play important role in proliferation, invasion and survival pathways. Therefore, synergistic co-inhibition of AXL-FN14 is a viable therapeutic strategy for treating NSCLC. In the present study, we compared the efficacy of inhibition of AXL and FN14 by siRNA (using nanoparticle delivery vehicles) and small-molecule inhibitors in A549 mouse xenografts. Our data further confirms that co-inhibition is effective and results in tumor volume reduction. The details of this study provide a rationale for enhanced treatment of drug resistant NSCLC.

Citation Format: Dhananjay Suresh, Soumavo Mukherjee, Ajit Zambre, Shreya Ghoshdastidar, Sairam Yadavilli, Anandhi Upendran, Raghuraman Kannan. Co-inhibition of AXL and FN14 by pharmacologic inhibitors or genetic inhibition in NSCLC mice models [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 4891.
Abstract 4904: PD-L1 pathway activation as an escape mechanism of resistance to MEK inhibitor treatment in human colorectal cancer models
Authors: Napolitano Stefania , Matrone Nunzia , Belli Valentina , Falco Vincenzo De , Martinelli Erika , Kopetz Scott , Ciardiello Fortunato , Troiani Teresa
Published: August 2018

Molecular mechanisms driving acquired resistance to anti-EGFR therapies in mCRC is complex and biochemically converge to activate the EGFR-RAS-MAPK pathway. In order to understand the mechanism underlying MEKi resistance, SW48 MEKi resistant cell lines (SW48-MR) have been established by in vivo selection. To expand our results, another CRC cell line sensitive to MEK blockade, such as HCT116, has been generated: HCT116-MR. Microarray analysis showed that several genes involved in the PD-L1 pathway were up regulated in SW48-MR versus SW48. Moreover, genes overexpressed in MEKi-resistant tumor were related to the gene signature of CMS4 subtype of CRC. In particular all the pathways that were up regulated in CMS4 tumors were up regulated in the resistant cell line, as confirmed by Western blot and Immunofluorescence analysis. The regulation of PD-L1 expression is regulated by multiple signaling pathways, including JAK/STAT. STAT3 gene is also up regulated in MEKi-resistant tumors. Inhibition of STAT3 expression by siRNA reduced PD-L1 expression and restored MEKi sensitivity. Moreover, a direct correlation has been demonstrated between EGFR and PD-L1. Exogenous activation of EGFR by TGFα stimulation in MEKi sensitive HCT 116 cell lines induced PD-L1 overexpression and resistance to MEKi. Once demonstrated the EGFR role in modulation of PD-L1 expression and subsequently development of acquired resistance to MEKi, we further evaluated the potential mechanism(s) responsible for EGFR activation. One major mechanism is the specific ligand stimulation. Gene expressions analysis revealed that all EGFR ligands were up regulated in MEKi resistant tumors compared to parental tumors. Moreover, we will perform an ELISA assay to confirm this data. To extend these in vitro findings, we will perform an in vivo study using MC38 and CT26 MEKi resistant syngeneic models that we have previously generated. These results suggest a strategy to potentially improve the efficacy of MEK inhibition by co-treatment with other agents and provide an additional therapeutic strategy via modulation of host immune responses in CMS4 CRC tumors.

Citation Format: Stefania Napolitano, Nunzia Matrone, Valentina Belli, Vincenzo De Falco, Erika Martinelli, Scott Kopetz, Fortunato Ciardiello, Teresa Troiani. PD-L1 pathway activation as an escape mechanism of resistance to MEK inhibitor treatment in human colorectal cancer models [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 4904.
Abstract 4912: Targeting cyclin-dependent kinase 7 in gastric cancer
Authors: Huang Jia-Rong , Zhang Wen-Ji , Jiang Qi-Wei , Yang Yang , Wei Meng-Ning , Shi Zhi
Published: August 2018

CDK7 is a member of the cyclin-dependent kinase family, which forms the CDK activating kinase (CAK) complex with Cyclin H and RING finger protein Mat1 to control cell cycle progression and transcription by phosphorylating other CDKs and RNA polymerase II. Upregulation of CDK7 frequently occurs in human gastric cancer (GC). In this study, we demonstrated that inhibition of CDK7 was able to induce cell growth inhibition, cell cycle arrest at G2/M phase and apoptosis with the increasing intracellular reactive oxidative species (ROS) levels. Pretreatment with ROS scavenger N-acety-L-cysteine partly reversed GC cell apoptosis induced by CDK7 inhibition. In the nude mice, inhibition of CDK7 also suppressed the growth of xenograft tumors of GC. Overall, our data showed that inhibition of CDK7 in GC presented outstanding anticancer effect in vitro and in vivo, suggesting that CDK7 is a potential therapeutic target for GC patients.

Citation Format: Jia-Rong Huang, Wen-Ji Zhang, Qi-Wei Jiang, Yang Yang, Meng-Ning Wei, Zhi Shi. Targeting cyclin-dependent kinase 7 in gastric 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 4912.
Abstract 4914: IFI16 is increased in response to chemotherapeutic agents in breast cancer cells
Authors: Lim Gayoung , Ka Na-Lee , Lee Mi-Ock
Published: August 2018

Breast cancer is the most common cancer in women worldwide and chemotherapy is one of the standard treatment options for breast cancer. Chemotherapeutic agents induce severe DNA damage in cancer cells, which have mutations in genes related with DNA repair, resulting in cancer cell death. Thus, controlling DNA damage response is an important factor in determining outcome upon chemotherapy. In this study, we identified IFN-γ inducible protein 16 (IFI16) as a regulator of DNA damage response in breast cancer cells. First, we found that the expression of IFI16 was increased in response to chemotherapeutic agents such as 5-fluouracil, doxorubicin and cisplatin. However, changes in mRNA level of IFI16 was not observed, suggesting that IFI16 expression could be regulated by post-translation modification. Second, nuclear fractionation and confocal analysis revealed that expression of nuclear IFI16 was increased and cytoplasmic IFI16 was decreased in response to doxorubicin. Taken together, IFI16 could be a possible candidate for regulating factor of DNA damage response upon chemotherapy in breast cancer cells.

Citation Format: Gayoung Lim, Na-Lee Ka, Mi-Ock Lee. IFI16 is increased in response to chemotherapeutic agents in 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 4914.
Abstract 4927: Plasma and CNS pharmacokinetics of the CHK-1 inhibitor prexasertib (LY-2606368) in mice bearing orthotopic group 3 medulloblastoma
Authors: Maharaj Anil , Davis Abigail , Zhong Bo , Roussel Martine F. , Stewart Clinton F.
Published: August 2018

Many children with brain tumors have limited treatment options and poor long-term survival. A key protein kinase involved in activating and maintaining the S and G2/M checkpoints is CHK1, which when inhibited in the absence of p53 leads to loss of DNA damage checkpoints and can enhance the activity of many DNA-damaging agents. Prexasertib (LY-2606368), a potent and selective small molecule inhibitor of CHK1 protein kinase activity in vitro (IC50 <1 nM), is currently under clinical evaluation for treatment of pediatric malignancies. To evaluate prexasertib in preclinical efficacy studies for pediatric brain tumor models, the ratio of free drug concentrations between the central nervous system (CNS) and plasma (Kpu,u) is typically computed as a surrogate to measure CNS penetration. To facilitate the prospective determination of preclinical CNS Kpu,u values, we evaluated the plasma pharmacokinetic (PK) disposition of prexasertib in a murine model of medulloblastoma. A plasma PK study was performed using CD1 nude mice orthotopically implanted with group 3 medulloblastoma to obtain initial prexasertib PK parameters. Tumor growth was assessed by bioluminescence imaging (D-Luciferin) and mice were enrolled in the study once tumors reached 1x10~8-9 photons/second. Prexasertib mesylate monohydrate was reconstituted in 20% Captisol and administered subcutaneously at a dosage of 10 mg/kg (free base). A population based study design was used in which 3 samples per mouse were obtained. Plasma samples were prepared using a liquid-liquid extraction technique, and analyzed for prexasertib using a validated LC-MS/MS method (LLOQ = 0.2 µg/L). A two-compartment PK model was fit to the prexasertib concentration-time data using nonlinear mixed effects modeling (NONMEM 7.3, ICON Development Solutions). Plasma PK parameters from this model were used with the D-optimality criterion as implemented in ADAPT (D'Argenio, 1981) to derive a limited sampling model (LSM) for use in upcoming cerebral microdialysis studies. The results of our plasma PK studies showed a Cmax, terminal half-life, and AUC in CD1 nude mice bearing group 3 medulloblastoma of 1015 µg/L, 4.5 hours, and 1773 µg*hr/L, respectively. The LSM for plasma PK that will be used in the cerebral microdialysis studies include the time points of 0.25, 6.6, and 24 hours. Now that the plasma PK and a LSM are defined, we are poised to begin cerebral microdialysis studies in CD1 nude mice bearing group 3 medulloblastoma.

Citation Format: Anil Maharaj, Abigail Davis, Bo Zhong, Martine F. Roussel, Clinton F. Stewart. Plasma and CNS pharmacokinetics of the CHK-1 inhibitor prexasertib (LY-2606368) in mice bearing orthotopic group 3 medulloblastoma [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 4927.
Abstract 493: Circulating biliary tract microRNA signature discriminates cholangiocarcinoma from pancreatic cancer
Authors: Puik Jisce R. , Meijer Laura L. , Large Tessa Y. Le , Heger Michal , Dijk Frederike , Funel Niccola , Garajová Ingrid , Giovannetti Elisa , Kazemier Geert
Published: August 2018

Accurate diagnosis of lesions in the pancreatic head is a challenge with substantial clinical consequences in daily practice. Located in the pancreatic head, both distal cholangiocarcinoma (CCA) and pancreatic ductal adenocarcinoma (PDAC) are highly lethal malignancies with limited but distinct therapeutic options. Given these different therapeutic considerations, discovery of novel biomarkers that can ensure differential diagnostic specificity is an unmet clinical need. This study aims to identify a circulating microRNA (miRNA) signature to accurately diagnose and discriminate distal CCA from PDAC. In the initial discovery phase, microarray profiling of a panel of 752 miRNAs was performed on seven distal CCA and seven age- and sex-matched healthy control plasma samples. Raw data were normalized to the global mean. Based on probability value and fold change, candidate miRNAs were selected for further validation. In the validation phase, selected candidate miRNAs were analyzed in an independent cohort of distal CCA (n = 22) and healthy controls (n = 32) using RT-qPCR. To further assess the origin of lesions, candidate miRNA expression profiles were analyzed in malignant disease (n = 90) compared to benign disease (n = 15), and distal CCA (n = 22) compared to PDAC (n = 30). Following an empirically based selection of potential reference genes, raw data were normalized to a combination of two normalizing reference miRNAs. In the discovery phase, microarray analysis revealed nineteen miRNAs that were significantly deregulated in patients with distal CCA compared to healthy controls. Thirteen miRNAs were selected for further validation. In the validation phase, using logistic regression analysis, a three-miRNA panel was identified as the most robust diagnostic signature to discriminate malignant from benign disease (AUC = 0.881). Remarkably, two miRNAs of this panel were validated as specific classifier for distal CCA, with an AUC of 0.814 in differentiating distal CCA from PDAC. In addition, a combination of miR-93 and miR-101 was used as bona fide normalizing reference gene for these analyses. In conclusion, a combined panel of plasma miRNAs shows promising diagnostic capability to serve as unique minimally invasive distal CCA biomarker and has the potential to help clinical decision making for patients affected by pancreatobiliary disease in the pancreatic head.

Citation Format: Jisce R. Puik, Laura L. Meijer, Tessa Y. Le Large, Michal Heger, Frederike Dijk, Niccola Funel, Ingrid Garajová, Elisa Giovannetti, Geert Kazemier. Circulating biliary tract microRNA signature discriminates cholangiocarcinoma from 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 493.
Abstract 4953: Metabolic targeting of chemoresistance perturbs clonal complexity in pancreatic cancer
Authors: Loponte Sara , Corti Denise , Seth Sahil , Poggetto Edoardo Del , Ho I-Lin , Li Chieh-Yuan , Jiang Shan , Marszalek Joseph R. , Francesco Maria Emilia Di , Genovese Giannicola , Draetta Giulio , Carugo Alessandro , Viale Andrea
Published: August 2018

A major barrier to achieving durable remission and definitive cure in oncology patients is the emergence of tumor resistance, a common outcome of different disease types independent from the therapeutic approach undertaken. Patients with pancreatic ductal adenocarcinoma (PDAC) continue to have a poor prognosis despite concerted efforts to advance new drugs to the clinic. One reason for this, in PDAC and other tumors, is that tumors are constantly adapting and evolving in response to external perturbations. To better investigate tumor evolution in response to therapy we developed a new clonal tracking platform that enables the in vivo study of long term self-renewing compartments and the generation of cohorts of patient-derived xenografts in which tumors are virtually identical and maintained by the same clones (clonal replica tumors), representing a unique tool to address fundamental questions about clonal dynamics in response to pharmacological treatment. Using this novel approach we demonstrate that standard of care in pancreatic cancer, despite inducing tumor regression, has minimal effect on the clonal composition of tumors that eventually relapse. Transcriptomic and metabolic characterization of residual tumor cells in patient derived xenograft models as well as in patients after chemoradiation shows that resistant cells that contribute to tumor relapse are metabolically rewired to upregulate mitochondrial respiration (OXPHOS). Combining a novel inhibitor of oxidative phosphorylation (IACS-010759), developed at the MD Anderson Institute for Applied Cancer Science and currently in phase I clinical trial in relapsed/refractory acute myelogenous leukemia and advanced solid tumors, with standard of care drugs drastically reduces tumor clonal complexity, underscoring the promise of inhibiting mitochondrial respiration as a new therapeutic strategy to prolong patient survival by eradicating resistant clones that survive chemoradiation.

Citation Format: Sara Loponte, Denise Corti, Sahil Seth, Edoardo Del Poggetto, I-Lin Ho, Chieh-Yuan Li, Shan Jiang, Joseph R. Marszalek, Maria Emilia Di Francesco, Giannicola Genovese, Giulio Draetta, Alessandro Carugo, Andrea Viale. Metabolic targeting of chemoresistance perturbs clonal complexity in 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 4953.
Abstract 4962: Metabolic rewiring of macrophages by CpG stimulates anti-tumor activity that overrides CD47-resistance in pancreatic cancer
Authors: Liu Mingen , O'Connor Roddy S. , Trefely Sophie , Snyder Nathaniel W. , Beatty Gregory L.
Published: August 2018

Although macrophages commonly support cancer growth, they can be induced with anti-tumor functions - such as the ability to phagocytose and kill tumor cells. This activity is governed by stimulatory as well as inhibitory signals (e.g. CD47). The role of central carbon metabolism in determining macrophage function, however, remains poorly understood. Here, we identify CpG-activated changes in the metabolism of macrophages to be a major determinant of their phagocytic and anti-tumor capabilities, including the ability to overcome inhibitory CD47 expressed by tumor cells. In a fully syngeneic model of murine pancreatic ductal adenocarcinoma (PDAC), systemic delivery of CpG re-directed macrophages to mediate anti-tumor activity against PDAC tumor cells independent of CD47 expressed by tumor cells. The anti-tumor potential stimulated by CpG was not associated with polarization of macrophages toward classical M1 or M2 phenotypes, but rather was dependent on a hybrid state of metabolic features found in M1 and M2 macrophages. These metabolic changes required shunting of tricarboxylic acid (TCA) cycle intermediates for de novo biosynthesis of lipids and elevated fatty acid oxidation (FAO) that was supported by glutamine anaplerosis. Coordination of these processes by carnitine palmitoyltransferase 1A (CPT1A) and ATP citrate lyase (ACLY) supported the ability of macrophages to bypass the CD47 inhibitory pathway and mediate anti-tumor functions. Our findings underscore a novel role for the metabolic state of macrophages in overriding barriers to anti-tumor activity, and reveal potential targets that may enhance CD47-based therapies.

Citation Format: Mingen Liu, Roddy S. O'Connor, Sophie Trefely, Nathaniel W. Snyder, Gregory L. Beatty. Metabolic rewiring of macrophages by CpG stimulates anti-tumor activity that overrides CD47-resistance in 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 4962.
Abstract 4963: Targeting glutamine metabolism as a means of enhancing antitumor T-cell responses
Authors: Leone Robert , Englert Judson , Sun Im-Meng , Oh Min-Hee , Alt Jesse , Sun Im-Hong , Tam Ada J. , Majer Pavel , Rais Rana , Slusher Barbara , Powell Jonathan
Published: August 2018

Tumors possess specialized metabolic programming that facilitates their rapid growth. We have developed a novel glutamine antagonist (JHU-083) that not only demonstrates single-agent efficacy in a variety of mouse tumor models but also synergizes with anti-PD-1, adoptive cellular therapy (ACT) and A2aR antagonism to overcome resistance to immunotherapy and promote durable cures. Interestingly, we have observed that JHU-083 markedly enhances endogenous antitumor T-cell responses even in the absence of additional immunotherapy. Flow cytometry analyses of tumor-infiltrating lymphocyte (TIL) reveal markedly increased infiltration of CD8+ T cells with increased proliferative index (Ki67) and the generation of robust memory phenotypes in JHU-083 treated mice. Interestingly, Gene Set Enrichment Analyses (GSEA) using RNA-sequencing data revealed that CD8+ TIL from vehicle treated mice showed significantly enhanced expression of apoptotic transcriptional programs compared with TIL from mice treated with JHU-083. Consistent with flow cytometry data, GSEA showed CD8+ TILs from mice treated with glutamine antagonism expressed transcriptional programs characteristic of long-lived memory cells. To this end, vaccination experiments in mice treated with JHU-083 demonstrate that many of the phenotypic changes observed in TIL can be generated outside the TME and are due to direct effects of glutamine blockade on effector T cells. Our studies show that these transcriptional changes are associated with profound remodeling of the histone epigenetic code and correlate with decreased intracellular levels of α-ketoglutarate in response to glutamine antagonism. Supplementation with dimethyl α-ketoglutarate, a cell-permeable form of α-ketoglutarate, partially reverses phenotypic and epigenetic changes in CD8 T cells undergoing activation in the setting of glutamine antagonism. These findings are consistent with inhibition of a family of enzymes known as α-ketoglutarate dependent dioxygenases, which are responsible for a broad range of demethylation reactions, including histone and DNA demethylation. Overall, our work demonstrates that glutamine antagonism can directly inhibit tumor growth and survival while also reprogramming maladaptive antitumor T-cell responses to enhance endogenous antitumor immunity. Such observations support the development of our novel glutamine antagonists as both monotherapy and in combination with immunotherapy.

Citation Format: Robert Leone, Judson Englert, Im-Meng Sun, Min-Hee Oh, Jesse Alt, Im-Hong Sun, Ada J. Tam, Pavel Majer, Rana Rais, Barbara Slusher, Jonathan Powell. Targeting glutamine metabolism as a means of enhancing antitumor T-cell responses [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 4963.
Abstract 4999: Identifying intercellular phenotypic stability factors for a hybrid epithelial-mesenchymal phenotype
Authors: Jolly Mohit Kumar , Bocci Federico , Tripathi Satyendra C. , Aguilar Mitzi , Hanash Samir M. , Levine Herbert , Onuchic Jose N.
Published: August 2018

Epithelial-mesenchymal transition (EMT) and its reverse Mesenchymal-epithelial transition (MET) are crucial for embryonic development, wound healing and cancer metastasis. Cells need not undergo full EMT or MET, rather they can maintain stably a hybrid epithelial/mesenchymal (E/M) phenotype and thus exhibit collective cell migration, forming clusters of circulating tumor cells - the primary drivers of metastasis. Mechanisms enabling cells to exhibit a hybrid E/M phenotype stably remain elusive. Here, using an integrated experimental-computational approach, we identify ‘phenotypic stability factors' (PSFs) that can stabilize a hybrid E/M phenotype by mediating cell-cell communication through Notch signaling pathway. Our mathematical model predicts that Numb or Numb-like (Numbl) can inhibit a full EMT and stabilize a hybrid E/M phenotype. This prediction is validated by the observation that knockdown of Numb in stably hybrid E/M cells H1975 results in a full EMT, thereby indicating that Numb behaves as a PSF. Generalizing the mathematical model to a multi-cell level, Numb is predicted to alter the balance of hybrid E/M versus mesenchymal cells in clusters, potentially resulting in a higher tumor-initiation ability. Our model also suggests another PSF – Nrf2 (nuclear factor E2-related factor 2) – that couples both to EMT and Notch signaling, and stabilizes a hybrid E/M phenotype too. Intriguingly, Nrf2 levels are maximally observed in a hybrid E/M phenotype as compared to purely epithelial or mesenchymal phenotypes. Finally, high levels of Numb or Nrf2 also correlate with a worse survival in multiple independent cancer datasets, strengthening the association of a hybrid E/M phenotype with increased cancer aggressiveness.

Citation Format: Mohit Kumar Jolly, Federico Bocci, Satyendra C. Tripathi, Mitzi Aguilar, Samir M. Hanash, Herbert Levine, Jose N. Onuchic. Identifying intercellular phenotypic stability factors for a hybrid epithelial-mesenchymal phenotype [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 4999.
Abstract 50: Elucidating the role of MUC5AC-MET axis in breast cancer brain metastasis
Authors: Nasser Mohd W. , Gautam Shailendra K. , Kanchan Ranjana , Atri Pranita , Siddiqui Jawed , Bhatia Rakesh , Carmicheal Joseph , Ghersi Dario , Kaur Sukhwinder , Mahapatra Siddharth , Jain Maneesh , Batra Surinder K.
Published: August 2018

Metastasis of breast cancer (BC) is the leading cause of mortality in American women. In particular, brain metastasis is a significant clinical problem present in 30-40% of triple receptor negative (TN) and epidermal growth factor receptor 2 (HER2) positive BC patients. These patients also have the worst prognosis of all breast cancers in women. Due to advances in clinical management at both the diagnostic and therapeutic levels, the lifespan or overall survival of these patients has increased, but the manifestation of brain metastasis at later stages of the disease remains an unfortunate burden for these patients. In addition, the blood-brain barrier (BBB) poses an additional hurdle for treatment of BC brain metastases, because the majority of chemotherapeutic drugs fail to cross the BBB. Furthermore, there is no single targeted therapy available for TNBC patients with brain metastasis, and thus there is an urgent requirement to discover novel biomarkers and therapeutics to address BC brain metastasis. In this regard, using a bioinformatics approach, we observed that several mucins, including MUC5AC, are significantly up-regulated in brain metastases of BC compared to primary tumors. However, we found the secretory mucin MUC5AC protein to be substantially high in brain-seeking BC (BSBC) cell lines and metastatic brain tissues. We also observed that knockdown of MUC5AC reduced proliferation and metastatic potential in these cell lines. Furthermore, we observed that MUC5AC colocalizes with activated MET receptor. MUC5AC knockdown in BSBC cell lines reduced MET activation and its downstream targets FAK and paxillin. Interestingly, MET and FAK inhibition using pharmacological inhibitors led to substantial decrease in MUC5AC expression. Overall, these studies suggest that MUC5AC-MET axis may regulate breast cancer brain metastasis. Understanding the MUC5AC-MET axis could be a novel approach for the development of better prognosis and prevention of life-threatening and treatment-resistant breast cancer brain metastasis.

Citation Format: Mohd W. Nasser, Shailendra K. Gautam, Ranjana Kanchan, Pranita Atri, Jawed Siddiqui, Rakesh Bhatia, Joseph Carmicheal, Dario Ghersi, Sukhwinder Kaur, Siddharth Mahapatra, Maneesh Jain, Surinder K. Batra. Elucidating the role of MUC5AC-MET axis in breast cancer brain metastasis [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 50.
Abstract 5009: Defined factors overcome T-cell exhaustion via abscopal effect
Authors: Khalil Danny , Campesato Luis Felipe , Budhu Sadna , Li Yanyun , Jones Caitlin , Suek Nathan , Liu Cailian , Gasmi Billel , Giese Rachel , Pourpe Stephane , Merghoub Taha , Wolchok Jedd D.
Published: August 2018

Irreversible exhaustion of tumor-associated T cells is an important factor limiting the efficacy of PD-1 blockade in a number of cancers. Recent data suggest that PD-1 blockade functions largely by augmenting T cell costimulation. Given that activated APCs provide necessary T-cell costimulation, we hypothesized that enforced activation of intratumoral APCs would prime tumor-specific T cells and synergize with PD-1 blockade to overcome T-cell exhaustion. After screening various agents in syngeneic murine models, we found that dual CD40/TLR4 activation within a single tumor triggers a systemic tumor-specific CD8 T-cell response that is dependent on BATF3+ dendritic cells. Remarkably, this approach abolishes exhausted PD-1+ intratumoral T cells in treated as well as distant tumors while sparing more proximal T cells outside the tumors. In addition to treating large established tumors, this approach also confers persistent immunity allowing animals to reject reimplanted tumors 90 days after treatment. Dual CD40/TLR4 activation within a single tumor is thus a promising method for overcoming tumor-associated T-cell exhaustion in a manner that may provide durable systemic control of metastatic human cancers while sparing healthy tissues.

Citation Format: Danny Khalil, Luis Felipe Campesato, Sadna Budhu, Yanyun Li, Caitlin Jones, Nathan Suek, Cailian Liu, Billel Gasmi, Rachel Giese, Stephane Pourpe, Taha Merghoub, Jedd D. Wolchok. Defined factors overcome T-cell exhaustion via abscopal effect [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 5009.
Abstract 5027: Heterocellular 3D scaffolds as biomimetic to recapitulate the tumor microenvironment of peritoneal metastases in vitro and in vivo
Authors: Wever Olivier De , Dejaeghere Emiel , Vlieghere Elly De
Published: August 2018

Peritoneal metastases cause cancer mortality and preclinical models are urgently needed to boost therapeutic progress. This study reports on a hybrid hydrogel-polylactic acid (PLA) scaffold that mimics the architecture of peritoneal metastases at the qualitative, quantitative and spatial level. Porous PLA scaffolds with controllable pore size, shape, and surface properties are functionalized by type I collagen hydrogel and co-seeding of cancer-associated fibroblast (CAF) increases cancer cell adhesion, recovery, and exponential growth by in situ heterocellular spheroid formation. Scaffold implantation at the peritoneal wall allows long-term follow-up (>14 weeks) and results in a time-dependent increase in vascularization, which correlates with cancer cell colonization in vivo. CAF, endothelial cells, macrophages and cancer cells show spatial and quantitative aspects as similarly observed in patient-derived peritoneal metastases. CAF provide long-term secretion of complementary paracrine factors implicated in spheroid formation in vitro as well as in recruitment and organization of host cells in vivo. In conclusion, the multifaceted heterocellular interactions that occur within peritoneal metastases are reproduced in this tissue-engineered implantable scaffold model.

Citation Format: Olivier De Wever, Emiel Dejaeghere, Elly De Vlieghere. Heterocellular 3D scaffolds as biomimetic to recapitulate the tumor microenvironment of peritoneal metastases in vitro and in vivo [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 5027.
Abstract 5041: Repair of azoxymethane-induced nuclear and mitochondrial DNA lesions in mouse liver
Authors: Torres-Ramos Carlos A. , Rodriguez-Munoz Adlin , Ballista-Hernandez Joan , Torres-Ortiz Ceidy , Ayala-Pena Sylvette
Published: August 2018

Background & Objectives: Azoxymethane (AOM) is a carcinogen that induces DNA lesions mainly repaired by the base excision repair (BER) pathway. Apurinic/apyrimidinic endonuclease 1 (APE1) is a central component of BER. Mice with homozygous deletion of the Apex1 gene (which encodes APE1) cannot survive development but heterozygous animals (Apex1+/-) have a normal lifespan. Our aim was to study the role of APE1 in the repair of DNA lesions and changes in mitochondrial DNA (mtDNA) abundance induced by AOM in the liver. Methods: We isolated DNA from 6-month-old C57BL/6 WT and Apex1+/- mice that were treated with a single AOM dose (10 mg/kg) and sacrificed 24, 48 and 72 hours after treatment. To quantify nuclear DNA (nDNA) lesions, mtDNA lesions and changes in mtDNA abundance in liver tissue we applied a PCR based assay (QPCR). Results: Our data show that 24h after AOM treatment both WT and Apex1+/- mice exhibit significant levels of nDNA and mtDNA damage. The lesion number decreases 48 and 72h after treatment but remains statistically elevated as compared to their respective controls. A two tailed ANOVA analysis shows no significant differences in DNA lesions between genotypes. However, Apex1+/- mice show a statistically significant 13% decrease in mtDNA abundance 72 hours after treatment. Conclusions: These results indicate that the liver is a major target of AOM and that APE1 plays a role in preventing AOM-induced loss of mtDNA abundance. Acknowledgments: Supported by R25GM061838, 2G12RR003051 and U54CA096297.

Citation Format: Carlos A. Torres-Ramos, Adlin Rodriguez-Munoz, Joan Ballista-Hernandez, Ceidy Torres-Ortiz, Sylvette Ayala-Pena. Repair of azoxymethane-induced nuclear and mitochondrial DNA lesions in mouse liver [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 5041.
Abstract 5045: Inflammation is upregulated in the normal colonic epithelium and stroma of Apc+/Min-FCCCmice with colon tumors
Authors: Leystra Alyssa A. , Vanderveer Lisa , Zhou Yan , Cooper Harry S. , Grivennikov Sergei , Clapper Margie L.
Published: August 2018

The Apc+/Min-FCCC mouse harbors a point mutation in the Apc tumor suppressor gene and is highly susceptible to the development of multiple colon tumors. Despite being maintained for many years as an inbred colony, Apc+/Min-FCCC mice are heterogeneous with respect to the time to colon tumor development and the number of colon tumors that ultimately form. This model therefore serves as a powerful tool with which to better understand the earliest events that lead to tumorigenesis. Identification of early alterations in these animals may provide targets for effective, long-term tumor prevention. The goal of this study was to characterize early changes in gene expression in the nonneoplastic colonic epithelium and stroma that may contribute to tumor formation in Apc +/Min-FCCC mice. Animals were defined as ‘tumor-bearing' if they harbored at least one gross colon tumor at necropsy. No lesions were detected at necropsy in the remaining animals (‘tumor-free'). Normal tissue was collected for analysis from the distal colon at least 1 cm away from the nearest tumor. Histologically-normal epithelial and stromal cells were microdissected from frozen tissue sections. Gene expression profiling was performed using the Agilent Mouse 4x44K microarray chip. Differentially expressed genes were selected using an FDR of 10% and a ≥ 2-fold change in gene expression. Gene set enrichment analyses confirmed the enrichment of stromal genes in samples of isolated stroma (p< 0.05). Numerous genes were differentially expressed in tumor-free vs. tumor-bearing ApcMin/+ mice, in both the epithelial and stromal compartments. Gene Ontology enrichment analyses revealed the associated biological processes that were dysregulated, including inflammation, immune cell migration, and response to bacterium (FDR<0.1%). These findings indicate that inflammatory processes are altered in the nonneoplastic colon well in advance of tumor formation and represent promising targets for preventive intervention.

Citation Format: Alyssa A. Leystra, Lisa Vanderveer, Yan Zhou, Harry S. Cooper, Sergei Grivennikov, Margie L. Clapper. Inflammation is upregulated in the normal colonic epithelium and stroma of Apc+/Min-FCCCmice with colon tumors [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 5045.