tbuckley/cabot-search huggingface dataset:
Abstract 155: Addition of RP6530, a dual PI3Kδ/γ inhibitor, accentuates Romidepsin activity in NHL cells <i>in vitro</i>
Authors: Srikant Viswanadha, Satyanarayana Eleswarapu, Seeta Nyayapathy, Swaroop Vakkalanka
Journal: Cancer Research (2017)

Abstract Background: Relapsed or refractory Non-Hodgkin’s Lymphoma (NHL) represents a significant unmet medical need. Novel treatment options currently being explored include Romidepsin, a histone deacetylase inhibitor, which has demonstrated significant preclinical and clinical activity. Because the PI3K pathway is often activated in NHL, combining Romidepsin with a PI3K δ/γ inhibitor may be a viable alternative to chemotherapy. RP6530 is a novel, potent, and selective PI3K δ/γ inhibitor that demonstrated high potency against PI3Kδ (IC50=25 nM) and γ (IC50=33 nM) enzymes with selectivity over α (&amp;gt;300-fold) and β (&amp;gt;100-fold) isoforms. Dose-escalation trials to evaluate the safety and efficacy of RP6530 in patients with hematological malignancies are currently underway at several sites in US and Europe. The objective of this study was to evaluate the effect of a combination of Romidepsin and RP6530 in NHL cells. Methods: Combination of Romidepsin and RPRP6530 was evaluated in the following NHL cell lines: OCI-LY-1, OCI-LY-10, RAJI, DAUDI, TOLEDO, JEKO, MAVER, and SU-DHL-1. Synergism or additivity between Romidepsin and RP6530 was determined using different concentrations of the compounds in a 5 x 5 grid and calculated based on the BLISS score. Effect on apoptosis and cell cycle were estimated by flow cytometry. Phosphorylated AKT and PARP were determined by Western Blotting. Results: Synergism or additivity between Romidepsin and RP6530 for anti-proliferative activity was noticed in all the cell lines tested with effect being most pronounced in OCI-LY-1, OCI-LY-10, JEKO, and SU-DHL-1. Incubating cells with 3-5 nM Romidepsin and 3 or 5 μM RP6530 resulted in a 2 to 5-fold increase in the total apoptotic population. Cell cycle analysis indicated that the combination of Romidepsin (1-5 nM) and RP6530 (1-5 μM) caused a G0/G1 arrest (2 to 3-fold) with a corresponding increase in percent of apoptotic cells. While Romidepsin had minimal effect of pAKT, treatment with RP6530 reduced expression by 40-90% across cell lines. Further, Romidepsin + RP6530 enhanced expression of cleaved PARP (8 to 60-fold) compared to either agent alone Conclusions: Addition of RP6530, a potent and selective dual PI3Kδ/γ inhibitor, potentiated Romidepsin activity in NHL cells. Findings provide a rationale for use of the combination in future clinical trials involving naïve or relapsed NHL patients thereby providing a safer alternative to currently available therapy. Citation Format: Srikant Viswanadha, Satyanarayana Eleswarapu, Seeta Nyayapathy, Swaroop Vakkalanka. Addition of RP6530, a dual PI3Kδ/γ inhibitor, accentuates Romidepsin activity in NHL cells in vitro [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 155. doi:10.1158/1538-7445.AM2017-155
Abstract 1922: S-SHIP promoter expression identifies a putative cancer stem cell population in C3(1)/Tag murine mammary tumors
Authors: Lu Tian, Chann Lagadec, Eric Adriaenssens, Emmanuel Bouchaert, Hélène Bauderlique-Le Roy, Xuefen LeBourhis, Roland P. Bourette
Journal: Cancer Research (2017)

Abstract Breast cancer is the most common cancer in women worldwide. The isolation and characterization of breast cancer stem cells (CSC) are crucial for understanding cancer biology and revealing potential therapeutic targets. One of the major issues in the study of CSC is the lack of reliable markers. A transgenic mouse model (Tg 11.5kb–GFP) was generated using the 11.5kb s-SHIP (stem-SH2-containing 5’-Inositol Phosphatase) promoter that specifically expressed enhanced green fluorescent protein (GFP) in embryonic and various tissue stem cells. In the mammary gland, previous experiments showed that GFP labels puberty cap cells and pregnancy basal alveolar bud cells, and it has been demonstrated that these mammary GFP+ cells are activated tissue stem cells. In order to determine if s-SHIP promoter expression could also mark mammary cancer stem cells, we generated a bi-transgenic mouse model by crossing Tg 11.5kb-GFP mice with Tg C3(1)/Tag mice. Tg C3(1)/Tag mice express SV40 T antigen under the regulatory control of the rat prostatic steroid binding protein C3(1) gene. In female mice, the transgene is expressed primarily in the mammary gland. Mice develop mammary hyperplasia by 3 months of age with subsequent development of mammary adenocarcinoma by 6 months of age. Here we show the presence of a rare population of GFP+ cells in mammary tumors of female bi-transgenic mice by using immunohistochemical and flow cytometry analysis. The GFP+ mammary cancer cells are also CD24+/CD49f+/CD29+. As compared to GFP- cells, GFP+ cells exhibit both a higher tumor sphere-forming potential, and a higher tumorigenicity when transplanted into SCID and FVB recipient mice. Moreover, upon subsequent transplantation, the GFP+ cells generated heterogeneous tumors that displayed properties similar to the primary tumor. Currently, we are conducting transcriptomic analysis of these GFP+ and GFP- cells in order to reveal differentially expressed genes. Altogether, these results suggest that s-SHIP promoter expression may be a new marker of mammary CSC that enables their identification and isolation via a single consistent parameter. Furthermore, our bi-transgenic C3(1)/Tag x 11.5kb-GFP mice provide a novel model for study of mammary cancer stem cells and investigation of potential therapeutic targets for breast cancer. Citation Format: Lu Tian, Chann Lagadec, Eric Adriaenssens, Emmanuel Bouchaert, Hélène Bauderlique-Le Roy, Xuefen LeBourhis, Roland P. Bourette. S-SHIP promoter expression identifies a putative cancer stem cell population in C3(1)/Tag murine mammary tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1922. doi:10.1158/1538-7445.AM2017-1922
Targeting the Myofibroblastic Cancer-Associated Fibroblast Phenotype Through Inhibition of NOX4
Authors: Christopher J Hanley, Massimiliano Mellone, Kirsty Ford, Steve M Thirdborough, Toby Mellows, Steven J Frampton, David M Smith, Elena Harden, Cedric Szyndralewiez, Marc Bullock, Fergus Noble, Karwan A Moutasim, Emma V King, Pandurangan Vijayanand, Alex H Mirnezami, Timothy J Underwood, Christian H Ottensmeier, Gareth J Thomas
Journal: JNCI Journal of the National Cancer Institute (2017)

Cancer-associated fibroblasts (CAFs) are tumor-promoting and correlate with poor survival in many cancers, which has led to their emergence as potential therapeutic targets. However, effective methods to manipulate these cells clinically have yet to be developed.
Abstract 242: The anti-tumor effect and the impact on gut microbiota of gypenosides Rd and Rb3 in ApcMin/+ mice
Authors: W.L. Wendy Hsiao, Guoxin Huang, Imran Khan, Lei Chen, Xiaoang Li
Journal: Cancer Research (2017)

Abstract Gut commensal microbiota (GM) is closely associated with health and diseases, including cancer. In our previous studies, we have demonstrated that Gynostemma pentaphyllum saponins (GpS) exert anticancer effects in both athymic and ApcMin/+ mice. Our recent study also showed that GpS exhibited prebiotic effect by up regulating the beneficial bacteria and improving the epithelial microenvironment of the gut. Gypenoside IV (Rb3) and gypenoside VII (Rd) are the two constituents of GpS. The Rd exhibits stronger anticancer effect than Rb3 in an in vitro cell model system established in our lab. Our aim is to confirm the antitumor effect of these two compounds, as well as their impact on GM in the ApcMin/+ mouse model. In the study, single daily dose of Rb3 or Rd at 20mg/kg, and solvent control were orally fed to the mice for 8 weeks, started at 6 weeks of age before the appearance of spontaneous intestinal polyps. The fecal microbiome was analyzed by enterobacterial repetitive intergenic consensus (ERIC)-PCR and 16S rRNA gene pyrosequencing technique. We found that Rd was more effective than Rb3 in reducing polyp formation, and improving the intestinal lining by up-regulating anti-inflammatory cytokines IL-4 and IL-10. Rd, not Rb3, might facilitate intestinal tissue repair by enhancing M2 marker while suppressing M1 markers expression. Moreover, Rd treatment greatly enhanced the ratio of Bacteroidetes vs Firmicutes, while profoundly reduced the abundance of cancer-cachexia-associated bacteria, such as Parabacteroides goldsteinii (22.25%), Dysgonomonas wimpennyi (48.54%) and Blautia coccoides (36.77%). These findings suggested that the level of antitumor effect of the gypenoside might be correlated with its ability to modulate the gut microbiota from dysbiosis state to symbiosis. This study was supported by Macau Science and Technology Development Fund 015/2014/A1. Citation Format: W.L. Wendy Hsiao, Guoxin Huang, Imran Khan, Lei Chen, Xiaoang Li. The anti-tumor effect and the impact on gut microbiota of gypenosides Rd and Rb3 in ApcMin/+ mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 242. doi:10.1158/1538-7445.AM2017-242
Abstract 4833: Perfusion-based bioreactor culture of primary cancer tissue maintains tumor microenvironment complexity and allow <i>in-vitro</i> testing of immune blockade therapy
Authors: Manuele Giuseppe Muraro, Simone Muenst, Celeste Manfredonia, Valentina Mele, Silvio Daester, Alexandar Tzankov, Luigi Terracciano, Walter Weber, Giulio C. Spagnoli, Giandomenica Iezzi, Ivan Martin, Savas D. Soysal
Journal: Cancer Research (2017)

Abstract In vitro culture of primary cancer tissue is still very limited and the generation of patient derived xenograft determine the loss of human-cancer associated stroma. In this context, the use of 3D in vitro systems based on human tissue may be an innovative system to be exploited for keeping the tumor microenvironment (TME) complexity of the tissue in vitro. Freshly excised colorectal (CRC) and breast cancer (BrCa) specimens were fragmented and cultured in 3D “sandwich-like format” between porous collagen scaffolds under perfusion flow (U-CUP, Cellec Biotek AG). The maintenance of tumor and immune-infiltrating cells, survival and phenotypic characterization were histologically assessed. In a second step cancer treatment were tested. U-CUP culture allowed the preservation, viability and expansion of tumor tissue with concomitant stromal and immune cells. Expanding cancer cells were viable after 10 and 21 days (CRC and BrCa, respectively). Administration of anti-ER treatment to Lumina A ER+ BrCa was associated with decreased expansion of cancer tissue into the scaffold after 21 days. The maintenance of immune-infiltrating cells allowed testing of immune blockade therapy. Administration of anti-PDL1 antibody, alone or in combination with anti-CTLA4, to the culture medium was associated with increased expression of markers of immune-activation (i.e. IFNγ) and decreased expression of immunosuppressive cytokine IL10. Preserving malignant, interstitial and immunocompetent cells comprised in surgically excised tumor specimens might allow a direct evaluation of the effects of various treatments on the complex TME. This engineered in vitro model could allow animal-free testing and it could be extended as a platform allowing the testing of innovative approaches for the treatment of human malignancies. Our findings shed the light on a promising system for selecting personalized treatment based on a patient’s tumor specific microenvironment. Note: This abstract was not presented at the meeting. Citation Format: Manuele Giuseppe Muraro, Simone Muenst, Celeste Manfredonia, Valentina Mele, Silvio Daester, Alexandar Tzankov, Luigi Terracciano, Walter Weber, Giulio C. Spagnoli, Giandomenica Iezzi, Ivan Martin, Savas D. Soysal. Perfusion-based bioreactor culture of primary cancer tissue maintains tumor microenvironment complexity and allow in-vitro testing of immune blockade therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4833. doi:10.1158/1538-7445.AM2017-4833
Abstract 2442: Simplified sequencing of full-length isoforms in cancer on the PacBio Sequel System
Authors: Meredith Ashby, Ting Hon, Elizabeth Tseng, Aparna Vedula, Tyson A. Clark
Journal: Cancer Research (2017)

Abstract Tremendous flexibility is maintained in the human proteome via alternative splicing, and cancer genomes often subvert this flexibility to promote survival. Identification and annotation of cancer-specific mRNA isoforms is critical to understanding how mutations in the genome affect the biology of cancer cells. While microarrays and other NGS-based methods have become useful for studying transcriptomes, these technologies yield short, fragmented transcripts that remain a challenge for accurate, complete reconstruction of splice variants. In cancer proteomics studies, the identification of biomarkers from mass spectroscopy data is often limited by incomplete gene isoform expression information to support protein to transcript mapping. The Iso-Seq™ protocol developed at PacBio offers the only solution for direct sequencing of full-length, single-molecule cDNA sequences needed to discover biomarkers for early detection and cancer stratification, to fully characterize gene fusion events, and to elucidate drug resistance mechanisms. Knowledge of the complete isoform repertoire is also key for accurate quantification of isoform abundance. As most transcripts range from 1 - 10 kb, fully intact RNA molecules can be sequenced using SMRT® Sequencing without requiring fragmentation or post-sequencing assembly. However, some cancer research applications have presented a challenge for the Iso-Seq protocol, due to the combination of limited sample input and the need to deeply sequence heterogenous samples. Here we report the optimization of the Iso-Seq library preparation protocol for the PacBio Sequel platform and its application to cancer cell lines and tumor samples. We demonstrate how loading enhancements on the higher-throughput Sequel instrument have decreased the need for size fractionation steps, reducing sample input requirements while simultaneously simplifying the sample preparation workflow and increasing the number of full-length transcripts per SMRT Cell. The results highlight the potential for broader application of the Iso-Seq method to more comprehensively characterize alternative splicing in cancer. Citation Format: Meredith Ashby, Ting Hon, Elizabeth Tseng, Aparna Vedula, Tyson A. Clark. Simplified sequencing of full-length isoforms in cancer on the PacBio Sequel System [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2442. doi:10.1158/1538-7445.AM2017-2442
Abstract 352: Myocardial Neuregulin-1/ErbB Signaling Is Impaired in the Setting of Post--Myocardial Infarction Heart Failure Complicated by Type 1 Diabetes Mellitus
Authors: Oghenerukevwe Odiete, Kathleen E Dennis, Douglas B Sawyer, Michael F Hill
Journal: Circulation Research (2012)

Background: Type 1 diabetes mellitus (DM) patients surviving myocardial infarction (MI) are at heightened risk for the subsequent development of heart failure (HF). Despite the worse outcomes, investigations into the pathophysiological mechanisms that contribute to the increased frequency of HF after MI in the type 1 DM heart remain scarce. Neuregulin-1 (NRG-1), along with the ErbB family of receptor tyrosine kinases through which NRG-1 ligands signal, have been shown to be intimately involved in mediating cardiac recovery after MI. However, the impact of type 1 DM on this signaling system post-MI remains to be elucidated. Therefore, in the present study, we examined myocardial NRG-1/ErbB signaling during post-MI HF in the presence of type 1 DM. Methods: Type 1 DM was induced in male Sprague-Dawley rats via a single intraperitoneal injection of streptozotocin (STZ) (65 mg/kg). Two weeks after induction of type 1 DM, MI was produced in DM and non-DM rats by ligation of the left anterior descending (LAD) coronary artery. Residual left ventricular (LV) function was assessed by echocardiography at 4 weeks post-MI. Following echocardiographic assessment, NRG-1, ErbB2, and ErbB4 protein expression was assessed in the remote, surviving LV myocardium of DM and non-DM rats using Western blot techniques. Results: LV Fractional Shortening (FS) and LV Ejection Fraction (EF) were significantly lower in the DM + MI group compared to the MI group ([LVFS: DM + MI, 17.9 ± 0.7 (n=6) vs. MI, 25.2 ± 2.2 (n=6), p &lt;0.05; LVEF: DM + MI, 35.5 ± 1.4 (n=6) vs. MI, 47.5 ± 3.5 (n=6), p &lt;0.05]), indicating an increased functional severity of HF in the diabetic post-MI group. The weight of myocardial scar caused by the infarction was not significantly different between the MI groups ([DM + MI, 0.19 ± 0.02 g (n=4) vs. MI, 0.20 ± 0.03 g (n=4), p =0.70]). ErbB2, ErbB4, and NRG-1 protein expression levels were all significantly lower in the DM + MI group compared to the MI group. Conclusions: These findings demonstrate that type 1 DM impairs myocardial NRG-1/ErbB signaling in response to MI, which may contribute to the accelerated progression of subsequent HF. Augmentation of NRG-1 or its downstream signaling pathways may represent a novel therapeutic strategy for ameliorating post-MI HF in the setting of type 1 DM.
Fatal Meningitis in Swine after Intrathecal Administration of Adeno-associated Virus Expressing Syngeneic Interleukin-10
Authors: Mark D. Unger, Josef Pleticha, James E. Collins, Anibal G. Armien, Jennifer L. Brazzell, Laura K. Newman, Lukas F. Heilmann, Jodi A. Scholz, Timothy P. Maus, Andreas S. Beutler
Journal: Molecular Therapy (2017)

Interleukin-10 (IL-10) delivered by intrathecal (i.t.) gene vectors is a candidate investigational new drug (IND) for several chronic neurological disorders such as neuropathic pain. We performed a preclinical safety study of IL-10. A syngeneic large animal model was used delivering porcine IL-10 (pIL-10) to the i.t. space in swine by adeno-associated virus serotype 8 (AAV8), a gene vector that was previously found to be nontoxic in the i.t. space. Unexpectedly, animals became ill, developing ataxia, seizures, and an inability to feed and drink, and required euthanasia. Necropsy demonstrated lymphocytic meningitis without evidence of infection in the presence of normal laboratory findings for body fluids and normal histopathology of peripheral organs. Results were replicated in a second animal cohort by a team of independent experimenters. An extensive infectious disease and neuropathology workup consisting of comprehensive testing of tissues and body fluids in a specialized research veterinary pathology environment did not identify a pathogen. These observations raise the concern that i.t. IL-10 therapy may not be benign, that previously used xenogeneic models testing the human homolog of IL-10 may not have been sensitive enough to detect toxicity, and that additional preclinical studies may be needed before clinical testing of IL-10 can be considered.
Abstract LB-119: Targeting DNA-PK via small molecule inhibitors of the Ku-DNA interaction
Authors: John J. Turchi, Navnath Gavande, Pamela VanderVere-Carozza, Tyler Vernon, Katherine Pawelczak
Journal: Cancer Research (2017)

Abstract DNA-PK, the DNA-dependent protein kinase, is a validated target for cancer therapeutics that drives the DNA damage response and plays a critical role in the nonhomologous end joining (NHEJ) DNA repair pathway. Inhibition of DNA-PK activity can elicit anticancer activity and also sensitize cancer cells to DNA damaging therapy. NHEJ is responsible for the repair of DNA double strand breaks (DSB), particularly those induced by ionizing radiation (IR). The generation of DNA DSBs is the mechanism of clinical efficacy of radiation therapy and numerous DNA damaging chemotherapeutic drugs used to treat various cancers. Modulating the pathway responsible for repairing these breaks has been shown to have a profound impact on the efficacy of IR or chemotherapy in the clinic. We have taken a completely unique and novel approach to inhibiting DNA-PK that is based on extensive knowledge regarding DNA-PK kinase activation and affords considerable advantages to current approaches in DNA-PK inhibition. DNA-PK activation and NHEJ pathway engagement requires binding of the Ku70/80 heterodimer to DNA ends and we are targeting this protein-DNA interaction. We have discovered and developed a series of highly potent and selective DNA-PK inhibitors that act via blocking the binding of Ku70/80 to DNA ends. The lead compound from this series of molecules inhibits DNA-PK catalytic activity at nanomolar concentrations, has single-agent anticancer activity in cancer cell lines, and potentiates cellular sensitivity to IR treatment and DSB-producing chemotherapeutic agents. These data demonstrate the utility of inhibiting DNA-PK via targeting the Ku-DNA interaction. Citation Format: John J. Turchi, Navnath Gavande, Pamela VanderVere-Carozza, Tyler Vernon, Katherine Pawelczak. Targeting DNA-PK via small molecule inhibitors of the Ku-DNA interaction [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-119. doi:10.1158/1538-7445.AM2017-LB-119
Abstract 4425: Analysis of host-Epstein-Barr virus microRNA expression in nasopharyngeal carcinoma
Authors: Wen-Hui Su, Jia-Hao Zhang, Kai-Ping Chang
Journal: Cancer Research (2017)

Abstract In the tumorigenesis process, miRNAs could act as oncogenes or tumor suppressor genes in the cancer cells by regulating miRNA target genes. Nasopharyngeal carcinoma (NPC) associated Epstein-Barr virus (EBV) can also influence the occurrence of NPC through its miRNAs. In this study, we used the next generation sequencing technology to analysis host-viral miRNA profiles in 24 NPC and 5 adjacent normal tissues. In average, 10.62% of mappable reads mapped to EBV genome in NPC tissue. Most of the small RNAs in NPC tissues which mapped to EBV genome were miRNAs (95.51%), rest of them were EBERs (1.80%) and v-snoRNA1 (2.68%). The percentages of EBV miRNA in total human and EBV mature miRNA reads were ranged from 4% to 51% (average 29.61%). For EBV miRNAs, the expression level of BART10-3p and BART9-3p were the top two in NPC tumors. BART20-5p, BART19-3p and BART21-5p were correlated to tumor stage. On the other hand, there were 11 and 34 human miRNAs differentially up- and down-regulated in NPC tissue, respectively. The present research provides the first evidence obtained through direct sequencing to systematically and comprehensively describe the expression of EBV and human miRNAs in NPC tissue. Citation Format: Wen-Hui Su, Jia-Hao Zhang, Kai-Ping Chang. Analysis of host-Epstein-Barr virus microRNA expression in nasopharyngeal carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4425. doi:10.1158/1538-7445.AM2017-4425