bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1872: Oleuropein reduces Prdx1 expression, cell proliferation and viability in K562 human leukemia cells
Authors: Fulco Natalie , Fagan Sophia , Phelan Shelley A.
Published: August 2019

Oleuropein is one of the two main phenolic compounds derived from olive leaves. Numerous studies have found that the polyphenols contained in olive leaves possess strong antioxidant properties, and increase apoptosis in many types of cancer cells, suggesting its possible efficacy as an anti-cancer treatments. The purpose of our study was to examine the effects of oleuropein extract on cell growth and viability of the K562 human leukemia cell line, and to investigate possible molecular mechanisms of action. Oleuropein of 98% purity was used for our studies. We first demonstrated that K562 cells treated with 200 and 400 μg/ml oleuropein exhibited a significant reduction in cell growth and viability after four days as compared to ethanol-treated controls. Cells treated with 200μg/ml oleuropein showed decreased cell proliferation and viability after two days, and exhibited approximately 90% reduction in cell density and viability after four days of treatment. A concentration of 400 μg/ml oleuropein resulted in 100% toxicity by day four. We also examined the effects of oleuropein on cellular cytotoxicity using an LDH release assay and found a two-fold elevation in LDH release within 24 hours of treatment with 200 μg/mL oleuropein. To begin to explore a mechanisms of action, we used western blotting to examine known antioxidant and signaling pathways for possible regulation by oleuropein in these cells. We examined expression of the Peroxiredoxin (Prdx) family of proteins, a group of thiol-specific antioxidants found to be elevated in many types of cancer. We found that 200 ug/ml oleuropein reduced Prdx1 expression by about 50% after eight and twenty-four hours, while having no significant effect on expression levels of Prdx-2, -3, -4, and -6. We are currently investigating the mechanism of cell death induced by oleuropein, as well as the role of Prdx1 in oleuropein-induced toxicity. Together with previously reported cancer-cell specific effects, this research provides a promising new avenue for the use of natural products as anti-cancer agents.

Citation Format: Natalie Fulco, Sophia Fagan, Shelley A. Phelan. Oleuropein reduces Prdx1 expression, cell proliferation and viability in K562 human leukemia cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1872.
Abstract 1877: The role played by SLUG, an epithelial-mesenchymal transition factor, in invasion and therapeutic resistance of malignant glioma
Authors: Lee Kyung-Hwa , Oh Se-Jeong , Ahn Eun-Jung , Jung Shin , Lee Jae-Hyuk , Kim Kyung-Keun , Kim Hangun , Moon Kyung-Sub
Published: August 2019

Purpose: In malignant gliomas, invasive phenotype and cancer stemness promoting resurgence of residual tumor cells, render treatment very difficult. Hence, identification of epithelial-mesenchymal transition (EMT) factors associated with invasion and stemness of glioma cells is critical.

Methods: To address the issue, we investigated several EMT factors in hypermotile U87MG and U251 cells, mouse xenograft model, and human glioma samples. Of several EMT markers, SLUG expression was notably increased at the invasive fronts of gliomas, both in mouse tumor xenografts and human glioma samples. The biological role played by SLUG was investigated using a colony-forming assay after chemotherapy and irradiation, and by employing a neurosphere culture assay. The effect of SLUG on glioma progression was examined in our patient cohort and samples, and compared to large public data from the REMBRANDT and TCGA.

Results: Genetic upregulation of SLUG was associated with increased levels of stemness factors and enhanced resistance to radiation and temozolomide. In our cohort, patients exhibiting lower-level SLUG expression evidenced longer progression-free survival (P=0.042). Also, in the REMBRANDT dataset, a group in which SLUG was downregulated exhibited a significant survival benefit (P<0.001). Although paired gliblastoma multiforme (GBM) samples from our patients did not showed significant increase of SLUG expression, increased mRNA levels of SLUG was found in recurred GBM from TCGA (P=0.052), and in TMZ-treated glioma cells and mouse xenograft.

Conclusion: SLUG may contribute to glioma progression by controlling invasion at infiltrating margins, associated with increased stemness and therapeutic resistance.

Citation Format: Kyung-Hwa Lee, Se-Jeong Oh, Eun-Jung Ahn, Shin Jung, Jae-Hyuk Lee, Kyung-Keun Kim, Hangun Kim, Kyung-Sub Moon. The role played by SLUG, an epithelial-mesenchymal transition factor, in invasion and therapeutic resistance of malignant glioma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1877.
Abstract 1882: Molecular context dictates the effects of eribulin on key EMT regulators: Snail and Slug
Authors: Kaul Roma , Risinger April L. , Mooberry Susan L.
Published: August 2019

Microtubule targeting agents (MTAs) are a mainstay in the treatment of breast cancer and a growing body of evidence demonstrates that they have non-mitotic effects that contribute to their anticancer actions. Even after decades of clinical use, there is much to be learned about the mechanisms of action these drugs, and differences among them, for optimal utility. MTAs rapidly alter microtubule dynamics, often within minutes, leading to significant changes in oncogenic cellular signaling. We evaluated the early effects of eribulin on key oncogenic signaling pathways following a 2 h incubation using clinically relevant concentrations and compared the effects to those initiated by other MTAs. These studies led to the identification of novel mechanisms by which MTAs disrupt oncogenic signaling and contribute to the reversal of epithelial to mesenchymal transition (EMT), including unanticipated differences among drugs. The TGF-β-mediated expression of the Snail transcription factor is a key driver pathway of EMT in breast cancer. The effects of eribulin and other MTAs on TGF-β-induced, Smad-dependent expression of Snail were evaluated. Our results show that eribulin and vinorelbine inhibit the ability of TGF-β to promote the transcriptional induction of Snail by impeding the nuclear transport of Smad2/3 proteins in 4 triple-negative breast cancer cell lines. The effects of microtubule destabilizers contrasted with those of paclitaxel, which had no effect on the expression of Snail. This study begins to explain how microtubule disruption might contribute to the eribulin-mediated EMT reversal. Slug is another member of the Snail family of transcription factors that plays a central role in breast cancer EMT. Although Snail and Slug are often grouped together due to functional similarities, it is now understood that they regulate non-overlapping sets of genes. In contrast to our findings with Snail, eribulin and vinorelbine, but not paclitaxel or ixabepilone, induced Slug over-expression in a subset of breast cancer cell lines. We have identified molecular contexts where eribulin and vinorelbine induced Slug expression, resulting in the identification of a potential biomarker for response to microtubule destabilization. This work highlights the multifaceted nature of MTA-mediated effects on EMT-associated signaling pathways in breast cancer cells. These studies are supported by Eisai Inc.

Citation Format: Roma Kaul, April L. Risinger, Susan L. Mooberry. Molecular context dictates the effects of eribulin on key EMT regulators: Snail and Slug [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1882.
Abstract 1884: Development of a novel VIM-RFP reporter line for colorectal cancer EMT study and drug discovery
Authors: Shu Weiguo , Douglas Diana , Kumari Sangeeta , Romero Luis , Rodriguez Luis G. , Zou Chaozhong , Newman Robert
Published: August 2019

Epithelial-to-Mesenchymal Transition (EMT) describes a dynamic and reversible process by which cells lose their epithelial characteristics and acquire mesenchymal properties. Accumulating evidence indicates that EMT displays an array of intermediate states, a phenotype referred to as “partial MET”. EMT is executed in response to signaling pathway molecules and microRNAs (miRNAs) that induce the expression of specific EMT associated transcription factors (EMT-TFs), including Zeb1/2, Snail1/2, and Twist. There is an ever-growing body of research and clinical evidence indicating that EMT plays an important role in cancer cell dissemination and distal metastasis. Therefore, targeting EMT is considered a novel opportunity in anti-cancer treatment and drug development.

Vimentin, a hallmark of mesenchymal cells, is of increasing interest as a novel anti-cancer therapeutic drug target. In colorectal cancer (CRC) patients, increased expression of the vimentin protein predicts a poor prognosis. Here, we developed a novel CRC HCT-116 VIM-RFP reporter line using CRISPR/Cas9 technology. In this cell line, the RFP reporter was incorporated into the endogenous vimentin gene just before the stop codon at the last exon, enabling real-time monitoring of EMT states in live cells. The VIM-RFP knock-in allele was confirmed at genomic, transcriptional, and translational levels. Bio-functional evaluation data revealed that miRNA-200 inhibitor treatment induced the increased expression of VIM-RFP, and decreased expression of E-cadherin, a hallmark of epithelial cells. The expression of EMT-TFs ZEB-1 and ZEB-2 was also upregulated upon induction. In addition, we showed that induced VIM-RFP cells displayed increased migration capacity. These data suggested that miRNA-200 inhibitor induced VIM-RFP cells have undergone EMT. Azacitidine, a clinically approved demethylating agent, has been extensively evaluated in a number of clinical trials as a treatment for CRC patients. Studies reported that azacitidine can induce MET in a number of cancer cell lines. We showed that azacitidine treatment of VIM-RFP cells can effectively induce RFP expression, suggesting a potential application of this VIM-RFP reporter line as a platform for drug evaluation and compound screening. Taken together, the HCT-116 Vim-RFP EMT reporter line could be a valuable tool for dissecting the molecular mechanisms underlying EMT and for evaluating or screening compounds targeting EMT in CRC.

Citation Format: Weiguo Shu, Diana Douglas, Sangeeta Kumari, Luis Romero, Luis G. Rodriguez, Chaozhong Zou, Robert Newman. Development of a novel VIM-RFP reporter line for colorectal cancer EMT study and drug discovery [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1884.
Abstract 19: Design and synthesis of biaryl indolyl benzamides as HDAC1-selective inhibitors via a fragment-based lead generation approach
Authors: Negmeldin Ahmed T. , Pflum Mary Kay H.
Published: August 2019

Histone deacetylase (HDAC) proteins are epigenetic regulatory enzymes that deacetylate protein substrates, leading to subsequent changes in cell function. HDAC proteins are implicated in cancers and several HDAC inhibitors have been approved by the FDA as anti-cancer drugs. Unfortunately, most of the FDA-approved drugs non-selectively inhibit most of the HDAC isoforms, which may lead to side effects in the clinic and limit their use on the bench top. Isoform-selective HDAC inhibitors may decrease or eliminate the side effects associated with non-selective inhibitors treatment. In addition, isoform-selective HDAC inhibitors can be used as molecular tools to study HDAC-related cancer biology. In this work, several biaryl indolyl benzamide compounds were designed and synthesized based on a fragment-based lead generation approach as potential HDAC1-selective inhibitors. The design was based on combining two different fragments from known HDAC1/2 and HDAC1/3 selective inhibitors. These fragments can discriminate against both HDAC2 and HDAC3 and possibly impart selectivity towards HDAC1. Docking studies were performed to validate the design of the inhibitors and to understand the binding of different isoform-selective inhibitors to HDAC1, HDAC2, and HDAC3. Potency and isoform selectivity of the new compounds were assessed via in vitro screening with different HDAC isoforms. All analogs showed modest selectivity for HDAC1 over HDAC2. Bnz-3 was identified as the best compound in terms of potency and selectivity. In particular, Bnz-3 displayed roughly 10- to 100-fold higher potency than the other compounds, with an IC50 value of 548 nM with HDAC1. In terms of selectivity, Bnz-3 showed about 10.6-fold selectivity for HDAC1 over HDAC2, with almost no inhibition of either HDAC3 or HDAC6. The new biaryl indolyl benzamide HDAC inhibitors represent potential lead compounds that can help to further develop more selective HDAC1 inhibitors.

Citation Format: Ahmed T. Negmeldin, Mary Kay H. Pflum. Design and synthesis of biaryl indolyl benzamides as HDAC1-selective inhibitors via a fragment-based lead generation approach [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 19.
Abstract 1919: Molecularly annotation of mouse avatar models derived from patients with colorectal cancer liver metastasis
Authors: Wang Jingyuan , Liu Wei , Gao Jing , Xing Baocai , Shen Lin
Published: August 2019

Background: Liver is the most common metastatic site in advanced colorectal cancer. Most patients with colorectal cancer liver metastasis (CRLM) do not benefit from current treatment. Patient-derived xenograft (PDX) with defined molecular signatures are attractive models for preclinical studies.

Methods: Successfully established PDXs were evaluated to elucidate their fidelity of patients’ biologic characteristics (pathologic, genetic and protein properties, together with chemosensitivity). The genomic variations of PDXs were analyzed by next-generation sequencing to explore the underlying molecular mechanism of metastasis and potential therapeutic targets.

Results: CRLM (N=73) showed a significantly higher successful PDX establishment rate than primary specimens (N=26; 76.7% vs. 57.7%). CRLM PDXs recapitulated the pathologic, genetic and protein properties of parental tumors, as well as chemosensitivity. Frequent altered genes in PDXs showed high consistency compared to patients’ genomic alterations and were enriched in MAPK, ErbB, cell cycle, focal adhesion pathways for CRLM PDXs, whereas primary tumor-derived PDXs only exhibited genomic variations involving ErbB and cell cycle. The genetic alterations showed high concordance between paired PDXs from primary and metastatic tissues, except for recurrent gene mutations (ARID1A, CDK8, ETV1, STAT5B and WNK3) and common copy number gains in chromosomes 20q (e.g., AURKA/TPX2). Several potential drug targets such as KRAS, HER2, and FGFR2 were validated using corresponding inhibitors. Additionally, PDX models could also be used in screening efficient regimens for patients with no druggable alterations.

Conclusion: This study has successfully established and validated a large panel of molecularly annotated platforms from patients with CRLM for preclinical studies.

Note: This abstract was not presented at the meeting.

Citation Format: Jingyuan Wang, Wei Liu, Jing Gao, Baocai Xing, Lin Shen. Molecularly annotation of mouse avatar models derived from patients with colorectal cancer liver metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1919.
Abstract 1920: PC9 AZD9291-resistant (PC9-OSIR) cell line, a useful cell model for novel drug discovery
Authors: Bai Zhaoshuai , Wang Guoqian , Zhou Xiaolin , Mu Zhongquan , Hao Feng , Ning Jinying
Published: August 2019

Non-small cell lung cancer (NSCLC) is the most common type of lung cancer and the leading cause of lung cancer-related mortality. Identification of EGFR as a driver gene of NSCLC has led to the development of EGFR inhibitors, such as Gefitinib, Erlotinib, Afatinib and Osimertinib (AZD9291). AZD9291, a third-generation EGFR inhibitor, has been approved as the first-line treatment for patients with EGFR T790M mutation-positive NSCLC. Resistance to AZD9291 was observed clinically and C797S mutation was reported to be a major mechanism for resistance to AZD9291.Our group has generated PC9 AZD9291-resistant (PC9-OSIR) cell line using a dose-escalation method after treatment of the cells with mutagenic agent. Specifically, PC9 cells was treated by AZD99291 with increasing dose for 2 weeks, and remaining live cells was retrieved and designated as PC9-OSIR. Cell survival assay showed that IC50 of PC9-OSIR was over 2μM, 100-fold more resistant than the parental PC9 cells. We explored the resistance mechanisms using whole exome sequencing (WES) and RNA-Seq, but did not find EGFR C797S mutation—the most common resistance mechanism to AZD9291—in PC9-OSIR, suggesting a likely novel resistance mechanism, possibly the activation of a bypass signaling pathway, which will be elucidated by further studies.The PC9-OSIR cell line can be a useful cell line model, in addition to cell lines with C797S mutation, in the development of future EGFR inhibitors. It can also be used to explore new mechanisms of resistance to AZD9291 resistance.

Note: This abstract was not presented at the meeting.

Citation Format: Zhaoshuai Bai, Guoqian Wang, Xiaolin Zhou, Zhongquan Mu, Feng Hao, Jinying Ning. PC9 AZD9291-resistant (PC9-OSIR) cell line, a useful cell model for novel drug discovery [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1920.
Abstract 1922: MYB mimic peptides targeting human and murine MYB-NFIB positive tumor cells
Authors: Jiang Yue , Forbes Lauren , Li Jianping , Freeberg Shelby , Cao Chunxia , Zajac-Kaye Maria , Licht Jonathan , Kentsis Alex , Kaye Frederic
Published: August 2019

Adenoid Cystic Cancer (ACC) is the most aggressive salivary gland tumor with no effective systemic therapy. MYB/MYBL1 fusion event underlies the etiology of ACC with few recurrent variants focused on NOTCH and chromatin remodeling pathways and low overall mutational tumor burden. No chemotherapy or targeted drugs have been approved for ACC. Therefore, there is an urgent need to develop novel therapeutic strategies. To study MYB biology and test new therapeutic agents we have developed and characterized the phenotype of a new murine conditional MYB-NFIB transgene model and have also generated a mammary tumor cell line expressing the human MYB fusion transgene with ACC-like features. We have also generated new human MYB-NFIB positive ACC tumor cell lines and xenografts from patient samples and have tested these tumor cells with newly developed MYB mimic peptides. We analyzed the phenotype of mice carrying conditional MMTV-CRE MYB-NFIB transgene alone or after crossing to heterozygosity with p16/ARF null alleles. We observed shortened survival in two different mouse transgene strains due to development of B-cell leukemia manifested by elevated WBC, low platelets, and marked splenomegaly. We also noted sporadic solid tumors including the development of a murine mammary tumor which generated a MYB-NFIB positive tumor cell line for testing new therapies. In addition, we have generated patient derived xenografts (PDX) and the successful generation of a human ACC tumor cell line from fresh ACC patient biopsies. Using shRNA reagents we observed reduced tumor cell growth and viability assayed by MTS assay following Notch1 and MYB knockdown. We also tested four candidate MYB inhibitors, celastrol, all-trans retinoic acid, and the peptidomimetic inhibitors, MYBMIM and CRYBMIM, against murine and human ACC tumor samples with immortalized normal cells as controls and will discuss these results.

Citation Format: Yue Jiang, Lauren Forbes, Jianping Li, Shelby Freeberg, Chunxia Cao, Maria Zajac-Kaye, Jonathan Licht, Alex Kentsis, Frederic Kaye. MYB mimic peptides targeting human and murine MYB-NFIB positive tumor cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1922.
Abstract 1926: Ba/F3 NTRK cell panel, a powerful cell model for NTRK inhibitor discovery and development
Authors: Hao Feng , Bai Zhaoshuai , Wang Guoqian , He Feng , Liu Changpeng , Ning Jinying
Published: August 2019

NTRK (Neurotrophic tyrosine kinase receptor)family, also named as TRK (Tropomyosin receptor kinase),is composedof three transmembrane protein receptors, TrkA, TrkB and TrkC,which play important rolesin neuronal differentiation and survival. Over-activation of NTRK1, NTRK2 and NTRK3 due to chromosomal rearrangement and mutations, was found in congenital fibrosarcoma, secretory breast carcinoma cancer and other types ofcancer. The identification of NTRK fusions as diver cancer genes has led to rapid development of anticancer therapeutic agents, including LOXO101, LOXO195 and TPX0005.Ba/F3 line is a popular system for studying both kinases and their inhibitors, because some protein kinases can render the Ba/F3 cells to be depended on the activation of the kinases instead of IL-3 supplement, while their inhibitors can antagonize the kinase-dependent growth effects. Our group used Ba/F3 cells togenerate a celllinepanel with 14 different NTRK fusion genes or mutations for NTRK inhibitor screening and development. Inaddition to use the cell lines for in vitro assays, thetransformed Ba/F3 linescan grow well in immune-deficient miceas in vivo models. LOXO101—a NTRK inhibitor—was used to test tumor growth inhibition, the results showedthat threewild-type fusion cell lineswere sensitive to LOXO101, but not other tumor models containing NTRK gatekeeper mutations. All the tumor models were sensitive to TPX0005, another NTRK inhibitor. Ourdata indicatethatthe celllinepanel of Ba/F3is apowerful cell model for new NTRK inhibitor discovery and development.

Citation Format: Feng Hao, Zhaoshuai Bai, Guoqian Wang, Feng He, Changpeng Liu, Jinying Ning. Ba/F3 NTRK cell panel, a powerful cell model for NTRK inhibitor discovery and development [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1926.
Abstract 1934: Development of spectrally distinct silica coated gold nanorods for detection of cancer using MSOT
Authors: Thomas Karl N. , Samykutty Abhilash , McNally Molly , McNally Lacey R.
Published: August 2019

Traditional cancer imaging devices are limited in their ability to screen for multiple contrast agents simultaneously in real time. Multispectral Optoacoustic Tomography (MSOT) is an emerging imaging modality capable of real-time imaging of numerous contrast agents with enhanced spatial resolution of 75µm at depths of 5 cm. The use of exogenous contrast agents in MSOT remains largely unexplored, so we developed two species of spectrally distinct gold nanorod as contrast agents for use in MSOT. Our goal was to evaluate the potential of MSOT to spectrally differentiate two exogenous contrast agents simultaneously. Two gold nanorod species were created using hydrogen peroxide (GNR-H2O2) or ascorbic (GNR-ASC) acid as reducing agents to modify the length of each species to create nanorods with individual light absorbance spectra in the IR range (680-900 nm). These gold nanorods were highly stabilized via encapsulation with mesoporous silica along with a subsequent chitosan capping. Human epidermal growth factor receptor 2 positive (HER2+) cells were specifically targeted by conjugating these mesoporous silica-coated chitosan capped gold nanorods (CMGs) to Trastuzumab resulting in TRA-CMG particles. Both TRA-CMG-ASC and TRA-CMG-H2O2 resulted in optoacoustic spectrally distinct signals when imaged in tissue phantoms both individually as well as mixed within the same well after multispectral processing using linear regression. Treatment of HER2+ breast cancer cell lines, DY36T2Q and SKBR3, with TRA-CMG-H2O2 resulted in 2.5x and 3.1x enhanced signal, respectively, as compared to HER2- MDA-MD468 cells. Treatment of DY36T2Q and SKBR3 cells with TRA-CMG-ASC demonstrated 3.7x and 6.9x, respectively, compared to MDA-MD468. In all three cell lines treated with a combination of TRA-CMG-H2O2’s and TRA-CMG-ASC’s clear and distinct signals were observed for each particle, demonstrating that each TRA-CMG possessed and maintained a detectibly distinct optoacoustic spectrum, in the IR range, allowing them to be detectable as separate contrast agents in MSOT while proximate to other targeted contrast agents. Both particles have demonstrated that they can be simultaneously administered and targeted at HER2+ cell while also maintaining distinct photoacoustic signals in MSOT upon consolidation. Each particle species, targeted to the same cells, were capable of being monitored individually in the presence of the other gold nanorod contrast agent.

Citation Format: Karl N. Thomas, Abhilash Samykutty, Molly McNally, Lacey R. McNally. Development of spectrally distinct silica coated gold nanorods for detection of cancer using MSOT [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1934.
Abstract 1938: Implantable nanosensor for non-invasive ovarian cancer biomarker detection with wearable interface
Authors: Heller Daniel A. , Williams Ryan M. , Levine Douglas A.
Published: August 2019

Patients with high-grade serous ovarian carcinoma (HGSC) exhibit poor 5-year survival rates, which may be significantly improved by early-stage detection. The US FDA-approved biomarkers for HGSC—CA-125 and HE4—do not generally appear at detectable levels in the serum until advanced stages of the disease. An implantable device placed proximal to disease sites, such as in or near the fallopian tube, ovary, uterine cavity, or peritoneal cavity, may constitute a feasible strategy to improve detection of HGSC. Furthermore, a sensor implant under the skin may allow real-time measurements after diagnosis, to monitor treatment for example, via interrogation using a wearable device. We engineered a prototype optical sensor composed of an antibody-functionalized carbon nanotube complex, which responds quantitatively to HE4 via modulation of the nanotube optical bandgap. The complexes measured HE4 with nanomolar sensitivity to differentiate disease from benign patient biofluids. The sensors were implanted into four models of ovarian cancer, within a semipermeable membrane, enabling the optical detection of HE4 within the live animals. We thus present the first in vivo optical nanosensor capable of noninvasive cancer biomarker detection in orthotopic models of disease. This technology may potentially be implanted locally in the uterine cavity, for point-of-care measurement, or under the skin, to then interface with a wearable device.

Citation Format: Daniel A. Heller, Ryan M. Williams, Douglas A. Levine. Implantable nanosensor for non-invasive ovarian cancer biomarker detection with wearable interface [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1938.
Abstract 1940: Comparison of anti-hepatocellular carcinoma efficacy between regorafenib and sorafenib using multimodality molecular imaging: A preclinical study
Authors: Du Yang , Tian Jie
Published: August 2019

Purpose: Sorafenib has been used as a targeted therapy for HCC for more than a decade, and regorafenib has recently been approved for HCC treatment and shown to prolong survival for advanced HCC patients after treatment failure with sorafenib. However, there has been no systematic study to directly compare the therapeutic effects of regorafenib and sorafenib against HCC. In this study, we systematically compared the therapeutic effects of sorafenib and regorafenib against HCC in vitro and in vivo using multimodality molecular imaging.

Experimental Design: we systemically investigated and compared the anti-tumorigenic and anti-angiogenic efficacy of regorafenib and sorafenib in vitro and in vivo using multi-modality molecular imaging as bioluminescence imaging (BLI), bioluminescence tomography (BLT), fluorescence molecular imaging (FMI), and computer tomography angiography (CTA) etc. Moreover, the safety and side effects of two drugs were simultaneously systemically evaluated.

Results: The results showed that compared with sorafenib treatment, regorafenib exerted stronger antitumor and antiangiogenic effects and significantly increased the survival rate of HCC model mice. Although both drugs caused side effects, their severity was usually higher with sorafenib, except that regorafenib caused hypertension, which was not observed with sorafenib.

Conclusions: The study suggests that multimodality molecular imaging can help sensitively evaluate the drug treatment efficacy against HCC before tumor anatomical changes occur and also helps understand the underlying antitumor mechanism of a drug at the molecular and cellular levels. Our data may provide an experimental basis for clinical application of HCC targeted treatment with regorafenib and sorafenib.

Citation Format: Yang Du, Jie Tian. Comparison of anti-hepatocellular carcinoma efficacy between regorafenib and sorafenib using multimodality molecular imaging: A preclinical study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1940.
Abstract 1943: Highly specific tumor-targeting near-infrared persistent luminescence nanomaterials (ZGO and LGGO) for lung cancer bioimaging
Authors: Chan Ming-Hsien , Liu Ru-Shi , Hsiao Michael
Published: August 2019

Lung cancer cells grow in-situ usually results in metastasis and is frequently recurrent after therapy. Moreover, treatment of lung cancer using surgery or chemotherapy is difficult to determine that all tumor tissue has been removed. ZnGa2O4/La3GaGe5O16 (ZGO and LGGO) persistent luminescence nanomaterials (PLNs) which are based on the biological characteristics of metastatic or remaining lung cancer cells, to promote the clinical application of these nanomaterials for long-term specific tracking. A silica shell-assisted synthetic route for mono-disperse, near infrared-PLNs, will be developed and applied in the, in vitro cell model and in vivo mouse model. The Cr3+-doped PLNs can hold the emitted light longer than several hours. These NIR-PLNs will absorb the energy of UV or X-ray light first and retain fluorescence emission. Moreover, we will identify the specific aptamer, MAGE-A3, and modify the PLN’s design accordingly, to increase the ability of cancer targeting. These targeted molecules can track the metastatic cell with specific binding. The advancement of nanoscale techniques in lung cancer is an urgent need. Hence, this research will deliver a credible diagnostic strategy and a novel method for therapeutic administration. To evaluate the luminescent effect in vitro and in vivo of NIR-PLNs, repeated in vivo NIR persistent luminescence images of a mouse after intravenous injection (I.V.) and intratracheal injection (I.T.) of NIR-PLNs. After the NIR-PLNs is irradiated with UV light for 120 seconds in vitro, the particles are entering in the mouse by I.V. and I.T. It is seen that NIR persistent luminescence signal could be repeatedly observed over the mouse’s lung tumor tissue for more than 6 hours. Interestingly, no significant NIR luminescence signal can be observed in the other parts of the mice. The major task is, providing a potential strategy for diagnostic in lung cancer. The current diagnostic approach does not easily distinguish the metastatic and remaining tumor cells. Moreover, we anticipate that our designed nanoplatform or accessed procedure

could be applied to other types of materials or cancers, leading NIR-PLNs nanomaterials to the exploration of gene-scale research.

Citation Format: Ming-Hsien Chan, Ru-Shi Liu, Michael Hsiao. Highly specific tumor-targeting near-infrared persistent luminescence nanomaterials (ZGO and LGGO) for lung cancer bioimaging [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1943.
Abstract 1954: Tracking of orally-administered particles within the gastrointestinal tract of murine models using multispectral optoacoustic tomography
Authors: Bhutiani Neal , Samykutty Abhilash , McMasters Kelly , Egilmez Nejat , McNally Lacey R.
Published: August 2019

While particle carriers have potential to revolutionize disease treatment, using these carriers requires knowledge of spatial and temporal biodistribution. The goal of this study was to track orally administered particle uptake and trafficking through the murine gastrointestinal (GI) tract using multispectral optoacoustic tomography (MSOT).

Polylactic acid (PLA) particles encapsulating AlexaFluor 680 (AF680) dye conjugated to bovine serum albumin (BSA) were orally gavaged into mice. Particle uptake and trafficking were observed using MSOT imaging with subsequent confirmation of particle uptake via fluorescent microscopy. Mice treated with PLA-AF680-BSA particles exhibited MSOT signal within the small bowel wall at 1 and 6 h, colon wall at 6, 12, and 24 h, and mesenteric lymph node 24 and 48h. Particle localization identified using MSOT correlated with fluorescence microscopy. Despite the potential of GI tract motion artifacts, MSOT allowed for teal-time tracking of particles within the GI tract in a non-invasive and real-time manner.

Future use of MSOT in conjunction with particles containing both protein-conjugated fluorophores as well as therapeutic agents could allow for non-invasive, real time tracking of particle uptake and drug delivery.

Citation Format: Neal Bhutiani, Abhilash Samykutty, Kelly McMasters, Nejat Egilmez, Lacey R. McNally. Tracking of orally-administered particles within the gastrointestinal tract of murine models using multispectral optoacoustic tomography [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1954.
Abstract 1967: Nab-paclitaxel chemotherapy of breast tumors upregulates chemokines that increase chemotaxis of myeloid-lymphatic progenitors
Authors: Volk-Draper Lisa D. , Ran Sophia
Published: August 2019

Introduction: Lymph node metastasis is the main prognostic indicator for poor survival of breast cancer patients. Metastasis is promoted by tumor-induced lymphangiogenesis mediated, in part, through recruitment of myeloid-derived lymphatic endothelial cell progenitors (M-LECP). We previously showed that differentiation of M-LECP is induced by activated Toll-like receptor-4 (TLR4). The goal of this study was to identify TLR4-induced chemokines produced by tumors alone or in response to nab-paclitaxel (nab-PXL) chemotherapy that enhance bone marrow (BM) differentiation and tumor recruitment of M-LECP.

Methods: Profile of BM composition for lymphatic-like, myeloid and progenitor cells was assessed by flow cytometry in normal and tumor-bearing mice treated either with saline or nab-PXL. We used two TLR4 overexpressing tumor models: MDA-MB-231 and HCC1806-TLR4+. RT-qPCR was used to determine differences in expression of 130 cytokines in cultured tumor cells and tumors from mice treated either with saline or nab-PXL. Targets with >2-fold upregulation were analyzed for protein expression by ELISA. ELISA-validated cytokines were tested for their ability to induce migration of M-LECP.

Results: Both MDA-MB-231 or HCC1806-TLR4+ tumors significantly increased the levels of differentiated M-LECP in the BM, and that was further enhanced by treatment with taxane chemotherapy. Expression of progenitor markers CD117, CD33, and Sca-1 was increased up to 7-fold in both tested tumor models, and further upregulated by nab-PXL treatment by additional 3 to 9-fold. Out of 23 cytokines measured in tumor lysates by ELISA, 40% were upregulated by nab-PXL. Migration assay showed that several of these cytokines increased chemotaxis of M-LECP with CXCL1 being the strongest. Neutralization of CXCL1 in the conditioned medium of nab-PXL-treated MDA-MB-231 cells inhibited migration of lymphatic progenitors by 75%.

Conclusion: Taxane chemotherapy acting presumably as a ligand for TLR4 significantly expanded M-LECP population in the BM. Our analysis suggests that these lymphatic precursors can mobilize to tumors by chemokines such as CXCL1 that are highly upregulated in TLR4-positive tumor cells, particularly when activated by paclitaxel. These data suggest that CXCL1 plays an important role in recruiting myeloid-lymphatic progenitors to breast tumors, which, in turn, promotes lymphatic vessel formation and metastasis. Blocking this cytokine might have clinical utility for prevention of tumor mobilization of M-LECP, and by extension, breast cancer metastasis to lymph nodes.

Citation Format: Lisa D. Volk-Draper, Sophia Ran. Nab-paclitaxel chemotherapy of breast tumors upregulates chemokines that increase chemotaxis of myeloid-lymphatic progenitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1967.
Abstract 1968: Tumor necrosis factor-α upregulation of CCR7 induces prostate cancer cell migration in lymphatic metastasis
Authors: Makino Tomoyuki , Izumi Kouji , Maolake Aerken , Natsagdorj Ariunbold , Iwamoto Hiroaki , Kadomoto Suguru , Naito Renato , Hiratsuka Kaoru , Kadono Yoshifumi , Mizokami Atsushi
Published: August 2019

Objectives: Prostate cancer (PCa) with lymph node (LN) metastasis has a poor patient prognosis. Broader understanding of the causes of LN metastasis and disease dissemination will assist in developing novel treatments. C-C chemokine receptor 7 (CCR7) and its ligand CCL21 are abundantly expressed in LN metastasis and during PCa progression. Tumor necrosis factor-alpha (TNF-α) is present at low levels within the tumor microenvironment. TNF-α and C-C chemokines interact in tumorigenesis. The study aim was to determine whether TNF-α might promote PCa dissemination from metastatic LN by activating the CCL21/CCR7 axis.

Methods: TNF-α and CCR7 mRNA and protein expression were assayed in PC-3, DU145, and LNCaP, and human PCa cells by reverse transcription-polymerase chain reaction and western blotting. Promotion of transmembrane migration by TNF-α was assayed in CCL21-pretreated PCa cells. Phosphorylation of several mitogen-activated protein kinases expressed downstream of TNF-α was assayed.

Results: Low-dose TNF-α induced CCR7 expression in PCa cells and promoted migration of CCL21-pretreated PCa cells. TNF-α-mediated CCR7 upregulation via the ERK signaling pathway. p38 was a downstream target of CCL21/CCR7 signaling and a key factor in the promotion of PCa cell migration.

Conclusions: This is the first demonstration of the influence of the TNF-α-CCL21/CCR7 axis on PCa cell migration in the microenvironment of LN metastasis. Because it is activated in PCa LN metastasis, the CCL21/CCR7 axis might be a novel therapeutic target and biomarker of PCa.

Citation Format: Tomoyuki Makino, Kouji Izumi, Aerken Maolake, Ariunbold Natsagdorj, Hiroaki Iwamoto, Suguru Kadomoto, Renato Naito, Kaoru Hiratsuka, Yoshifumi Kadono, Atsushi Mizokami. Tumor necrosis factor-α upregulation of CCR7 induces prostate cancer cell migration in lymphatic metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1968.
Abstract 1972: CXCR5-CXCL13 signaling contributes to CLL relapse and patient survival
Authors: Griffen Tiara , Young Corey , Dill Courtney D. , Carey Kaylin , Adebayo Olayinka , Nunez Sha'kayla , Lillard James
Published: August 2019

Chronic Lymphocytic Leukemia (CLL) is characterized by the appearance of mature-appearing B cells in the blood, bone marrow, and secondary lymphoid organs. There is an average of 19,000 new cases, in the United States annually, but the molecular pathogenesis of CLL is incompletely understood. Chemokines and their receptors have been shown to contribute to the progression and metastasis of several types of cancers. Chemokine receptor 5 (CXCR5) and chemokine ligand 13 (CXCL13) are produced by cells in the microenvironment of secondary lymphoid organs such as lymphatic endothelial cells, which promote disease progression. There is a gap in understanding the molecular mechanisms and functions resulting from CXCL13-CXCR5 interactions during CLL. This gap is important to fill, because these molecules could be additional therapeutic targets to treat this disease. Together this provided the rationale to test the hypothesis CXCR5-CXCL13 signaling contributes to the growth, metastasis, and survival of CLL. The role of CXCR5, CXCL13, and associated genes was determined by conducting Weighted Gene Network Co-expression (WGCNA) and upstream regulator, canonical pathway, and gene interaction analyses of 297 International Cancer Genome Consortium (ICGC) CLL patients. Our previous studies show CXCR5-CXCL13 signaling activates CTNNB1, JUN, and VAV2 transcriptional factors. These factors promote the expression of GNA, GRK, PIK3, RAC and RHO family genes, which were significantly associated with the WGCNA-designated Yellow signaling module. Interestingly, the WGCNA-designated Grey module contained independently expressed genes, e.g., CXCL13, LTB, IGHV, ZAP70, which were significantly associated (p = 0.00003) and correlated (R2 = 0.29) with relapse. Genes in this module significantly correlated with CLL relapse and survival. Taken together, our data demonstrate the clinical significance of the CXCL13-CXCR5 pathway in CLL progression.

Citation Format: Tiara Griffen, Corey Young, Courtney D. Dill, Kaylin Carey, Olayinka Adebayo, Sha'kayla Nunez, James Lillard. CXCR5-CXCL13 signaling contributes to CLL relapse and patient survival [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1972.
Abstract 1977: Chemokines expression in an in vitro model of melanoma brain metastasis
Authors: Santos Renata , Sampaio Maria Clara , Albuquerque Amanda , Pereira Michelly , Pitta Maira , Rêgo Moacyr
Published: August 2019

Melanoma brain metastasis is still a great challenge. This work aimed to characterize the secretion profile and understand the possible role of chemokines in a xenogeneic in vitro model of melanoma brain metastasis. Brain primary mixed cultures from cerebral cortex were prepared from the neonates Wistar rats and maintained at 37°C, 5% CO2until 80% of confluency and maturity. Melanoma lineages used were UACC62 and SKMEL-28. Co-cultures were performed to study the direct interactions between melanoma and cortical cells. The indirect effects in the cerebral microenvironment were evaluated through conditioned medium (CM) from brain cells on melanoma lineages. It was used an early CM collected at 24h (CM24) and a late CM collected after 48h (CM48), whose both were incubated during more 24h (24CM24 and 24CM48) or 48h (48CM24 and 48CM48) on melanoma cultures. The chemokines IP10, CCL5 and IL8 were studied through Cytometric Bead Array (CBA),MIG and CCL2 also, however their values were undetectable. After the characterization of the model, IP10 was silenced to evaluate its influence in the model. CBA demonstrated that in SKMEL-28 the late soluble factors during 48h (48CM48) were capable of rase IP10 levels (p=0,0286). On the other hand, direct and indirect contact with brain cells reduced IL8 levels, compared to SKMEL28 alone with respectively p = 0.02 and p=0,0286, where both chemokines were not detected. On UACC62, IP10 levels were higher only in the co-culture (p=0,02), suggesting the need of contact between the different cells. Also, IL8 was decreased in all conditions compared to UACC62. After IP10 silencing, SKMEL-28 presented higher levels of IL8 in 48CM48. On UACC62, the IP10 silencing resulted in elevation of IL8 levels when the cells were in co-culture. RANTES did not present alterations in its levels in any of the conditions after IP10 knock-down. The IP10 silencing was also capable of impair the transwell invasion of SKMEL-28 in the presence of CM24 and CM48. Together, these results confirmed that IP10 alters IL8 levels, but no RANTES in a mimetic microenvironment of melanoma brain metastasis. Furthermore, IP10 is capable of influencing SKMEL-28 migration in this xenogeneic model. More studies to better understand the role of IP10 and IL8 in the establishment of melanoma brain metastasis in this model are in progress.

Citation Format: Renata Santos, Maria Clara Sampaio, Amanda Albuquerque, Michelly Pereira, Maira Pitta, Moacyr Rêgo. Chemokines expression in an in vitro model of melanoma brain metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1977.