bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1956: ICG pHLIP: A novel agent for fluorescence-guided surgery
Authors: Crawford Troy , Moshnikova Anna , Roles Sean , Carter Lukas M. , Lewis Jason S. , Engelman Donald M. , Andreev Oleg A. , Reshetnyak Yana K.
Published: August 2019

Introduction: Fluorescence imaging has applications in medicine for many image-guided procedures such as mapping and visualization of lymph nodes, targeting and marking cancerous lesions and assessment of tumor margins. Several fluorescent molecules are approved for clinical use including the ICG (indocyanine green) near infrared (NIR) fluorescent dye. We conjugated ICG to a pH Low Insertion Peptide (pHLIP®), which senses and targets the acidity at the surfaces of cancer cells, and evaluated ICG pHLIP® as a targeted imaging agent for tumor visualization. Our construct is progressing toward clinical trials.

Experimental Procedures: ICG pHLIP® was manufactured for proof-of-concept tumor-targeting imaging studies in mice, and toxicity evaluation in mice, dogs and rats. Tumor targeting was assessed in various human and murine tumor models. Biodistribution was evaluated at different time points from 5 min to 48 hours after a single intravenous administration of 0.5 mg/kg of ICG pHLIP®. The distribution of the agent within tumors was evaluated in tumor sections.

Results: The fluorescence of ICG is enhanced about 15 times when pHLIP® tethers it to a cell membrane, significantly improving the contrast index. No toxicity is seen at 25-30x of the proposed human dose, evaluated in mice and dogs. ICG pHLIP® demonstrates excellent tumor targeting, including targeting of 1-3 mm sized tumors in breast. About 10-15% ID/g was observed within tumors at 4 hours p.i., and the signal persists for up to 24 hours with a slight decline at 48 hours. The background signal in major organs decayed after agent administration. The highest uptake, of about 50% ID/g, was observed in liver at 4 hrs p.i. and decayed after that. An excellent correlation was established between ICG pHLIP® NIRF imaging and H&E histopathology at the indicated tumor location.

Conclusions: ICG pHLIP® is a promising new imaging agent for fluorescence-guided surgical applications. It is on the path to clinical translation for accurate resection of breast tumors. Since pHLIP® targeting of tumors is based on the general principle of cancer cell surface acidity, ICG pHLIP® is an agent that may be used in a range of fluorescence-guided surgical interventions.

Citation Format: Troy Crawford, Anna Moshnikova, Sean Roles, Lukas M. Carter, Jason S. Lewis, Donald M. Engelman, Oleg A. Andreev, Yana K. Reshetnyak. ICG pHLIP: A novel agent for fluorescence-guided surgery [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 1956.
Abstract 1957: Novel substrates for NanoLuc luciferase with improved brightness and signal duration for bioluminescence imaging in vivo
Authors: Walker Joel R. , Park Yunhee , Lin Michael , Kirkland Thomas A. , Hall Mary P. , Encell Lance P. , Oh Younghee , Liu Lan
Published: August 2019

NanoLuc® luciferase, and its substrate furimazine, are becoming a preferred bioluminescent system in cellular and biochemical assays due to its brightness and glow-type kinetics. The construct Antares, made by fusing NanoLuc to the cyan-excitable orange-red fluorescent protein CyOFP, combines the high catalytic rate of NanoLuc with the high quantum yield and red-shifted emission of CyOFP (584 nm), which is well suited for in vivo imaging. Transgenic mice were created to constituently express Antares and these mice were used to screen coelenterazine analogues for a substrate that showed appropriate distribution and brightness in vivo. Our new Antares (NanoLuc) substrates are administered i.p., have improved solubility over furimazine, exhibit long (>30 min) signal kinetics, and are brighter than D-luciferin/Luc2 expressed on the same promoters.

Citation Format: Joel R. Walker, Yunhee Park, Michael Lin, Thomas A. Kirkland, Mary P. Hall, Lance P. Encell, Younghee Oh, Lan Liu. Novel substrates for NanoLuc luciferase with improved brightness and signal duration for bioluminescence imaging in vivo [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 1957.
Abstract 1966: Obesity and prostate cancer progression: Role of the chemokine CXCL12
Authors: Ahn Songyeon , Saha Achinto , Kolonin Mikhail G. , DiGiovanni John
Published: August 2019

Obesity is a significant risk factor for several cancers in both men and women. A number of studies have shown that obesity is associated with increased prostate cancer (PCa) progression and higher mortality. However, the mechanism(s) for obesity-driven PCa remain unclear. In earlier studies, we showed that obesity enhances PCa progression in the HiMyc mouse model by changing the surrounding tumor microenvironment, especially the white adipose tissue (WAT). WAT consists of many cells including inflammatory cells, mature adipocytes and adipose stromal cells (ASCs). Here, we further investigated the role of ASCs and ASC derived CXCL12 in obesity-driven PCa progression. In HMVP2 cells, a PCa cell line derived from HiMyc mice expressing high levels of CXCR4, we observed activation of MAPKs (JNK and ERK), AKT, STAT3 and NFkB following treatment with CXCL12. These CXCL12-induced signaling pathways were inhibited by treating with AMD3100, a CXCR4 antagonist, and also by CXCR4 knock-down. To confirm this in vivo, we compared PCa tumor growth in lean and obese mice using the HMVP2 allograft model. Our data demonstrates that obesity promotes EMT in tumor cells and increases invasion of surrounding WAT into the tumor stoma. Treatment of mice with AMD3100 significantly inhibited the growth of HMVP2 tumors and suppressed EMT in both the obese and control diet groups. Furthermore, D-CAN, a hunter-killer peptide developed for targeting ASCs, also suppressed obesity-induced EMT and tumor growth in HMVP2 tumor allografts. Lastly, AMD3100 in combination with D-CAN, showed a significant reduction in PCa growth in obese mice compared to either agent used alone. These results provide evidence that ASCs activate the CXCL12/CXCR4 signaling as well as other pathways playing an important role in obesity-driven PCa progression. Our data suggests this pathway as a novel therapeutic target for inhibiting obesity-driven PCa growth and progression. Research supported by NCI Grant CA196259.

Citation Format: Songyeon Ahn, Achinto Saha, Mikhail G. Kolonin, John DiGiovanni. Obesity and prostate cancer progression: Role of the chemokine CXCL12 [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 1966.
Abstract 198: The T3R domain of thrombospondin-1 delays tumor growth and improves tumor response to chemotherapy by remodeling the tumor vasculature
Authors: Pinessi Denise , Resovi Andrea , Morosi Lavinia , Borsotti Patrizia , Berndt Alexander , Giavazzi Raffaella , Zucchetti Massimo , Taraboletti Giulia
Published: August 2019

The disorganized and inefficient nature of the tumor vasculature affects the delivery and activity of antineoplastic treatments. Certain antiangiogenic agents induce a transient normalization of tumor vessels, hence representing a promising strategy to improve the delivery and efficacy of cytotoxic therapy. Our previous studies demonstrated the antiangiogenic activity of the type III repeats (T3R) domain of thrombospondin-1 (TSP-1), a major endogenous inhibitor of angiogenesis. Aim of this study was to investigate the role of the T3R domain in tumor growth and tumor response to chemotherapy. Ectopic tumor cell expression of T3R delayed in vivo growth in two tumor models, the human ovarian carcinoma A2780-1A9 and endometrial adenocarcinoma HEC-1B. This was associated with vascular remodeling, decreased cell proliferation and increased apoptosis. T3R was active in preventing tumor growth also when expressed by the host cells (rather than the tumor cells), following pretreatment of mice with adeno-associated virus (rAAV)-T3R before tumor cell implantation. Expression of T3R also resulted in improved tumor response to paclitaxel (PTX 10 mg/kg Q4x3, i.v.) and cisplatin (DDP 4 mg/kg Q4x3, i.v.). Through the pharmacokinetic study, conducted by HPLC and MALDI imaging mass spectrometry analysis, a higher concentration and a more homogeneous drug distribution were observed in tumors expressing T3R 4h after a single administration of PTX (60 mg/kg, i.v.). This was associated with a marked reorganization of the tumor vasculature that presented decreased vessel area and vessel diameter and increased number of CD31-positive vessels. These findings indicate the ability of T3R to reduce tumor growth and improve tumor response to chemotherapy through the remodeling of tumor vasculature. This sets the basis for the development of new therapeutic strategies based on T3R to be used in combination therapies.

Supported by AIRC. DP is recipient of the AIRC fellowship “Lina e Giovanni Giasini”

Citation Format: Denise Pinessi, Andrea Resovi, Lavinia Morosi, Patrizia Borsotti, Alexander Berndt, Raffaella Giavazzi, Massimo Zucchetti, Giulia Taraboletti. The T3R domain of thrombospondin-1 delays tumor growth and improves tumor response to chemotherapy by remodeling the tumor vasculature [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 198.
Abstract 1981: M2-macrophage derived exosomes mediates tumor progression via GATA3-IL-4-IL-13 axis in ovarian cancer
Authors: Kanlikilicer Pinar , El-Arabey Amr Ahmed , Denizli Merve , Bayraktar Recep , Ozpolat Bulent , Lopez-Berestein Gabriel , Salama Salama Abdu , Abd-Allah Adel Rashad
Published: August 2019

Tumor microenvironment is the key factor for tumor progression. Stromal cells which consist of, tumor associated macrophages(TAMs), tumor associated fibroblast as well as cancer stem cells has been implicated in the acquisition of oncogenic phenotype via cell-cell communication. Exosomes are the major players in tumor microenvironment that contributes to tumorigenesis. Exosomes carry genetic information from host cells to another cell to reprogram the recipient cell. Exosomes from TAMS has emerged as important mediators of tumor growth. Here, we report that GATA3 is released abundantly from TAM cells via exosomes and promote oncogenesis. GATA3 acts as an oncogenic protein and silencing GATA3 by siRNA led to decrease in tumor cell viability. Our data showed that silencing GATA3 increased the release of IL-4 and IL-13 from EOC cells, which leads to M2 cell polarization from M0 cells. Our results suggest that GATA3 released from macrophage exosomes contributes to tumor growth by affecting intracellular processes as well as tumor microenvironment. P.K. and A. El-A. contributed equally.

Citation Format: Pinar Kanlikilicer, Amr Ahmed El-Arabey, Merve Denizli, Recep Bayraktar, Bulent Ozpolat, Gabriel Lopez-Berestein, Salama Abdu Salama, Adel Rashad Abd-Allah. M2-macrophage derived exosomes mediates tumor progression via GATA3-IL-4-IL-13 axis in ovarian cancer [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 1981.
Abstract 1991: Exploring the potential of exosomes for cancer diagnosis and management: Comparative analysis of isolation methods for optimum yield purity and downstream applications
Authors: Patel Girijesh K. , Khan Mohammad Aslam , Zubair Haseeb , Srivastava Sanjeev Kumar , Khushman Moh’d , Singh Seema , Singh Ajay Pratap
Published: August 2019

Exosomes have received significant attention due to their involvement in several diseases, including cancer, and are being explored as a tool for disease diagnosis and management. As a result, several kits based on different principles are now available in the market for exosome isolation. In this study, we have compared the relative efficacy of four commercial kits from Invitrogen, 101Bio, Wako and iZON along with conventional ultracentrifugation-based method for exosome yield, purity and quality for downstream applications. Cancer cell culture supernatant was used as an abundant and uniform source of exosomes for all methods and exosome quantity, size distribution, zeta potential, marker expression, RNA (qRT-PCR) and protein (mass spectrometry) quality was determined. The Invitrogen method showed the highest yield but the preparation showed broader size distribution likely due to co-precipitation of microvesicles and had the least magnitude of zeta potential (dispersion stability). Exosome preparations from other methods showed accepted size range (<150 nm) with a variable zeta potential (-12mV to -29mV) with good purity. The gel filtration method; however, showed a broader size distribution in the lower size range, which could be due contamination of aggregates of proteins and vitamins molecules. Quality of RNA isolated from all preparations was comparable; however, proteins isolated from Invitrogen method-based exosomal preparation showed contamination of polyethylene glycol (PEG) in mass spectrometry analyses. Similarly, other chemical impurities coming from the precipitant in Invitrogen kit could also be the reason for toxicity of resultant exosomal preparation in biological growth measurement assay. Together, these findings from comparative analyses should serve as a guide for choice and further optimization of exosome isolation methods for their desired downstream applications in cancer biology, therapy and/or biomarker discovery.

Citation Format: Girijesh K. Patel, Mohammad Aslam Khan, Haseeb Zubair, Sanjeev Kumar Srivastava, Moh’d Khushman, Seema Singh, Ajay Pratap Singh. Exploring the potential of exosomes for cancer diagnosis and management: Comparative analysis of isolation methods for optimum yield purity and downstream applications [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 1991.
Abstract 1996: Epigenetic reactivation of BAI1/ADGRB1 suppresses tumor invasion by preventing TGFβ1-induced mesenchymal switch in glioblastoma
Authors: Osuka Satoru , Yang Liquan , Zhu Dan , Hashimoto Hideharu , Devi Narra S. , Meir Erwin G. Van
Published: August 2019

Glioblastoma (GBM) is the most common and lethal type of malignant brain tumor in adults. GBM cells are highly invasive and diffusely infiltrate throughout the brain, which strongly restricts multimodal therapies. Acquiring a better knowledge of molecular defects underlying GBM invasion is essential for the development of effective therapies.

Brain-specific Angiogenesis Inhibitor 1 (BAI1/ADGRB1) is a transmembrane receptor of the adhesion GPCR family widely expressed in normal brain, but its expression is lost in the majority of glioblastoma through epigenetic silencing and restoration of its expression can inhibit glioma growth (Zhu D. et al, Cancer Res, 2012). Recently, we reported that BAI1 protects p53 from Mdm2-mediated degradation and regulate tumor growth in medulloblastoma (Zhu D. et al, Cancer Cell, 2018). However, it is unclear whether BAI1 loss is important for tumor invasion and the mesenchymal phenotype in GBM.

Microarray analysis of the GBM TCGA dataset revealed that low BAI1 mRNA expression correlates with poor outcome and with expression of many key mesenchymal genes, including Fibronectin1, SLUG, and TWIST1. Restoration of BAI1 expression in human GBM cells suppresses mesenchymal gene expression in culture, and dramatically decreases brain tumor invasion in mice xenografts. Mechanistically, we found that the N-terminal thrombospondin type 1 repeat (TSR#1) of BAI1 inhibits the maturation process of TGFβ1, a key growth factor involved in EMT. BAI1 is silenced epigenetically in GBM cells by methylated CpG-binding protein MBD2, and its expression can be reactivated by KCC-07, a blood-brain barrier permeable MBD2 inhibitor. We found that restoration of BAI1 expression by KCC-07 treatment dramatically reduced tumor cell invasion in orthotopic xenografts model of GBM cells.

These experiments demonstrate that epigenetic silencing of BAI1 is important for activation of the GBM invasive phenotype through TGFβ1 pathway activation. Epigenetic targeting of this process by KCC-07 can reduce GBM invasion and improve patient survival.

Citation Format: Satoru Osuka, Liquan Yang, Dan Zhu, Hideharu Hashimoto, Narra S. Devi, Erwin G. Van Meir. Epigenetic reactivation of BAI1/ADGRB1 suppresses tumor invasion by preventing TGFβ1-induced mesenchymal switch in glioblastoma [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 1996.
Abstract 200: The MCP-1/CCR2 axis enhances VEGF-A-dependent angiogenesis by directly downregulating miR-29c expression in oral squamous cell carcinoma
Authors: Lien Ming-Yu , Tang Chih-Hsin
Published: August 2019

Oral squamous cell carcinoma (OSCC), the sixth most common malignancy worldwide, has a poor prognosis due to its aggressiveness and potential for metastasis. Activation of monocyte chemoattractant protein-1 (MCP-1) and its receptor, CC chemokine receptor 2 (CCR2), induces angiogenic activation of endothelial cells, inflammatory responses and aberrant MCP-1/CCR2 expression in malignant tissues. However, the effect of MCP-1 and CCR2 in OSCC angiogenesis is unclear. Our study sought to determine how the MCP-1/CCR2 axis mediates VEGF-A expression in OSCC and results in angiogenesis. Immunohistochemistry (IHC) analysis identified higher concentrations of VEGF-A protein expression in tissue obtained from patients with OSCC compared with tissue from healthy controls; the degree of VEGF-A tumor expression was significantly associated with the clinical disease stage. Our in vitro investigations found that MCP-1 dose-dependently regulated VEGF-A expression through the CCR2, ILK and MEK signal transduction pathways. Following MCP-1 application, conditioned medium from the human OSCC cell line SCC4 significantly promoted endothelial progenitor cell (EPC) migration and tube formation; these events were attenuated and VEGF-A production was suppressed when we stimulated the OSCC cells with the pathway inhibitors RS102895, KP392, and PD98059. We also showed that microRNA (miR)-29c expression was negatively regulated by MCP-1 via the CCR2/ILK/MEK cascade, tying in with an analysis of the cancer genome atlas (TCGA) database showing lower levels of miR-29c expression in tissues from patients with oral cancer compared with samples from healthy controls. According to our results, the MCP-1/CCR2 axis promotes VEGF-A expression in OSCC via the ILK and MEK signal transduction pathways and downregulates miR-29c expression. Thus, MCP-1/CCR2 mediation of VEGF-A mobilizes EPCs required for OSCC and is important for tumor angiogenesis. MCP-1/CCR2 may be an appropriate target in the treatment of OSCC angiogenesis.

Citation Format: Ming-Yu Lien, Chih-Hsin Tang. The MCP-1/CCR2 axis enhances VEGF-A-dependent angiogenesis by directly downregulating miR-29c expression in oral squamous cell carcinoma [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 200.
Abstract 2008: CENP-E kinesin facilitates chemokine ligand CCL18 translocation to promote migration in triple negative breast cancer
Authors: Mullen McKay M.
Published: August 2019

Our research is focused on the molecular mechanisms responsible for enhanced cellular proliferation and migration as it relates to triple negative breast cancer (TNBC). Advances in visualization techniques and technologies work to provide an in-depth view of various cellular mechanisms, allowing us to capture molecular level protein-protein interactions previously unknown. There are more than 40 kinesin motor proteins found within the spindle assembly checkpoint, classified into 14 families, which each play essential roles in microtubule-dependent intracellular trafficking. This assembly is a critical component to the cell cycle control complex that acts during mitosis to prevent chromosome missegregation and aneuploidy. Centromere associated protein E (CENP-E) is a kinesin motor protein that shows aberrant overexpression in TNBC which facilitates exponential cell growth and eventual metastasis that occurs in TNBC. Growing awareness through scientific discoveries sheds light on the fact that it is a rare occurrence for biological functions to be attributed to a single mutated macromolecule. Most physiological processes arise from a complex orchestration of cellular events occurring between multiple molecules acting in concert. The nature of protein-protein transactions that drive gene activation appear to be dynamic in TNBC cells thus enabling uncontrolled proliferation. We believe that a pleiotropic network of small molecules may be working cooperatively with CENP-E to facilitate the exponential cell growth and eventual metastasis that occurs in TNBC cells. Currently, we are carrying out spectral imaging and biophysical force measurements to delineate CCL18-elicited cell mechanics and breast cancer metastasis. To address this, we have incorporated syntelin, a small inhibitory molecule targeted to CENP-E in an effort to analyze the resultant effect it has on this network of proteins we believe may be involved in the CENP-E signaling cascade. Taken together, these results indicate that syntelin may comprise a novel targeted chemotherapeutic intervention for TNBC.

Citation Format: McKay M. Mullen. CENP-E kinesin facilitates chemokine ligand CCL18 translocation to promote migration in triple negative breast cancer [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 2008.
Abstract 2009: Leronlimab, a humanized monoclonal antibody to CCR5, blocks breast cancer cellular invasion and enhances cell death induced by DNA damaging chemotherapies
Authors: Jiao Xuanmao , Wang Min , Pestell Richard G.
Published: August 2019

Purpose of the study. To assess binding and functional interaction of the humanized monoclonal antibody to CCR5 (Leronlimab) with human breast cancer cell lines. The G protein coupled receptor CCR5, is normally expressed on a subset of T cells and serves as a co-receptor for HIV infection. During malignant transformation CCR5 expression is known to increase in a number of cancers (breast cancer (BCa), prostate cancer, colon cancer, melanoma). CCR5 targeted cancer clinical trials using small molecular inhibitors opened to accrual in late 2018. CCR5 is expressed in >50% of human BCa, primarily in triple negative BCa. Its expression in human BCa correlates with poor outcome and CCR5+ BCa epithelial cells have characteristics of cancer stem cells, forming mammospheres and initiating tumors with >60-fold greater efficiency in mice. Reintroduction of CCR5 expression into CCR5 negative BCa cells promotes tumor metastases and induces DNA repair gene expression and activity. The CCR5 inhibitor Leronlimab has been used for treatment of >550 patients HIV, including meeting its primary endpoints in a phase III study, without significant adverse events reported.

Results. Leronlimab bound to CCR5 expressed in human breast cancer cell lines with 98% efficiency. Leronlimab abrogated CCL5 induced Ca+2 flux and blocked 3-d matrigel invasion of MDA-MB-231 cells. Leronlimab also augmented cell killing by DNA damage inducing agents Doxorubicin.

Conclusions. Leronlimab binds CCR5 in BCa cells, blocking breast cancer cellular invasion and augmenting cell killing by DNA damage inducing chemotherapies. As CCR5 augments DNA repair and is expressed selectively on cancerous but not normal breast epithelial cells, Leronlimab may enhance the tumor specific activities of DDR-based treatments, allowing a reduction in dose of standard chemotherapy and radiation.

Citation Format: Xuanmao Jiao, Min Wang, Richard G. Pestell. Leronlimab, a humanized monoclonal antibody to CCR5, blocks breast cancer cellular invasion and enhances cell death induced by DNA damaging chemotherapies [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 2009.
Abstract 201: The mechanism of vascular mimicry in clear cell renal cell cancer
Authors: Yang Jun-Ping , Peng Xing-Si , Qian Chao-Nan
Published: August 2019

Although anti-angiogenic drugs were used as first-line treatment in metastatic clear cell renal cell cancer (ccRCC), some patients with metastatic ccRCC still had limited survival benefit from anti-angiogenic therapy. The main reason was resistance to anti-angiogenesis treatment. Tumor blood vessel could be divided into endothelial cell-derived or tumor cell-derived vessels. Tumor cell-derived vascular channel was called as tumor vascular mimicry (VM). PAS-CD34 staining in ccRCC tissues revealed that the expression status of VM was different in ccRCC patients. And the progression-free survival and overall survival were significantly shorter in ccRCC patients with high VM expression level. To explore the effect of VM on the resistance to anti-angiogenesis treatment and the molecular mechanism of VM, we successfully established a three-dimensional culture model for the formation of tumor cell-derived vessel-like structures. We found the formation of blood vessel-like structures in kidney cancer cells was not affected by the anti-angiogenic drug, such as Sunitinib, suggesting that VM was closely associated with resistance to anti-angiogenic therapy. Moreover, we had discovered the “molecular signature of vascular mimicry in renal cancer”. Using this signature, 531 ccRCC patients in TCGA could be divided into two groups with different prognosis. More importantly, we found that HGF/c-MET signaling pathway regulated the expression of Serpinb2 through the downstream AKT pathway, and thus affected the formation of vascular mimicry in SN12C, 786-O cells. In addition, we also had found that HGF could promote the formation of blood vessel-like structure in lung and bladder cancer cells, and the " molecular signature of vascular mimicry in renal cancer " also could differentiate patients with renal papillary carcinoma, bladder cancer, endometrial cancer, lung adenocarcinoma cancer, and pancreatic cancer. These finding suggested that the molecular mechanism of vascular mimicry in renal cancer was universal, to some extent. This project partially elucidated the molecular mechanism of vascular mimicry in renal cancer and its clinical significance, laying a theoretical foundation for the development of more effective anti-angiogenic drugs.

Citation Format: Jun-Ping Yang, Xing-Si Peng, Chao-Nan Qian. The mechanism of vascular mimicry in clear cell renal cell cancer [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 201.
Abstract 2011: Potassium usnate inhibits invasion and metastasis in colorectal cancer: Implication of clinical application of a water-soluble form of usnic acid in anticancer therapy
Authors: Yang Yi , Bae Woo Kyun , Lee Kyung Hwa , Moon Kyung-Sub , Park So-Yeon , Zhou Rui , Taş Isa , Gamage Chathurika , Lee Sueun , Jung Chaeyong , Kim Kyung Keun , Ha Hyung-Ho , Kim Hangun
Published: August 2019

Usnic acid (UA), a lichen secondary substance, has considerable anticancer activity in vitro, whereas its effect in vivo is limited. Here, potassium usnate (KU) was prepared by the salinization of UA to enhance its water solubility. KU showed increased bioavailability compared with UA in the tumor, liver, and plasma of a CT26 syngeneic mouse tumor xenograft model after oral administration, as determined by LC-MS/MS analysis. KU exhibited potent anticancer effects on colorectal cancer cells and inhibited liver metastasis in an orthotopic murine colorectal cancer model. KU treatment downregulated the epithelial-mesenchymal markers Twist, Snail, and Slug and the metastasis-related genes CAPN1, CDC42, CFL1, IGF1, WASF1, and WASL in cells and tumor tissues. The present results suggest the potential application of the water-soluble form of UA, KU, in anticancer therapy.

Citation Format: Yi Yang, Woo Kyun Bae, Kyung Hwa Lee, Kyung-Sub Moon, So-Yeon Park, Rui Zhou, Isa Taş, Chathurika Gamage, Sueun Lee, Chaeyong Jung, Kyung Keun Kim, Hyung-Ho Ha, Hangun Kim. Potassium usnate inhibits invasion and metastasis in colorectal cancer: Implication of clinical application of a water-soluble form of usnic acid in anticancer therapy [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 2011.
Abstract 2018: Cannabinoid receptor 2 agonist JWH-015 inhibits growth and metastasis of triple negative breast cancers through regulation of autophagy mechanism
Authors: Chatterjee Nabanita , Das Subhadip , Ahirwar Dinesh K. , Mishra Sanjay , Ganju Ramesh
Published: August 2019

Metastasis to distant organs is the major cause of mortality associated with breast cancer. Triple-negative breast cancer (TNBC) subtype has been shown to be associated with worst patient survival due to high metastasis rate and lack of targeted therapies. Therefore, there is an utmost need to develop novel therapeutic strategies against TNBC growth and metastasis. Cannabinoids are known to possess anti-cancer activity. However, the associated psychotropic activity limits the therapeutic use of Cannabinoids in breast cancer patients. Here, we used synthetic cannabinoid JWH-015 which activates cannabinoid receptor 2 (CB2) and is devoid of psychotropic activity. In vitro molecular analysis revealed that JWH-015 induces TNBC cell death by activating autophagy associated apoptosis. In addition, we have further shown using in vivo tumor models, that JWH-015 potentially inhibits TNBC growth and metastasis through activation of autophagy mechanisms. Further analysis showed that JWH-015 treated tumors recruited the reduced number of macrophages compared to vehicle control treated tumors. In addition, there were more anti-tumor M1-type macrophages present in JWH-015 treated tumors. Conversely, vehicle control treated tumors possessed a higher number of pro-tumor M2-type macrophages. These observations have established the anti-metastatic potential of JWH-015 in different TNBC pre-clinical mouse models. The mechanistic studies have elaborated the role of JWH-015-induced autophagy in promoting tumor cell death to inhibit TNBC growth and metastasis.

Citation Format: Nabanita Chatterjee, Subhadip Das, Dinesh K. Ahirwar, Sanjay Mishra, Ramesh Ganju. Cannabinoid receptor 2 agonist JWH-015 inhibits growth and metastasis of triple negative breast cancers through regulation of autophagy mechanism [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 2018.
Abstract 203: Pseudosemiglabrin inhibits tumor angiogenesis and tumor growth of human colon cancer in xenograft mouse models by downregulating endothelial functions, Notch and VEGF pathways
Authors: Hassan Loiy E. Ahmed , Seeni Azman , Majid Amin M. Abdul
Published: August 2019

Many antineoplasic agents were identified from natural product. One such agent is (-)-pseudosemiglabrin extracted from aerial parts of Tephrosia apollinea, while significant progress has been made in understanding the antineoplasic and anti-inflammatory of prenylated flavone (-)-pseudosemiglabrin (SSG), little is known about its antiangiogenic and antitumor in vivo. Using rat aortic ring assay and zebrafish model, SSG dramatically suppressed angiogenesis in vitro, ex vivo and in vivo models, SSG profoundly inhibited the outgrowth of new vessels from explants rat aortic ring, moreover it impaired human endothelial cells (HUVEC) migration and formation of capillary-like structure in vitro in none toxic doses. In this study we hypothesize that SSG can hampered tumor angiogenesis and tumor growth of colorectal cancer in ectopically and orthotopically implanted in nude mice, oral administration of SSG (25- 100mg/kg) significantly inhibited colon tumors growth in nude mice. Histological and Immunohistochemistry analysis further revealed that the stained blood vessels and expression of CD31 in xenograft were remarkably decreased in tumors excised from animals treated with SSG. Mechanistically the compound inhibits tumor angiogenesis by downregulation of Notch downstream (NCID); also SSG was found to inhibit VEGF secretion from human cancer cells as well as endothelial cells. In conclusion, the results demonstrated an original role of (-)-pseudosemiglabrin in inhibiting tumor angiogenesis and tumor growth via inhibition Notch and VEGF pathways. This study also validates the central role of Notch in tumor angiogenesis and suggests that (-)-pseudosemiglabrin could be a potential therapeutic candidate for cancer and other angiogenesis related diseases.

Note: This abstract was not presented at the meeting.

Citation Format: Loiy E. Ahmed Hassan, Azman Seeni, Amin M. Abdul Majid. Pseudosemiglabrin inhibits tumor angiogenesis and tumor growth of human colon cancer in xenograft mouse models by downregulating endothelial functions, Notch and VEGF pathways [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 203.
Abstract 2030: Inhibiting the GAS6/AXL axis suppresses tumor progression by blocking the interaction between cancer-associated fibroblasts and cancer cells in gastric carcinoma
Authors: Lee Dagyoeng , Woo Jongsu , Bae Cheong A , Ham In-Hye , Oh Hye Jung , Son Sang-Yong , Han Sang-Uk , Kim Tae-Min , Yoon Jung Hwan , Park Won Sang , Hur Hoon
Published: August 2019

Cancer-associated fibroblasts (CAF) are a major component of tumor stroma and their effect on the progression of gastric carcinoma (GC) has recently been demonstrated. However, agents targeting the interaction between CAF and GC cells have not been applied in a clinical setting. The aim of this study was to investigate the effect of an AXL inhibitor on CAF-induced GC progression. We examined growth arrest specific factor 6 (GAS6) expression and phosphorylation of AXL in various cell lines, including GC cells and non-cancerous cells, by qPCR, western blot, and immunocytochemistry. We investigated the role of CAF-derived GAS6 on the migration and proliferation of GC cells using migration and cell survival assays. The effect of the AXL inhibitor, Bemcentinib, on the CAF-induced aggressive phenotype of GC cells was also investigated. Finally, we performed immunohistochemistry to measure the phosphorylation of AXL in a tissue microarray composed of 175 GC tissues, and evaluated its correlation with the prognosis of GC patients. qPCR and western blot analysis showed that GAS6 expression was higher in CAF relative to other cells. We found that CAF increased the phosphorylation of AXL (P-AXL), differentiation into a mesenchymal-like phenotype, and cell survival in GC cell lines. When the expression of AXL was inhibited in GC cells, the effect of CAF was reduced. In addition, Bemcentinib, a small molecule inhibitor of AXL, suppressed the effects of CAF on GC cell lines. In GC tissues, P-AXL positively correlated with the expression of vimentin (P < 0.001) and high levels of P-AXL were significantly associated with poor overall survival (P = 0.022).These findings suggest that CAF is a major source of GAS6 in the GC microenvironment and actively promotes an aggressive phenotype in GC cells through AXL activation. We conclude that an AXL inhibitor may be a novel agent for GC treatment.

Citation Format: Dagyoeng Lee, Jongsu Woo, Cheong A Bae, In-Hye Ham, Hye Jung Oh, Sang-Yong Son, Sang-Uk Han, Tae-Min Kim, Jung Hwan Yoon, Won Sang Park, Hoon Hur. Inhibiting the GAS6/AXL axis suppresses tumor progression by blocking the interaction between cancer-associated fibroblasts and cancer cells in gastric carcinoma [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 2030.
Abstract 2033: Bidirectional paracrine IL-33-TGF-beta signaling underlies a tumor-promoting stem cell niche
Authors: Oshimori Naoki , Oshimori Sachiko , Elhance Ajit , Leitenberger Justin , Kumar Sushil
Published: August 2019

Malignant tumor progression and aggressive tumor recurrence after treatment are driven by subsets of cancer stem cells (CSCs), but targeting CSCs remains a major challenge due to their plasticity. CSCs in squamous cell carcinoma reside at the tumor-stromal interface, a paramount tumor microenvironment (TME) for interactive crosstalk between cancer cells and various other cells via soluble mediators, and tumor-infiltrating immune cells are pivotal regulators of tumor progression. However, the molecular link between tumor-associated immune-inflammatory responses and the regulation of CSCs are largely unknown. To define a framework of the CSC niche, we have been focusing on crosstalk between TGF-beta-activated CSCs and their neighboring immune cells. By transcriptomic analysis, we identify interleukin-33 (IL-33) as the most upregulated cytokine in TGF-beta-activated CSCs. We find that IL-33 is secreted in response to the activation of antioxidant responses, and IL-33 mobilizes a subset of tumor-promoting macrophages that express the IL-33 receptor in the adjacent stroma. Tumor-specific IL33 knockdown in vivo dampens both invasive carcinogenesis and paracrine TGF-beta signaling in tumor cells, indicating that IL-33 has a crucial role in the formation of TGF-beta-rich tumor-promoting TME by affecting tumor-infiltrating bone marrow-derived monocytes. Together, these findings on a bidirectional paracrine signaling axis provide novel insights into mechanisms of the formation a CSC privilege niche, which may hold great potential to drastically improve the therapeutic efficacy of current and future cancer treatments.

Note: This abstract was not presented at the meeting.

Citation Format: Naoki Oshimori, Sachiko Oshimori, Ajit Elhance, Justin Leitenberger, Sushil Kumar. Bidirectional paracrine IL-33-TGF-beta signaling underlies a tumor-promoting stem cell niche [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 2033.
Abstract 2047: Identification of endometrial cancer specific immuno-therapeutic targets by comprehensive In Silico analysis
Authors: Ikeda Yuji
Published: August 2019

Background; Immune-therapeutic targets specific to endometrial cancer has been unclarified. In term of HLA allele type, about 60% and 40% of Japanese has HLA-A*24 and -A*02,respectively. Development of new immune-therapeutics taking account of these types is required. We herein aimed to identify immune-therapeutic targets specific to endometrial cancer and commonly available for Japanese patients using in Silico analysis, which can search comprehensively peptides binding to HLA molecule in an allele-specific manner. Methods; mRNA sequencing data obtained from 582 samples of endometrial cancer and 53 control samples from 31 normal organs were extracted from cancer genome atlas (TCGA) database and Genotype-Tissue Expression project, respectively. After filtering of protein coding genes, 8 to 12 peptides with high binding affinity to HLA-A*24 and -A*02 alleles were identified by an artificial neural network soft trained for peptide-MHC class I binding. Results; Based on mRNA sequencing data, we identified 71 genes that are expressed in endometrial cancer but not in control organs. Of them, top ten genes with high mRNA levels in the cancer are selected. Analysis of binding affinity revealed that 21 and 146 peptides derived-from ten genes of interest showed high binding affinity to HLA-A*24 and -A*02, respectively. Especially, one target gene showed the highest expression levels in endometrial cancer, and possessed 15 and 58 candidate peptides in HLA-A*24 and -A*02, respectively. Conclusion; By comprehensive analysis using mRNA data, we identified reliable candidates target of immune-therapeutics for HLA-A*02 and -A*24 patient with endometrial cancer.

Citation Format: Yuji Ikeda. Identification of endometrial cancer specific immuno-therapeutic targets by comprehensive In Silico analysis [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 2047.
Abstract 2050: Cell penetrating peptide, ST101, disrupts ATF5 regulation of anti-apoptotic Bcl-2 family proteins, resulting in induction of cancer cell death in vitro and tumor growth inhibition/regression in vivo
Authors: Rotolo Jim A. , Ramirez Rick , Koester Mark , Leong Siok , Ghamsari Lila , Merutka Gene , Kappel Barry J.
Published: August 2019

Anti-apoptotic B cell lymphoma 2 (Bcl-2) family proteins are frequently overexpressed across a variety of tumors, resulting in tumor cell survival and resistance to therapy. Inhibition of the expression or activity of these survival factors is an attractive approach for cancer therapy. Activating transcription factor 5 (ATF5) regulates gene transcription of anti-apoptotic Bcl-2 family proteins in neural progenitor cells and a wide range of human cancer cells. Here, we describe ST101, a rationally designed, synthetic, D-amino acid, cell penetrating peptide therapeutic designed to disrupt the protein-protein interactions driving ATF5-regulated gene transcription. Exposure of HL60 promyelocytic leukemia cells and MCF7 breast adenocarcinoma cells to low micromolar concentration of ST101 resulted in a decrease in MCL-1, BCL-2 and BIRC5 (Survivin) mRNA expression at 4 and 24 hrs post exposure. Further, exposure to ST101 resulted in a dose-dependent loss of viability across a panel of human cancer cells, including MCF7, HL60, U251 glioblastoma, A375 melanoma, DU145 prostate cancer, and A549 lung adenocarcinoma, characterized by an increase in annexin V and PI staining by flow cytometry peaking 48 hrs post exposure, resulting in a median half maximal effective concentration (EC50) value of 4.0 micromolar. In contrast, normal human peripheral blood mononuclear cells and bone marrow mononuclear cells were resistant to ST101-mediated cell death, with >80 micromolar EC50 values. In mouse xenograft experiments, 25mg/kg ST101 administered three times per week for three weeks resulted in significant tumor regression in MCF7 and U251 subcutaneous tumors as well as tumor growth delay in HL60 subcutaneous tumors. Tumor growth remained significantly inhibited weeks after the last treatment in the MCF7 and U251 models. In summary, ST101 selectively kills cancer cell lines by decreasing BCL-2 family gene expression, resulting in significant reductions in tumor growth in mouse models. Taken together, these data validate ST101 as a potent peptide therapeutic candidate for a variety of solid tumor and hematologic malignancies.

Citation Format: Jim A. Rotolo, Rick Ramirez, Mark Koester, Siok Leong, Lila Ghamsari, Gene Merutka, Barry J. Kappel. Cell penetrating peptide, ST101, disrupts ATF5 regulation of anti-apoptotic Bcl-2 family proteins, resulting in induction of cancer cell death in vitro and tumor growth inhibition/regression in vivo [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 2050.
Abstract 2053: Targeting GRK2 increases chemotherapy-induced apoptosis in medulloblastoma
Authors: Pathania Anup S. , Ren Xiuhai , Mahdi Min , Shackleford Gregory M. , Erdreich-Epstein Anat
Published: August 2019

Background: G protein receptor kinase-2 (GRK2; ADRBK1) belongs to the family of G protein-coupled receptor (GPCR) kinases (GRKs). GRK2 phosphorylates GPCR and non-GPCR targets including cell surface receptors, cytoskeletal and mitochondrial proteins and transcription factors. It also functions in embryonic development, heart function, metabolism, and some cancer models. Here we hypothesized that targeting GRK2 will sensitizes cancer cells to therapy. We tested this in medulloblastoma, the most common malignant brain tumor in children.

Methods: siRNA transfections, shRNA viral infections, SDS-PAGE, western blotting, flow cytometry. Cells: UW228 and DAOY medulloblastoma.

Results: We found that GRK2 inhibition promotes anti-tumor effects of chemotherapeutic drugs cisplatin and etoposide in UW228 and DAOY medulloblastoma cell lines. GRK2 silencing increased apoptosis induced by cisplatin and etoposide and GRK2 overexpression decreased apoptosis induced by these drugs in UW228 and DAOY. Time dependent studies revealed that GRK2 overexpression delayed cisplatin- and etoposide-induced apoptosis measured by the induction of Annexin V positive population and by PARP-1 cleavage. GRK2 mutants, GRK2 K220R (kinase deficient mutant) and GRK2 S670A (cannot be phosphorylated) were similar to wildtype GRK2 in delaying the cisplatin and etoposide induced apoptosis. GRK2 knockdown antagonized serum induced AKT and ERK phosphorylation in both lines whereas GRK2 overexpression promoted phosphorylation, suggesting that GRK2 is important in medulloblastoma survival pathways.

Conclusions: These results demonstrate that targeting GRK2 enhances cisplatin- and etoposide-induced apoptosis in medulloblastoma cell lines and suggests that GRK2 inhibitors may enhance responsiveness of medulloblastoma to chemotherapy.

Citation Format: Anup S. Pathania, Xiuhai Ren, Min Mahdi, Gregory M. Shackleford, Anat Erdreich-Epstein. Targeting GRK2 increases chemotherapy-induced apoptosis in medulloblastoma [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 2053.