bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4155: Interstitial fluid flow-induced hepatocellular carcinoma cell invasion requires MEK/ERK signaling
Authors: Shah Arpit D. , Bouchard Michael , Shieh Adrian C. 
Published: August 2015

Abstract
Background: Worldwide hepatocellular carcinoma (HCC) is the third leading cancer-related cause of death. Within the United States, HCC maintains a rising incidence rate of 3% each year since 1992 with a 5 year survival rate of less than 15%. The American Cancer Society has estimated 33,190 new cases off HCC and 23,000 HCC related deaths in the year 2014 within the United States. Our work aims to determine the role of biomechanical forces such as interstitial fluid flow (IFF) in HCC cell invasion. We hypothesized that HCC cells utilize the hypothesized ‘autologous chemotaxis’ mechanism to invade via secretion of the chemokine CXCL12 and expression of its receptor CXCR4. Autologous chemotaxis occurs when a biologically significant chemokine gradient is established by IFF, driving downstream invasion. In an effort to elucidate signaling pathways involved in IFF-induced HCC invasion, we have identified a potential role for MEK/ERK signaling.
Materials and Methods: HCC cells or primary rat hepatocytes (PRH) were seeded into in a 3-D matrix composed of Matrigel and type I rat tail collagen, then exposed to static or interstitial flow conditions for 24 hours. The functional role of CXCR4 was tested using the CXCR4-antagonist AMD3100. MEK and ERK were inhibited using U0126 (25 μM) and FR180294 (10 μM), respectively. Phospho-MEK/ERK was assessed using western blot.
Results and Discussion: The 3D invasion assay demonstrated that the HCC cells, Huh7 and Hep3B, invaded significantly more in response to IFF (5.5 fold increase in invasion compared to their respective static condition), while PRHs showed no response. AMD3100 was used to block the CXCR4 receptor which resulted in reduced invasion in Huh7 and Hep3B cells. Furthermore when U0126 and FR180204 were incorporated into the 3D invasion assay a significant reduction in flow-induced invasion was observed in Huh7 cells. However, interstitial flow or CXCR4 inhibition did not affect pMEK/ERK, suggesting that MEK/ERK are not modulated directly by IFF and are not downstream of CXCR4.
Conclusion: Our results demonstrate that interstitial fluid flow stimulates HCC cell invasion. CXCR4/CXCL12 signaling and MEK/ERK signaling both are required for invasion, but appear to operate via separate mechanisms. Further investigation into these two separate mechanisms would provide better insight into flow-induced HCC invasion. This information could potentially enhance current cancer therapies and provide patients with better treatment options.
Citation Format: Arpit D. Shah, Michael Bouchard, Adrian C. Shieh. Interstitial fluid flow-induced hepatocellular carcinoma cell invasion requires MEK/ERK signaling. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4155. doi:10.1158/1538-7445.AM2015-4155
Abstract 4173: Endoglin regulation of Smad2/3 function mediates beclin1 expression and endothelial autophagy during angiogenesis
Authors: Lee Nam Y. , Pan Christopher , Kumar Sanjay 
Published: August 2015

Abstract
Autophagy is the targeted degradation of proteins and organelles critical for homeostasis and cell survival. Transforming growth factor beta (TGF-beta) is known to differentially regulate autophagy under distinct physiological conditions by inducing inhibitory, stimulatory, or a combination of these opposing signaling pathways. Importantly, how TGF-beta signaling controls autophagic responses in the endothelial cell (EC) system during angiogenesis remains unknown. Here we identified endoglin, an EC-specific TGF-beta co-receptor essential for angiogenesis, as a key determinant of autophagy. We found that Smad2/3 are major transcriptional regulators of basal autophagy that specifically target beclin1 (BECN1) gene expression. Selectively blocking ALK5 activation or silencing endogenous Smad2/3 markedly increased BECN1 expression whereas altering ALK1/Smad1/5/8 signaling had no effect. Endoglin potentiates basal autophagy by suppressing ALK5/Smad2/3 activation. Notably, increased beclin1 levels upon Smad2/3 knockdown directly correlated with enhanced autophagy during angiogenesis. Taken together, these results establish endoglin as a critical mediator of autophagy and demonstrate a new cellular mechanism by which Smad2/3 inhibits angiogenesis.
Citation Format: Nam Y. Lee, Christopher Pan, Sanjay Kumar. Endoglin regulation of Smad2/3 function mediates beclin1 expression and endothelial autophagy during angiogenesis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4173. doi:10.1158/1538-7445.AM2015-4173
Abstract 4190: A mouse model of pRb2/p130 in prostate cancer
Authors: Boffo Silvia , Klein-Szanto Andres J. , Rizzolio Flavio , Giordano Antonio 
Published: August 2015

Abstract
INTRODUCTION
Prostate Cancer (PCa) is the most commonly diagnosed cancer in men and it is responsible for the second highest number of cancer-related deaths in the United States. There is an urgent need for new treatments and better experimental animal models to study PCa development, progression, and metastasis.
Prostate cancer is a heterogeneous and multifocal disease that arises from several genetic and epigenetic aberrations. Among different altered pathways, the Retinoblastoma is inactivated in more than fifty percent of PCa. Despite a plentiful literature, the activity of pRb members (pRb, pRb2/p130 and p107) is not fully elucidated.
From a previously published mouse model, we know that the inactivation of all the pRb members induces aberrant proliferation and it is sufficient for tumor initiation.
Here, we describe a novel double ko (Rb, pRb2/p130) mouse model, which offers a new opportunity to dissect prostate tumor initiation and development.
MATERIALS AND METHODS
To study the effect of pRb2/p130 inactivation on prostate tumorigenesis and progression, we knocked out pRb and/or pRb2/p130 in the prostate tissue (Probasin-CRE). The genotypes of each gene were determined by PCR. To analyze the effects of pRb2/p130 loss, we collected prostate tissues at different time points (3, 6, 9, 12 and 18 months). A portion of each prostate sample was fixed in formalin and embedded in paraffin to perform immunohistochemistry analyses and another portion was flash-frozen for molecular studies.
The histological classification of each prostate samples was made on H&E-stained sections by A.K.S., a veterinary pathologist with extensive experience in murine prostate pathology.
RESULTS
As reported in literature, pRb family disruption in the prostate epithelium induces prostatic intraepithelial neoplasia. The prostate of triple ko mice showed extensive abnormal gland architecture beginning as early as 2 months of age with a focal hyperplasia. By 3 months of age, all the triple ko mice displayed characteristic traits of murine prostatic intraepithelial neoplasia (mPIN). Double ko (pRb and pRb2/p130) mice developed hyperplasia after 6 months and focally invasive well-differentiated adenocarcinomas by 12-18 months.
CONCLUSIONS
Our preliminary findings suggest that the inactivation of pRb and pRb2/p130 leads to the development of a prostate tumor phenotype and highlight the importance of the function of pRb2/p130 in the prostatic epithelium.
This model will be useful to clarify the molecular mechanism of pRb2/p130 function and represents a new opportunity for cancer therapy.
Citation Format: Silvia Boffo, Andres J. Klein-Szanto, Flavio Rizzolio, Antonio Giordano. A mouse model of pRb2/p130 in prostate cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4190. doi:10.1158/1538-7445.AM2015-4190
Abstract 420: The role of tumor microenvironment in therapy resistance and melanoma progression
Authors: Somasundaram Rajasekharan , Zhang Gao , Wagner Stephan N. , Fukunaga-Kalabis1 Mizuho , Herlyn Meenhard 
Published: August 2015

Abstract
Melanoma patients develop resistance to both chemo- and targeted-therapy drugs. Promising pre-clinical and clinical results with immune checkpoint inhibitors using antibodies directed against CTLA4 and PD1 have re-energized the field of immune-based therapies in melanoma. However, similar to chemo- or targeted-therapies only subsets of melanoma patients respond to immune checkpoint blockade. Studies from our laboratory have indicated that the tumor microenvironment (TME) might play a role in the emergence of therapy resistant tumor subpopulations. The TME actively recruits a number of immune and non-immune cells. A recent study describes immune cells as an important component of TME. About 33% of the infiltrating immune cells are of ‘B-cell’ lineage and yet, there is very little information on the role of these cells in melanoma cell biology. Our results suggest that B cells isolated from the tumor tissues of melanoma patients show higher inflammatory cytokine expressions (IGF-1, IL-1, PDGF and VEGF) when compared to circulating B cells. Presence of inflammatory cytokines in the TME results in the induction of heterogeneous melanoma subpopulation, down modulation of melanoma associated antigens (MAA), and thus, therapy resistance of melanoma. Here, we report a novel mechanism of acquired drug resistance in melanoma induced by tumor-associated B cells involving FGF-2/FGFR-3 and IGF-1 signaling. B-cell derived IGF-1 modulates the emergence of therapy resistant heterogeneous melanoma subpopulations. Neutralization of IGF-1 reverses the induction of heterogeneous melanoma subpopulations. We are currently evaluating the mechanism of down modulation of MAA in presence of inflammatory cytokines. Our studies confirm the important role of the TME in the induction of therapy resistant tumor subpopulations and offer a new combination treatment approach of targeting melanoma (Supported by AMRF and NIH grant CA114046).
Citation Format: Rajasekharan Somasundaram, Gao Zhang, Stephan N. Wagner, Mizuho Fukunaga-Kalabis1, Meenhard Herlyn. The role of tumor microenvironment in therapy resistance and melanoma progression. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 420. doi:10.1158/1538-7445.AM2015-420
Abstract 4201: Development of a syngeneic metastatic mouse model of malignant lymphoma
Authors: Matsumoto Takuro , Suetsugu Atsushi , Shibata Yuhei , Nakamura Nobuhiko , Aoki Hitomi , Kunisada Takahiro , Shimizu Masahito , Tsurumi Hisashi , Hoffman Robert M. 
Published: August 2015

Abstract
We have developed a mouse model of highly malignant lymphoma. The EL4 cell line was established from a lymphoma induced in a C57/BL6 mouse by 9,10-dimethyl-1,2-benzanthracene. Red fluorescent protein (RFP)-EL4 lymphoma cells were established. EL4-RFP cancer cells were injected in the tail vein of C57/BL6 GFP transgenic mice. EL4-RFP metastasis was observed in the lymph nodes of the superior mediastinum and in the liver 28 days later. The EL4-RFP tumors were visualized in the liver with the Olympus SZX7 microscope. Large EL4-RFP liver metastases in C57/BL6 GFP mice had GFP stromal cells derived from the host animal around the edge of EL4-RFP tumors. In addition, EL4-RFP lymphoma cells metastasis was formed in perigastric lymph node. EL4-RFP lymphoma cells circulated in the peripheral blood. Furthermore, EL4-RFP lymphoma cells were observed in bone marrow of C57/BL6 GFP transgenic mice. In summary, lymph node metastasis, liver metastasis and bone marrow metastasis were found approximately 4 weeks after transplantation. This malignant lymphoma model can be used to study early tumor development, metastasis, the role of stroma and discovery and evaluation of novel therapeuticus for this treatment-resistant disease.
Citation Format: Takuro Matsumoto, Atsushi Suetsugu, Yuhei Shibata, Nobuhiko Nakamura, Hitomi Aoki, Takahiro Kunisada, Masahito Shimizu, Hisashi Tsurumi, Robert M. Hoffman. Development of a syngeneic metastatic mouse model of malignant lymphoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4201. doi:10.1158/1538-7445.AM2015-4201
Abstract 4216: Targeting NANOG: genes, proteins and response to viral RNAi in preclinical models
Authors: Jessup J. Milburn , Mattoo Abid R. , Korokhov Nikolay 
Published: August 2015

Abstract
Purpose: Since the NANOG family controls stemness and neoplastic progression in human colorectal carcinoma (CRC), our postulate is that targeting the dominant NANOG and NANOGP8 members with viral vector delivered shRNA will inhibit CRC progression in preclinical models and that replicating viral vectors are more active than non-replicating vectors.
Methods: Cells were human CRC lines (Clone A, CX-1 and LS174T) and mouse fibroblasts (3T3). shRNA specific to NANOG (shNG-1), NANOGP8 (shNp8-1) or a control (shNEG) were created in lentivirus (LV) and chimeric adenovirus 5/3 (Ad5/3). CRC and 3T3 were cultured in monolayer with serum-containing medium or in suspension in ultra low attachment plates without serum. Nanostring 770 gene cancer panel was analyzed in 72 hr cultures with NMF clustering and GSEA analysis. Annexin V expression and caspase-3, 8 or 9 activation in suspension culture assessed Apoptosis. LV or Ad5/3 shRNAs were added to spheroids in 3-D suspension cultures to test therapy in vitro. Intratumoral injection of LV or Ad5/3 shRNA into 5-10 mm sub cutis nodules in groups of 5-10 NOD/SCID mice measured activity in vivo.
Results: LV shNp8-1 in both Clone A and CX-1 caused significant inhibition of TGF-beta, EGFR, NRF2, E2F3 and the Cyclin D1 pathways by NMF and GSEA analysis in monolayer culture. LV shNp8-1 inhibited growth of LS174T, Clone A and CX-1 by 50% in 3 days in 3-D suspension cultures without further decrease. Apoptosis was due to Caspase 9 activation. In contrast, Ad5/3-shNp8-1 decreased spheroid growth by up to 94% at 9 -14 days while Ad5/3 decreases spheroid growth by 80%. Ad5/3-shNp8-1 but nit Ad5/3 was active in co-cultures with 3T3 to a 2:1 CRC:3T3 ratio. Transduction level was ∼50% in LV treated cells but ∼90% in Ad5/3 treated cells. Intratumoral injection of LVshNG-1 or LVshNp8-1, decreased tumor size by 50% compared to LVshNEG or untreated CX-1 tumors for 11 days after which tumor growth rebounded to match controls. Ad5/3-shNp8-1 injection into 5-9 mm nodules inhibited CX-1 and LS14T growth by ∼50% until euthanized.
Conclusions: Targeting NANOGP8 more than NANOG inhibits proliferation and activates apoptosis through multiple pathways. Nonreplicating LV delivered shRNA inhibited tumor growth in in vitro and in vivo preclinical models but replicating oncolytic Ad5/3 infects more cells and promotes shRNA efficacy. Replicating vectors targeting NANOG are important for further development.
Citation Format: J. Milburn Jessup, Abid R. Mattoo, Nikolay Korokhov. Targeting NANOG: genes, proteins and response to viral RNAi in preclinical models. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4216. doi:10.1158/1538-7445.AM2015-4216
Abstract 4217: Targeting head and neck cancer-initiating cells with combinatorial therapeutics through pharmacological inhibition of ROS scavenger and conventional chemotherapy
Authors: Chang Ching-Wen , Lo Jeng-Fan 
Published: August 2015

Abstract
Background: Cancer-initiating cells (CICs) contribute to tumor initiation, progression and therapy resistance through critical physiology mechanisms. Redox homeostasis plays an important role in the regulation of cancer stemness. We recently have reported that cancer cells containing differential level of ROS display distinct physiological properties of stemness, proliferative activity, chemoresistance and tumorigenicity. Therefore, to uncover the regulatory physiological mechanisms by which to sustain the low lever of ROS in CICs warrants an important study for future therapy development.
Results: In this study, we demonstrated that lower ROS levels in head and neck CICs (HN-CIC) enriched population are associated with increased expression of ROS scavenger such as catalase and SOD2. Pharmacological and genetic inhibition of SOD2 or catalase caused the increased expression of differentiation marker, cytokeratin 18, but the decreased expression of CIC marker in HN-CICs. In addition, the sphere formation abilities of HN-CICs under genetic inhibition of SOD2 or catalase were also significantly abolished. Notably, combinatorial treatment with ROS scavenger inhibitors and conventional anticancer drugs synergistically reduced the stemness properties and tumorigenicity both in vitro and in vivo. Using the Oncomine database, we found associations between SOD2 and catalase expression and increasing head and neck cancer recurrence, tumor invasion and resistance to Gemcitabine treatment.
Conclusion: Together, these data demonstrate the importance of antioxidant defense to maintain the low ROS level, cell viability and chemosensitivity of HN-CICs. Pharmacological inhibition of ROS scavenger (SOD2 and catalase) might be a future alternative HNSCC treatment in combination with conventional chemotherapy
Citation Format: Ching-Wen Chang, Jeng-Fan Lo. Targeting head and neck cancer-initiating cells with combinatorial therapeutics through pharmacological inhibition of ROS scavenger and conventional chemotherapy. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4217. doi:10.1158/1538-7445.AM2015-4217
Abstract 4218: Phenotypic selection screening reveals cancer stem cell therapeutic targets
Authors: Hurt Elaine M. , Flynn Matt , Thomas Suneetha , van Vlerken-Ysla Lilian , Sandercock Alan , Rust Steven , Ralph Minter , Hollingsworth Robert 
Published: August 2015

Abstract
Cancer stem cells (CSCs) represent an important subpopulation of tumor cells that are untouched and perhaps even expanded after chemotherapy and radiation. This population has been shown in murine models of cancer to be solely responsible for the regrowth of tumors following chemotherapy, underscoring the importance of therapeutically targeting this population. Therefore, we sought to find novel targets against CSCs using a phenotypic selection screen. Using a designed ankyrin repeat (DARpin) library, we panned cells disaggregated from tumor spheres from three patient-derived xenograft models of late stage pancreatic cancer to enrich for DARPins that bind CSCs. Once released from the cells, the DARPins were further analyzed for binding to both normal, pancreatic cancer total tumor cells and pancreatic CSCs in order to choose targets that showed either pancreatic cancer-specific or pancreatic CSC-specific binding. These DARPins were then tested in a sphere formation assay, an EZH2-based high content imaging screen, and a standard proliferation assay. Those DARPins showing good cancer-specific binding profiles were also tested for internalization and killing using a saporin-conjugated secondary antibody. Using this approach, we identified 30 DARPins of interest for either direct inhibitory antibody approaches or for potential antibody drug conjugate or nanoparticle delivery approaches. Identification of the proteins bound by each of the DARPins was done using either immunopreciptation followed by liquid chromatography tandem-mass spectrometry or through Retrogenix's cell expression microarray technology. Identified targets included known CSC antigens such as integrin alpha6 (CD49f) as well as other potentially novel targets. These results highlight the possibilities of finding therapeutic targets for CSCs and opens up an exciting possibility for novel therapies.
Citation Format: Elaine M. Hurt, Matt Flynn, Suneetha Thomas, Lilian van Vlerken-Ysla, Alan Sandercock, Steven Rust, Minter Ralph, Robert Hollingsworth. Phenotypic selection screening reveals cancer stem cell therapeutic targets. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4218. doi:10.1158/1538-7445.AM2015-4218
Abstract 422: The role of CCL2 in the breast cancer tumor microenvironment and metastasis
Authors: Lavender Nicole , Sai Jiqing , Yang Jinming , Novitskiy Sergey V. , Richmond Ann 
Published: August 2015

Abstract
Background: Although many effective breast cancer treatments exist, metastasis remains a major problem. Increasing evidence suggests that leukocyte-associated chemokines may play an important role in tumor growth and metastasis. For instance, recent data shows that reduction of CCL2 diminishes the recruitment of inflammatory monocytes to the tumor microenvironment (TME) and inhibits metastatic seeding to lung and bone marrow. Therefore, our goal is to examine the impact of CCL2 on the TME and pre-metastatic niche (i.e., lung).
Methods: We are investigating how delivery of this chemokine affects breast tumor growth and metastasis by treating mice possessing highly metastatic and poorly metastatic tumors with CCL2, a CCL2 receptor (CCR2) antagonist, or vehicle for two weeks. Treatment will start 7 days after tumor implantation. On days 7, 14 or 21, tumors, lungs, and regional lymph nodes will be harvested and tumor size, number of metastases, and leukocyte infiltrate in both the tumor and the metastatic niche will be analyzed by FACS. This will allow us to fully characterize how intranasal delivery of CCL2 will affect the presence of mesenchymal stem cells (CD44+, CD73+, CD90+, CD146+/NOTCH3+, CD19-, CD45-), subsets of T cells, and myeloid cells in the TME or pre-metastatic niches. We will evaluate the intracellular cytokine and enzymatic profile (IL-4, IL-13, IL-17, ARG1 or IFNγ, IL-12, iNOS2) of Brefeldin-treated leukocytes to reveal whether they are pro- or anti-tumorigenic, respectively.
Results: Our preliminary studies demonstrate that CCL2 enhances the release of hydrogen peroxide from neutrophils, a mechanism utilized by these cells to kill tumors cells. Furthermore, the addition of CCL2 to co-cultures of neutrophils and tumor cells increases the ability of neutrophils to kill tumor cells.
Conclusions & Future Directions: Our data shows that delivery of CCL2 in vitro enhances the activity of neutrophils and their ability to kill tumor cells. Ongoing studies are examining the effects of in vivo and ex-vivo delivery of CCL2 or a CCR2 antagonist on primary tumor growth and metastasis. We will also investigate how administration of CCL2 or a receptor antagonist will affect the presence or absence of leukocytes in the TME. These findings will determine whether the addition of CCL2 during treatment of the primary tumor will provide a mechanism for reducing the potential of future breast cancer metastases.
Citation Format: Nicole Lavender, Jiqing Sai, Jinming Yang, Sergey V. Novitskiy, Ann Richmond. The role of CCL2 in the breast cancer tumor microenvironment and metastasis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 422. doi:10.1158/1538-7445.AM2015-422