bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4025: An old foe with a new function: JAM-A regulates expression of EphB4 and inhibitor of apoptosis proteins in breast cancer
Authors: Vellanki Sri HariKrishna , Cruz Rodrigo G. , Richards Cathy E. , Brennan Kieran , Hopkins Ann M. 
Published: August 2018

Abstract
Over-expression of Junctional Adhesion Molecule-A (JAM-A) in breast tumors has been linked with increased risk of metastasis. Since JAM-A is co-expressed with the receptor tyrosine kinase (RTK) HER2 and regulates HER2 protein expression, we hypothesized that JAM-A regulates the expression and function of other RTKs. Using a joint RT-PCR array/western blot approach in three breast cell lines with modified JAM-A expression, we identified EphB4 as a downstream target of JAM-A. Interestingly, EphB4 gene silencing also cross-regulated the protein expression of JAM-A. Immunofluorescence analysis revealed colocalisation of JAM-A and EphB4 at the cell membrane. Since JAM-A contains a PDZ-binding motif and EphB4 contains a PDZ domain, we speculated that this might underlie the basis of an interaction. Accordingly, overexpression of a JAM-A mutant lacking the PDZ domain failed to upregulate EphB4 expression to the same extent as wild type JAM-A. Next we investigated the signalling events downstream of JAM-A and EphB4 and found downregulation of the tight junction protein ZO-1 in breast cells silenced for JAM-A or EphB4. Using a bioinformatics approach, we identified the transcription factor c-FOS as a potential regulator of JAM-A and EphB4 expression. In JAM-A- or EphB4-silenced cells, c-FOS expression was not localized in the nucleus (relative to cells transfected with negative control siRNAs). Furthermore, ERK5 phosphorylation was inhibited in JAM-A- or EphB4-silenced cells, suggesting a mechanism for reduced c-FOS phosphorylation on Serine-32 and its consequent exclusion from the nucleus. In the context of a proposed block on c-FOS transcriptional activity, we examined the genes affected and show for the first time downregulation of inhibitor of apoptosis proteins (IAPs) in JAM-A- or EphB4-silenced cells. In conclusion, our data are consistent with a model whereby JAM-A regulates EphB4 expression in a PDZ-dependent manner and influences apoptosis via a pathway involving ERK5 and c-FOS. Our data suggest that the pharmacological antagonism of JAM-A in JAM-overexpressing breast cancers merits consideration by virtue of its predicted ability to downregulate EphB4 expression and remove a blockade on apoptosis.
Citation Format: Sri HariKrishna Vellanki, Rodrigo G. Cruz, Cathy E. Richards, Kieran Brennan, Ann M. Hopkins. An old foe with a new function: JAM-A regulates expression of EphB4 and inhibitor of apoptosis proteins in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4025.
Abstract 4037: Interactions between STAT3 and TGFβ in the regulation of hyaluronic acid synthesis and signaling in immortalized human mammary epithelial cells
Authors: Trevizo-Barresi Brenda G. , Welsh JoEllen 
Published: August 2018

Abstract
Evidence suggests that growth of cancer stem cells (CSCs) drives intratumor heterogeneity, a major clinical problem that results in drug resistance, recurrence of tumors, and metastatic progression of breast cancer. Therapeutic strategies to eliminate CSCs require identification of signaling pathways that facilitate the emergence and/or expansion of CSCs. These signaling pathways are implicated in altering cell behavior, as well as promoting the expression of genes involved in epithelial-mesenchymal transition (EMT), cell migration and invasion. Previous work demonstrated that a cooperative STAT3/SMAD3 gene transcription program increased the expression of Hyaluronan Synthase 2 (HAS2), an enzyme that produces Hyaluronic Acid (HA) which serves as a CD44 ligand and facilitates EMT. We hypothesize that STAT3 is a critical regulator of HA synthesis and signaling which enhances EMT and stemness of human mammary epithelial cells, driving tumor progression. We tested this hypothesis in a TGFβ triggered model of EMT. Immortalized human mammary epithelial cells (HMLE) expressing either a control vector (pWZL-GFP) or a vector containing TGFβ1 (pWZL-TGFβ1-GFP) were characterized with respect to STAT3 signaling, HA synthesis, and mammosphere formation. Our data revealed that HMLE-pWZL-TGFβ1-GFP cells have significantly higher expression of active STAT3 (Y705) compared to control cells, but total STAT3 gene and protein expression were similar in the two cell lines. HMLE-pWZL-TGFβ1-GFP cells exhibited a significant increase in expression of HAS2 and HAS2-AS1, a long non-coding RNA that enhances the transcription of HAS2. Preliminary results show that overexpression of TGFβ1 increased HA secretion in media compared to control cells. HMLE-pWZL-TGFβ1-GFP cells demonstrated significantly higher mammosphere forming ability compared to control cells, suggesting that TGFβ increases the percentage of stem cells in the population. Initial results demonstrated changes in mammosphere size and morphology of HMLE-pWZL-TGFβ1-GFP cells treated with the HA synthesis inhibitor 4-Methylumbelliferone (4MU). Together, these results suggest STAT3 and TGFβ1 co-regulate HAS2 expression and HA production in a model of EMT supporting the use of the HMLE-pWZL cell series as a promising model to study STAT3 regulation of the HA-CD44 signaling pathway.
Citation Format: Brenda G. Trevizo-Barresi, JoEllen Welsh. Interactions between STAT3 and TGFβ in the regulation of hyaluronic acid synthesis and signaling in immortalized human mammary epithelial cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4037.
Abstract 4039: PAX8 increases migration and metastasis of high grade serous ovarian cancer through upregulation of Rho GTPases
Authors: Hardy Laura , Pergande Melissa , Hernandez Angel , Cologna Stephanie , Burdette Joanna 
Published: August 2018

Abstract
High grade serous ovarian cancer, the most lethal subtype of ovarian cancer, can originate in either the fallopian tube epithelium (FTE) or ovarian surface epithelium (OSE). PAX8 is a lineage specific transcription factor that is ubiquitously expressed in HGSOC. We have shown that knockdown of PAX8 using shRNA in multiple ovarian tumor cells lines leads to apoptosis, suggesting that PAX8 plays an essential role in cancer survival. In this study, we used CRISPR technology to delete PAX8 from the OVCAR8 cell line. PAX8 deletion led to a decrease in migration and invasion in vitro and an increase in survival and a reduction in tumor volume in vivo. Previous work using RNA-sequencing and ChIP-sequencing identified cell adhesion as a top differentially expressed gene between malignant ovarian cancer and benign fallopian tube cell lines. We performed quantitative proteomic analysis of the OVCAR8-PAX8-/- cell line to define PAX8 altered proteins. We also performed quantitative proteomics and transcriptomic analyses on a previously generated murine OSE cell line with forced PAX8 expression (MOSE-PAX8). These analyses identified several genes that contribute to an increase in migration and EMT. Specifically, our data indicates PAX8 upregulates key drivers involved in altering cell morphology including the GTPases: RhoA, Cdc42, and Ras. Inhibition of RhoA led to a greater decrease in migration for both OVCAR8-PAX8-/- and MOSE-PAX8 when compared to control. Inhibition of Ras had a greater effect in OVCAR8-PAX8-/- while inhibition of Cdc42 had a greater effect in MOSE-PAX8. These data provide a mechanistic explanation for the role of PAX8 on increasing migration and metastasis in ovarian cancer.
Citation Format: Laura Hardy, Melissa Pergande, Angel Hernandez, Stephanie Cologna, Joanna Burdette. PAX8 increases migration and metastasis of high grade serous ovarian cancer through upregulation of Rho GTPases [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4039.
Abstract 4043: A novel role for Hedgehog signaling in macrophage-mediated immune evasion
Authors: Hanna Ann , Chen Dongquan , Chandrashekar Darshan S. , Varambally Sooryanarayana , Samant Rajeev S. , Shevde Lalita A. 
Published: August 2018

Abstract
In the tumor microenvironment, breast cancer cells participate in crosstalk with the surrounding stroma. This tumor-stromal interaction forms a balance that dictates tumor suppressing or promoting response mechanisms. Macrophages in the tumor microenvironment are plastic and can mediate several functions depending on their activation states. Tumor-associated macrophages co-exist as two major phenotypes: anti-tumorigenic and immune-eliciting classically activated M1 as well as tumor-promoting and immune-suppressive alternatively activated M2 macrophages. Alternatively activated macrophages are associated with more aggressive stages and poor clinical outcomes in breast cancer patients as they suppress the tumoricidal properties of the immune system, thus facilitating tumor cell proliferation and dissemination. Hedgehog (Hh) signaling is a critical developmental pathway; its activation modulates vital cellular processes such as cell proliferation, stem-cell differentiation, and angiogenesis. While the Hh signaling pathway is normally tightly controlled, it can often be dysregulated which fosters tumorigenesis and tumor progression. Aberrant Hh signaling activation is particularly implicated in breast cancer progression and metastasis. In this study, we investigate the role of Hh signaling in polarizing breast cancer-associated macrophages toward the detrimental M2 subtype. We report that targeting different steps of the Hh signaling cascade attenuates the cytokine profile associated with M2 macrophages. Additionally, we demonstrate that Hh signaling enables the molecular mechanism responsible for alternative macrophage polarization. We furthermore identify significant shifts in immune cell populations infiltrating the primary tumor when we administer an FDA-approved Hh inhibitor to an in vivo mammary tumor model. This altered immune profile is dynamically characterized by a reduction in immune-suppressive cells concomitant with enhanced antigen presentation properties and cytotoxic immune cell infiltration, overall culminating in reduced metastasis. Thus, we describe a novel role for Hh signaling in enabling pro-tumorigenic immunity by alternatively polarizing breast cancer-associated macrophages. This study identifies a novel strategy, which has potential clinical implications to treat breast cancer through targeting aberrant tumor cell Hh activation and eliciting a robust anti-tumorigenic immune response.
Citation Format: Ann Hanna, Dongquan Chen, Darshan S. Chandrashekar, Sooryanarayana Varambally, Rajeev S. Samant, Lalita A. Shevde. A novel role for Hedgehog signaling in macrophage-mediated immune evasion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4043.
Abstract 4058: Race-associated breast cancer disparities: The role of CRYBB2 and its pseudogene
Authors: Barrow Maya A. , Martin Megan E. , Reaves Denise K. , Jeffcoat Breanna , Smith Ginger R. , Tarpley Michael , Williams Kevin P. , Fleming Jodie M. 
Published: August 2018

Abstract
Research demonstrates young African American (AA) women have higher mortality rates for triple-negative breast cancer (TNBC), despite lower incidence rates, when compared to Caucasian American (CA) women. These disparate poor patient outcomes may be attributed to access to quality care, socioeconomic status, education, or geographic location. However, numerous other studies have demonstrated that gene expression differences may also contribute to the higher mortality rates experienced by AA women, suggestive of biological differences among these two patient populations. Consequently, CRYBB2 is one gene that has consistently been associated with AA race in normal and malignant tissues. Although, its basic function in these tissues is currently unknown, higher expression of CRYBB2 is significantly correlated with poor outcome in breast, colorectal and prostate cancers in AAs. We analyzed all publically available gene expression datasets from studies focused on breast cancer health disparities and discovered that all probes detected CRYBB2 as well as its pseudogene, CRYBB2P1. It is becoming increasingly clear that pseudogenes have the ability to modify the mechanisms of their functional parental genes. However, it remains unclear whether the high expression of CRYBB2, CRYBB2P1, or both, affect cell behaviors to alter disease progression. Herein, we investigated the biological consequences of CRYBB2 and CRYBB2P1 in three basal-like TNBC cell lines (SUM159, MDA-MB-231, Hs578t) using CRISPR/Cas9 and overexpression technologies. The functional consequence of CRYBB2 and CRYBB2P1 expression were evaluated using in vitro and biochemical assays, as well as in vivo tumorigenesis studies. Our findings demonstrate overexpression of CRYBB2 and CRYBB2P1 alter cell proliferation, cytokine and growth factor production, and tumorigenesis. This data highlights the potential molecular mechanisms by which CRYBB2 and CRYBB2P1 contribute to poor patient outcome.
Citation Format: Maya A. Barrow, Megan E. Martin, Denise K. Reaves, Breanna Jeffcoat, Ginger R. Smith, Michael Tarpley, Kevin P. Williams, Jodie M. Fleming. Race-associated breast cancer disparities: The role of CRYBB2 and its pseudogene [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4058.
Abstract 4059: Vaccination causes a clear subtype shifting of tumor infiltrate Treg in mouse breast cancer homografts
Authors: AN Xiaoyu , Ouyang Davy Xuesong , Liu Jinping , Chen Li , Li Henry Qixiang 
Published: August 2018

Abstract
Tumor-infiltrating lymphocytes (TIL), including CD8+ TIL-Teff (effector T-cells) and TIL-Treg (regulatory T-cells with immune suppression on self-restricted Teff), are believed to play a significant, but somewhat conflicting, role in the cancer prognosis and sensitivity to immune-oncology (I/O) treatments, including immune checkpoint inhibitors (ICI). Forkhead box P3 (FoxP3+) has long been considered a key marker of Treg while recent research has indicated significant heterogeneity of Treg with different levels of FoxP3 and the associated functional variations. The heterogeneity may explain some of the conflicting observations. The influence of the Treg heterogeneity on ICI treatment has yet to be investigated. We have previously created a mouse tumor homograft, mBR6004, by orthotopic-implanting breast adenocarcinoma spontaneously developed in MMTV-PyVT transgenic mice (GEMM) to the syngeneic FVB/N mice. The homograft has the similar growth, metastasis and histopathology as seen for the original GEMM tumors, and it responds to the CTLA-4 but not to the PD1/PDL1 inhibitors. Further, we also found that a prior vaccination treatment seemed to potentially render the tumor sensitive to PD1/PD-L1 inhibitors with increased CD8+ TIL in the treated tumors. To explore the underlying mechanism of vaccination, we are performing extensive immunophenotyping of TILs in the vaccinated tumors. Interestingly, our analysis clearly displayed a drastic shift of TIL-Treg from FoxP3lo to FoxP3hi population in the tumors with the prior vaccination as compared to the tumor without vaccination, which likely corresponds to the reported naïve FoxP3lo Treg and FoxP3hi effector Treg (eTreg). This FoxP3hi -shifted Treg is also largely absent in peripheral blood, even in the vaccinated mice, consistently to those seen in patients. Our data seem to suggest that the TIL-eTreg status, not naïve Treg, could play a critical role in tumor response to the checkpoint inhibitor treatment, and that the methods, such as vaccination to change TIL-eTreg, could be an important strategy to enhance checkpoint inhibitor treatment. In addition, the elucidation of TIL-Treg in animal model could also greatly help to investigate I/O mechanisms in the experimental settings.
Citation Format: Xiaoyu AN, Davy Xuesong Ouyang, Jinping Liu, Li Chen, Henry Qixiang Li. Vaccination causes a clear subtype shifting of tumor infiltrate Treg in mouse breast cancer homografts [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4059.
Abstract 406: A proteome-transcriptome-miRnome integrated analysis identifies similarity between UV-exposed skin and wounding skin
Authors: Nguyen Tran N. , Rajapakshe Kimal , Avdieiev Stanislav , Nicholas Courtney , Chitsazzadeh Vida , Welsh Eric , Fang Bin , Koomen John , Coarfa Cristian , Einspahr Janine , Tsai Kenneth Y. 
Published: August 2018

Abstract
Cutaneous squamous cell carcinoma (cuSCC) is the second most common skin cancer, for which long term UV exposure and chronic wounding are the dominant risk factors. Despite these clinically established connections, little is understood about the early molecular response of human skin to UV exposure and its connection to acute wounding and cuSCC. Thus, our goal is to find common and specific signatures driven by UV-exposure and wounding as a means of developing new approaches for treating and preventing cuSCC. Here, we perform integrated analyses of proteomic, RNA-seq and miR-seq on 3 datasets: (1) UV-unexposed and acute UV-exposed human skin, (2) public dataset on acute wound healing and (3) our previously published dataset on normal skin and cuSCC from humans. We find that biological signatures and processes regulated by acute UV exposure and wounding has profound similarity. miR-seq data shows that miR-223, miR-132 and miR-142 are significantly upregulated in both acute events. Combined gene set enrichment analysis shows that G-protein-coupled-receptors (GPCRs) pathways are upregulated, possibly through Gαi activation. While ECM remodeling is significantly enriched in all three datasets, gene expression regulated by PPARα is suppressed. Interestingly, upregulation of matrisome components is observed among all three datasets. This suggests that these changes are important early events that regulated by both UV-exposure and wounding which eventually can promote cuSCC initiation. Thus, our findings suggest that these common signatures can be potentially validated as chemopreventive targets for cuSCC.
Citation Format: Tran N. Nguyen, Kimal Rajapakshe, Stanislav Avdieiev, Courtney Nicholas, Vida Chitsazzadeh, Eric Welsh, Bin Fang, John Koomen, Cristian Coarfa, Janine Einspahr, Kenneth Y. Tsai. A proteome-transcriptome-miRnome integrated analysis identifies similarity between UV-exposed skin and wounding skin [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 406.
Abstract 4070: The relationship between members of the canonical NF-κB pathway, components of the tumor microenvironment and cancer-specific survival in colorectal cancer patients
Authors: Quinn Jean A. , Bennett Lindsay , Patel Meera , Loi Lynette , Frixou Mikaela , Roseweir Antonia , Park James H. , Horgan Paul G. , McMillan Donald C. , Edwards Joanne 
Published: August 2018

Abstract
The canonical NF-κB pathway regulates the transcription of many genes which may be involved in processes such as inflammation and proliferation, suggesting a potential link between the tumor, the tumor microenvironment and cancer-specific survival in colorectal cancer patients.
Immunohistochemistry was used to assess the expression of the upstream kinase TAK1/pTAK1 and four members of the canonical NF-κB pathway in tissue microarrays. Protein expression was determined using the weighted histoscore method. BRaf status, tumor stroma percentage, local inflammation, systemic inflammation and cancer-specific survival were examined in patients with colorectal cancer.
Cytoplasmic IKKβ was significantly associated with the inflammatory cell infiltrate (p=0.0015), systemic inflammation (p=0.03) and cancer-specific survival (p=0.046). In contrast, no significant association was found with the other three members. Cytoplasmic pTAK1 was associated with the tumor microenvironment (p=0.045) and cancer-specific survival (p=0.032). On multivariate analysis only cytoplasmic IKKβ associated with survival (HR1.75, 95%CI 1.05-291, p=0.033) and this was independent of TNM stage and markers of the tumor microenvironment. When cytoplasmic IKKβ was stratified with BRaf status (wild type (n=151) or mutant (n=48)), it was no longer associated with cancer-specific survival. However, when cytoplasmic pTAK1 was stratified with BRaf status its association with cancer-specific survival was strengthened. Cytoplasmic pTAK1 was significantly associated with cancer-specific survival in patients with wild type BRaf (p=0.014). In patients with BRaf mutations there was no significant association with cancer-specific survival (p=0.105), however the numbers were low and this is currently being investigated in a larger cohort.
The results of the present study show that high expression of cytoplasmic IKKβ was associated with decreased cancer-specific survival and with markers of the tumor microenvironment in patients with colorectal cancer. Expression of cytoplasmic pTAK1 was associated with cancer-specific survival and this was enhanced in patients with tumors expressing wild type BRaf.
Citation Format: Jean A. Quinn, Lindsay Bennett, Meera Patel, Lynette Loi, Mikaela Frixou, Antonia Roseweir, James H. Park, Paul G. Horgan, Donald C. McMillan, Joanne Edwards. The relationship between members of the canonical NF-κB pathway, components of the tumor microenvironment and cancer-specific survival in colorectal cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4070.