bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4816: Development of an integrated analysis platform of circulating melanoma cells for PD-L1 expression as a predictive biomarker
Authors: Gibbs Benjamin K. , Strotman Lindsay N. , Thiede Stephanie M. , Sperger Jamie M. , Casavant Benjamin P. , Berry Scott M. , Beebe David J. , Lang Joshua M. 
Published: May 2015

Abstract
Novel therapies that target the PD-1/PD-L1 axis have shown great potential to improve survival for patients with advanced melanoma. However, there are limited predictive biomarkers to identify patients who may respond to this class of agents. Previous studies suggest that a subset of responders to PD-1/PD-L1 targeting therapies have expression of PD-L1 in tumor biopsies. The development of these predictive biomarkers is complicated by intratumoral heterogeneity across primary and metastatic lesions for PD-L1 expression. Circulating tumor cells (CTCs) are one potential source of tumor samples for serial analysis. An integrated CTC capture and analysis technology known as the VERSA (Versatile Exclusion-based Rare Sample Analysis) platform permits evaluation of CTCs at the protein, DNA and mRNA level for biomarkers of both therapeutic response and resistance. Using this platform, we have developed a method to isolate circulating tumor cells from patients with advanced melanoma. Using a cocktail of capture monoclonal antibodies including targets of CD146, GD2 and NG2, we have recovered cells across multiple cell lines with high efficiency. Using single antibody capture of SK-Mel-28 cells, capture was 52%, 54%, and 45% with CD146, GD2 and NG2 targets respectively. However, when using a cocktail of capture antibodies containing CD146, GD2 and NG2, capture increased to 92%. Similar results were obtained with other melanoma cell lines including, 624 and M21 cell lines. Cells were also stained for PD-L1 as a potential predictive biomarker of response to PD-L1 and PD-1 targeting immunotherapies. We are now collecting patient samples with this capture platform to identify CTCs expressing PD-L1 as a potential predictive biomarker to PD-1/PD-L1 targeting therapies.
Citation Format: Benjamin K. Gibbs, Lindsay N. Strotman, Stephanie M. Thiede, Jamie M. Sperger, Benjamin P. Casavant, Scott M. Berry, David J. Beebe, Joshua M. Lang. Development of an integrated analysis platform of circulating melanoma cells for PD-L1 expression as a predictive biomarker. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4816. doi:10.1158/1538-7445.AM2014-4816
Abstract 4824: Evaluation of CTCs enriched with a flexible microspring array device as a prognostic biomarker in lung adenocarcinoma
Authors: Harouaka Ramdane , Liu Xin , Khan Waleed , Khan Tasleema , Asiaii Ayeh , Christ Christine , Miller Rebecca , Rice Shawn , Belani Chandra , Zheng Siyang 
Published: May 2015

Abstract
The dissemination of circulating tumor cells (CTCs) implicated in the metastatic spread of cancer accounts for the majority of cancer-related deaths. CTCs have been established as a prognostic biomarker and are associated with worse survival outcomes in various cancer types. However, the implication of CTCs as a prognostic marker in non-small cell lung cancer (NSCLC) has not yet been established. We have employed a flexible micro spring array (FMSA) device for label-free mechanical enrichment of CTCs from whole blood samples obtained from patients with advanced and metastatic lung adenocarcinoma. CTCs were enriched and analyzed qualitatively and quantitatively based on immunocytochemical determination of phenotype and morphological characteristics. Follow up samples were analyzed during the course of therapy and CTC results were correlated to patient survival and tumor burden. FMSA enriched CTCs obtained from a blood sample may prove to be a prognostic biomarker for advanced lung cancer.
Citation Format: Ramdane Harouaka, Xin Liu, Waleed Khan, Tasleema Khan, Ayeh Asiaii, Christine Christ, Rebecca Miller, Shawn Rice, Chandra Belani, Siyang Zheng. Evaluation of CTCs enriched with a flexible microspring array device as a prognostic biomarker in lung adenocarcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4824. doi:10.1158/1538-7445.AM2014-4824
Abstract 4826: Mobilization of tumor cells into the circulation during radiation therapy for lung cancer
Authors: MacManus Michael , Anderson Robin , Russell Prudence , Cox Ashley , Swierczak Agnieszka , Doherty Judy P. , Jacobs Daphne , Smith Jai , Siva Shankar , Daly Patricia E. , Ball David L. , Martin Roger F. , Martin Olga A. 
Published: May 2015

Abstract
Thousands of patients die annually with distant metastasis after curative-intent or “radical” radiation therapy (RT). Because non-small cell lung cancer (NSCLC), the most common cause of cancer-related mortality, exhibits an especially high rate of distant metastasis after radical RT or chemo-RT for locoregionally-advanced disease, it represents a suitable model to investigate the relationship between RT and metastasis. We hypothesized that disruption of tumour architecture during RT could result in the release of viable tumor cells into the peripheral circulation.
We enumerated circulating tumor cells (CTCs) by fluorescence microscopy of blood samples immunostained with conventional CTC markers. We measured their DNA damage levels using γ-H2AX, a biomarker for radiation-induced DNA double-strand breaks, either by fluorescence-activated sorting (FACS) or by immunofluorescence microscopy. Twenty seven RT-treated NSCLC patients had blood samples analyzed by one or more methods. We identified increased CTC numbers after commencement of RT in 7 of 9 patients treated with palliative RT, and in 4 of 8 patients treated with curative-intent RT. CTCs were also identified, singly and in clumps in large numbers during RT by cytopathologic examination (in all 5 cases studied). Elevated γ-H2AX signal in post-RT blood samples signified the presence of CTCs derived from irradiated tumors. Blood taken after the commencement of RT contained tumor cells that proliferated extensively in vitro (in all 6 cases studied), and first experiments revealed that these cells survive in bloodstream of NOD/SCID mice for several months. CTCs formed γ-H2AX foci in response to ex vivo irradiation providing further evidence of their viability.
Although of concern, these findings could represent an opportunity to monitor and target CTCs during RT. They provide a rationale for the development of strategies to reduce the concentration of viable CTCs by modulating RT fractionation or by co-administering systemic therapies.
Citation Format: Michael MacManus, Robin Anderson, Prudence Russell, Ashley Cox, Agnieszka Swierczak, Judy P. Doherty, Daphne Jacobs, Jai Smith, Shankar Siva, Patricia E. Daly, David L. Ball, Roger F. Martin, Olga A. Martin. Mobilization of tumor cells into the circulation during radiation therapy for lung cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4826. doi:10.1158/1538-7445.AM2014-4826
Abstract 4827: Investigation of the survival of circulating breast tumor cells using biomimetic microfluidic circulation system
Authors: Wang Yang 
Published: May 2015

Abstract
Distant metastases are the cause of about 90% of breast cancer patient death and the overall survival rate for metastatic breast cancer is only about 16%. Circulating tumor cells (CTCs) shed from the primary tumor, travel through the blood to distant anatomic sites and cause metastasis. The survival of CTCs in blood vascular system is a challenging pre-requisite of successful metastasis, as the biochemical and mechanical environment of circulation is vastly different from the primary tumor sites. Studies related to the fate of CTCs in circulation are mainly limited by lack of well-controlled and realistic experimental models.
We have developed a microfluidic device that mimics the human blood circulatory system in order to study the mechanism underlying the survival of cancer cells in the blood circulation. The circulation device integrates a network of channels with a self-contained micropump can generate pulsatile fluid flow pattern and corresponding fluidic forces, which can represent the nature of blood flow and haemodynamic forces in the vasculature.
The survival rate of breast cancer cell line MCF7 in circulation was measured via this microfluidic platform. MCF-7 cells in circulation device had a much lower survival rate compared with those in control device. The survival rate of MCF-7 cells after circulating 1, 2 and 4 hours are respectively 48.8%, 36.3% and 10.7%, while the survival rate in control device after 1, 2 and 4 hours are 93.7%, 77.6% and 94.30%. We further compared the survival rate of MCF-7 cells with SUM-159 cells. We hypothesize that by assessing the differential survival ability of breast cancer cells with varied EMT traits can reveal the key signatures of the metastatic tumor cells.
Citation Format: Yang Wang. Investigation of the survival of circulating breast tumor cells using biomimetic microfluidic circulation system. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4827. doi:10.1158/1538-7445.AM2014-4827
Abstract 4831: Breast cancer cells metastasize to bone through E-selectin+ vascular gateways
Authors: Price Trevor , Burness Monika , Sivan Ayelet , Cheng Renee , Magnani John L. , Sipkins Dorothy A. 
Published: May 2015

Abstract
The bone marrow (BM) has been suggested to serve as a protective environment for disseminated breast cancer cells (BCCs). The precise molecular signals that circulating BCCs use to identify and adhere to BM vasculature are unknown, yet these interactions are essential for the development of bone metastases. To address this question, we used in vivo fluorescence microscopy to track the interactions of estrogen receptor (ER)+ and ER- BCC lines with bone in real time in an intracardiac injection xenograft mouse model. Our data show that circulating tumor cells enter the bone through unique vascular niches where endothelial cell expression of the adhesive molecule E-selectin and of the chemokine SDF-1 are both upregulated. These vascular niches are the same sinusoidal capillary beds that serve as the site of entry for peripherally circulating hematopoietic stem and progenitor cells (HSPCs) returning to the BM. Similar to HSPCs, BCCs aberrantly express the major cell surface molecules known to regulate hematopoietic cell entry and exit through the BM, including CXCR4 (SDF-1 receptor) and multiple E-selectin ligands. While neither CXCR4 blockade nor general chemokine signaling inhibition in BCCs prevented BM dissemination, E-selectin blockade by the specific glycomimetic antagonist GMI-1271 decreased BCC entry in the BM by approximately three fold at 2 and 24 hours after treatment. Moreover, in studies comparing FACS isolated stem and non-stem MCF-7 BCCs, GMI-1271 inhibition of E-selectin specifically targeted CD44+CD24- breast cancer stem cell homing. Non-stem CD44+CD24+ BCCs demonstrated significantly less BM homing compared to stem BCCs, and their BM trafficking was less E-selectin dependent. These data reveal for the first time a molecular mediator of bone metastases at the point-of-entry for BCCs in the BM. In addition, the ability of BCCs to hijack HSPC trafficking pathways to metastasize to bone correlates with E-selectin ligand expression and may be a specific characteristic of the BCC stem cell subpopulation.
Citation Format: Trevor Price, Monika Burness, Ayelet Sivan, Renee Cheng, John L. Magnani, Dorothy A. Sipkins. Breast cancer cells metastasize to bone through E-selectin+ vascular gateways. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4831. doi:10.1158/1538-7445.AM2014-4831
Abstract 4840: DNA methyltransferase 3B as a mediator in cancer associated inflammation and progression
Authors: Skrypek Nicolas G. , Achyut B R. , Hollander Christine , Yang Li 
Published: May 2015

Abstract
For a long time ignored, the tumor microenvironment is now known as an important contributor in tumor progression and metastasis. However, the molecular mechanisms as how cancer-associated inflammation promotes metastasis remain to be investigated. Previous work in our lab showed an inflammatory and immune suppressive microenvironment in the pre-metastatic lung prior to tumor cell arrival. Recently, we found that inflammation is responsible for down-regulation of cyclin-dependent kinase inhibitors p15, p16, and p21 in a mouse model of invasive squamous cell carcinoma, in which gene encoding TGFb receptor 2 (Tgfbr2) was deleted in the stromal fibroblasts (Tgfbr2fspKO). There was an induction of inflammation and significant DNA damage in the neighboring epithelia of the forestomach due to the stromal deletion of Tgfbr2. We found that an up-regulation of DNA methyltransferase 3B (DNMT3B) was responsible for these epigenetic alterations. Based on these studies, we hypothesize that the cancer-associated inflammation could induce DNMTs activity leading to the repression of genes important for inhibition of tumor progression. We are currently investigating the impact of inflammation on DNMTs expression using 4T1 mammary tumor models. We hope that our studies will provide insight for molecular mechanisms underlying tumor-host interaction in the metastatic process and identify the novel therapeutic targets.
Citation Format: Nicolas G. Skrypek, B R. Achyut, Christine Hollander, Li Yang. DNA methyltransferase 3B as a mediator in cancer associated inflammation and progression. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4840. doi:10.1158/1538-7445.AM2014-4840
Abstract 4856: Survivin packaging into exosomes is lipid raft-dependent
Authors: Valenzuela Malyn May Asuncion , Bennit Heather R. Ferguson , Osterman Carlos Joel Diaz , Khan Salma , Casiano Carlos , Wall Nathan R. 
Published: May 2015

Abstract
We have shown that Survivin is exported out of the cancer cell via exosomes despite its lack of a secretory signaling peptide. However, the mechanism of Survivin exosomal loading is not yet known. Exosomal protein sorting can be endosomal sorting complex required for transport (ESCRT)-dependent or ESCRT-independent. ESCRT dependent protein sorting requires monoubiquitination of the target protein, which serves as an exosome sorting signal. The ESCRT independent mechanism is through preferential aggregation of the proteins which is based on intrinsic physical properties to lipid-rafts present in exosomes. Our lab has recently shown that exosomal Survivin is associated with both Hsp70 and Hsp90, as well as PRDX1 which have also been shown associated with detergent-resistant lipid rafts. Here, we determine whether Survivin is lipid-raft dependent and whether Hsp70 and Hsp90 and/or PRDX1 are required for exosomal Survivin loading. Exosomes were collected from conditioned media taken from HeLaS cells and isolated by centrifugation. Colocalizations of Survivin with Hsp70, Hsp90 and PRDX1 in the cell and exosomes were detected by immnocytochemistry visualized by confocal microscopy and immunoprecipitation. HeLaS cells were treated with increasing concentrations of methyl-β-cyclodextrin to disrupt lipid rafts as a way to determine protein lipid raft association. Hsp70, Hsp90 and PRDX1 knock-down using siRNAs will be accomplished to verify the requirement of these proteins for Survivin exosome loading and lipid-raft dependents. Here we show that Survivin colocalizes with Hsp70, Hsp90 and PRDX1 in HeLaS cells as well as their exosomes. Increasing concentrations of methyl-β-cyclodextrin disrupts lipid rafts in the HeLaS cells and releases exosomes which also subsequently contain decreasing PRDX1 and Survivin protein amounts. The release of Survivin into the tumor microenvironment is an important finding as this IAP may contribute to the aggressiveness of the cancer as well as the development of chemoresistance. Despite lacking a secretory signaling peptide, Survivin is being released into the extracellular space via exosomes using chaperones. Knowing how Survivin is exosomally packaged may prove useful in selecting patient specific treatment regimens.
Citation Format: Malyn May Asuncion Valenzuela, Heather R. Ferguson Bennit, Carlos Joel Diaz Osterman, Salma Khan, Carlos Casiano, Nathan R. Wall. Survivin packaging into exosomes is lipid raft-dependent. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4856. doi:10.1158/1538-7445.AM2014-4856
Abstract 4857: Clonal expansion of Lkb1-deficient stromal cells underlies polyp development in mouse models of Peutz-Jeghers syndrome
Authors: Ollila Saara , Laajanen Kaisa , Wong Iris , Vaahtomeri Kari , Mäkelä Tomi P. 
Published: May 2015

Abstract
While the role of LKB1 mutations in the Peutz-Jeghers polyposis syndrome (PJS) is uncontroversial, the originating cell type remains unclear as Lkb1 mutations in both epithelial cells and stromal smooth muscle cells (SMCs) have been proposed as tumor drivers. Since SMCs do not represent a major fraction of stromal cells in polyps, altered signaling from Lkb1-deficient SMCs to epithelium has been suggested as a possible mechanism for polyposis. Here we investigate the cell type origin and tumor development mechanism of PJS type polyps in mice by targeting Lkb1 in either early mesenchymal progenitors (Twist2-Cre) or in stromal fibroblasts (Fsp1-Cre). Remarkably, both Twist2 (Twist2-Cre;Lkb1flox/+) and Fsp1-driven (Fsp1-Cre;Lkb1flox/+ and Fsp1-Cre;Lkb1flox/flox) Lkb1 deletion led to formation of PJS-type gastrointestinal polyps predominantly in the stomach, as noted before for Lkb1+/- mice. Furthermore, lineage-tracing experiments demonstrated that Lkb1 deletion results in early local expansion of Lkb1-deficient myofibroblast-like alpha smooth muscle actin-expressing cells between gastric glands, and subsequent clonal expansion filling the stroma of the forming polyps. The simultaneous expansion of the adjacent epithelium seems to be secondary to the stromal growth, based on lack of noticeable genetic alterations in the epithelial cells. Immunohistochemical and mRNA analyses demonstrate that the polyps arising from the stromal deletion models are indistinguishable from Lkb1+/- mice and PJS patient polyps. These results indicate that polyps in the Twist2-Cre and Fsp1-Cre mice represent stromal tumors, and suggest that the identified tumorigenic mechanism is shared in PJS syndrome.
Citation Format: Saara Ollila, Kaisa Laajanen, Iris Wong, Kari Vaahtomeri, Tomi P. Mäkelä. Clonal expansion of Lkb1-deficient stromal cells underlies polyp development in mouse models of Peutz-Jeghers syndrome. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4857. doi:10.1158/1538-7445.AM2014-4857
Abstract 4873: IKKα bridges central tolerance to innate immunity and inflammation
Authors: Zhu Feng , Chen Zhisong , Willette-Brown Jami , Lomada Dakshayani , Davis Sean R. , Back Timothy , Yoshimura Teizo , Sun Zhonghe , Wu Xiaolin , Wiltrout Robert , Richie Ellen , Siebenlist Ulrich , Trichieri Giorgio , Hu Yinling 
Published: May 2015

Abstract
IKKα has emerged as a tumor suppressor in squamous cell carcinomas (SCCs) of skin, head and neck, and lung. Our recent studies indicate that inflammation is involved in lung SCCs in kinase-dead knock-in IkkαKA/KA mice. However, how IKKα prevents inflammation has not been fully understood yet. In the present studies, we showed that IkkαKA/KA mice developed reduced thymic medullary regions, systemic inflammation, and severe skin damage, indicating that IKKα plays an essential role linking central tolerance to innate immunity and inflammation. In line with previous studies of the role of NF-κB pathway in thymic medulla formation and the establishment of central tolerance, we found that NF-κB pathway was inactivated in the presence of kinase inactive form of IKKα in thymic epithelial cells. Depleting the lymphocytes, thymus, T cells, macrophages, and/or reintroducing transgenic IKKα into the skin or thymus rescued the severe skin phenotypes. Transferring IkkαKA/KA T cells, but not wild-type T cells, reconstituted the severe skin phenotypes and SCCs in the lungs and forestomach of IkkαKA/KA/Rag-/- mice. We found that the self-reactive T cells provided TGFβ signaling, which enhanced the expression of monocyte chemoattractant protein-1 (MCP-1) in keratinocytes. As a result, increased level of MCP-1 led to robust recruitment of macrophages to the skin, resulting in skin inflammation and hyperproliferation. This study reveals an important IKKα/TGFβ/MCP-1 signaling axis that orchestrates central tolerance and inflammation in the maintenance of tissue homeostasis.
Citation Format: Feng Zhu, Zhisong Chen, Jami Willette-Brown, Dakshayani Lomada, Sean R. Davis, Timothy Back, Teizo Yoshimura, Zhonghe Sun, Xiaolin Wu, Robert Wiltrout, Ellen Richie, Ulrich Siebenlist, Giorgio Trichieri, Yinling Hu. IKKα bridges central tolerance to innate immunity and inflammation. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4873. doi:10.1158/1538-7445.AM2014-4873