bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5228: Multi-modal in vivo imaging of breast cancer cells expressing dual reporters
Authors: Urban Konnie , Ang Angel , Kuo Chaincy , Francis Kevin , Singh Raj , Roskey Mark , Kim Jae-Beom 
Published: June 2012

Abstract
We previously engineered tumor cell lines with bioluminescent markers and fluorescent proteins for whole animal imaging. In addition to conventional non-invasive tumor growth detection, these cells when harvested from animals can also be used to analyze cell-cell interactions by immunohistochemistry. Moreover, fluorescent cells can be isolated using fluorescent activated cell sorting (FACS) for gene expression profiling. In this study, we used a mouse mammary gland tumor cell line 4T1 that was labeled with firefly luciferase and tdTomato fluorescent protein (4T1-luc2-tdTomato). The generated cell line was tested for in vitro signal stability with respect to bioluminescence and fluorescence prior to injection into the animals. Tumor cells were implanted subcutaneously and monitored for tumor development by acquisition of bioluminescent and fluorescent images. In a separate study, 4T1-luc2-tdTomato cells were implanted orthotopically into mammary fat pads. Tumor growth was monitored and signals from the implantation sites were reconstructed into three dimensional images. In order to non-invasively detect bone metastases by 4T1-luc2-tdTomato cells, we applied both microCT and optical imaging. For this, an interchangeable imaging platform between an optical imager and microCT machine was developed. 4T1-luc2-tdTomato cells were injected into nu/nu mice by intracardiac injection. Bioluminescent imaging was performed immediately to verify the injection of cells into the left ventricle. Fluorescent signals were not detected, likely due to the small number of cells in the secondary tumor sites. Animals demonstrating substantial whole body bioluminescence were selected for further monitoring until post injection day 9. We found that bioluminescent signals were located in the knee joint regions. For these animals, bioluminescent and X-ray images were also taken without changing the positions of the animals using the interchangeable platform. Bioluminescent images in the knee joints were reconstructed using a diffused luminescence imaging tomography algorithm. These images were then co-registered with microCT images. The results showed that bioluminescent signals were colocalized within the joint area obtained by microCT. Moreover, high resolution images of the joints from these mice revealed bone erosion in the tibia induced by 4T1-luc2-tdTomato cells. These findings demonstrate that multimodal imaging can pinpoint the tumor lesions non-invasively. In addition, low dose microCT imaging can be used to acquire high resolution skeletal images longitudinally without inducing adverse effects on the animals.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5228. doi:10.1158/1538-7445.AM2011-5228
Abstract 523: Modeling the crosstalk between head and neck carcinomas and fibroblasts that underlies malignant invasion and drug resistance
Authors: Räsänen Kati A. , Basu Devraj , Montone Kathleen , Zhang Gao , Rustgi Anil , Herlyn Meenhard 
Published: June 2012

Abstract
There is an ongoing need to advance in vitro models of carcinoma invasion and drug resistance, which are both processes likely to be shaped by the complex crosstalk between normal and malignant cells at stromal interfaces. Epithelial to mesenchymal transition (EMT) generates a mesenchymal-like malignant subpopulation within carcinomas with enhanced invasive potential. Here this subpopulation is hypothesized to play a critical role in a tumor-fibroblast crosstalk that enhances invasion and resistance to chemotherapy. To assess the biologic and therapeutic significance of this subpopulation, we have defined head and neck squamous cell carcinoma (HNSCC) cell lines where ongoing EMT sustains an “Ecad-lo” subset; these cells are distinguished by a mesenchymal-like overall gene expression profile and also display key markers typically associated with activated fibroblasts. The mesenchymal-like (Ecad-lo) subset is separable from epithelial-like (Ecad-hi) cells based on loss of surface E-cadherin and expression of vimentin. As an in vivo correlate of Ecad-lo cells, we identified a subpopulation of vimentin-positive malignant cells in primary advanced stage human HNSCCs. In vitro, the Ecad-lo subset showed resistance to cytotoxic and EGFR-targeted therapy; comparable findings were seen in vivo, where treatment led to enrichment of vimentin positive cells in direct xenografts of human HNSCCs. Interactions of stroma with Ecad-hi versus Ecad-lo cells were compared using collagen embedded tumor spheroids and organotypic 3D reconstructs. Here Ecad-lo cells were shown necessary to create an invasive front into collagen matrix. Ecad-lo cells were also needed to drive fibroblast migration and formed a direct interface with fibroblasts in the collagen layer of organotypic 3D cultures. Activated stromal fibroblasts subsequently participated in a feedback loop by promoting EMT in the epithelial-like Ecad-hi subpopulation of cancer cells, increasing the size of the invasive, treatment-resistant Ecad-lo subset. These results define systems for modeling the coordinated acquisition of mesenchymal-like features, invasiveness, and drug resistance by HNSCC cells via active crosstalk with stromal fibroblasts. In this capacity, the models will serve as a platform for identifying targetable mechanisms underlying these central features of malignant behavior.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 523. doi:10.1158/1538-7445.AM2011-523
Abstract 5233: Abnormal lymphatic drainage and function from vascular endothelial growth factor (VEGF)-C overexpressing B16F10 melanoma
Authors: Kwon Sunkuk , Robinson Holly , Wu Grace , Lachance Pier-Anne , Sevick-Muraca Eva 
Published: June 2012

Abstract
The lymphatic system provides a major transport route for initial cancer cell dissemination for many cancers. Tumor lymphangiogenesis (i.e., the new formation of tumor-associated lymphatic vessels) is associated with metastatic potential and is an emerging therapeutic target to arrest metastasis. Recent studies show that the major lymphangiogenic growth factor, vascular endothelial growth factor (VEGF)-C induces tumor lymphangiogenesis and enhances tumor spread to the regional lymph nodes (LNs) and distant sites in mouse models of breast cancer and melanoma. Yet despite the important role of lymphatics as a critical pathway for cancer metastasis, relatively little is known about lymphatic function during tumor progression and metastasis, due mainly to the lack of lymphatic imaging approaches. Recently, we have developed in vivo dynamic near-infrared (NIR) fluorescence imaging methods for quantifying propulsive lymph flow. Herein, we employed a dynamic, functional lymphatic imaging technique and longitudinally imaged spatial and temporal changes in lymphatic function and architecture in mice bearing vascular endothelial growth factor (VEGF)-C overexpressing B16F10 (VEGF-C-B16F10) or mock-transduced B16F10 (mock-B16F10) melanoma tumors. C57BL6 mice were injected with 10ul of indocyanine green (ICG) or FITC-Dextran in the base of the tail and were dynamically imaged for baseline information before implantation of VEGF-C-B16F10 or mock-B16F10 in the hindlimb every three to four days for up to 27 days after inoculation. Our dynamic NIR fluorescence imaging data show that ICG-laden lymph accumulates into a VEGF-C-B16F10 tumor at 3 days post implantation as confirmed with FITC-Dextran leakage from lymphatic vessels; thereafter lymphatic leakage is visualized only in peritumoral lymphatic vessels. Our longitudinal imaging data also show dilated and tortuous lymphatic vessels and reduced or loss of propulsive lymphatic flow in the process of VEGF-C-B16F10 growth. However, mock-B16F10 tumors do not show lymphatic vessel leakage during tumor progression. NIR fluorescence imaging technology can be used to non-invasively and quantitatively detect functional lymphatic changes associated with cancer, which may enable accurate nodal staging of cancer patients and provide new approaches to diagnose and treat cancer metastasis.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5233. doi:10.1158/1538-7445.AM2011-5233
Abstract 5237: Development of organ-selective neuroblastoma cell lines to identify genes mediating bone marrow and liver colonization
Authors: Neiron Zillan , Jankowski Kacper , Murray Jayne , Champion Sophia , Norris Murray D. , Haber Michelle , Fletcher Jamie I. 
Published: June 2012

Abstract
Neuroblastoma is a pediatric tumor with a high propensity for metastasis to bone marrow, cortical bone and liver, and almost 50% of patients have metastatic disease at diagnosis. This study aims to identify genes contributing to neuroblastoma colonization of the bone marrow and liver through a combination of in vivo cell selection techniques and gene expression analyses. Ultimately, it aims to identify potential therapeutic targets for this disease.
When the human neuroblastoma cell line SK-N-BE(2) is xenografted into immunodeficient mice, metastases arise in the adrenal medulla, liver and bone marrow. In contrast, an established subclone forms metastases limited to the bone marrow, suggesting that the original line contains populations with different abilities to colonise each site. We xenografted luciferase positive SK-N-BE(2) cells into immunodeficient mice and isolated pure tumour cell populations from the bone marrow and liver. Increased selectivity for the organ of origin was confirmed for some of these populations using bioluminescent imaging following engraftment into secondary recipients.
We conclude that neuroblastoma cell populations with selectivity for particular organs can be generated using in vivo selection methods. Whole genome expression analyses will now be used to identify candidate genes underlying this selectivity. These genes will subsequently be validated in xenograft models.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5237. doi:10.1158/1538-7445.AM2011-5237
Abstract 524: Stromal fibroblasts diminish the growth-inhibitory effect of the epidermal growth factor receptor antagonist cetuximab in head and neck squamous cell carcinoma cell cultures
Authors: Johansson Ann-Charlotte , Ansell Anna , Östman Arne , Roberg Karin 
Published: June 2012

Abstract
There is a growing body of evidence that components of the tumor microenvironment, including stromal fibroblasts, may modulate the treatment sensitivity of tumor cells. Thus, we aimed to investigate the possible influence of stromal fibroblasts on the cetuximab sensitivity of head and neck squamous cell carcinoma (HNSCC) cells.
Stromal fibroblasts isolated from 7 HNSCC patients were co-cultured with HNSCC cells in a transwell system. Mono- and co-cultures were exposed to the epidermal growth factor receptor (EGFR) antagonist cetuximab (30 nM) and cell proliferation was determined by crystal violet staining. The level of EGFR, Akt, Erk (p42/p44 MAPK), and Met activation was assessed by western blot analysis.
Cetuximab elicited an anti-proliferative response in tumor cell monocultures while the growth rate of stromal fibroblasts remained unchanged. When tumor cells were co-cultured with fibroblasts or grown in media conditioned from fibroblast cultures the anti-proliferative effect of cetuximab was diminished. Furthermore, stromal fibroblasts desensitized tumor cells to treatment with gefitinib, an EGFR tyrosine kinase inhibitor, excluding the possibility that fibroblast-derived factors confer resistance by interfering with the EGFR-cetuximab interaction. Results from experiments using size-fractionated conditioned media suggest that at least two soluble factors, 50-100 kDa and greater than 100 kDa in size, confer treatment resistance. In cetuximab-treated cultures, phosphorylation of EGFR and Erk was reduced, while the level of Akt phosphorylation remained unchanged. The cetuximab resistance observed in co-cultures could not be explained by changes in either EGFR or Erk phosphorylation. However, there was a higher level of activation of the hepatocyte growth factor (HGF) receptor Met in co-cultures, and when recombinant HGF was added to tumor cell monocultures the response to cetuximab was reduced.
These results identify a previously unrecognized fibroblast-dependent modulation of cetuximab sensitivity, and also suggest fibroblast-derived HGF as one of the mediators of this paracrine cross-talk. The results also suggest targeting of fibroblast-derived factors, possibly including HGF, as a novel strategy to improve the therapeutic effects of cetuximab.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 524. doi:10.1158/1538-7445.AM2011-524
Abstract 5244: Genistein's anti-invasion and anti-metastatic effects are linked to decreases in the expression of several different gene in Human prostate cancer that regulate metastatic behavior
Authors: Huang Xiao Ke , Bergan Raymond 
Published: June 2012

Abstract
Genistein has been shown by our group to inhibit Human prostate cancer (PCa) cell invasion in vitro and metastasis in vivo. Related studies demonstrate that endoglin (ENG), a TGF b superfamily receptor, suppresses invasion via activation of Smad 1, and that Genistein therapeutically compensated for endoglin loss by activating Smad 1 signaling. The mechanisms underlying these effects by Genistein are not clear. We hypothesized that they resulted from effects by genistein upon multiple genes that together regulate metastatic behavior.
Methods: PC3M cells were treated or not with 10uM Genistein × 3 days, RNA was isolated, treated with DNAse, and effects upon gene expression were screened for using SAbiosciences metastasis specific and extracellular matrix and adhesion molecule specific gene expression array platforms. Array experiments were performed 2-3 separate times. Positive array findings were then evaluated by real time qRT/PCR, using gene-specific primer/probe sets. These experiments were performed 3 separate times. A student's T-test P-value&lt;/= 0.05 was required.
Results: Genistein significantly decreased MMP2, MMP10 and MMP 13 by 2.81, 3.98 and 4.87 fold, respectively, on the metastasis array, and decreased MMP1, MMP10, MMP13 and SPARC (osteonectin) by 2.29, 3.25, 5.36 and 3.57 fold, respectively, on the matrix /adhesion array. With qRT/PCR, Genistein decreased MMP1, MMP2, MMP10, MMP13 and SPARC by 2.13, 2.17, 4.43, 6.82 and 2.57 fold, respectively. We then treated previously characterized stably engineered ENG human PCa cell lines. Hi-ENG, vector control (VC) and NO-ENG, with Genistein or not, followed by qRT/PCR, ENG expression did not alter gene expression in a consistent fashion. Importantly, ENG status had no effect upon Genistein's ability to decrease the expression of any of the five genes.
Conclusions: We demonstrate for the first time that Genistein inhibit MMP1, MMP2, MMP10, MMP13 and SPARC gene expression. Also, Genistein's effect upon these genes is not dependent upon the endoglin pathway. Further, endoglin does not appear to regulate these genes. Finally these data support the notion that Gensitein's anti-invasion and anti-metastatic actions result from effects upon several genes that together affect invasive and metastatic behavior.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5244. doi:10.1158/1538-7445.AM2011-5244
Abstract 5248: Quantum dot multiplexing of prognostic marker and stem cell marker expression in colorectal cancer circulating tumor cells
Authors: Das Avisnata , Allen Joshua E. , Dicker David T. , Peters Kristi L. , Joudeh Jamal , El-Deiry Wafik S. 
Published: June 2012

Abstract
Circulating tumor cells (CTCs) reflect disease burden in metastatic colorectal cancer (mCRC) and show reduced numbers in patients who are responding to chemotherapy. Current analysis is limited to counting CTCs and little additional marker analysis. We sought to develop methodology to allow multiplexing of marker detection in individual CTCs to assess tumor cell heterogeneity and to gain insight into relationships of prognostic and therapy response markers. This ongoing study is evaluating expression of up to 8 markers in a single CTC by a multispectral multiplex imaging technique using nano-particle type quantum dot (Q-Dot) dyes. We hypothesize that CTCs that are extremely rare (∼ 1 in 10 million RBCs) in the blood of cancer patients play an important role in metastasis. In view of the small number of such cells in the blood of mCRC patients (prognostication based on &lt;3 versus &gt;3 CTCs/7.5 ml of blood) there is a need to derive more molecular correlations from single CTCs. We designed a clinical protocol to collect blood from mCRC patients receiving chemotherapy with the intent to evaluate multiplexed molecular marker expression in CTCs in responders versus non-responders using the FDA-approved Veridex CellSearch technology. We are evaluating CD26, CD133, EGFR, Ki 67, p27, c-Myc, ALDH1, Bcl-Xl, Mcl-1, phosho-Akt, phospho-ERK, TRAIL receptors, IAPs, and p53 using antibodies conjugated to different Q-Dots. Ongoing studies include prognostic marker, stem cell marker, and cell proliferation/death response marker analysis in human tumor xenografts of HCT116, p53-null HCT116, DLD1, SW620, and SW480. Initial profiling of tumor cell lines revealed that mutant p53 and KRAS-expressing SW620 derived from a patient with mCRC has CD26 and CD133 expression including a sub-population of tumor cells that co-express these two stem cell markers. HCT116 showed positive EGFR, CD26, but little or no CD133 expression. We are evaluating correlations between the primary tumors in mice as compared to the CTCs in mice and similar experiments are part of the clinical protocol. The studies should unravel the complexity of CTC heterogeneity, and define informative subsets of markers that can be routinely included in studies to assess prognosis and response to therapy. The ease with which CTCs can be isolated makes these studies of comparisons with primary tumors critical in order to establish and validate new methods for assessing tumor behavior and response to therapy in the clinic through this CTC “liquid biopsy” technique.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5248. doi:10.1158/1538-7445.AM2011-5248
Abstract 5280: PET imaging of proteolysis: Evaluation of 68Ga-DOTA and 68Ga-NODAGA chelates of an uPAR-specific peptide in a human glioblastoma xenograft model
Authors: Persson Morten , Madsen Jacob , Østergaard Søren , Ploug Michael , Kjaer Andreas 
Published: June 2012

Abstract
Background & Aim: The urokinase-type plasminogen activator receptor (uPAR) has been shown to facilitate cancer cell invasion and metastasis. uPAR is over-expressed in various cancers, including human breast, prostate and colorectal cancer, and over-expression has been shown to correlate with poor prognosis. The established correlation between uPAR expression and disease progression provides an opportunity to develop an in vivo imaging agent, which could identify cancer patients at risk. In the present study, we produced and compared two new PET tracers based on 68Ga, the chelators DOTA and NODAGA and the uPAR antagonist peptide AE105 in a human glioblastoma xenograft model using microPET/CT.
Method: Both compounds were synthesized using standard Fmoc solid-phase peptide chemistry and subsequently N-terminal conjugated with either DOTA or NODAGA. The conjugated peptides were radiolabeled with 68Ga using a 68Ge/68Ga generator system, which render the production of the tracers independent of a onsite cyclotron. The compounds were evaluated in vitro by surface plasmon resonance against human uPAR and in vivo using micro-positron emission tomography in a mouse model bearing human glioblastoma U87MG xenografts. All findings were validated using uPAR ELISA on tumor tissue and uPAR immnohistochemistry on tumor slides.
Results: Both compounds were labeled under mild conditions with high yield and radiochemical purity (&gt;95%). Both compounds showed excellent uPAR binding (IC50 values of 10-15 nM) in vitro, which was identical to the un-conjugated peptide AE105. In vivo, they displayed different tumor uptake, with 68Ga-DOTA-AE105 and 68Ga-NODAGA-AE105 having an uptake of 0.48±0.3 and 2.58±0.02 %ID/g 0.5 hr post injection, respectively. Besides tumor uptake, high uptake in the kidneys was also observed for both compounds. Dynamic PET scans illustrated higher stability in vivo for 68Ga-NODAGA-AE105 compared to 68Ga-DOTA-AE105, thus generating the best tumor-to-background contrast.
Conclusion: A new 68Ga-based PET tracer for tumor imaging of human uPAR has been developed. Based on head-to-head comparison, NODAGA seems to be the optimal chelator, giving the highest uptake in tumor tissue. The use of generator-produced 68Ga could further support translation into the clinic.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5280. doi:10.1158/1538-7445.AM2011-5280