bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3969: Stromal depletion of transforming growth factor receptor 2 promotes the development of forestomach squamous cell carcinoma
Authors: Achyut B R. , Bader David A. , Yan Hannah H. , Li Chaoyang , Pang Yanli , Yang Li 
Published: February 2011

Abstract
Deletion or attenuation of transforming growth factor beta (TGFβ) signaling pathway has significant impact on human tumor development and progression. Specific deletion of transforming growth factor receptor 2 gene (Tgfbr2) in a subset of fibroblast (Tgfbr2fspKO, cre-mediated DNA recombination driven by fibroblast specific protein, FSP) specific promoter, leads to the development of squamous cell carcinoma of the forestomach in 100% of Tgfbr2fspKO mice by 6 Wks of Age. Here we dissect the underlying mechanisms. We first confirmed that deletion of Tgfbr2 is primarly restricted to the stromal components but not epithelial cells using TBR2 (TGFβR2) immunofluorescence and immunohistochemistry, which was done in both tumor tissue sections and cancer epithelial cells derived from tgfbr2fspKO mice. The functionality of TGFβ signaling was further examined in vivo by p-smad2 expression and nuclear localization which was clearly observed in the epithelial but not the stromal components. There was also a clear nuclear localization of p-smad2 protein in cultured cancer epithelial cells when treated with TGFβ1 (10 ng/ml). We further observed that stromal deletion of Tgfbr2 resulted in elevated proliferation of fibroblasts. In addition, there was a significantly increased inflammation in forestomach lesion which was featured by increased CD45+ leukocyte infiltration and elevated expression of inflammatory mediators COX2, NFkB, Akt. Interestingly, the alterations in epithelial cell compartment in Tgfbr2fspKO mice is very evident compared with floxed control littermates Tgfbr2fspKO mice, showing a significant enrichment of epithelial cancer stem cells. Our data demonstrate the importance of stromal TGFβ signaling mediated inflammation and carcinogenesis.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3969.
Abstract 3970: DRAK2 inhibits the TGF-B signaling pathwsy through interaction with TGF-B receptors
Authors: Yang Kyung-Min , Kim Seong-Jin 
Published: February 2011

Abstract
Transforming growth factor β (TGF-β) controls a diverse set of cellular responses, including proliferation, differentiation, migration, apoptosis and immune modulation. In recent years, research has begun to unravel the mechanisms that tune the TGF-β signaling pathway. These mechanisms include interactions and modifications that regulate the stability of TGF-β's signaling machinery, receptors and Smads. In addition, they are known to be of major importance for maintaining proper physiological TGF-β signaling. In this study, we have identified a novel TGF-β receptor binding protein called DRAK2 (DAP kinase-related apoptosis-inducing kinase 2). Here, we show that DRAK2 inhibits TGF-β-induced Smad2/3 phosphorylation. To identify the interaction between TGF-β receptors and DRAK2, we performed immunoprecipitation, immunofluorescence staining and in situ proximity ligation assay. DRAK2 specifically bound to the TGF-β receptors. Although ectopic expression of DRAK2 did not inhibit the interaction between TGF-β receptor type I and type II, transient transfection of DRAK2 reduced TGF-β-dependent reporter activity. In stable cell lines that overexpress DRAK2, DRAK2 suppressed the phosphorylation of Smad2 and Smad3 and activity of Smad2-specific (ARE) and Smad3-specific (CAGA) reporters. Furthermore, the expression of DRAK2 attenuated TGF-β-induced translocation of Smad2 and Smad3 from the cytoplasm to nucleus. Conversely, the knockdown of DRAK2 enhanced the TGF-β signaling pathway including TGF-β-dependent promoter activity and phosphorylation of Smad2 and Smad3. Thus, these results suggest that DRAK2 may be a negative regulator in the TGF-β signaling pathway.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3970.
Abstract 3971: The type III tgf-β receptor mediates bmp signaling in normal and cancerous mammary epithelial cells
Authors: Gatza Catherine E. , Blobe Gerard C. 
Published: February 2011

Abstract
BMP signaling has been shown to enhance breast cancer invasion and bone metastasis, while inhibition of BMP signaling abrogates bone metastasis in an in vivo model. The type III TGF-β receptor (TβRIII) is known to mediate BMP signaling and can inhibit migration, invasion, and metastasis in breast cancer, however the role of TβRIII in regulating BMP signaling in breast cancer has not been examined. The MDA-MB-231, MCF7, and mouse 4T1 breast cancer cell lines, which express low levels of TβRIII, are responsive to BMP2 and 4 treatment as demonstrated by increased Smad1/5/8 phosphorylation, cell migration, and invasion. Restoring TβRIII expression in these cell lines or treatment of MDA-MB-231-Neo cells with recombinant soluble TβRIII (sTβRIII) or with conditioned media from MDA-MB-231-RIII cells inhibited BMP2 and BMP4 induced Smad1/5/8 phosphorylation (pSmad1/5/8) and BMP mediated cell migration. MDA-MB-231-RIII cells also exhibited enhanced growth arrest in response to BMP2 compared to MDA-MB-231-Neo cells, mediated through the up-regulation of p21. In reciprocal studies, shRNA-mediated knockdown of TβRIII expression in the normal mammary epithelial cell lines HMEC and MCF10A, abrogated BMP mediated induction of pSmad1/5/8, demonstrating that TβRIII mediates BMP signaling in normal mammary epithelial cells. Additionally, treatment with both recombinant sTβRIII and conditioned media from MDA-MB-231-RIII cells inhibits BMP mediated induction of pSmad1/5/8, suggesting that sTβRIII is able to inhibit BMP signaling in normal mammary epithelial cells by sequestration of ligand. BMP2 treatment induces cell migration in HMEC cells, which was attenuated by knockdown of TβRIII or treatment with sTβRIII. These data demonstrate that TβRIII mediates BMP signaling in normal mammary epithelial cells and breast cancer cell lines, regulating cell migration, invasion, and proliferation. The effects of sTβRIII suggest that the ratio of membrane bound versus soluble TβRIII plays an important role in mediating BMP signaling and cellular effects in normal and cancer mammary epithelial cells. Further work is underway to elucidate the mechanism by which TβRIII regulates BMP signaling in breast cancer in vitro and in vivo.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3971.
Abstract 3975: CLIC4 induces a Smad7 variant to enhance TGF-β signaling in keratinocytes
Authors: Shukla Anjali , Yang Yihan , Madanikia Sara , Ho Yan , Yuspa Stuart H. 
Published: February 2011

Abstract
CLIC4 (Chloride intracellular channel 4) is a member of a family of intracellular chloride channel proteins that are ubiquitously expressed in multiple tissue types. It is a dimorphic protein that exists in both soluble and membrane-bound forms. CLIC4 is multifunctional, traffics between the cytoplasm and nucleus, and participates in cell cycle control and differentiation in keratinocytes and other cell types. CLIC4 expression is lost in multiple human epithelial cancers. We have recently shown that nuclear CLIC4 enhances transforming growth factor beta (TGF-β) signaling through stabilization of phospho-Smad2/3. The TGF-β signaling pathway is a major growth regulator of multiple cell types, and its regulation is altered in many human cancers suggesting that CLIC4 could participate in altering TGF-β regulation in tumor cells. Here we show that in addition to stabilizing phospho-Smads 2/3, CLIC4 alters Smad7, a feedback inhibitor of the TGF-β pathway. Elevated levels of CLIC4 in primary keratinocytes lead to the expression of a truncated form of Smad7 (Smad7Δ), which is detectable as a PCR product. The Smad7Δ variant contains a missing region of 94bp in exon 4 of Smad7 and appears to be an alternately spliced form. Absence of these 94bp leads to a frameshift mutation and an early protein truncation causing the Smad7Δ variant to lack the MH2 domain of Smad7, the domain responsible for inhibition of TGF-β signaling. While wild-type Smad7 expression inhibits TGF-β signaling, exogenously expressed Smad7Δ variant does not inhibit TGF-β signaling, based upon analysis of TGF-β dependent proliferation, reporter assays and phosphorylation of Smad proteins. Together, these data suggest that CLIC4 regulates TGF-β signaling by multiple mechanisms. In light of this, CLIC4 may serve as a target for modifying TGF-β function in cancer and other conditions where this pathway is involved in pathological changes.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3975.
Abstract 3976: A novel violet-laser excitable ratiometric probe for the detection of membrane asymmetry breakdown during apoptosis
Authors: Hicks Justin , Bradford Jolene 
Published: February 2011

Abstract
Apoptosis is carefully regulated and essential part of normal tissue development and homeostasis. Regulatory changes in the apoptotic pathway have been implicated in many diseases; most notable are many types of cancers and autoimmune disorders. Normal cells exhibit a remarkable asymmetry in lipid distribution between the outer and inner cell membranes characterized by phosphatidyl-serine (PS) and phosphatidylethanolamine (PE) normally located on the inner leaflet of the cell membrane. During apoptosis translocation of PE and PS to the external cellular environment facilitates recognition and elimination of these cells by macrophages. The Violet Ratiometric Membrane Asymmetry Probe, 4′-N, N-diethylamino-6-(N, N, N-dodecyl-methylamino-sulfopropyl)-methyl-3-hydroxyflavone (F2N12S), is a novel violet excitable dye for the detection of membrane asymmetry changes during apoptosis. The dye exhibits an excited-state intramolecular proton transfer (ESIPT) reaction resulting in a dual fluorescence with two emission bands corresponding to 530 nm and 585 nm, producing a two-color ratiometric response to variations in surface charge. This ratiometric probe is a self-calibrating absolute parameter of apoptotic transformation, which is independent of probe concentration, cell size, and instrument variation. The two-color ratiometric response of the F2N12S reagent will detect apoptotic cells in samples that vary in cell concentration by as much as 100 fold. The Violet Ratiometric Membrane Asymmetry Probe rapidly targets the plasma membrane requiring only a 5 minute room temperature incubation without additional buffers or wash steps reducing the chance of cell death that can occur from prolonged sample processing. The Violet Ratiometric Membrane Asymmetry Probe will work in both suspension and adherent cell lines and its efficacy is confirmed by caspase and mitochondrial apoptosis markers. Annexin V staining produced a larger population of apoptotic cells during early time points in both suspension and adherent cell lines that do not correlate with caspase and mitochondrial apoptosis markers. The violet 405 nm excitation of F2N12S expands the capability of multi-parameter apoptosis assays to study the apoptotic process while reducing or eliminating the need to create complex compensation controls by taking advantage of the time and spectral separation a violet-excitable reagent provides.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3976.
Abstract 3985: New quantification tools for perfusion and VEGFR2 Expression in tumors
Authors: Foster F. Stuart , Coulthard Tonya , Needles Andrew , Theodoropoulos Catherine 
Published: February 2011

Abstract
Microbubble contrast agents are used to enhance the visualization of the microcirculation in human and mouse tumors. Specifically, contrast agents allow for the relative quantification of tissue perfusion and vascular volume. The in vivo expression of endothelial cell surface markers can also be studied using targeted microbubbles. In this study a nonlinear contrast agent imaging technique, employing amplitude modulation on a linear array-based micro-ultrasound system was used to visualize and quantify time to peak enhancement (TP) and vascular volume (VV) in subcutaneous tumors, as well as to assess expression levels of VEGFR2.
A Vevo 2100 high resolution ultrasound imaging system (VisualSonics Inc, Toronto, Canada) was used and all images were acquired with the MS-250 (center operating frequency of 20MHz, axial resolution 75um) probe. Tumor flow was assessed using MicroMarker contrast agents. A bolus delivery was employed to generate time-intensity curves of contrast wash-in; prototype analytical software was used to perform curve fit analysis and to generate parametric images visually depicting VV and TP as an overlay on the anatomical images. Additionally, Target-Ready MicroMarker contrast agents were conjugated to VEGFR2 antibodies to assess the expression level of this endothelial cell surface receptor. Again, prototype analytical software was used to quantify the relative expression of this target and to generate parametric images of its expression pattern within the tumor. All images were acquired from athymic nude mice implanted with hepatocellular carcinoma cells (Hep3B-Luc-C4) subdermally in the hind limb 4 to 5 weeks before imaging.
Results showed heterogeneity in TP and VV in all tumors. TP ranged from 3.5 seconds in well perfused regions to 12 seconds in poorly perfused tumor regions. The graphical appearance of the associated parametric images readily revealed these differences. Assessment of VEGFR2 expression completed in 2D and 3D showed 2-3 times enhancement in VEGFR2 binding over that observed with the isotype control antibody. These results are consistent with previous methods using linear contrast processing. Overall this study shows the utility of nonlinear processing of contrast signals in assessment of various flow related parameters of the tumor microcirculation.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3985.
Abstract 3989: Ovarian cancer protein biomarker identification from the tumor tissue microenvironment
Authors: Hoskins Ebony R. , Hood Brian , Conrads Tom , Edwards Robert P. 
Published: February 2011

Abstract
Objective: In order to circumvent the challenges associated with serum-based biomarker discovery for ovarian cancer screening development, we utilized a novel sampling strategy to harvest shed and secreted proteins from tissue interstitial fluid (TIF) from the ovarian tumor tissue microenvironment. The motivating hypothesis is TIF, the proximal fluid that bathes ovarian cancer cells and stroma, is rich in cancer-specific proteins that are readily identifiable by mass spectrometry (MS)-based proteomics.
Materials: Ovarian tumor tissue and ascites were collected within 30 minutes of removal from the patient, at the time of debulking surgery. Samples used in the analysis were from four chemotherapy naïve patients with pathology-proven papillary serous ovarian cancer. We implemented a standardized sampling strategy, involving incubation of tumor tissue in saline for 1 hour immediately following resection resulting in protein rich TIF. The TIF and ascites samples were analyzed by MS for protein sequence identification and relative quantitation.
Results: Over 2700 total proteins were identified in TIF across all patient samples, while ascitic fluid contained over 2000 proteins across all patient samples. 13 proteins were identified in all TIF samples that were 3-fold or greater in quantity compared to ascitic fluid samples. For comparison, 14 proteins that were 3-fold greater in ascitic fluid compared to TIF, was also identified. Interestingly, these select proteins from two microenvironments had contrasting function and location. TIF samples contained largely intracellular proteins, which are secreted and/or shed while ascitic fluid sample contained extracellular proteins, similar to serum, thus confirming the utility of TIF representing the tumor microenvironment, see figure. The results demonstrate the ability to readily identify hundreds of proteins in TIF, which are at higher abundance compared to those, identified in ascitic fluid from the same patients.
Conclusions: We developed a standard workflow for the discovery of candidate biomarkers from the ovarian cancer tissue microenvironment. The TIF proteome contains more lower abundant and cancer specific proteins compared to ascitic fluid. We are prospectively validating candidate biomarkers from this microenvironment in patient serum by western blot analysis and immunohistochemistry. Ultimately we will expand our standard protocol to a cohort of patients with likely early stage cancers in order to translate to a serum-based screening test.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3989.
Abstract 399: A special childhood T-ALL model of NOD/SCID mice can recapitulate the clinical patient disease
Authors: Zhang Amy , Huang Liang , Ma Shuyan , Wang Na , Liu Jingping , Song Xiaoming , Chen Taiping , Wery Jean-Pierre , Chen Yiyou , Zhou Jianfeng , Zhou Pengfei 
Published: February 2011

Abstract
Despite recent advances in the management of childhood acute lymphoblastic leukemia (ALL), the risk of relapse remains about 30 percent. Here we report an interesting childhood T-acute lymphoblastic leukemia (T-ALL) patient case, engrafting into NOD/SCID mice successfully, recaptured the clinical course accurately including the rapid remission induction, minimal residual disease and relapsing. Immunophenotypes of primary, secondary and third xenografts indicated the cells expressing CD3/CD5/CD7/CD4, as well as the genetic lesions, were unaltered compared with the original patient sample. Immunohistochemistry (IHC) studies showed more than 90% infiltration of leukemic cells in spleen, liver, lung and variable level in other organs include brain within 4 weeks in primary xenografts and 2 weeks in secondary xenografts. The response to Vinscristine and Dexamesathone of primary engrafts was similar to what the original patient did in the clinic. A minimal residual disease phase and relapsing model had also been established in this model after complete remission induced by Vincristine plus Dexamesathone. In addition, Dexamesathone treatment alone was able to prolong the survival of the T-ALL hosts for more than two weeks in the presence of over 90% cancer cell infiltration in the peripheral blood. This study supports previous finding that the NOD/SCID model of childhood T-ALL is an accurate representation of the patient disease and can be used in the screening of anticancer drugs for childhood ALL. This model also warrants further investigation in T-ALL at relapse and potentially in the study of minimal residual disease for chemotherapy-resistant T-ALL subgroups.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 399.
Abstract 3990: Methods for Fine Needle Aspiration (FNA) biopsy sampling and “ex vivo” signal transduction analysis of murine xenografted human tumors
Authors: Aguero Martin F. , Jeffries Emiko , Schayowitz Adam , Clark Douglas P. 
Published: February 2011

Abstract
Fine Needle Aspiration (FNA) biopsy is a common procedure for sampling human tumors to enable diagnosis and, increasingly, molecular diagnostics on fixed samples. However, FNA is rarely used to sample murine xenografted human tumors. FNA sampling of xenografted tumors offers several advantages over surgical excision: 1) efficiency; 2) enables serial sampling of individual tumors over time; 3) provides live cells to perform “ex vivo” analysis of signal transduction pathways. Objectives: To define a protocol for FNA sampling of murine xenografted tumors using an HCT-116 human colon carcinoma model; to characterize the cellular and molecular composition of such samples and to present proof-of-concept studies for “ex vivo” analysis from FNA samples. Methods: FNA biopsies were performed on HCT-116 xenografts using a standardized protocol that will be defined. Samples were assessed by microscopic analysis of direct smears stained with the Diff-Quik or Papanicolaou stains. Tumor cell number was determined by hemocytometer count and cell viability by trypan blue exclusion. Samples were lysed and total protein and RNA were extracted and quantitated by spectrophotometry. RNA quality was determined by RIN calculation. “Ex vivo” stimulation was performed by treating tumor cells obtained from FNA with TPA and EGF and subsequently analyzing phosphoprotein levels relative to baseline using Western blots. Results: FNA sample: Cellular composition: 25 % tumor cells, 50% macrophages, 20% lymphocytes, and 5% degenerate cells (n=5). Tumor cell viability median: after 1 hour post-FNA: 42% (range 22-88%) n=15. Cell viability decreases by 50% after 2 h post-FNA, (n=5). Total Protein median/FNA: 555 ug/ml (range 36-4222 ug/FNA) n=33; Total RNA median 14.80 ug/FNA (range 3.24-26.37 ug) n=18; RIN number median/FNA: 8.40 (range 3.10-9.50) n=15. “Ex vivo” analysis of signal transduction: a 60%, transient increase in phospho-ERK was identified within the first 5 min. post-FNA procedure, which returned to baseline. No increase in p-AKT or p-JNK was noted during this time period. Following “ex vivo” EGF or PMA stimulation, FNA samples showed a 1-3 fold increase -respectively- in p-ERK relative to baseline. Conclusions: Murine xenografts can be efficiently sampled by FNA biopsy. Such samples yield sufficient high-quality biomolecules for molecular analysis. FNA samples also enable live cell analysis, such as “ex vivo” stimulation of signal transduction pathways.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3990.