bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1394: CD44 isoform expression illuminates multiple CSC subpopulations in PC3
Authors: Hernandez James R. , Mooney Steven M. , Torga Gonzalo , Verdone James E. , Pienta Kenneth J.
Published: August 2015

The concept of the cancer stem cell (CSC) is neither new nor without controversy. It is important to study these cells since they are often hypothesized to be androgen insensitive and resistant to chemotherapeutics. Therefore, finding reliable markers for identifying and targeting these cells has become increasingly attractive. The hyaluronic acid receptor, CD44, is associated with a wide variety of cell types, including CSCs. CD44 has many different isoforms which can be categorized as belonging to one of two groups; one expressing different combinations of 10 variant extracellular exons (CD44v), and the other only expressing standard exons found in all isoforms, but none of the 10 variant exons (CD44s). CD44s is typically associated with the mesenchymal stem cell niche, while the various CD44v forms are related to hematopoietic stem cells and increased cell adhesion. However, both CD44s and CD44v have been linked to the CSC niche and cancer progression. Similar to CD44, the aldehyde dehydrogenases (ALDH) superfamily expression is also associated with CSCs and cancer progression. Previously, our laboratory demonstrated that epithelial cancer cells undergo an EMT upon exposure to M2 macrophages. In order to obtain a purely epithelial PCa population, our laboratory isolated a single cell clone of PC3 that had high E-cadherin expression, denoted PC3-Epi. PC3-Epi cells were then incubated with M2 macrophages, which caused a stable EMT to occur after only a few days in culture, denoted PC3-EMT. Our finding demonstrate that these PC3 clones exhibit numerous CSC characteristics, such as high ALDH activity, CD44 expression and cell plasticity. FACS analysis of CD44 isoform expression show distinct CSC subpopulations within these PC3-Epi and PC3-EMT cell types. These subpopulations showed the ability to recapitulate the presorted parental population after being returned to culture. This data suggests not only that the PC3 cell line displays numerous CSC characteristics, but also that distinct CSC subpopulations may exist.

Note: This abstract was not presented at the meeting.

Citation Format: James R. Hernandez, Steven M. Mooney, Gonzalo Torga, James E. Verdone, Kenneth J. Pienta. CD44 isoform expression illuminates multiple CSC subpopulations in PC3. [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 1394. doi:10.1158/1538-7445.AM2015-1394
Abstract 1397: Identification of a hepatocellular carcinoma cell line capable of monitoring the differentiation of CD13(+)CD166(-) cancer stem cells and effects of sorafenib
Authors: Yamada Takeshi , Abei Masato , Danjoh Inaho , Shirota Ryoko , Yamashita Taro , Endo Shinji , Hyodo Ichinosuke , Nakamura Yukio
Published: August 2015

Background: Recent cancer stem cell (CSC) research has highlighted the necessity to develop drugs targeting CSCs. However, only few in vitro models are available which show CSC hierarchy and allow monitoring of CSC differentiation and screening of drugs targeting CSC.

Aim: In the present study, we aimed to identify such hepatocellular carcinoma (HCC) cell line(s).

Methods: Based on a hypothesis that the proportion of CSCs would decrease during culture because of their differentiation into progenitors (population switch), we tested the positivity of several markers (CD13, EpCAM, CD133, CD44, CD90, CD24, CD166) in HCC cell lines (HuH-7, Li-7, PLC/PRF/5, HLF, HLE) before and after 2 months culture.

Results: Only Li-7 cells showed relevant population switch: the proportion of CD13 decreased, while that of CD166 increased during the culture. High tumorigenicity of Li-7 was lost after the population switch. CD13(+)/CD166(-) cells showed slow growth and reconstructed the bulk Li-7 populations composed of CD13(+)/CD166(-), CD13(-)/CD166(-) and CD13(-)/CD166(+) fractions, while CD13(-)/CD166(+) cells showed rapid growth but could not reproduce any other cell populations. CD13(+)/CD166(-) cells showed high ALDH activity, spheroid forming ability and resistance to 5FU. Microarray analysis demonstrated higher expression of stemness-related genes in CD166(-) than CD166(+) fraction. These results indicated the hierarchy of Li-7 cells, in which CD13(+)/CD166(-) and CD13(-)/CD166(+) cells serve as slow growing CSCs and rapid growing progenitors, respectively. Interestingly, sorafenib selectively targeted CD166(-) fraction including CD13(+) CSCs, which exhibited higher mRNA expression of FGF3 and FGF4, candidate biomarkers for sorafenib. 5-FU followed by sorafenib inhibited the growth of bulk Li-7 cells more effectively than reverse sequence or either alone.

Conclusions: We identified an useful cell line Li-7 capable of monitoring the CD13 (+) CSC differentiation and screening the drugs targeting the CSCs of HCC and found that sorafenib preferentially targeted CD13(+) CSCs in vitro. Screening cell lines for “population switch” may be useful to identify novel candidate model systems for screening drugs targeting CSCs.

Citation Format: Takeshi Yamada, Masato Abei, Inaho Danjoh, Ryoko Shirota, Taro Yamashita, Shinji Endo, Ichinosuke Hyodo, Yukio Nakamura. Identification of a hepatocellular carcinoma cell line capable of monitoring the differentiation of CD13(+)CD166(-) cancer stem cells and effects of sorafenib. [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 1397. doi:10.1158/1538-7445.AM2015-1397
Abstract 1429: Contribution of membrane-bound carboxypeptidase M to tumor growth and metastasis by regulating epithelial mesenchymal transition in esophageal squamous carcinoma
Authors: Kudo Yuji , Fukuda Junki , Harigai Ritsuko , Kato Kazunori
Published: August 2015

Carboxypeptidase M (CPM) is a membrane-bound exopeptidase that recognizes the arginine (Arg) and lysine (Lys) residues in a C-terminal of polypeptide chain or protein. The cleavage of some soluble factors such as SDF-1 and EGF by CPM leads to the alteration of binding affinity to their receptors. In this study, we examined the properties of CPM for tumor growth and metastasis in human esophageal carcinoma cells. In order to investigate the expression of CPM on human tumor cells, we established the monoclonal antibody specific for human CPM and found the constitutive expression of CPM on various human carcinomas by flow cytometry and confocal laser microscopy. Treatment of cancer cells with EMT-mediated cytokines, including TGF-beta, IL-6 and OSM, increased CPM expression on their cell surface followed by enhancement of cell migration and change of cell shapes from epithelial to mesenchymal type. Silencing CPM expression by small interfering RNA (siRNA) specific for CPM in esophageal and breast carcinoma cells were confirmed by flow cytometry. We found that CPM-silenced carcinoma exhibited reduced cell proliferation not only in 2D culture flask but also in 3D spheroid culture plate. Importantly, the treatment of siRNA for CPM inhibited the expression of EMT-related genes such as snail and slug which are important for cancer motility. Consistently, we found that CPM-silenced carcinoma exhibited reduced motility of cell in wound healing assay. Taken together, these data indicate that CPM contributes to cell growth and EMT and might be a therapeutic target in esophageal carcinoma.

Citation Format: Yuji Kudo, Junki Fukuda, Ritsuko Harigai, Kazunori Kato. Contribution of membrane-bound carboxypeptidase M to tumor growth and metastasis by regulating epithelial mesenchymal transition in esophageal squamous carcinoma. [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 1429. doi:10.1158/1538-7445.AM2015-1429
Abstract 1440: Mitochondrial superoxide dismutase (Sod2) modulates ovarian clear cell carcinoma transcoelomic metastatic pathway
Authors: Hemachandra L.P.Madhubhani P. , Dier Usawadee , Hempel Nadine
Published: August 2015

Ovarian clear cell carcinoma (OCCC) is a highly aggressive histological subtype that represents approximately 10% of all ovarian cancer cases and displays high expression of mitochondrial superoxide dismutase (Sod2), a major mitochondrial antioxidant enzyme. In the female body ovarian cancer cells metastasize via the transcoelomic route, where cells detach from the primary tumor into the intraperitoneal (IP) cavity and survive as unattached single cells or spheroid aggregates until they get carried via ascites fluid to the omental wall and surrounding organs for metastatic colonization. Ovarian cancer spheroids evade anoikis and display strong resistant to most anti-proliferative cancer drugs. This may contribute to the high recurrence rate and chemoresistance of ovarian cancer. The present study suggests that Sod2 is critically important for the key events of this transcoelomic metastatic route of OCCC. Levels of Sod2 are further increased in OCCC cell lines when these are cultured as anchorage-independent spheroid aggregates, suggesting that Sod2 may play a major role in spheroid stability and survival. When Sod2 levels are reduced using RNA interference, spheroid morphology of ES-2 OCCC cell line is compromised, suggesting a potential correlation between anoikis resistant and Sod2 levels in OCCC. Sod2 knockdown compromises mitochondrial metabolism, as a consequence of reduced superoxide scavenging, and enhances spheroid sensitivity to cisplatin. In addition, Sod2 is necessary for spheroid attachment and outgrowth on extracellular matrix components, representing the invasion of spheroids into the omental wall. Further, tumor cell metastasis is attenuated with reduced Sod2 levels when ES-2 spheroids are placed on the vasculature of the Chorioallantoic membrane (CAM). Scavenging H2O2 by exogenous addition of catalase inhibits the migration of OCCC and our data suggest that high levels of Sod2 in OCCC shift the intracellular redox balance to drive H2O2-dependent pro-metastatic signaling. These results indicate that Sod2 plays a major role in OCCC tumor metastasis, by regulating survival and chemoresistance of tumor spheroids in the IP cavity and by supporting attachment and migration at secondary metastatic sites. Inhibition of Sod2 can be a potential therapeutic target against OCCC transcoelomic metastasis.

Citation Format: L.P.Madhubhani P. Hemachandra, Usawadee Dier, Nadine Hempel. Mitochondrial superoxide dismutase (Sod2) modulates ovarian clear cell carcinoma transcoelomic metastatic pathway. [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 1440. doi:10.1158/1538-7445.AM2015-1440
Abstract 1445: Overexpression of angiotensin II type 1 receptor induces epithelial-mesenchymal transition and promotes tumorigenesis of human breast cancer cells
Authors: Oh Eunhye , Kim Ji Young , Cho Youngkwan , Lee Nahyun , An Hyunsook , Kim Jun Suk , Seo Jae Hong
Published: August 2015

Propose: Accumulating evidence suggests that angiotensin II type 1 receptor (AGTR1) is involved in tumorigenesis and metastasis in many types of cancer, however little is known about its role in breast cancer. Our objective of the present study is to investigate the effect of AGTR1 overexpression on cell proliferation, epithelial-mesenchymal transition (EMT), migration, invasion, angiogenesis and tumorigenesis of human breast cancer.

Methods: To determine the role of AGTR1, MCF7 cells were stably transfected with an expression construct carrying the GFP cDNA for AGTR1. MTS assay, Matrigel invasion assay, wound healing assay, Western blotting and immunofluorescence were used. For in vivo study, parental MCF7 or AGTR1-MCF7 cells were subcutaneously injected into the left and right flanks of nude mice. Tumor growth and angiogenesis were determined.

Results and conclusion: AGTR1-MCF7 cells, expressing high level of AGTR1 exhibited significant increases in cell proliferation, migration and invasion capacities compared with parental MCF7 cells. Furthermore, AGTR1-MCF7 cells displayed the mesenchymal phenotype, as evidenced by elongated spindle morphology, decreased E-cadherin level, increased N-cadherin and snail, and nuclear accumulation of smad3 and smad4. Interestingly, blockade of smad4 using small interfering RNA strongly induced mesenchymal to epithelial transition (MET) and restoration of E-cadherin expression in AGTR1-MCF7 cells, suggesting that AGTR1-induced EMT is ultimately mediated by smad4 activation. Finally, we showed that AGTR1 overexpression enhanced tumor volume (p<0.001, n = 10) and weight (p<0.01) as well as increased angiogenesis in human breast cancer xenograft in vivo. In summary, we demonstrate that AGTR1 overexpression promotes cell proliferation, EMT, angiogenesis and tumorigenic properties of breast cancer cells. Our findings suggest that AGTR1 could be a useful target of therapy in breast cancer.

Citation Format: Eunhye Oh, Ji Young Kim, Youngkwan Cho, Nahyun Lee, Hyunsook An, Jun Suk Kim, Jae Hong Seo. Overexpression of angiotensin II type 1 receptor induces epithelial-mesenchymal transition and promotes tumorigenesis of human breast cancer cells. [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 1445. doi:10.1158/1538-7445.AM2015-1445
Abstract 1487: Rotating 3D projections of prostate lesions imaged by Cu-64-VPAC1 ligand PET and CT
Authors: Wampole Matthew E. , Paudyal Bishnuhari , Trabulsi Edouard J. , Thakur Mathew L. , Wickstrom Eric
Published: August 2015

Surgeons are obliged to examine multiple planar images of fine needle biopsies, CT, and PET images for diagnosis and evaluation of prostate cancer. We hypothesize that optimal comprehension of prostate lesions in the prostate, as they present in the surrounding tissue, will be achieved by 3D registration and presentation of all available images. In particular, PET imaging of a cancer cell surface marker maximizes specificity, relative to imaging of F-18-deoxyglucose uptake. We have demonstrated specific PET imaging in patients of vasoactive intestinal peptide and pituitary adenylate cyclase activating peptide receptor 1 (VPAC1), which is overexpressed on breast cancer and prostate cancer cells. Our PET agent is Cu-64-TP3805, a peptide derived from vasoactive intestinal peptide. From our ongoing prostate cancer patient imaging study, we segmented and co-registered CT image slices and TP3805 PET image slices, then created moving 3D images. Viewing the rotating 3D images from all sides revealed clear delineation of prostate lesions from the adjoining bladder. The lesion images colocalized in space with intraprostatic tumor foci identified from H&E staining of whole mount slices after prostatectomy. Thus, rapid 3D presentation of anatomical, histological, and molecular images could be a useful preparative tool to minimize complications during prostate cancer surgery. This result supports proceeding to 3D analysis of a larger prostate cancer cohort.

Citation Format: Matthew E. Wampole, Bishnuhari Paudyal, Edouard J. Trabulsi, Mathew L. Thakur, Eric Wickstrom. Rotating 3D projections of prostate lesions imaged by Cu-64-VPAC1 ligand PET and CT. [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 1487. doi:10.1158/1538-7445.AM2015-1487
Abstract 150: Long non-coding RNA are differentially expressed in breast cancer clinical subtypes
Authors: Kumar Surendra , Seiler Michael , Bjørklund Sunniva , Børresen-Dale Anne-Lise , Kristensen Vessela , Bhanot Gyan , Ganesan Shridar
Published: August 2015

A primary benefit of RNA-Seq over traditional DNA microarrays is the ability to interrogate expression in unannotated areas of the transcriptome. Methods for identifying novel genes in genomic DNA sequence can use features such as an open reading frame and conserved functional protein elements. Non-coding RNA structures are more difficult to detect, and most of our knowledge of them comes from expressed sequence tags (ESTs), which are large, sequenced contigs from transcriptome cDNA libraries. Carninci et al. showed that over half of these sequences appear to originate from non-coding sources, though very few are annotated. Moreover, many of these non-coding sources are poorly conserved over multiple species indicating that these transcripts may not be functional, but instead, may be the result of “random” transcription. As more studies focus on these “non-coding” regions of the genome evidence of conservation and functionality is emerging. To identifying possibly functional transcripts, we took the approach of identifying only non-coding transcripts which were differentially expressed between breast cancer tumor subtypes, and tumor versus non-tumor samples by RNAseq analysis. We reason that such ncRNA transcription is more likely to be functional because of its association with a specific disease phenotype and the possibility that they affect some regulatory pathway. We have identified known long-non coding tumor suppressors, such as H19 and MEG3, as expressed exclusively in non-tumor samples. In addition we have identified 59 intergenic regions representing a total of 132 300 bp which were differentially expressed between ER+/HER2- and ER-/HER2- breast tumor subtypes. This resulted in 4 potential non-coding regions associated with ER-/HER2- tumors, and 6 potential non-coding regions associated with ER+/HER2- tumors. These transcripts are of great interest, and should be studied in more detail to elucidate the function of these non-coding RNA molecules.

Citation Format: Surendra Kumar, Michael Seiler, Sunniva Bjørklund, Anne-Lise Børresen-Dale, Vessela Kristensen, Gyan Bhanot, Shridar Ganesan. Long non-coding RNA are differentially expressed in breast cancer clinical subtypes. [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 150. doi:10.1158/1538-7445.AM2015-150
Abstract 152: The long non-coding RNA HOTAIR affects exosome-mediated bladder cancer progression
Authors: Berrondo Claudia , Flax Jonathan , Messing Edward M. , Beckham Carla
Published: August 2015

Exosomes are 30-100nM membrane-bound vesicles that participate in intercellular communication and facilitate tumor microenvironments. The mechanisms by which exosomes modify tumor niches are not fully understood. One possibility is that exosomes deliver biologically active molecules like mRNA, protein, miRNA and long non coding RNA (lncRNA) that alter the metabolism of recipient cells to produce pathological phenotypes. Exosomes may also activate the expression of genes involved in tumor progression in recipient cells. Overexpression of the lncRNA HOTAIR is associated with poor prognosis and affects tumor progression, in part, by recruiting PRC2 and LSD1 chromatin modifying complexes to thousands of target genes involved in tumor progression processes such as epithelial-to-mesenchyme transition (EMT). We show that HOTAIR is enriched in bladder cancer (BC) patient tumors and urinary exosomes. BC cell lines also express elevated levels of HOTAIR. Knockdown of HOTAIR in BC cell lines significantly reduces migration and invasion in trans-well and 3D culture, which correlates with EMT gene expression changes in these cells. Exosomes isolated from shHOTAIR vs shScramble cells fail to facilitate migration and invasion in 3D culture of a recipient BC cell line. Selected bands were isolated from a coomassie gel of shHOTAIR and shScramble exosomes and sent for mass spectrometry. Hundreds of putative proteins were identified including proteins involved in RNA binding, apoptosis, vesicle trafficking and integrins. These data support HOTAIR as an important player in exosome-mediated BC tumor progression possibly through affects on gene expression in exosome producer cells. For example, in shHOTAIR cells the loss of PRC2 and/or LSD1 complexes from key genes may ultimately affect the proteins available for packaging into exosomes. Exosomes have important roles in the tumor microenvironment and these data support a role for HOTAIR in the modulation of exosome-mediated tumor progression.

Citation Format: Claudia Berrondo, Jonathan Flax, Edward M. Messing, Carla Beckham. The long non-coding RNA HOTAIR affects exosome-mediated bladder cancer 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 152. doi:10.1158/1538-7445.AM2015-152
Abstract 1522: Survival of skin cancer tumor initiating cells requires the Ezh2 polycomb group protein
Authors: Adhikary Gautam , Grun Dan , Kerr Candace , Balasubramanian Sivaprakasam , Rorke Ellen , Xu Wen , Eckert Richard
Published: August 2015

Epidermal squamous cell carcinoma is among the most common cancers in humans. These tumors are comprised of phenotypically diverse of cells that display varying potential for proliferation and differentiation. Polycomb group (PcG) proteins, including Ezh2, are important candidate stem cell maintanence proteins. Our previous study showed that Ezh2-enriched epidermal cancer stem cells (ECS cells) represent a minority of cells in tumors, yet drive aggressive tumor formation. In the present study, we show that Ezh2 is required for ECS cell survival, migration, invation and tumor formatiom and this is associated with increased histone H3 trimethylationon lysine 27, a mark of Ezh2 action. We also show that Ezh2 knockdown or treatment with Ezh2-selective inhibitors, GSK126 or EP-6438, reduce Ezh2 level and activity, leading to reduced spheroid formation, migration, invasion and tumor formation. Use of these inhibitors was associated with reduced ECS cell vaibility and induction of apoptosis. These studies indicate that epidermal squamous cell carcinoma cells contain a subpopulation of cancer stem cells-like tumor initiating cells that are enriched in Ezh2 expression and that Ezh2 is required for optimal survival of these cells and tumor formation. These studies suggest that treatment with Ezh2 inhibitors may be a strategy for reducing epidermal cancer stem cell survival and supressing tumor formation.

Citation Format: Gautam Adhikary, Dan Grun, Candace Kerr, Sivaprakasam Balasubramanian, Ellen Rorke, Wen Xu, Richard Eckert. Survival of skin cancer tumor initiating cells requires the Ezh2 polycomb group protein. [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 1522. doi:10.1158/1538-7445.AM2015-1522
Abstract 1534: The microbiome of cancer as seen through the lens of public genome sequence data
Authors: Robinson Kelly M. , Sieber Karsten B. , Anderson Kathleen E. , Riley David R. , Dunning Hotopp Julie C.
Published: August 2015

There are 10 times more bacterial cells in the human body than human cells and various bacteria are known to influence carcinogenesis. Therefore we sought to investigate if publicly available whole genome and whole transcriptome sequencing data generated by large public cancer genome efforts, like The Cancer Genome Atlas (TCGA), could be used to examine the microbial composition of tumors. The Burrows-Wheeler Aligner (BWA) was used to align a selection of Illumina paired-end sequencing data from TCGA data to the human reference genome and all complete bacterial genomes in the RefSeq database. Reads were filtered for low complexity and duplicates and then assigned an Operational Taxonomic Unit (OTU). Through careful consideration of all of the bacterial taxa present in the cancer types investigated, we found an increased abundance of Acinetobacter spp. in acute myeloid leukemia (AML) samples, Mycobacterium tuberculosis complex in ovarian serous cystadenocarcinomas (OV), and Pseudomonas spp. in stomach adenocarcinoma (STAD) samples. Further investigation prompted us to conclude that Mycobacterium tuberculosis complex read pairs were found in the OV samples due to sequencing center batch effects. The same analysis of the AML and STAD samples suggests that Acinetobacter spp. are truly present in AML samples and that Pseudomonas spp. are actually present in the STAD samples investigated. Upon completion of this analysis, we determined that it is possible to identify bacteria in human tumor samples. However, care must be taken to assess bacteria that arrived in the sample via various forms of contamination. It is also important to note that the sequencing library method heavily biases the bacterial taxa identified. One sample sequenced with RNA-Sequencing, whole exome sequencing (WXS), and whole genome sequencing (WGS) had extensively different bacteria reflected in the RNA-Seq data compared to that of WGS and WXS. In the future, a comprehensive approach should be used when evaluating the microbiome of tumor samples via sequencing data. The methods used in this study are valid and useful for identifying the presence of bacteria in human samples, but may not be an exhaustive picture. More weight should be given to this approach in the future when bacterial associations with diseases are suspected.

Citation Format: Kelly M. Robinson, Karsten B. Sieber, Kathleen E. Anderson, David R. Riley, Julie C. Dunning Hotopp. The microbiome of cancer as seen through the lens of public genome sequence data. [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 1534. doi:10.1158/1538-7445.AM2015-1534
Abstract 1535: Omentin: A novel adipokine linking visceral obesity to ovarian cancer progression
Authors: Au Yeung Chi Lam , Co Ngai Na , Onstad Michaela , Yeung Tsz-Lun , Leung Cecilia S. , Schmandt Rosemarie , Lu Karen H. , Mok Samuel C.
Published: August 2015

Advanced stage serous ovarian cancer metastasizes preferentially to the omentum, which is a major site of intra-abdominal fat accumulation, suggesting that the omental microenvironment is a favorable niche for ovarian cancer cells. The 5-year survival drops to below 50% for the cancer cases that spread beyond the pelvis to the omentum. It has also become apparent that obesity contributes to a poor clinical outcome. The mechanisms by which omental adipose tissue promotes tumor growth and disease progression are not entirely clear.

Using transcriptome profiling analysis on the microdissected adipose tissue from patients with benign gynecologic diseases and from patients with advanced high-grade serous ovarian cancer (HGSOC), we found that a novel adipokine called omentin (Intestinal Lactoferrin Receptor ITLN1) was significantly down-regulated in ovarian cancer associated adipose tissue compared with the normal adipose tissue. Survival correlation studies demonstrated that patients with serum ITLN1 levels of >350 ng/mL at the time of first treatment experienced longer survival times than those with lower levels of ITLN1. We showed that ITLN1 suppressed the ovarian cancer cell migration ability and invasion potential in vitro. To delineate the underlying molecular mechanisms, RNA sequencing and pathway analyses were performed on ITLN1 treated ovarian cancer cells and we identified MMP1 as one of the potential mediators. It has been shown that MMP1 expression was induced by lactoferrin, which is abundant in ascites. We hypothesized ITLN1 could abrogate the effect of lactoferrin on ovarian cancer motility and invasion potential. Our preliminary data suggested that secreted ITLN1 may sequester lactoferrin in the ascites, thereby preventing it from binding to the low-density-lipoprotein-receptor-related-protein-1 (LRP-1) on ovarian cancer cell surface and thus inactivate downstream signaling pathways that control MMP1 expression.

The study provides the first evidence that ovarian cancer cells modify the visceral adipose tissue through down-regulation of omentin to facilitate their growth in the omental microenvironment.

Citation Format: Chi Lam Au Yeung, Ngai Na Co, Michaela Onstad, Tsz-Lun Yeung, Cecilia S. Leung, Rosemarie Schmandt, Karen H. Lu, Samuel C. Mok. Omentin: A novel adipokine linking visceral obesity to ovarian cancer 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 1535. doi:10.1158/1538-7445.AM2015-1535
Abstract 1547: The clinical implication of MMP-1 and TIMP-1 on prognosis in patients with non-small cell lung cancer
Authors: An Ho Jung , Kang Jin-Hyoung , Lee Yoon-Jin , Hong Soon Uk , Lee Kyo Young , Kim Jeong-Oh
Published: August 2015

Introduction: The interaction between tumor cells and tumor micro-environment (TME) plays important role in tumor initiation, progression, treatment resistance, and prognosis in solid tumors including non-small cell lung cancer (NSCLC). The aim of this study is to evaluate the prognostic role of TME-related molecules in NSCLC patients who received surgical resection. To find out a possibility of a potential serum biomarker, protein expression in tissue from surgical specimen and pre-operative serum were investigated together.

Methods: For 85 NSCLC patients who underwent surgical resection from August 2006 to September 2010 and had pre-operative serum samplings, we screened the serum levels of 60 TME-related molecules with angiogenesis array kit for 8 patients who had recurrences and 4 patients who did not. For matrix metalloproteinase (MMP)-1 and tissue inhibitor of matrix metalloproteinase (TIMP)-1 in which significant difference was observed between recurrence and recurrence-free groups, protein expressions were investigated using ELISA in serum and immunohistochemistry in tumor tissue.

Results: High serum MMP-1 was related to carcinoembryonic antigen (CEA) level≥5 at diagnosis (P = 0.034), higher frequency of the presence of vascular invasion (P = 0.041). High expression of MMP-1 in tumor cells (intensity score≥2) was related to tumor differentiation (P = 0.057), whereas high MMP-1 expression in stroma was significantly associated with lymph node metastasis and TNM stage (P = 0.021 and P = 0.022, respectively). In the case of TIMP-1, expression in tumor cells was higher for adenocarcinoma but higher in stromal cells for squamous cell carcinoma (P = 0.046). There was no correlation between the tissue and serum levels of MMP-1 and TIMP-1. High serum MMP-1 showed a trend for short overall survival (P = 0.080). When serum MMP-1 was combined with CEA level or presence of vascular invasion, the prognostic implication for overall survival was statistically significant. Neither tumor nor stromal positivity for MMP-1 and TIMP-1 was associated with survival outcome; however, a tumor/stroma TIMP-1 intensity ratio≥1 was strongly associated with early recurrence in multivariate analysis (HR = 13.1, P = 0.008).

Conclusion: In surgically resected NSCLC patients, serum MMP-1 showed a possibility as a prognostic biomarker. In addition, the tumor/stroma TIMP-1 ratio (≥1) in tissue may be a useful to predict tumor recurrence.

Citation Format: Ho Jung An, Jin-Hyoung Kang, Yoon-Jin Lee, Soon Uk Hong, Kyo Young Lee, Jeong-Oh Kim. The clinical implication of MMP-1 and TIMP-1 on prognosis in patients with non-small cell lung 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 1547. doi:10.1158/1538-7445.AM2015-1547
Abstract 1562: Proteomic changes observed in colon cancer tumors by label-free differential profiling of paired tumor versus normal FFPE samples
Authors: Agrawal Lokesh , McAlister Fiona , Roy Sushmita , Unger Florian , David Kerstin , Juhl Harmut , Chelsky Daniel , Moore Helen
Published: August 2015

Colorectal cancer (CRC) is the third most common cancer and if caught early enough has a good prognosis. Mass screening of the adult population is advisable to reduce CRC mortality rate as well as reduce the cost of treating advanced CRC cases. A blood-based biomarker panel with the ability to diagnose CRC as well as determine prognosis and predict the response to drug therapy is desired. However, blood biomarker discovery is hampered by problems of complexity and dynamic range of protein abundance.

Formalin-fixed and paraffin-embedded (FFPE) tissue is an ideal sample type for cancer biomarker analysis due to its ubiquitous use in cancer diagnosis in the clinic and consequent availability in oncology hospitals and clinics as well as biobanks. Biomarker discovery to date using FFPE have used only very few patient samples, making clinically relevant biomarker discovery difficult due to large patient-to-patient variation. As a part of our Biospecimen Research Network (BRN) program we funded a proteomic biomarker study of a large number of colorectal cancer and adjacent normal tissue samples. The investigators employed two platforms for biomarker discovery: label-free intensity based discovery using high resolution tandem mass spectrometry and targeted quantitation using multiple reaction monitoring. These two techniques allowed them to dig deeper into the proteome and to profile phosphopeptides. Paired tissue from 40 patients was profiled and of the 2875 proteins quantified, 367 proteins changed significantly in patients whose tumor size was greater than 50%. Large changes in secreted proteins were observed and many of these proteins were also observed in blood, offering promise for a potential blood-based biomarker for colorectal cancer. A blood-based biomarker assay for colorectal cancer could ultimately reduce the number of colonoscopies required, reducing cost and discomfort.

Citation Format: Lokesh Agrawal, Fiona McAlister, Sushmita Roy, Florian Unger, Kerstin David, Harmut Juhl, Daniel Chelsky, Helen Moore. Proteomic changes observed in colon cancer tumors by label-free differential profiling of paired tumor versus normal FFPE samples. [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 1562. doi:10.1158/1538-7445.AM2015-1562
Abstract 1563: Stool proteomics reveals novel candidate biomarkers for colorectal cancer screening
Authors: Bosch Linda JW , de Wit Meike , Hiemstra Annemieke C. , Piersma Sander , Pham Thang , Oudgenoeg Gideon , Scheffer George , Mongera Sandra , Komor Malgorzata , Terhaar Sive Droste Jochim , Oort Frank A. , van Turenhout Sietze , Ben Larbi Ilhame , Mulder Chris JJ , Carvalho Beatriz , Fijneman Remond JA , Jimenez Connie , Meijer Gerrit A.
Published: August 2015

Introduction and objectives: Early detection of colorectal cancer (CRC) and its precursor lesions is an effective approach to reduce CRC mortality rates. The fecal immunochemical test (FIT) is a non-invasive CRC screening test that detects small traces of the blood protein hemoglobin. Although beneficial in its current form, the FIT test characteristics leave room for improvement. The aim of the present study was to identify and validate novel protein biomarkers in stool that complement or outperform the current hemoglobin-based test, to improve its diagnostic accuracy.

Methods: Proteins isolated from stool from 10 subjects without any signs of colorectal neoplasia (controls) at colonoscopy and from 12 CRC patients were analyzed by GeLC-MS/MS (discovery set). Data were analyzed by comparing protein abundancies, measured as spectral counts. Analysis of differential proteins was performed using the beta-binomial test. Findings were validated by mass spectrometry (Q-Exactive) in an independent series of 292 stool samples obtained from control subjects (n = 109) and subjects with adenomas (n = 55), advanced adenomas (n = 53), or CRCs (n = 75).

Results and Discussion: In total 830 human proteins were identified in the discovery set, of which 134 were significantly enriched in CRC. These included 78 proteins that were significantly more enriched in FIT-negative CRC stool samples compared to controls. Preliminary analysis of the validation set indicates that more than half of these markers are significantly more abundant in CRC samples compared to controls.

Conclusion: Proteome profiling of stool samples revealed novel candidate biomarkers to improve current CRC screening tests. More data analysis is currently ongoing to select most promising protein biomarkers for clinical assay development.

Citation Format: Linda JW Bosch, Meike de Wit, Annemieke C. Hiemstra, Sander Piersma, Thang Pham, Gideon Oudgenoeg, George Scheffer, Sandra Mongera, Malgorzata Komor, Jochim Terhaar Sive Droste, Frank A. Oort, Sietze van Turenhout, Ilhame Ben Larbi, Chris JJ Mulder, Beatriz Carvalho, Remond JA Fijneman, Connie Jimenez, Gerrit A. Meijer. Stool proteomics reveals novel candidate biomarkers for colorectal cancer screening. [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 1563. doi:10.1158/1538-7445.AM2015-1563
Abstract 1578: Identification of type X collagen as a pan-cancer serum biomarker
Authors: Prendes Maria J. , Chapman Karen B. , Tam Orlena , Wagner Joseph
Published: August 2015

We previously reported elevated expression of Type X Collagen (COL10A1) in solid tumors of diverse origin (Future Oncology 2012 Aug;8(8): 1031-40). This elevated expression in tumors was primarily localized to vasculature, while COL10A1 expression in most adult tissues was negligible. Given this pattern of expression, we hypothesized that COL10A1 might demonstrate utility as a blood-born tumor biomarker. Serum samples from healthy volunteers and from patients with active cancers of the breast, lung, pancreas, colon, kidney, and bladder were obtained from multiple existing bio-banks. COL10A1 serum levels were assessed in over 400 total patient samples using a polyclonal COL10A1 sandwich ELISA. The overall utility of COL10A1 levels in determining presence of cancer was judged by calculating ROC-AUC scores for each individual cancer type. In general, COL10A1 levels were increased in cancer patients, although this increase varied as a function of cancer type as well as by the bio-bank the sera was obtained from. Given the sensitivity and specificity limitations of polyclonal ELISA technology, even greater performance of COL10A1 as a tumor biomarker could be attainable using high-specificity monoclonal antibodies. We are currently characterizing such tests in prospectively collected sera samples from breast cancer patients. These data suggest that COL10A1 is a potential blood-born pan-cancer biomarker that could prove useful in non-invasive cancer screening as well as in monitoring tumor progression and treatment response.

Citation Format: Maria J. Prendes, Karen B. Chapman, Orlena Tam, Joseph Wagner. Identification of type X collagen as a pan-cancer serum biomarker. [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 1578. doi:10.1158/1538-7445.AM2015-1578
Abstract 1579: Circulating cell-free DNA as a diagnostic marker for epithelial ovarian cancer and an association with epithelial-mesenchymal transition
Authors: Koo Yu-Jin , Min Kyung-Jin , Hong Jin-Hwa , Lee Jae-Kwan
Published: August 2015

Objectives: Although technologies continue to develop, there are still many limitations on detecting circulating tumor cells in blood. In addition, the great genetic heterogeneity in ovarian cancer is one of the major concerns. Therefore, we aimed to investigate the cell-free DNA (cfDNA) as a diagnostic marker for epithelial ovarian cancer (EOC) and evaluate the cfDNA level of epithelial-mesenchymal transition markers.

Methods: Blood was preoperatively collected from women who were assigned to surgery for pelvic mass. DNA was extracted using Qiagen DNA Mini kit and quantification of total cfDNA was performed using real-time PCR with TaqMan Assay with primers directed to the following genes:

GAPDH, β-globin, β-actin, E-cadherin, and fibronectin.

Results: A total of 32 patients with EOC and 17 patients of benign ovarian disease (controls) were included. The median cfDNA level of β-globin was 0 (range, 0-0.35)GE/ml in EOC and 0.41 (range, 0-33.5)GE/ml in control (p = 0.04).

The sensitivity of β-globin cfDNA (>0.4 GE/ml) was 51.9%, which was lower than that of CA-125 (96.3%, if CA-125 >35IU/ml). cfDNA was significantly higher in advanced stage of EOC than early stage EOC (p = 0.02). Plasma E-cadherin level was significantly related with β-globin cfDNA (coefficient 0.886, p<0.001). However, there was no significant association with cfDNA amount and survival outcomes.

Conclusions: Circulating cfDNA can be a promising target for early detection of EOC. The levels of epithelial-mesenchymal transition markers showed a different pattern in plasma and were associated with other cfDNA amounts.

Citation Format: Yu-Jin Koo, Kyung-Jin Min, Jin-Hwa Hong, Jae-Kwan Lee. Circulating cell-free DNA as a diagnostic marker for epithelial ovarian cancer and an association with epithelial-mesenchymal transition. [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 1579. doi:10.1158/1538-7445.AM2015-1579
Abstract 1585: Application of a graphene oxide based microfluidic device (GO Chip) to prostate cancer circulating tumor cell capture and analysis
Authors: Kozminsky Molly , Yoon Hyeun Joong , Palanisamy Nallasivam , Cooney Kathleen , Hussain Maha , Alva Ajjai , Morgan Todd , Nagrath Sunitha
Published: August 2015

One in seven men in the United States will be diagnosed with prostate cancer in his life time, in part because of the prevalence of screening for serum levels of prostate specific antigen (PSA). However, this has led to the overtreatment of the disease, leading to recommendations against frequent PSA testing and establishing the need for a more informative biomarker in prostate cancer. Circulating tumor cells (CTCs) are rare cells present in the blood stream of cancer patients at the low frequency on the order of one CTC in one billion normal blood cells. The graphene oxide chip (GO Chip) is a microfluidic CTC capture device with increased sensitivity and purity due to its use of the nanomaterial graphene oxide to present a capture antibody against the epithelial cellular adhesion molecule (EpCAM). In this study, whole blood was collected from 50 prostate cancer patients, including those with metastatic, localized, castrate resistant, and castrate sensitive disease, and analyzed using the GO Chip. Captured cells were stained for cytokeratin, CD45, and DAPI, with nucleated cytokeratin positive cells being denoted as CTCs. CTCs were detected in 47 out of 50 patients, and an average of 5.94 CTCs/mL was recovered from each sample. Correlation of CTCs with surrogate endpoints could improve the utility of CTCs as a liquid biopsy, potentially advancing their application in translational research.

Citation Format: Molly Kozminsky, Hyeun Joong Yoon, Nallasivam Palanisamy, Kathleen Cooney, Maha Hussain, Ajjai Alva, Todd Morgan, Sunitha Nagrath. Application of a graphene oxide based microfluidic device (GO Chip) to prostate cancer circulating tumor cell capture and analysis. [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 1585. doi:10.1158/1538-7445.AM2015-1585
Abstract 1591: Comprehensive molecular profiling of circulating tumor cells from metastatic breast cancer patients
Authors: Magbanua Mark Jesus Mendoza , Hauranieh Louai , Roy Ritu , Pendyala Praveen , Sosa Eduardo , Scott Janet , Rugo Hope , Park John
Published: August 2015

Background: Recent enumeration studies on CTCs have demonstrated the clinical value of these cells. The molecular biology of CTCs, however, remains poorly understood.

Methods: We performed RNA and DNA profiling of CTCs from metastatic breast cancer (MBC) patients. 244 blood samples from 162 MBC patients were subjected to immunomagnetic enrichment and fluorescence activated cell sorting (IE/FACS) to isolate highly pure CTCs. Microfluidic-based multiplex QPCR analysis was utilized to determine the expression levels of 64 cancer-related genes. Parallel genome-wide copy number analysis by array comparative genomic hybridization was also performed on CTCs isolated from the same enriched blood samples.

Results: Transcriptional analysis of CTCs was successfully performed in 151 samples from 105 patients. We observed the up-regulation of several genes including CCND1, EPCAM, KRT7 and MUC1 and the down-regulation of hematopoietic-related genes including PTPRC (CD45) and CD68 (adjusted p <0.001) in CTCs. Clustering analysis of expression profiles revealed three CTC clusters, one containing both subtypes basal (ESR1-low) and normal-like (ESR1-low, CAV1/VIM-high) and two other clusters containing luminal subtype (ESR1-high). A subset of CTCs expressed epithelial-mesenchymal transition genes, however, genes for stem cellness were not generally expressed. Serial CTC expression profiling in 28 patients demonstrated changes in expression over time. Exploratory single CTC expression analysis revealed cell-to-cell heterogeneity. Parallel genome-wide copy number analysis in matched CTCs from 49 patients revealed gains (e.g. 1q and 8q), losses (e.g., 8p and 16q), and focal amplifications (e.g. on 8q and 11q including CCND1) that are frequently observed in primary breast cancers. We also observed three copy number clusters with varying levels of genomic aberrations. Up-regulation of CCND1 and ERBB2 in CTCs was associated with increased level of genomic instability.

Conclusions: Parallel gene expression and copy number profiling provided novel insights on the biology of CTCs. Molecular characterization of CTCs may shed further light on the biology of metastasis and disease progression.

Citation Format: Mark Jesus Mendoza Magbanua, Louai Hauranieh, Ritu Roy, Praveen Pendyala, Eduardo Sosa, Janet Scott, Hope Rugo, John Park. Comprehensive molecular profiling of circulating tumor cells from metastatic breast cancer patients. [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 1591. doi:10.1158/1538-7445.AM2015-1591