bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5142: A naturally occurring model for gastric cancer
Authors: AbdelMageed Manar A. , Betancur-Boissel Monica , Foltopoulou Parthena , Muthupalani Sureshkumar , Fox James G. , McNiel Elizabeth A.
Published: August 2015

Gastric cancer is the fourth most common cancer and the second leading cause of cancer related deaths. While there are increasing data characterizing the molecular features and defining potential targets for gastric carcinoma, preclinical animal models that recapitulate the human disease are limited. We have developed a number of resources to permit the characterization of spontaneously occurring canine gastric cancer which may provide a useful resource to understand gastric cancer carcinogenesis, its prevention and treatment.

While gastric carcinoma is reportedly uncommon in dogs, certain breeds have significantly increased risk. To study this disease, we have established a database and tissue repository. Our goal has been to characterize canine gastric carcinoma clinically, histologically, and molecularly to determine its comparative value as a model for the human disease. Recently, we have performed a histologic review of 50 canine gastric carcinomas and have found that the canine disease is comprises largely of diffuse type gastric carcinoma and is enriched in the signet ring variant.

We report for the first time a technique for culturing primary gastric cancer from affected dogs. Gastric cancer tissue specimens collected from spontaneously occurring tumors in dogs were disaggregated with collagenase and cells were grown in mammary epithelial growth media with or without 5% FBS. Four different cell lines were derived from tumors of a Labrador mix (GC1), a Basset hound (GC2), a Bouvier de Flandres (GC3) and a mixed breed (GC4). Early passages of all cell lines were able to form colonies when grown in soft agar.

Different growth patterns were observed when cells were cultured with versus without serum. Serum treated cells tend to be fast growing, fibroblastic in appearance and senesce within 10 to 18 passages and these may represent cancer associated fibroblasts or mesenchymal differentiation of cancer cells. However, cells grown in mammary epithelial growth media without serum tend to form spheres that grow well when transferred to ultra-low attachment plate and form extensions when grown in matrigel. That semi-attached growth behavior is consistent with that reported for some human gastric cancer cell lines.

Canine gastric cancer cell lines provide a new resource for the characterization of this spontaneous model that recapitulates diffuse, signet ring cell gastric carcinoma.

Citation Format: Manar A. AbdelMageed, Monica Betancur-Boissel, Parthena Foltopoulou, Sureshkumar Muthupalani, James G. Fox, Elizabeth A. McNiel. A naturally occurring model for gastric 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 5142. doi:10.1158/1538-7445.AM2015-5142
Abstract 5143: Rats heterozygous for Pten develop fibroadenoma and adenocarcinoma arising from the mammary gland
Authors: McCoy Aaron M. , Manickam Bala , Reid Lara , Weinstein Edward , Cui Xiaoxia
Published: August 2015

To further characterize the function of the tumor suppressor Pten in the role of tumorigenesis we have created a colony of Pten mutant Sprague Dawley (SD) rats. Utilizing zinc finger nuclease technology we introduced a 7bp deletion in Exon 7 of the Pten gene, creating a frameshift mutation and an early stop codon. Pten homozygous null animals are not viable so a colony of heterozygous animals is maintained. Spontaneous tumors of the mammary gland arise in these animals and were removed for staining and pathological analysis. We describe briefly the creation of this model and preliminary genotypic and proteomic characterization. Further we describe pathological observations made on tumor tissues removed from aging Pten heterozygous animals.

We have confirmed a reduction in Pten expression by Western Blot utilizing a monoclonal antibody targeting a central locus in the protein. Heterozygous Pten animals develop spontaneous tumors of the mammary gland which were removed if animals demonstrated deteriorating health or any condition within our Humane Care and Use guidelines. Thirty six tissues from 16 animals were submitted for histopathological analysis. Eight of the animals were male, 8 of the animals were female and had a mean lifespan of 159 days (165 male, 153 female). Remaining consistent with human breast cancer incidence in women below 30 years of age, the most common lesion observed is a benign fibroadenoma (30/36 samples). The malignant adenocarcinoma was observed in 5/36 samples submitted. Fibroadenoma only lesions were observed in 12 of the 16 rats while both fibroadenoma and adenocarcinoma was observed in 4 of the 16 rats.

Taken together we believe that a Pten heterozygous rat provides an additional tool for the elucidation of the role of Pten in human cancers. This model may also be useful in the study of neurological disorders such as Autism, Parkinson's disease, Alzheimer's disease, obesity, cardiovascular disease, and diabetes.

Mutations in Pten strongly correlate with several human disorders including Cowden syndrome, Bannayan-Zonana syndrome, and Lhermitte-Duclos disease. These disorders result in a tumor pathologies including an increased risk for breast cancers.

Citation Format: Aaron M. McCoy, Bala Manickam, Lara Reid, Edward Weinstein, Xiaoxia Cui. Rats heterozygous for Pten develop fibroadenoma and adenocarcinoma arising from the mammary gland. [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 5143. doi:10.1158/1538-7445.AM2015-5143
Abstract 5147: Canine spontaneous head and neck squamous cell carcinomas represent their human counterparts at the molecular level
Authors: Liu Deli , Xiong Huan , Ellis Angela E. , Northrup Nicole C. , Shin Dong M. , Zhao Shaying
Published: August 2015

Spontaneous canine head and neck squamous cell carcinoma (HNSCC) represents an excellent model of human HNSCC but is greatly understudied. To better utilize this valuable resource, we performed its first genome-wide characterization by investigating 12 canine HNSCC cases, of which 9 are oral, via high density array comparative genomic hybridization and RNA-seq. The analyses reveal that these canine cancers faithfully recapitulate many key molecular features of human HNSCC. These include similar genomic copy number abnormality landscapes, analogous sequence mutation patterns, and recurrent alteration of known HNSCC genes (e.g., MYC, CDKN2A) and pathways (e.g., cell cycle, mitogenic signaling, TGFβ signaling). Amplification or overexpression of protein kinase genes, matrix metalloproteinase genes, and epithelial-mesenchymal transition genes TWIST1 and SNAI are also prominent in these canine tumors.

The strong dog-human molecular homology fundamentally justifies the use of our novel dog-human comparison strategy for driver-passenger discrimination for HNSCC. This cross-species comparison will significantly advance beyond the current findings with the massive genomic data of the cancer genome atlas (TCGA), as exemplified by our pilot study on human 8q, a recurrently amplified site in human HNSCC.

HNSCC represents the sixth leading cancer by incidence in humans; thus developing effective therapeutic interventions is important. While our understanding of the biology of HNSCC has been greatly advanced over past decades, translating these findings into clinical success has been frustratingly slow. A significant obstacle is the lack of translational model that can bridge the gap between preclinical research and human clinical trials. Our study indicates that spontaneous canine HSCC can effectively bridge this gap, significantly accelerating anti-HNSCC drug discovery.

Note: This abstract was not presented at the meeting.

Citation Format: Deli Liu, Huan Xiong, Angela E. Ellis, Nicole C. Northrup, Dong M. Shin, Shaying Zhao. Canine spontaneous head and neck squamous cell carcinomas represent their human counterparts at the molecular level. [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 5147. doi:10.1158/1538-7445.AM2015-5147
Abstract 5151: Yap reprograms glutamine metabolism and supports growth during liver development and tumorigenesis
Authors: Cox Andrew G. , Hwang Katie L. , Evason Kimberley , Brown Kristin K. , Beltz Sebastian , O'Connor Keelin , Galli Giorgio G. , Yimlamai Dean , Chhangawala Sagar , Lien Evan , Camargo Fernando D. , Asara John , Houvras Yariv , Stainier Didier Y. , Goessling Wolfram
Published: August 2015

Hepatocellular carcinoma is a global health problem with poor prognosis and limited therapeutic options. Recently, the Hippo pathway has emerged as a master regulator of organ size control and tumorigenesis. However, the metabolic impact of the pathway is poorly understood. Using a transgenic zebrafish model with liver-specific activation of the Hippo pathway effector Yap, we have shown that Yap promotes hepatomegaly and liver dysplasia. In addition, we demonstrate that the Yap transgenics are highly susceptible to chemically-induced hepatocarcinogenesis. Transcriptomic and metabolomic profiling reveals that Yap enhances glutamine synthase (GS) expression and elevates steady-state levels of glutamine, respectively. Intervention studies with the GS inhibitor methionine sulfoximine established that elevated GS activity contributes to the rapid liver growth observed during Yap-driven hepatomegaly. Finally, studies in cultured human cancer cells identify GS as a bone-fide Yap target gene, confirming that the GS regulation by Yap is evolutionarily conserved. We conclude that Yap regulates GS expression and reprograms nitrogen metabolism, which contributes to liver growth during development and tumorigenesis. We hypothesize that Yap integrates the anabolic demands of rapid cell proliferation by increasing the flux of glutamine into nucleotide biosynthesis.

Citation Format: Andrew G. Cox, Katie L. Hwang, Kimberley Evason, Kristin K. Brown, Sebastian Beltz, Keelin O'Connor, Giorgio G. Galli, Dean Yimlamai, Sagar Chhangawala, Evan Lien, Fernando D. Camargo, John Asara, Yariv Houvras, Didier Y. Stainier, Wolfram Goessling. Yap reprograms glutamine metabolism and supports growth during liver development and tumorigenesis. [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 5151. doi:10.1158/1538-7445.AM2015-5151
Abstract 5171: EpCAM expression governs tumor growth or distant metastasis of human endometrial cancer: grow or go theory
Authors: Wen Kuo-Chang , Wang Peng-Hui , Sung Pi-Lin , Lee Mong-Hong , Wu Cheng-Wen
Published: August 2015

Endometrial cancer is the most common gynecologic malignancy in United States. The prognosis is generally better because most patients are diagnosed at early stage. However, the overall survival rate declines to 10∼20% at advanced stage. There are no effective treatment in the management of patient with late stage of endometrial cancer or distant metastasis. EpCAM (also called TACSTD1 or CD326), a transmembrane glycoprotein, can mediate cell-cell adhesion. Its overexpression negatively correlates with clinical survival in gallbladder cancer, ampullary carcinoma, and squamous cell carcinoma of head &. neck. However, a controversial role of EpCAM is reported in Lynch syndrome and rectal cancer as a tumor suppressor gene. To study the role of EpCAM in endometrial cancer, we manipulate the EpCAM expression in loss-of-function experiment in vitro; the results demonstrate that decrease of EpCAM expression is significantly related to increase invasion ability and diminish cell proliferation. These finding can be reversed after gain-of-function in EpCAM knock-down endometrial cancer cells. EpCAM knock-down is indeed linked to the cell signaling pathway of proliferation (regulation of cell proliferation GO:0042127, regulation of cell death:GO:0010941, regulation of apoptosis: GO:0042981) and migration (biological adhesion:GO:0022610, cell adhesion:GO:0007155, cell migration:GO:0016477) from microarray analysis. Significant more number of lymph node metastasis can be found in vivo at mice with IUI (intra-uterine injection) of EpCAM knock-down cells; moreover, smaller tumor size is observed in subcutaneously injected mice. Among the patients with endometrial cancer, the clinicopathologic characteristics, such as stage, lymph node metastasis, CA-125 value, and overall survival, are significantly related to the low EpCAM expression. Conclusively, diminishing EpCAM can promote endometrial cancer invasion; furthermore, the EpCAM expression is an significant prognostic factor in human endometrial cancer. These data has important implication that development targeting EpCAM in the treatment of endometrial cancer is the new application in future.

Citation Format: Kuo-Chang Wen, Peng-Hui Wang, Pi-Lin Sung, Mong-Hong Lee, Cheng-Wen Wu. EpCAM expression governs tumor growth or distant metastasis of human endometrial cancer: grow or go theory. [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 5171. doi:10.1158/1538-7445.AM2015-5171
Abstract 5198: Contribution of inflammatory cytokines to CD44-mediated breast cancer metastatic potential
Authors: Covert Hunter , Mellor Liliana
Published: August 2015

The purpose of this study was to elucidate the relationship between the inflammatory cytokine oncostatin M (OSM) and cell migration-promoting CD44 in breast cancer metastasis. OSM-induced CD44 expression was studied in multiple human breast cancer cell lines, including triple negative (TNBC) MDA-MB-231 and estrogen receptor/progesterone receptor-positive (ER+/PR+) T47D and MCF-7 cells. Western blot analysis and RT-PCR demonstrated that OSM is capable of upregulating the expression of the standard form of CD44 (CD44s), as well as inducing multiple CD44 variants produced by alternative splicing. Confocal microscopy was performed and displayed highly organized CD44 around the edge of the cell after OSM treatment. OSM treatment also increased the breast cancer stem cell (BCSC) population (CD44+/CD24−), as measured by flow cytometry. To examine the importance of OSM-induced CD44 in tumor cell migration, we conducted migration assays with and without siCD44 and treatment with OSM. These results verified that OSM-induced CD44 is important for cellular migration. In conclusion, our studies demonstrate that OSM increases the expression of CD44s and its variants, as well as induce a BCSC phenotype. Furthermore, we show that OSM-induced CD44 plays a role in migration, which is an essential characteristic of breast cancer metastasis.

Citation Format: Hunter Covert, Liliana Mellor. Contribution of inflammatory cytokines to CD44-mediated breast cancer metastatic potential. [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 5198. doi:10.1158/1538-7445.AM2015-5198
Abstract 5205: Docosahexaenoic acid (DHA) alters breast cancer exosome-mediated microRNA signaling
Authors: Hannafon Bethany N. , Carpenter Karla , Berry William , Janknecht Ralf , Dooley William , Ding Wei-Qun
Published: August 2015

DHA is a long-chain omega-3 polyunsaturated fatty acid that has anticancer properties, including the ability to suppress tumor angiogenesis. However, the precise mechanism of DHAs anti-angiogenic activity is not well defined. It is recognized that the intercommunication between cancer cells and their microenvironment is essential to tumor angiogenesis. Exosomes are nanometer-sized extracellular vesicles that are important mediators of intercellular communication and in recent years, the role of tumor-derived exosomes in cancer progression has been realized. However, ways to limit or alter their influence on cancer progression has not been demonstrated. At this time, very little is known about the role of breast cancer derived exosomes in contributing to breast cancer progression and whether these exosomes mediate DHAs anticancer activity. To investigate, breast cancer exosomes were collected from the conditioned media of MCF7 and MDA-MB-231 breast cancer cells engineered to express GFP-tagged CD63 after treatment with DHA. We observed an increase in exosome secretion from the DHA-treated cells. Total RNA was extracted from the DHA-treated and control MCF7-derived exosomes and analyzed by small RNA sequencing. The expression of 83 exosome microRNAs was altered by DHA (>2-fold) and several of the most abundant exosomal microRNAs (miR-23b, miR-27a/b, miR-21, and miR-320b) are known to have anti-angiogenic activity. When DHA-treated MCF7 cells were co-cultured with or the collected exosomes were directly applied to endothelial cell cultures, we observed an increase in the expression of these microRNAs in endothelial cells. Furthermore, overexpression of miR-23b and miR-320b in endothelial cells decreased the expression of their pro-angiogenic target genes (PLAU, AMOTL1, NRP1 and ETS2) and significantly inhibited tube formation by endothelial cells, an effect that could be reversed by inhibition of exosome secretion via Rab27A knockdown. These results suggest that the microRNAs transferred by exosomes mediate DHAs anti-angiogenic action. Our data demonstrates that DHA alters breast cancer exosome secretion and microRNA contents that leads to the inhibition of endothelial tube formation. These results provide insight into the future possibility of developing new cancer therapeutic strategies targeting exosome secretion and content transmission.

Citation Format: Bethany N. Hannafon, Karla Carpenter, William Berry, Ralf Janknecht, William Dooley, Wei-Qun Ding. Docosahexaenoic acid (DHA) alters breast cancer exosome-mediated microRNA signaling. [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 5205. doi:10.1158/1538-7445.AM2015-5205
Abstract 5208: Neovascularization in brain metastasis is through both angiogenesis and vasculogenesis
Authors: Mok Stephanie , Ang Lee-Cyn , Howlett Christopher J. , Khan Zia A.
Published: August 2015

Metastatic brain tumors are the most common brain tumor in adults with an incidence 10 times greater than primary brain tumors. A variety of malignancies eventually spread to the brain, with a majority arising from cancers of the lung, breast, kidney, and skin. Experimental studies using breast and melanoma cell lines have recently shown that growth of metastatic brain tumors may occur by utilizing pre-existing blood vessel, or by co-opting rather than inducing new vessel formation. Whether the same mechanisms of vascular utilization and expansion play a role in human metastatic brain tumors is unknown and is the focus of the present study.

We examined the immuno-phenotype of blood vessels in primary and metastatic human brain tumors to determine the presence of new blood vessels formation. Glioblastoma multiforme (n = 12) and secondary tumors from melanoma (n = 10), breast (n = 10), kidney (n = 5), and lung (n = 13) all showed extensive immunoreactivity for endothelial marker CD31. We also found a significant number of endothelial cells expressing Ki-67, most notably among breast carcinoma, glioblastoma multiforme and melanoma cases, indicating microvessel proliferation and angiogenesis. Interestingly, both primary and secondary metastatic tumors also showed vascular endothelial cells concomitantly expressing stem cell markers, such as Oct4, suggesting that some of these microvessels are derived through vasculogenesis. These findings show that both proliferation of endothelial cells and de novo endothelial differentiation may underlie metastatic tumor growth to different degrees depending on their site of primary origin. Currently, our laboratory is determining the exact contribution of angiogenesis and vasculogenesis in brain metastasis.

Our findings show that neo-vessels are evident in brain metastasis. Elucidating the source of these vessels may uncover new treatment targets for patients with brain metastasis.

Citation Format: Stephanie Mok, Lee-Cyn Ang, Christopher J. Howlett, Zia A. Khan. Neovascularization in brain metastasis is through both angiogenesis and vasculogenesis. [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 5208. doi:10.1158/1538-7445.AM2015-5208
Abstract 521: Connexin 43 and E-cadherin modulate prostate cancer cell migration
Authors: Aloliqi Abdulaziz , Zhang Ao , Brett-Morris Adina , Lathia Justin , Fraizer Gail C.
Published: August 2015

Connexins play a major role in cell-cell communication and form a bidirectional signaling pathway to assemble gap junctions and, along with cadherins, alter cell behaviors, such as motility. To assess the effect of Connexin 43 (Cx43) on migration of prostate cancer cells we performed two types of migration assays using shRNA Cx43 transfected PC3 cells with suppressed Cx 43. Wound healing and transwell chamber assays were used to assess the migratory potential of stably transfected shCx43-PC3 cells. When we reduced Cx 43 we saw approximately a two-fold reduction in migration of shCx43 transfected PC3 cells compared to those transfected with scramble controls. Since suppression of Cx43 reduced migration of the highly migratory PC3 cells, we wondered whether this reflected a partial loss of mesenchymal traits (e.g., motility) and acquisition of epithelial markers, (e.g.,E-cadherin expression). Using quantitative RT-PCR, E-cadherin expression was assessed in shCx43-PC3 and scramble control cells. Suppression of Cx43 increased E-cadherin mRNA levels 5-fold or greater in shCx43-PC3 cells than in scramble control cells. An alternative approach used to transcriptionally up-regulate E-cadherin mRNA in PC3 cells (via suppression of a transcriptional repressor) resulted in a similar approximately 2.5-fold suppression of migration as measured by wound healing and transwell chamber assays. Thus, an inverse relation of Cx 43 and E-cadherin was observed in prostate cancer cells and alterations in either led to aberrant migration. These results were consistent with our hypothesis that differential connexin expression in prostate cancer cells contributes to different metastatic potential. To correlate differential connexin subunit expression with migratory potential, we quantified gene expression of 13 different Connexin subunits in non-invasive LNCaP and in invasive PC3 cells using qRTPCR. In LNCaP cells we observed low levels of Cx43 along with high levels of Cx32 and E-cadherin expression and the converse in PC3 cells. Overall, these results suggest that reducing high levels of Cx43, associated with enhanced E-cadherin expression and reduced migratory behavior could potentially block lethal metastatic tumor progression of prostate cancer cells.

Citation Format: Abdulaziz Aloliqi, Ao Zhang, Adina Brett-Morris, Justin Lathia, Gail C. Fraizer. Connexin 43 and E-cadherin modulate prostate cancer cell migration. [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 521. doi:10.1158/1538-7445.AM2015-521
Abstract 522: Involvement of Zyxin in aggressiveness of human prostate cancer
Authors: Yamamoto Kei , Hori Yasuki , Seo Jieun , Hashimoto Saho , Endo Yoshihiro , Nittami Tadashi , Ito Yusuke , Ishiguro Hitoshi , Uemura Hiroji , Watanabe Masatoshi
Published: August 2015

Many epidemiological studies have shown that abdominal obesity was associated with an increased risk in several malignancies. The association between prostate cancer risk and obesity has been also investigated, but has been in dispute. The involvement of adipocytokines as putative molecular mediators between prostate cancer and obesity has been also investigated. We have already shown effects of conditioned medium (CM) from adipocyte culture in behaviors of prostate cancer cells (DU145). In this study, Phospho-peptide in DU145 cells cultured in CM was analyzed using Liquid chromatography-tandem mass spectrometry (LC-MS/MS). Zyxin was identified as a CM-related phosphor-peptide. Zyxin is a conserved protein that has two major domains, an N-terminal half containing proline-rich sequences and a C-terminal LIM domain region. Its functions are involved in regulation of cell adhesion and motility. This protein also appears to have tumor suppressor activity with a few reports, however in dispute. We analyzed the involvement of Zyxin in behavior of DU145 cells. Functional analysis of Zyxin in cell migration, invasive potential and others of DU145 cells was conducted using Zyxin siRNA. Knockdown of Zyxin by siRNA in DU145 cells inhibited cell migration and invasion. As for clinical paired samples, Zyxin expression in tumor tissues was higher than in normal tissues by real-time PCR. These results suggest that Zyxin may play an important role in aggressiveness of human prostate cancer.

Citation Format: Kei Yamamoto, Yasuki Hori, Jieun Seo, Saho Hashimoto, Yoshihiro Endo, Tadashi Nittami, Yusuke Ito, Hitoshi Ishiguro, Hiroji Uemura, Masatoshi Watanabe. Involvement of Zyxin in aggressiveness of human prostate 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 522. doi:10.1158/1538-7445.AM2015-522
Abstract 5225: Endothelial-specific Jagged1 blockade prevents solid tumor growth in pre-clinical models
Authors: Duarte Antonio , Pedrosa Ana-Rita , Trindade Alexandre , Carvalho Catarina , Graça José , Carvalho Sandra , Peleteiro Maria C. , Adams Ralf H.
Published: August 2015

Angiogenesis is an essential process required for tumor growth and progression. The Notch signaling pathway has been identified as a key regulator of the neo-angiogenic process. Jagged-1 (Jag1) is a Notch ligand required for embryonic and retinal vascular development. Although its role as a pro-angiogenic ligand has been thoroughly studied in developmental contexts, the real contribution of this Notch ligand to tumor angiogenesis remains unknown.

The current study addresses the role of endothelial Jagged1-mediated Notch signaling in the context of tumoral angiogenesis in two different mouse tumor models: subcutaneous Lewis Lung Carcinoma (LLC) tumor transplants and the autochthonous Transgenic Adenocarcinoma of the Mouse Prostate (TRAMP).

The role of endothelial Jagged1 in tumor growth and neo-angiogenesis was investigated with endothelial-specific Jag1 gain- and loss-of-function mouse mutants (eJag1OE and eJag1cKO). We show that the tumor vasculature of eJag1OE mutants presents increased vessel density, branching, and perivascular maturation, causing increased tumor perfusion. Ultimately, this leads to increased proliferation and survival of tumor cells, and increased epithelial-to-mesenchymal transition (EMT) resulting in increased LLC tumor growth as well as development and progression of prostate adenocarcinoma in mice. In contrast, in eJag1cKO mutants the vasculature is characterized by decreased vessel density, branching, and perivascular maturation, which restricts tumor vascular perfusion and increases vascular extravasation. This vascular phenotype leads to decreased proliferation and increased death of tumor cells, reduced EMT culminating in decreased LLC growth as well as inhibited development and progression of prostate adenocarcinoma in mice. Therefore, endothelial Jagged1 may constitute a novel promising target for solid cancer therapy.

Note: This abstract was not presented at the meeting.

Citation Format: Antonio Duarte, Ana-Rita Pedrosa, Alexandre Trindade, Catarina Carvalho, José Graça, Sandra Carvalho, Maria C. Peleteiro, Ralf H. Adams. Endothelial-specific Jagged1 blockade prevents solid tumor growth in pre-clinical models. [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 5225. doi:10.1158/1538-7445.AM2015-5225
Abstract 5226: Glioma stem cells internalize perivascular bevacizumab via a non-canonical pathway and target it for recycling or degradation
Authors: Muller-Greven Gaelle , Carlin Cathleen , Lathia Justin , Prayson Richard , Ahluwalia Manmeet , Toms Steven , Bredel Markus , Rich Jeremy , Hamerlik Petra , Gladson Candece L.
Published: August 2015

Antiangiogenic therapy shows great promise for treatment of cancer. Bevacizumab is a humanized mAb that blocks VEGF-A, thereby inhibiting angiogenesis. It has received FDA approval for patients with recurrent glioblastoma (GBM); however, approximately 30% of patients are non-responsive and the underlying mechanism for the lack of response is not known. It has been assumed that Bevacizumab solely targets circulating VEGF-A; however, VEGF-A found in the perivascular space of GBM tumors could also be an important target. We hypothesized that Bevacizumab is transcytosed across brain endothelial cells (ECs), gains access to the perivascular niche containing glioma stem cells (GSCs), and that differences in GSC endocytosis and trafficking of Bevacizumab to either a recycling or degradation compartment determines a patients’ response. We found that Bevacizumab is internalized by normal and tumor-isolated brain ECs in a time-dependent manner. ECs transcytose 20% of Bevacizumab after 30 minutes, and 40% after 120 minutes. Importantly, we found that 95% of GSCs internalize Bevacizumab within 30 minutes. Internalized Bevacizumab in GSCs can be recycled to the extracellular space via a Rab4+ fast recycling compartment, or degraded via trafficking to the LAMP1+ compartment (late endosome/lysosome). Furthermore, Bevacizumab internalization in GSCs was partially inhibited by a macropinocytosis inhibitor (EIPA) suggesting macropinocytosis is one mechanism of internalization. GSCs do not internalize Bevacizumab via the canonical IgG internalization pathway since they do not express the neonatal Fc Receptor (FcRn). Supporting our in vitro data, we found a gradient of Bevacizumab extending from the vessel into the tumor in an orthotopic xenograft mouse model of GBM administered Bevacizumab, and Bevacizumab was readily detected within perivascular tumor cells. Our data suggest that the transcytosis of Bevacizumab by brain and tumor-associated ECs, and the endocytosis and trafficking of Bevacizumab by perivascular GSCs influences Bevacizumab effectiveness in patients with GBM.

Citation Format: Gaelle Muller-Greven, Cathleen Carlin, Justin Lathia, Richard Prayson, Manmeet Ahluwalia, Steven Toms, Markus Bredel, Jeremy Rich, Petra Hamerlik, Candece L. Gladson. Glioma stem cells internalize perivascular bevacizumab via a non-canonical pathway and target it for recycling or degradation. [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 5226. doi:10.1158/1538-7445.AM2015-5226
Abstract 527: Ectopic expression of truncated IFIT2 at C-terminal tetratricopeptide repeats enhances the migration activity of oral squamous cell carcinoma cells
Authors: Regmi Prabha , Lee Te-Chang
Published: August 2015

Interferon-induced protein with tetratricopeptide repeats 2 (IFIT2) belongs to one of interferon-stimulated gene family that comprise of tetratricopeptide repeats (TPR). TPR repeats are thought to mediate protein-protein interaction. Our previous report suggests that depletion of IFIT2 enhances cell migration and metastatic activity in oral squamous cell carcinoma (OSCC) cells via activation of atypical PKC signaling. However, the exact mechanism by which IFIT2 depletion activates the aPKC signaling pathway also remains unknown. Since IFIT2 comprises of nine TPR repeats, this study aims to explore the role of TPR in functioning of IFIT2. To address the above questions, different stable cell lines of truncated forms of IFIT2 were subjected to cell migration assay. Interestingly, C-terminal depleted cells exhibited increased migration ability and also showed loss of cell polarity thus leading to random cell orientation. Furthermore, these depleted cells also showed altered expression of junction proteins such as Claudin-1 and E-cadherin in comparison to IFIT2 overexpressed cells. Taken together, C-terminal depleted cells exhibit features of loss of function of IFIT2 suggesting the possible role of terminal TPR domain in maintaining the functional role of IFIT2. To identify specific TPR domain, different mutation construct are generated and its functional role is under study.

Citation Format: Prabha Regmi, Te-Chang Lee. Ectopic expression of truncated IFIT2 at C-terminal tetratricopeptide repeats enhances the migration activity of oral squamous cell carcinoma 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 527. doi:10.1158/1538-7445.AM2015-527
Abstract 5278: Mutation in protein tyrosine phosphatase receptor type K (PTPRK) enhances progression of colon cancer through WNT signaling
Authors: Tojo Masayuki , Konishi Kazuo , Shinjo Keiko , Ohka Fumiharu , Katushima Keisuke , Hatanaka Akira , Ichimura Norihisa , Nozawa Hisako , Shimazaki Tomoe , Yoshida Hitoshi , Kondo Yutaka
Published: August 2015

Recent comprehensive genome analyses revealed the accumulation of genetic alterations in colorectal cancers (CRCs). However, it is still unknown which genetic alterations are closely associated with certain phenotype of CRCs, such as tumors with highly invasive character regardless of its small size. Here we examined the small invasive CRCs using high-throughput sequencing in order to clarify the common molecular features in those CRCs.

Seven small invasive CRCs (types 0-IIa+IIc or 0-Is+IIc, less than 20mm in diameter and with submucosal massive invasion) were examined by exon sequencing. Seven thousands six variants (SNVs and indels) were detected. Among them, PTPRK was frequently mutated (3 cases, 43%; 1 nonsense and 2 missense mutations). We further examined public database (TCGA and COSMIC) and found that 50 of 617 (8.17%) CRCs contain PTPRK mutation. Those cases are frequent in proximal colon, extensively methylated (CIMP-H), microsatellite instability-high (MSI-H) and pathologically mucinous type. Consistent with clinical behavior, invasion assay in CRC cell lines revealed that CRCs with PTPRK domain D1 mutation showed high invasiveness via WNT signaling activity. These data indicate that PTPRK domain D1 mutation contributes to the invasiveness and progression of proximal CRCs, especially CIMP-H and MSI-H cases.

Citation Format: Masayuki Tojo, Kazuo Konishi, Keiko Shinjo, Fumiharu Ohka, Keisuke Katushima, Akira Hatanaka, Norihisa Ichimura, Hisako Nozawa, Tomoe Shimazaki, Hitoshi Yoshida, Yutaka Kondo. Mutation in protein tyrosine phosphatase receptor type K (PTPRK) enhances progression of colon cancer through WNT signaling. [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 5278. doi:10.1158/1538-7445.AM2015-5278
Abstract 5296: FOXP1 expression is associated with tumor progression and poor prognosis in cervical cancer
Authors: Cho Hanbyoul , Yang Woo Kyeom , Kim Sol , Shin Ha-Yeon , Lee Eun Ju , Kim Jae-Hoon
Published: August 2015

Background: The forkhead box protein 1 (FOXP1) is considered as both a tumor suppressor candidate and a potential oncogene. Here, we investigated FOXP1 expression in cervical cancer, and the clinical significance of FOXP1 and it's mechanism of action in cervical cancer.

Methods: To investigate the potential correlation between FOXP1 and various clinicopathologic parameters, we assessed the expression of FOXP1 in archival tumor tissue specimens from 158 patients with cervical cancer and 280 with cervical intraepithelial neoplasia as well as 378 matched nonadjacent normal tissues by immunohistochemical (IHC) staining. Subsequently, we studied the FOXP1's functional role by small interfering RNA (siRNA) approaches.

Results: IHC staining demonstrated that FOXP1 expression increased during the normal to tumor transition of cervical carcinoma (P<0.001), and this increased expression was significantly associated with tumor stage (P = 0.009) and tumor grade (P<0.001). In multivariate analysis, FOXP1+ (P = 0.031) and tumor stage (P = 0.032) were independent prognostic factors for overall survival. FOXP1 knockdown by siRNA decreased cell proliferation, migration, and tumorigenesis in HeLa and CaSki cells.

Conclusions: Taken together, our data indicate that FOXP1 has a crucial role in cervical cancer progression, and its overexpression is associated with poor prognosis, supporting that FOXP1 may be used as a promising novel target for therapeutic interventions.

Citation Format: Hanbyoul Cho, Woo Kyeom Yang, Sol Kim, Ha-Yeon Shin, Eun Ju Lee, Jae-Hoon Kim. FOXP1 expression is associated with tumor progression and poor prognosis in cervical 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 5296. doi:10.1158/1538-7445.AM2015-5296
Abstract 5297: Chromosomal instability as a prognostic marker in cervical cancer
Authors: How Christine , Bruce Jeff , So Jonathan , Pintilie Melania , Haibe-Kains Benjamin , Hui Angela , Clarke Blaise , Hedley David , Hill Richard , Milosevic Michael , Fyles Anthony , Yip Kenneth W. , Liu Fei-Fei
Published: August 2015

Previous studies have demonstrated that chromosomal instability (CIN) is a consistent feature of the majority of solid tumors. In this current study, we sought to examine the published CIN70 gene signature in a cohort of cervical cancer patients treated at the Princess Margaret (PM) Cancer Centre (n = 79) and an independent cohort of The Cancer Genome Atlas (TCGA) cervical cancer patients (n = 130). Patients with a high CIN70 score had a higher number of copy number alterations (Spearman's correlation coefficient (r) = 0.28, p<0.001), and a higher percentage of genome altered (r = 0.19, p = 0.016). According to Kaplan-Meier analysis, the CIN70 signature achieved borderline significance for para-aortic nodal or distant relapse, with a hazard ratio of 3.02 and Wald p-value of 0.05, but not significant for overall, disease-free survival, or local relapse. In summary, these findings demonstrate that chromosomal instability plays an important role in cervical cancer, and is significantly associated with patient outcome. For the first time, this CIN70 gene signature provided prognostic value for patients with cervical cancer.

Citation Format: Christine How, Jeff Bruce, Jonathan So, Melania Pintilie, Benjamin Haibe-Kains, Angela Hui, Blaise Clarke, David Hedley, Richard Hill, Michael Milosevic, Anthony Fyles, Kenneth W. Yip, Fei-Fei Liu. Chromosomal instability as a prognostic marker in cervical 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 5297. doi:10.1158/1538-7445.AM2015-5297
Abstract 5301: Integrative protein-marker prognostic model for early-stage endometrioid endometrial carcinoma
Authors: Liang Han ,   UT MD Anderson RPPA/TCGA working group
Published: August 2015

Endometrioid endometrial carcinoma (EEC) is the major histological type of endometrial cancer, the most prevalent gynecologic malignancy in USA. EEC recurrence or metastasis is associated with an abysmal prognosis. Early-stage EEC is generally curable, but a subset has high risk of recurrence or metastasis. Prognosis estimation for early-stage EEC mainly relies on clinicopathological characteristics, but is unreliable. We aimed to identify patients with high-risk early-stage EEC who are most likely to benefit from more extensive surgery and adjuvant therapy by building a prognostic model that integrates clinical variables and protein markers. We employed two large, independent EEC datasets as training and validation cohorts, and generated the levels of 186 proteins and phosphoproteins using reverse-phase protein arrays. Using the training samples, we developed a predictive model containing two clinical factors and 18 protein markers and optimized the risk group classification. Kaplan-Meier survival analyses in the validation cohort confirmed an improved prognostic power. Compared with clinical variables (stage, grade, and patient age), only the risk groups defined by the integrative model were consistently significant in both univariate and multivariate analyses across both cohorts. Our prognostic model is potentially of high clinical value for stratifying patients with early-stage EEC and optimizing their treatment strategies.

Note: This abstract was not presented at the meeting.

Citation Format: Han Liang, UT MD Anderson RPPA/TCGA working group. Integrative protein-marker prognostic model for early-stage endometrioid endometrial 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 5301. doi:10.1158/1538-7445.AM2015-5301
Abstract 5307: Analysis of telomere length in classic and variant hairy cell leukemia
Authors: Edelman Daniel C. , Arons Evgeny , Stevenson Holly , Gomez Allison , Petersen David , Zhou Hong , Wang Yonghong , Waterfall Joshua J. , Meltzer Paul S. , Kreitman Robert
Published: August 2015

Telomeres are ribonucleoprotein structures which protect the ends of chromosomes from aberrant recombination events and have been reported to be relatively short in several hematologic malignancies; adverse prognosis has been documented in chronic lymphocytic leukemia and several non-B-cell malignancies. However, their association with prognosis of classic and variant hairy cell leukemia (HCL and HCLv) has not been described. To study telomeres in HCL and HCLv, DNA from leukemic cells of patients was measured by monochrome multiplex quantitative PCR (MMQPCR) to determine relative telomere length (RTL). Of 46 patients in the cohort, 5 of 27 HCL (19%) vs. 12 of 19 HCLv (63%) patients had unmutated (>98% homology to germline sequence) immunoglobulin rearrangements (p = 0.0045). IGHV4-34+ rearrangements were present in 12 of the 17 (70%) unmutated cases, including five HCL and seven HCLv. RTL was shown to be shorter in unmutated HCL (p = 0.03) and in patients who had died of their disease (p = 0.03). RTL negatively correlated with IGHV homology to germline sequence (r = -0.331; p = 0.025), and age (r = -0.315; p = 0.035). Shorter RTL was associated with death from disease in either HCL (p = 0.0063) or HCLv (p = 0.0034), in patients with mutated IGHV rearrangements (p = 0.005), or in all patients combined (p = 0.0008). Median survival from diagnosis was less in time in patients with shorter RTL (p = 0.016) and this significant difference persisted when examining just the 19 patients with HCLv (p = 0.029). Gene expression profiling identified several genes related to telomere maintenance with significantly different expression levels between mutated and unmutated cases. This first of kind study in HCL/HCLv suggests that telomeres play a role in the relative pathology of HCL and HCLv and RTL could be used as a prognostic biomarker in gauging the indolent course of these cancers.

Citation Format: Daniel C. Edelman, Evgeny Arons, Holly Stevenson, Allison Gomez, David Petersen, Hong Zhou, Yonghong Wang, Joshua J. Waterfall, Paul S. Meltzer, Robert Kreitman. Analysis of telomere length in classic and variant hairy cell leukemia. [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 5307. doi:10.1158/1538-7445.AM2015-5307
Abstract 5320: Targeting HER3 and EGFR in NRG1 positive and HER3 mutated lung squamous cell carcinoma
Authors: Pinzon-Ortiz Maria C. , Rong Xianhui , Versace Richard , Sheng Qing , Cao Z. Alexander
Published: August 2015

HER3 is a member of the ErbB family of receptor tyrosine kinases. Aberrant activation of HER3 as a result of HER2 amplification or neuregulin 1 (NRG1) over-expression has been demonstrated to mediate constitutive activation of downstream oncogenic signals. LJM716 is a fully human IgG1 anti-HER3 monoclonal antibody. It locks HER3 in an inactive conformation and prevents HER3 dimerization with other ErbB family members. This unique mode of action enables LJM716 to block both ligand dependent and ligand independent HER3 activation. In search of new indications that may benefit from anti-HER3 therapies, we noted that squamous cell carcinomas (SCC) tend to have relatively higher NRG1 expression as compared to their adenocarcinoma counterparts. Highest NRG1 expression was observed in the lung SCC cell lines in cell line encyclopedia (CLE). A subset of the lung SCC cell lines with high NRG1 expression are moderately sensitive to LJM716, with HER3 activation detected in all of the sensitive lines. Additionally, EGFR activation is consistently observed in our panel of squamous lung carcinoma models. In some cases, activation of HER2 and/or HER3 is also noted. We hypothesize that squamous lung carcinomas may be dependent on signaling through EGFR, HER2 and HER3. To completely block EGFR, HER2-HER3 signaling, we set to test the treatment of LJM716 and Cetixumab in a number of NGR 1 positive squamous lung carcinomas tumor models. Enhanced anti tumor activity of LJM716 and Cetixumab is observed in four out nine tumor models. Analysis of broad activation status of receptor tyrosine kinases (RTK) reveals that a subset of responsive models have activated pEGFR, pHER2 and pHER3. These data suggest that activated HER3 signaling may be important in a subset of lung squamous cell carcinoma. Combination of anti-HER3 and anti EGFR treatment is effective at inhibiting growth in these tumors.

Note: This abstract was not presented at the meeting.

Citation Format: Maria C. Pinzon-Ortiz, Xianhui Rong, Richard Versace, Qing Sheng, Z. Alexander Cao. Targeting HER3 and EGFR in NRG1 positive and HER3 mutated lung squamous cell 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 5320. doi:10.1158/1538-7445.AM2015-5320