bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5073: C5a receptor promotes invasive ability of gastric cancer
Authors: Kaida Takayoshi , Nitta Hidetoshi , Kitano Yuki , Yamamura Kensuke , Inoue Risa , Arima Kota , Higashi Takaaki , Taki Katunobu , Hayashi Hiromitsu , Imai Katsunori , Hashimoto Daisuke , Chikamoto Akira , Beppu Toru , Baba Hide
Published: July 2016

Background: C5a is one of the important factors produced via the complement pathway and a strong chemokine. The C5a receptor (C5aR) is expressed in various cancer cells and is linked to C5a-induced promotion of cancer cell invasion. However, the role of C5aR in gastric cancer (GC) is still mostly unknown.

Aim: To investigate the clinical role of C5aR-expression in human GC.

Method: 1) We investigated the C5aR expression for 237 GC patients who underwent gastrectomy from Jan. 2004 to Dec. 2011 by immunostaining using monoclonal human C5aR-antibody and analyzed the association between C5aR expression and clinicopathological factors. 2) We investigated the association between C5aR expression and invasiveness of GC cells, in vitro.

Result: 1) From immunostaining analysis, High-expression of C5aR was observed in 86 patients (36.8%) and was significantly related to depth of invasion, stage and vascular invasion. The 5-years RFS and OS rates of the patients with high-expression of C5aR were significantly worse than those of the patients with low-expression. 2) We found that several GC cell lines had high-expression of C5aR with western blotting. By using AGS and NUGC3 cell, the cell line which had low-expression of C5aR, We produced AGS (AGS/C5aR) and NUGC3 (NUGC3/C5aR) which had the overexpression of C5aR. Matrigel chamber assay revealed that C5a recombinant enhanced the cell invasion of AGS/C5aR and NUGC3/C5aR cell. Migration assay by using the timelapse imaging revealed that C5a recombinant enhanced the cell motility of AGS/C5aR and NUGC/C5aR cell. In addition, C5aR-antagont decreased the cell invasion and motility of those cell lines. The immunofluorescence microscopy revealed that C5a recombinant enhanced the cytoskeletal rearrangement, such as stress fiber, fillipdia and lamellipodia, in NUGC/C5aR cell. Then, C5a recombinant enhanced RhoA activity of NUGC3/C5aR.

Conclusions: The stimulation of C5a receptor may enhance the cell motility via RhoA activity, then it may enhance the cell invasion ability. In addition, High-expression of C5aR was associated with poor prognosis for GC patients. Therefore, C5a receptor might be a target of medical treatment.

Citation Format: Takayoshi Kaida, Hidetoshi Nitta, Yuki Kitano, Kensuke Yamamura, Risa Inoue, Kota Arima, Takaaki Higashi, Katunobu Taki, Hiromitsu Hayashi, Katsunori Imai, Daisuke Hashimoto, Akira Chikamoto, Toru Beppu, Hide Baba. C5a receptor promotes invasive ability of gastric cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5073.
Abstract 5077: The insulin-like growth factor-2 (IGF-2): A potential therapeutic target for triple-negative breast cancer metastasis
Authors: Hamilton Nalo , Marquez-Garban Diana , Pietras Richard
Published: July 2016

Introduction: Triple-negative breast cancers (TNBC) occur in 15-17% of patients, yet this subtype accounts for almost half of all breast cancer (BC) deaths. This breast cancer sub-type often occur in younger and African American (AA) women. TNBCs cannot be treated with current chemotherapies. Although initially responsive to chemotherapy, TNBCs tend to relapse and metastasize early. As there are no specific treatments for TNBCs, we aim to identify the therapeutic potential of insulin-like growth factor- 2 (IGF-2) in TNBC management.

Methods: Using immunohistochemistry archival breast tissue from women with metastatic TNBC was evaluated and scored for IGF-2 expression. To assess the ability of IGF-2 to stimulate cell proliferation and migration we used commercially available TNBC cell lines validated to be ER-alpha-negative (ER-), progesterone receptor-negative (PR-) and Her2 overexperssion-negative (HER2).

Results: We have found IGF-2 to be highly expressed archival breast tissue of women with metastatic TNBC. Using TNBC cell lines MDA-MB-231, HCC 38, HCC 1806 and HCC 1937, our in vitro studies demonstrate the ability of IGF-2 to stimulate TNBC proliferation and migration (P = 0.05). Further, the combination of NVP-AEW541, an insulin-like growth factor-1 receptor (IGF-1R) inhibitor, and BMS-754807, an inhibitor of IGF-1R and the insulin receptor (IR), cause a significant (P < 0.05) reduction in TNBC proliferation and migration. Investigation into the cellular characteristics of the TNBC cell lines used, suggest that BRCA1 mutations increase cell susceptibility to IGF-1R inhibitors.

Conclusion: Our data indicate that IGF-2 plays a role in TNBC metastasis, an action that can be inhibited by targeting multiple IGF-2 mediated receptors.

Citation Format: Nalo Hamilton, Diana Marquez-Garban, Richard Pietras. The insulin-like growth factor-2 (IGF-2): A potential therapeutic target for triple-negative breast cancer metastasis. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5077.
Abstract 5081: hnRNP A18: an emerging novel target for cancer therapy
Authors: Parekh Palak R. , Chang Elizabeth , Yang Qingyuan , Carrier France
Published: July 2016

Heterogenous ribonucleoprotein A18 (hnRNP A18) increases de novo protein translation during cellular stress. Although hnRNP A18 has recently been associated with several cancers, the exact role of it during the multifactorial process of cancer growth and progression is still unknown. Accordingly, we examined the dependability of various malignant cancer cell lines on hnRNP A18. Down-regulation of hnRNP A18 impaired the proliferative, invasive and migratory properties of melanoma and triple negative breast cancer cell lines in vitro, while overexpression of hnRNP 18 caused an increase in invasiveness. Consistent with these observations, we detected remarkable reduction of tumor growth in two mouse xenograft models, melanoma and breast cancer. Moreover, down-regulation of hnRNP A18 increased the sensitivity of melanoma cells and breast cancer cells to paclitaxel and 5-fluoro uracil, respectively. In conclusion, hnRNP A18 plays a key role in tumor growth and progression. It may serve as a novel target for the treatment of cancer.

Citation Format: Palak R. Parekh, Elizabeth Chang, Qingyuan Yang, France Carrier. hnRNP A18: an emerging novel target for cancer therapy. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5081.
Abstract 5088: ACVRIB-dependent regulation of extracellular matrix protein in squamous cell carcinoma progression
Authors: Loomans Holli A. , Quast Laura L. , Andl Claudia A.
Published: July 2016

Squamous cell carcinomas (SCC) of the head and neck and esophagus pose a substantial public health risk globally. As SCC is often diagnosed in late stage, therapeutic options are limited. A more thorough understanding of the mechanisms involved their progression is needed. One potential mode by which SCCs progress is via Activin A signaling. The downstream effects of Activin A signaling have been primarily characterized as growth inhibiting and anti-angiogenic. However, Activin A is commonly overexpressed in SCC. Therefore, we hypothesized that SCC become insensitive to Activin A stimulation through the dysregulation of the signaling pathway. To model this dysregulation we utilized CRISPR-mediated regulation of ACVRIB (ACVRIB-kd), the necessary type IB receptor to transduce Activin A signaling in three SCC cell lines. Downregulation of ACVRIB enhanced invasiveness in the OSC19 SCC three-dimensional organotypic cultures. Additionally, ACVRIB-kd cells showed significant changes in protein expression, when compared to control. In particular, the correlative markers of SCC aggressiveness and putative downstream Activin A targets, podoplanin and fibronectin, were substantially altered in ACVRIB-kd cells. Integrin expression, particularly integrins β3, α5, and αv, was markedly changed in ACVRIB-kd cells, indicating additional ACVRIB-mediated effects to ECM proteins. We are currently analyzing ACVRIB-kd SCC tumor growth and metastasis in vivo using tongue orthotopic xenografts and subcutaneous flank injections. In conclusion, SCC-specific disruption of Activin A signaling promotes an aggressive phenotype. Re-activation of the canonical Activin A pathway may provide a novel therapeutic strategy to combat SCC.

Citation Format: Holli A. Loomans, Laura L. Quast, Claudia A. Andl. ACVRIB-dependent regulation of extracellular matrix protein in squamous cell carcinoma progression. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5088.
Abstract 5094: Acid-induced collagen remodeling promotes cancer progress as a result of niche engineering competition between cancer and stroma cells
Authors: Damaghi Mehdi , Chen Tinjan , Johnson Josef , Gillies Robert J.
Published: July 2016

Early carcinogenesis is an avascular and hypoxic disease that makes tumor cells export metabolically derived acid through glycolysis. Even in presence of oxygen cancer cells keep the glycolytic phenotype (Warburg effect) that acidifies the environment more and constantly. Hence the physical microenvironment of solid tumors is profoundly acidic. Pre-malignant cells within this niche must adapt to acidosis in order to survive this hostile microenvironment. We think pre-malignant cells adapt to acidosis to not only survive in this hostile microenvironment but also to acquire an advantage that helps them to outcompete the others and eventually help them to invade. Recently many evidences have been provided showing the collaboration of tumor associated stroma cells such as Fibroblasts (TAF) with tumor cells in cancer progress. Here we also show how these cells use acidification and acid adaptation as a strategy to construct and engineer their niche in their own favor for growth and proliferation. In this study we focused on collagen as major proteins of extra cellular matrix (ECM) and even arguably the most dominant one. The unique mechanical properties of fibrillar collagen are mainly controlled by its structure and any change in it can lead to ECM induced tumor growth. ECM unique compositions and topographies are generated through a dynamic biochemical and biophysical interplay between the various cells and the evolving microenvironment. In this study we developed the window chamber and intra vital microscopy to study the collagen remodeling lively. Later we used 3D cell culture and co-culture to validate our findings. Finally we used proteomics to understand molecular mechanism behind the changes in acid adapted versus non-adapted cancer cells and TAfs. We found cancer cells take different strategy than TAFs, although both changes promote cancer progression.

In conclusion, there are evolutionary advantages behind acid production and adapting to it for cancer and cancer associated cells. We discuss how acquiring these advantages can helps us to design new therapeutics against these unique phenotypes.

Citation Format: Mehdi Damaghi, Tinjan Chen, Josef Johnson, Robert J. Gillies. Acid-induced collagen remodeling promotes cancer progress as a result of niche engineering competition between cancer and stroma cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5094.
Abstract 5100: Interplay between ECM stiffness, miR-203 and mammographic density
Authors: Dean Ivory , Acerbi Irene , Au Alfred , Chen Yunn-Yi , Hwang Shelley , Weaver Valerie
Published: July 2016

High mammographic density (MD) is associated with a greater risk of breast cancer and is characterized by high extracellular matrix (ECM) collagen. Elevated collagen increases ECM stiffness and can promote the malignant transformation of oncogenically-primed mammary epithelial cells in culture and in vivo. Whether or not high MD reflects a breast stroma that is stiff and if ECM stiffness accounts for the elevated breast cancer risk in high MD tissue has yet to be resolved. To address this question we are conducting a comprehensive biophysical and molecular analysis of BIRADS status in prophylactic mastectomies in women with low (BIRADS 1) versus high MD (BIRADS 4). Data revealed that the intra-lobular ECM associated with the terminal end-buds contains anisotropic compliant and relaxed collagen fibrils. By contrast, the inter-lobular ECM contains stiff, oriented collagen fibrils. Preliminary data suggest that the ECM associated with the terminal end-buds in the upper outer quadrant may be stiffer in women with high MD as compared to low MD. These same tissues also expressed lower levels of the microRNA miR-203, which has been implicated in epithelial-to-mesenchymal transition (EMT) inhibition. Consistently, in vitro studies revealed that miR-203 was repressed in MECs cultured on a stiff ECM. We also found that a stiffened mammary ECM decreased miR-203 expression and promoted an EMT. Additionally, inhibiting ECM stiffening restored miR-203 levels and reduced an EMT. Moreover and importantly, we showed that triple negative breast tumors that often express EMT markers have reduced miR-203 levels. Studies are underway to elaborate these preliminary findings and to assess their clinical relevance. (This work is supported by the: NIH NSRA T32 CA 108462-11 to Ivory Dean, Susan G Komen PDF12230246 to Irene Acerbi, W81XWH-13-1-0216, USMRAA DOD-BCRP BC122990, NIH R01 CA138818-01A1 and NIH R01 CA192914-01 to Valerie Weaver.)

Citation Format: Ivory Dean, Irene Acerbi, Alfred Au, Yunn-Yi Chen, Shelley Hwang, Valerie Weaver. Interplay between ECM stiffness, miR-203 and mammographic density. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5100.
Abstract 5101: Tumor-secreted acids alter macrophage phenotype
Authors: El-Kenawi Asmaa , Ibrahim-Hashim Arig , Luddy Kimberly , Ruffell Brian , Pilon-Thomas Shari , Gatenby Robert , Gillies Robert
Published: July 2016

Within tumors, tumor-associated macrophages (TAMs) usually adopt an M2 phenotype with tumor-promoting characteristics, as opposed to the M1 phenotype responsible for host protection. As acidic pH can affect the characteristics of immune cells, we aimed to test whether low pH as a common stress factor in the tumor microenvironment could be partially responsible for the phenotypic characteristics and/or functional activity of macrophages in prostate cancer. To test this hypothesis, we allowed TRAMP (TRansgenic Adenocarcinoma of the Mouse Prostate) mice to develop neoplastic lesions of variable histologic severity. Thereafter, we isolated tumor-associated macrophages from control tumors and analyzed their expression of the M1 and M2 markers; iNos, Il-6, Cd206 and Arg1. Gene expression analysis confirms previous studies which reported higher expression of both M1 and M2 markers in TAMs compared with peritoneal macrophages. Treatment of TRAMP mice with sodium bicarbonate to continuously buffer acids secreted by neoplastic cells, decreased TAMs infiltration into the stromal compartment of prostate. In addition, sodium bicarbonate decreased iNos, Arg1, and Cd206 but not Il-6 expression in TAMs isolated from subcutaneously growing TRAMP-C2 tumors, a TRAMP-derived prostate tumor cell line injected into syngeneic male hosts. Since decreased expression of Arg1 and iNos has been shown to mediate the suppressive activity of myeloid cells, tumor acidity might promote the suppressive microenvironment by upregulating both enzymes. In conclusion, extracellular acidosis increases the density of macrophages and their L-arginine metabolic potential; coincident with delayed tumor progression in prostate tumors.

Citation Format: Asmaa El-Kenawi, Arig Ibrahim-Hashim, Kimberly Luddy, Brian Ruffell, Shari Pilon-Thomas, Robert Gatenby, Robert Gillies. Tumor-secreted acids alter macrophage phenotype. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5101.
Abstract 5110: The role of autophagy in the tumor microenvironment
Authors: Dower Christopher M. , Wang Hong-Gang
Published: July 2016

The tumor microenvironment (TME) is a complex and dynamic breeding ground for the selection of aggressive tumor cells with an advantage of survival and metastatic potential. Autophagy has emerged as a powerful cellular process that can greatly influence how cancer cells respond to nutrient deprivation and hypoxic stress, the hallmarks of the TME. However, the physiological importance of hypoxia-regulated autophagy remains unknown. Here, we used a novel hypoxia-dependent reversible dominant-negative strategy, which allowed for physiological regulation of autophagy at the cellular level during tumor growth and metastasis and creates a heterogeneous population comprised of autophagy-competent and autophagy-deficient tumor cells. We demonstrate that the suppression of autophagy by HIF1-α dependent ULK1 inhibition within the hypoxic tumor microenvironment enhances the metastatic potential of breast cancer cells both in vitro and in vivo. Functional proteomic and transcriptome analysis indicated that the loss of hypoxia-regulated autophagy promotes metastasis through extracellular remodeling pathways, primarily through induction of the fibronectin-integrin signaling axis, along with LOX-mediated extracellular remodeling and pre-metastatic niche formation. In this way, the role of physiological hypoxia regulated autophagy was assessed for the first time in a triple negative breast cancer model. Currently, we are exploring the interplay between autophagy-competent and autophagy-deficient tumor cells within the TME, and how it relates to metastasis. Although the importance of autophagy in tumor-stromal interactions has recently become appreciated, the role autophagy plays in tumor cell-tumor cell interactions remains unknown. Through investigation of these intra-tumor cell networks, we can elucidate the precise mechanism of how physiologically-regulated autophagy in tumor cells influences the surrounding TME and tumor progression.

Citation Format: Christopher M. Dower, Hong-Gang Wang. The role of autophagy in the tumor microenvironment. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5110.
Abstract 5115: Pre-clinical development of next generation inhibitors of the enzymes indoleamine 2,3-dioxygenase 1 and tryptophan 2,3-dioxygenase as cancer immunotherapies
Authors: Wise Alan , McGuinness Barry E. , Trewick Sarah C. , Brown Thomas J. , Cowley Phillip M.
Published: July 2016

The enzymes tryptophan 2,3-dioxygenase (TDO) and indoleamine 2,3-dioxygenase (IDO1) catalyse oxidation of the essential amino acid tryptophan (Trp) leading to the formation of immunosuppressive kynurenine (Kyn) pathway metabolites that dampen the immune response in the tumor microenvironment. Both IDO1 and TDO have been shown to be up-regulated in a variety of cancers and blockade of their activity has been shown to stimulate the anti-tumour immune response in pre-clinical animal models. We have discovered and optimized multiple novel chemical series of both highly selective and dual-acting inhibitors of IDO1 and TDO. We describe the characterization of molecules that we are progressing into the clinic.

We have characterized the IDO1 and TDO-selective inhibitors and dual-acting inhibitors which demonstrate nM potencies. In IFNγ-stimulated A172 glioblastoma cells expressing both enzymes, the dual-acting molecules fully inhibit Kyn production, whilst selective inhibitors yield only a partial response, demonstrating the utility of dual enzyme blockade in cancer cells. The compounds also relieve inhibition of T-cell proliferation in cancer cell conditioned media/T-cell co-culture assays and demonstrate highly favourable physico-chemical and pharmacokinetic properties. These properties translate to superior PK/PD effects following oral dosing in rodents with profound and sustained modulation of plasma Kyn levels, combined with an exceptional safety profile. In a mouse model of LPS-induced IDO1 activation, our IDO1 inhibitors fully ablate elevation of Kyn in both plasma and lung in a dose-dependent manner. Following oral administration, the IOmet IDO1 inhibitors provide excellent single-agent tumor growth control in PAN02 pancreatic adenocarcinoma syngeneic models. In addition, combining IOmet IDO1 inhibitors with standard-of-care chemotherapy (gemcitabine/abraxane) in the PAN02 model completely inhibits tumor growth.

In summary, we describe the characterization of novel potent, selective and dual-acting IDO1 and TDO inhibitors. These candidate molecules demonstrate excellent rodent in vivo PK/PD, safety and anti-tumor profiles, supporting their progression into the clinic.

Citation Format: Alan Wise, Barry E. McGuinness, Sarah C. Trewick, Thomas J. Brown, Phillip M. Cowley. Pre-clinical development of next generation inhibitors of the enzymes indoleamine 2,3-dioxygenase 1 and tryptophan 2,3-dioxygenase as cancer immunotherapies. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5115.
Abstract 5129: Delineating immune networks in colorectal cancers to predict effects of immune checkpoint inhibitors using CANScript platform technology preserving tumor immune microenvironment
Authors: Majumder Pradip K. , Brijwani Nilesh , Karandikar Nikita N. , Radhakrishna Vinod D. , Oliyarasi Muthusamy , Pinto Dency D. , Balakrishnan Babu , Sekar Vasathakumar , Rajappa Manoj , Mehrotra Debapriya G. , Chevur Priyanka , Chevur Priyanka , Ghosh Arkasubhra , Thyiagarajan Saravanan , Radhakrishnan Padhma , Majumder Biswanath
Published: July 2016

Colorectal cancer (CRC) is one of the leading causes of cancer related mortality. Recent findings from clinical studies showed that immune checkpoint inhibitors are making good progress towards clinical practise. However, existing biomarkers and 3D platforms are not equipped to offer reliable prediction of response of these drugs. We recently engineered a personalized ex vivo systems biology based platform using patient tumors called CANScriptTM (Majumder B et al Nat. Commun. 2015, Goldman A et al Nat. Commun. 2015) and a companion in vivo patient derived immune reconstituted xenograft model (Mi-HTX) preserving multiple critical phenotypic profiles orchestrating tumor-immune cross talk and dysfunction. We further performed baseline characterization of immune markers (CD4, CD8, CD68, CXCR4 and CD45-RO). Intratumor heterogeneity of both M1 and M2 phenotypes were observed irrespective of clinico-pathological features. Baseline CD68 enrichment status was found to be correlated with therapy failure for some patients where clinical outcome (PERCIST) was available. Flow cytometry and microarray analysis showed preservation of diverse sets of functional CD4, CD8 and NK phenotypes along with degranulation markers (CD107a), IFN-γ and FoxP3 in both in vivo and ex vivo tumors reconstituted with ex vivo primed PBMC. The comparative response profiling of these models following treatment with immune checkpoint inhibitors suggested enhanced antitumor effects (as measured by multiple functional parameters like viability, proliferation, morphology and apoptosis). Together, these findings highlight the strengths of immune-competent functional testing tools to stratify patients for precision immunotherapy where direct response prediction biomarkers are still elusive.

1. Majumder B et al. Nat. Commun.6:6169 (2015).

2. Goldman A et al. Nat. Commun. 6:6139 (2015).

Citation Format: Pradip K. Majumder, Nilesh Brijwani, Nikita N. Karandikar, Vinod D. Radhakrishna, Muthusamy Oliyarasi, Dency D. Pinto, Babu Balakrishnan, Vasathakumar Sekar, Manoj Rajappa, Debapriya G. Mehrotra, Priyanka Chevur, Priyanka Chevur, Arkasubhra Ghosh, Saravanan Thyiagarajan, Padhma Radhakrishnan, Biswanath Majumder. Delineating immune networks in colorectal cancers to predict effects of immune checkpoint inhibitors using CANScript platform technology preserving tumor immune microenvironment. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5129.
Abstract 5149: A novel regulatory circuit involving STIM1 and HIF-1 mediates hypoxia-driven hepatocarcinogenesis
Authors: Li Yongsheng
Published: July 2016

Hypoxia and intracellular Ca2+ transient are fundamental traits of cancer. Signaling cascades initiated or regulated by HIF-1 is essential for hypoxic responses. Stromalinteraction molecule 1 (STIM1) is well known as a Ca2+ sensor on endoplasmic reticulum (ER) that mediates SOCE activation and promotes tumor invasion and migration. However, there was no general consensus about the relationship between HIF-1 and STIM1. Also, the significance of STIM1, as well as the Ca2+ mobilization route in cancer cells during hypoxia remains unknown. Our results show that STIM1 correlates with elevated hypoxia-inducible factor-1 alpha (HIF-1α) in hypoxic hepatocarcinoma cells (HCC) and is upregulated during hepatocarcinoma growth. HIF-1 directly transcripts STIM1 and contributes to store-operated Ca2+ entry (SOCE), while STIM1-mediated SOCE is also required for HIF-1 accumulation in hypoxic HCC, via activating Ca2+/calmodulin-dependent protein kinase II and p300. Administration of YC-1, a HIF-1 inhibitor, or knockdown of HIF1A significantly diminishes hypoxia-enhanced STIM1 and suppresses tumorigenesis. Moreover, ectopic expression of STIM1 or HIF-1α partially reverses the impaired growth of the tumor treated with YC-1. These results suggest a mutual dependency and regulation of STIM1 and HIF-1 in controlling Ca2+ mobilization and hypoxic tumor growth, and highlight a potential target for early hypoxia-related intervention.

Citation Format: Yongsheng Li. A novel regulatory circuit involving STIM1 and HIF-1 mediates hypoxia-driven hepatocarcinogenesis. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5149.
Abstract 5156: The roles of x-linked Midline 1 interacting protein 1 (MID1IP1) in gender disparity and development of hepatocellular carcinoma
Authors: Chiu Elley Yung Tuen , Ng. Irene Oi Lin
Published: July 2016

HBV-associated hepatocellular carcinoma (HCC) has a male predominance, with a male-to-female ratio of about 5:1, suggesting gender difference is one of the major risk factors for HCC development. However, up till now, no concrete evidence is obtained to elucidate the male predominance of HCC. It is believed that the gender-specific signaling pathway is critically involved and targeting this pathway may provide a significant treatment outcome.

To target candidates that are implicated in the male predominance in HCC, aberrant expression of sex-linked genes between HCC tumors and non-tumorous livers (NT-L) draws particular attention. Previously, we performed transcriptome sequencing on sixteen pairs of human HCC tumors and NT-Ls. Our preliminary data show that a potential target, Midline 1 interacting protein 1 (MID1IP1) which is involved in hepatic lipogenesis and microtubule stabilization, was remarkably overexpressed in HCC tumors as compared to the NT-Ls (fold change = 2.73; p = 0.017). Moreover, the expression of MID1IP1 was significantly higher in male non-tumorous liver tissue than that of female (fold change = 2.63; p = 0.001). In addition, the upregulation of MID1IP1 correlated with less tumor encapsulation, more tumor microsatellite formation, more venous and direct liver invasion, and poorer tumor stage, features of a more aggressive biological behavior in our clinicopathological correlation study. Moreover, upregulation of MID1IP1 in the tumors was associated with a poorer overall survival. In its functional study, upon knockdown of MID1IP1, HCC cells (BEL7402, SMMC7721 and HepG2) had reduced proliferation rates in the cell proliferation assay and colony formation assay. Here, we also showed that the migratory abilities of the HCC cell lines with MID1IP1 knockdown were remarkably reduced in the transwell assay. Furthermore, the STRING interaction network analysis suggests that MID1IP1 interacts with Suppressor of Fused (Sufu), which is a regulator of Gli in the hedgehog (HH) pathway. To examine its regulatory pathway, we showed that the mRNA expression of the negative regulator of HH pathway, Gli3, was increased in the MID1IP1 knockdown HCC cells. The downstream target of Gli, cyclin D, was also downregulated upon MID1IP1 knockdown. In summary, our data suggest that the MID1IP1 plays a significant role in the male predominance of human HCC. MID1IP1 may also play a novel role in the regulation of hedgehog signaling.

Citation Format: Elley Yung Tuen Chiu, Irene Oi Lin Ng. The roles of x-linked Midline 1 interacting protein 1 (MID1IP1) in gender disparity and development of hepatocellular carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5156.
Abstract 5160: Establishment of in vitro carcinogenic model of high-grade serous ovarian carcinoma using immortalized fallopian tubal secretory epithelial cell
Authors: Nakamura Kohei , Nakayama Kentaro , Kiyono Tohru , Ishikawa Noriyoshi , Ishikawa Masako , Katagiri Hiroshi , Minamoto Toshiko , Ishibashi Tomoka , Sato Emi , Sanuki Kaori , Yamashita Hitomi , Iida Kouji , Sultana Razia , Kyo Satoru
Published: July 2016

High-grade serous ovarian carcinoma (HGSOC) is the most lethal gynecologic malignancy in the world. Recent studies suggest that fallopian tubal secretory epithelial cells (FTSECs) are candidates for the origin of HGSOC. Several genetic alterations involved in the carcinogenesis have been reported in HGSOC, but their minimal requirement and the specific combinations remained unclear. To clarify these points, we established an in vitro stepwise carcinogenesis model using immortalized FTSECs with overexpressed cyclinD1/cdk4 and hTERT, in which we additionally mimicked selected genetic abnormalities frequently observed in clinical samples. Mutational analysis using clinical samples of HGSOCs identified frequent p53 mutations (96%). Furthermore, RAS/PI3K pathway is important signaling pathway and was deregulated in 45% of HGSOCs. PI3K/AKT pathway is also important pathway in HGSOCs. Thus, we introduced dominant negative form of p53 alone or in combination with oncogenic mutant KRAS allele into immortalized cells, but both failed to exhibit tumorigenic phenotypes. Additional introduction of genetic factors were attempted, and overexpression of c-Myc or phosphorylated Akt (p-Akt) were independently found to confer tumorigenic phenotypes. Importantly, all transformed FTSECs exhibited high-grade serous carcinomas that were grossly, histologically, and immunohistochemically similar to human HGSOCs. Thus, p53/KRAS/c-Myc or p53/KRAS/PI3K-AKT pathways were determined to be minimal required for carcinogenesis. The strength of our study is the use of immortalized FTSECs that have original characteristics of fallopian tube as well as the introduction of genetic mutations clinically observed in human HGSOCs. This experimental model provides a foundation for future studies of early events in carcinogenesis of HGSOCs and the identification of candidate targets for drug development and secreted biomarkers for early detection. It is also likely that a similar model can be broadly applied to studying the normal biology of other epithelial cells where precursor lesions and malignant counterparts are not entirely understood.

Citation Format: Kohei Nakamura, Kentaro Nakayama, Tohru Kiyono, Noriyoshi Ishikawa, Masako Ishikawa, Hiroshi Katagiri, Toshiko Minamoto, Tomoka Ishibashi, Emi Sato, Kaori Sanuki, Hitomi Yamashita, Kouji Iida, Razia Sultana, Satoru Kyo. Establishment of in vitro carcinogenic model of high-grade serous ovarian carcinoma using immortalized fallopian tubal secretory epithelial cell. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5160.
Abstract 5163: Mitochondrial mutations and gene expression analysis in colorectal adenopolyps
Authors: Wallace LaShanale M. , Mehrabi Sharifeh , Yao Xuebiao , Aikhionbare Felix
Published: July 2016

Colorectal cancer (CRC) is the third most common diagnosed and cause of cancer related deaths in both men and women in the United States. Numerous studies have analyzed mitochondria DNA mutations in CRC and other tumors. Results from these studies have detected high mutation rates which may lead to mitochondrial deregulation and tumor progression. Most CRCs develop from adenopolyps via the adenoma-carcinoma sequence. Therefore, analysis of mitochondrial mutations and gene expression may provide a mechanism for inhibition of this tumoral sequence in individuals with a high risk of developing CRC. In the present study, PCR based sequencing and reverse transcription-quantitative PCR (RT-qPCR) were used to determine if mutations in mitochondrial encoded genes and levels of expression of these genes could influence the progression of the adenoma- carcinoma tumoral sequence. Genes analyzed included MT-RNR1, MT-COI, MT-ATP6, MT- MT-CYB, and mitochondrial ND genes that are involved in the normal metabolism of mitochondria. Measurements were made for 34 tissue sample pairs obtained from various types of colorectal adenomas and corresponding normal surrounding tissues. Additionally, mitochondrial complexes I (NADH: ubiquinone oxidoreductase) and III (CoQH2-cytochrome c reductase) protein was analyzed. There was progressive differential expression of mt-genes and complexes I and III proteins among the colorectal tumor stages relative to their paired normal samples. The level of complexes I and III were higher in tumor tissues relative to normal surrounding tissues. Noticeably, the expression of MT-COI was higher in late stage carcinomas among; all studied transcripts. We detected 54 point mutations in one region ranging from 11871-11877. The frequency of these mutations in all stages was as followed; 71.5% in tubular adenoma, 57% tubulovillous, and 43% in villous and carcinoma. Our results suggest that alterations in mt-gene expression play a role in the transformation of the colorectal tumoral stages.

Citation Format: LaShanale M. Wallace, Sharifeh Mehrabi, Xuebiao Yao, Felix Aikhionbare. Mitochondrial mutations and gene expression analysis in colorectal adenopolyps. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5163.
Abstract 5164: Protein kinase C epsilon is a mediator of KRAS-driven lung tumorigenesis
Authors: Garg Rachana , Kapoor Veena , Abba Martin , Feldser David M. , Albelda Steven M. , Kazanietz Marcelo G.
Published: July 2016

Exposure to a number of environmental carcinogens, such as polycyclic aromatic hydrocarbons (PAHs), are the major causative agents of lung cancer, the leading cause of cancer related deaths in the U.S. Oncogenic mutations in KRAS, one of the most common alteration found in non-small cell lung cancer (NSCLC), are induced in high frequency by PAHs and environmental carcinogens. Studies from many laboratories, including ours, established key roles for PKCϵ, a member of the protein kinase C (PKC) family, in mitogenesis and survival of cancer cells. Notably, PKCϵ is up-regulated in several epithelial cancers, including human NSCLC, suggesting a role in the development and/or maintenance of the malignant phenotype. We observed that PKCϵ inhibition/depletion blunt the proliferative, motile, and invasive properties of lung cancer cells with KRAS mutation (PLoS One;7(2):e31714,2012). More importantly, silencing PKCϵ reduced the ability of human NSCLC cells with KRAS mutation to form tumors in nude mice as well as impaired their metastatic potential (Oncogene 31(20):2593-600, 2011). As Ras-transformed cells have enhanced DAG levels and signaling, we hypothesized that genetic targeting of PKCϵ reverses lung tumorigenesis driven by oncogenic Ras. To address this, we intercrossed lung-specific mutant Ras mice (KrasLSL-G12D/+) with PKCϵ knockout mice. Notably, significant inhibition occurred in the formation of lung tumors upon loss of either one or both PKCϵ gene (PRKCE) alleles, with significant extension of mice lifespan, suggesting a role for PKCϵ in the initiation of lung tumorigenesis driven by oncogenic mutant KRas. Furthermore, in silico database analysis of KRAS mutated human lung adenocarcinomas revealed a significant association between high PRKCE expression and poor patient outcome. As environmental lung carcinogens (PAHs) induce mutations in Ras, we next intended to test the hypothesis that PKCϵ is implicated in the action of these carcinogens. Interestingly, alveolar hyperplasia as well as pulmonary adenomas induced by benzo(a)pyrene, a prototypical PAH, were significantly reduced upon loss of one or two PKCϵ alleles, suggesting that genetic ablation of PKCϵ impairs chemically-induced lung carcinogenesis. Overall, our results indicate that PKCϵ is a novel effector of KRAS in lung cancer that may represent a promising target for disease treatment.

Citation Format: Rachana Garg, Veena Kapoor, Martin Abba, David M. Feldser, Steven M. Albelda, Marcelo G. Kazanietz. Protein kinase C epsilon is a mediator of KRAS-driven lung tumorigenesis. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5164.
Abstract 5201: Identification of growth factors that promote the proliferation and drug resistance of human primary acute B lymphoblastic cells
Authors: Li Peng , Jiang Zhiwu
Published: July 2016

Bone marrow mesenchymal stem cells (BMMSCs) contribute the survival, expansion, and drug resistance of leukemic cells. However, how BMMSCs promote the proliferation and drug resistance of leukemia remains to be poorly understood. Here, we derived a BM-derived stromal cell line that supported unlimited growth of human primary acute B lymphoblastic leukemic cells (B-ALL) by physical contacts with B-ALL cells and secreting growth factors. Nevertheless, its capacity of promoting primary B-ALL proliferation was abolished once the cell line was induced to differentiate into adipocytes. In xenograft models, we demonstrate that the presence of adipocytes suppressed the engraftment of primary B-ALL cells while the decrease of adipocytes enhanced the reconstitution of B-ALL. Consistently, we observed that the increase of numbers and diameters of adipocytes was correlated with remission after chemotherapy treatments in B-ALL patients. By comparing the global transcription profiles between the BM stromal cells and the adipocytes that were derived from the cell line, we identified a combination of cytokines and adherent proteins that were able to support the long-term growth of human primary B-ALL cells and desensitize them to chemotherapy drugs in a stromal-free culture condition. Furthermore, we showed that the PI3K/AKT pathways were involved in chemotherapy-induced resistance because targeting these signaling pathways increased the chemotherapy sensitivity of B-ALL in xenografts. Collectively, our results uncovered the mechanisms of BMMSCs supporting the survival, expansion, and drug resistance of human B-ALL cells.

Citation Format: Peng Li, Zhiwu Jiang. Identification of growth factors that promote the proliferation and drug resistance of human primary acute B lymphoblastic cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5201.
Abstract 5205: Changes in trends in colorectal cancer incidence rate by anatomic site between 1978 and 2004 in Japan
Authors: Nakagawa Hiroko , Ito Hidemi , Hosono Satoyo , Oze Isao , Mikami Haruo , Hattori Masakazu , Nishino Yoshikazu , Sugiyama Hiromi , Nakata Kayo , Tanaka Hideo
Published: July 2016

Although colorectal cancer (CRC), a major cancer worldwide, has shown a proximal or right-sided shift in subsite distribution in western countries, trends in subsite incidence in Asian countries remain unclear. Here, we evaluated subsite-specific trends in CRC incidence rate between 1978 and 2004 in Japan using large data from 10 population-based cancer registries. The colorectal sites (C18-20) were categorized into three groups, proximal colon (C18.0-18.5), distal colon (C18.6-C18.7) and rectum (C19.9 and C20.9). Trends in age-standardized incidence rates (ASRs) were characterized by Joinpoint regression analysis. A total of 474,651 colorectal cancer cases were analyzed. Overall, ASRs increased remarkably until 1993, with an annual percent change (APC) of 4.9%, and then stabilized thereafter. By subsite, however, ASRs of proximal colon significantly increased, with APCs of 7.1% (1978∼1991), 3.8% (1991∼96) and 0.9% (1996∼2004); distal colon showed an initial significant increase, with an APC of 7.6%, but stabilized from 1991 until the end of observation; and rectal cancer showed an initial significant increase, with APCs of 1.9% (1978∼88) and 5.6% (1988∼92), but then decreased abruptly in 1992, the year colorectal cancer screening was introduced nationwide, with an APC of -1.0%. Thus, we revealed that changes in incidence trends for the three anatomic sites apparently began to differ in the 1990s, and this is a first report to our knowledge. We also discuss factors that could explain these changes in Japanese. Careful monitoring is necessary to confirm whether these trends are changing in the Japanese population.

Citation Format: Hiroko Nakagawa, Hidemi Ito, Satoyo Hosono, Isao Oze, Haruo Mikami, Masakazu Hattori, Yoshikazu Nishino, Hiromi Sugiyama, Kayo Nakata, Hideo Tanaka. Changes in trends in colorectal cancer incidence rate by anatomic site between 1978 and 2004 in Japan. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5205.
Abstract 5208: Trends in fatal prostate cancer incidence by race among US men
Authors: Kelly Scott P. , Rosenberg Philip S. , Anderson William F. , Cleary Sean D. , Younes Naji , Andreotti Gabriella , Cook Michael B.
Published: July 2016

Background: Prostate-specific antigen testing (PSA) has dramatically changed the composition of prostate cancer, making it difficult to assess incidence trends. By defining fatal prostate cancer as the underlying cause of death from disease within 10 years of diagnosis, we have been able to conduct an in-depth analysis of “clinically relevant” prostate cancer incidence trends.

Methods: We extracted incident prostate cancer cases, causes of death and survival among men aged 45-84 years using the US Surveillance, Epidemiology, and End Results (SEER) database (1975-2002). In addition to standard descriptive analyses, the ability to distinguish prostate cancer cases that were ultimately fatal enabled us to fit age-period-cohort models.

Results: Among 51,682 fatal prostate cancer cases, incidence increased 1% per year prior to 1992, declined 15% per year from 1992-1995, and further declined by 5% per year through 2002. Age-specific incidence rates of fatal disease decreased over 2% per year among men aged 55 years and older, yet increased up to 1% per year among younger men. Fatal disease rates were more than 2-fold higher in black men compared with white men, a racial disparity that increased to 3-6 fold among younger men.

Conclusion: In the United States, after widespread use of PSA screening and advances in prostate cancer treatment, fatal prostate cancer rates have substantially declined. Yet, rates of fatal prostate cancer among younger men have remained relatively stable, suggesting the need for additional attention to early-onset prostate cancer that has the potential to result in death. This study also highlights the persistent black-to-white racial disparity in fatal prostate cancer, underscoring the need for a greater understanding of the causes of this difference so that strategies may be implemented to reduce, and ultimately eliminate, racial disparities in prostate cancer.

Citation Format: Scott P. Kelly, Philip S. Rosenberg, William F. Anderson, Sean D. Cleary, Naji Younes, Gabriella Andreotti, Michael B. Cook. Trends in fatal prostate cancer incidence by race among US men. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5208.