bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3961: Effectiveness of the combined treatment with hsp90 inhibitor and heavy ion irradiation
Authors: Okayasu Ryuichi , Masaoka Aya , Hirakawa Hirokazu , Fujisawa Hiroshi , Sunada Shigeaki , Hirayama Ryoichi , Noguchi Miho , Li Huizi , Fujimori Akira
Published: May 2015

Hsp90 inhibitors have been studied as effective sensitizers for radiation therapy. Some encouraging results have been observed and the mechanism of radio-sensitization has also been investigated mostly with X-or gamma-rays. There has not been any study for Hsp90 inhibitors with heavy ion radiation. In this report we show the data with the combined effect with hsp90 inhibitors including 17AAG and PU-H71 with high linear energy transfer (LET) heavy ion radiation. As in proton radiation treatment, heavy ions can be targeted to tumor tissues due to its physical properties, while significantly lower radiation doses are delivered in surrounding normal tissues. In addition, with heavy ions higher biological effectiveness, i.e, more tumor cell killing, can be attained than with proton or conventional photon radiation. We are finding that if tumor cells were pre-treated with an hsp90 inhibitor and then exposed to heavy ion (carbon) radiation, very effective cell killing were observed using SQ5 human lung tumor cells. Another interesting feature of hsp90 inhibitors is that normal cells are not much influenced by the drug. Our in vivo mouse xenograft studies using nude mice also indicate a better tumor control with the combination of hsp90 inhibitor and local carbon irradiation as compared with carbon irradiation alone. Further studies show that the mechanism behind this radio-sensitization may be attributed to inhibition of DNA double strand breaks (DSBs) by hsp90 inhibitors; particularly homologous recombination pathway of DSB repair might be affected. Our flow cytometry study also reveals some new aspect of tumor cell killing by this combination therapy. Our data suggest that hsp 90 inhibitors are good candidates for the combined cancer therapy using heavy ion irradiation.

Citation Format: Ryuichi Okayasu, Aya Masaoka, Hirokazu Hirakawa, Hiroshi Fujisawa, Shigeaki Sunada, Ryoichi Hirayama, Miho Noguchi, Huizi Li, Akira Fujimori. Effectiveness of the combined treatment with hsp90 inhibitor and heavy ion irradiation. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3961. doi:10.1158/1538-7445.AM2014-3961
Abstract 3963: MYCN-dependent expression of sulfatase2 regulates neuroblastoma cell survival
Authors: Solari Valeria , Borriello Lucia , Fernandez G. Esteban , Shimada Hiroyuki , Sposto Richard , Asgharzadeh Shahab , Yates Edwin A. , Turnbull Jeremy E. , Declerck Yves A.
Published: May 2015

Despite the well-demonstrated role of MYCN in the pathogenesis of neuroblastoma, the mechanisms underlying its oncogenic function are not entirely understood and there is evidence that its function is in part dependent on the tumor microenvironment (TME). Heparan sulfate proteoglycans (HSPG) play a critical role in the interaction between tumor cells and the TME in great part because of their ability to retain cytokines, chemokines and growth factors and to control their bioavailability. This function of HSPG is dependent on the sulfation pattern that itself is controlled by sulfotransferases that add sulfate groups to the repeating disaccharide units and sulfatases (SULF) that selectively remove 6-O-sulfates. An unbiased analysis of the expression of 12 of these enzymes by RT-PCR in 8 human neuroblastoma cell lines revealed higher levels of expression of SULF2 in cell lines with amplification of the MYCN oncogene (MYCN-A), an observation that was confirmed at the protein level by Western blot analysis. We then demonstrated that the knock down (KD) of SULF2 in MYCN-A cells by siRNA resulted in a significant decrease in cell viability caused by a decrease in cell cycle entry and in apoptosis as documented by increase in caspase 3/7 activity, PARP cleavage and annexin-V expression. Evidence was provided that SULF2 is a downstream target of MYCN by demonstrating that overexpression of MYCN in CHLA-255 neuroblastoma cells increased SULF2 expression whereas downregulation of MYCN expression in SHEP-21N cells was followed by a downregulation of SULF2. Underlying the importance of SULF2 in neuroblastoma cell survival independently of MYCN, we demonstrated that overexpression of SULF2 in MYCN-NA cells increased cell viability without increasing MYCN expression. Finally, an analysis of SULF2 protein expression in 65 primary human neuroblastoma tumors indicated a statistically significant higher level of expression in MYCN-A tumors along with an almost complete absence of expression in MYCN-non amplified (NA) tumors, whether they were of favorable or unfavorable histology. The data thus identify SULF2 as a new downstream target of MYCN and a key contributor to its oncogenic function in human neuroblastoma.

Citation Format: Valeria Solari, Lucia Borriello, G. Esteban Fernandez, Hiroyuki Shimada, Richard Sposto, Shahab Asgharzadeh, Edwin A. Yates, Jeremy E. Turnbull, Yves A. Declerck. MYCN-dependent expression of sulfatase2 regulates neuroblastoma cell survival. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3963. doi:10.1158/1538-7445.AM2014-3963
Abstract 3975: PID1, a new growth-inhibitory gene, sensitizes brain tumor cell lines to chemotherapy
Authors: Xu Jingying , Ren Xiuhai , Tran Anthony , Shackleford Gregory M. , Erdreich-Epstein Anat
Published: May 2015

BACKGROUND: Phosphotyrosine Interaction Domain containing protein-1 (PID1) is a gene discovered in 2006 in the context of obesity, which we recently showed inhibits growth of brain tumor cell lines and its mRNA level is directly correlated with survival of patients with gliomas and medulloblastomas (Erdreich-Epstein et al, Clin Cancer Res, In Press). PID1 mRNA was lower in more aggressive medulloblastomas and gliomas, and patients whose tumor had higher PID1 mRNA had longer survival. Moreover, overexpression of PID1 markedly decreased 2D colony formation in cell lines of medulloblastomas, GBM and atypical teratoid rhabdoid tumors (ATRT). The growth-inhibitory effect was due to both decreased proliferation and increased cell death.

METHODS/RESULTS: COSMIC showed no mutations of PID1 in brain tumors, suggesting that PID1 was not a formal tumor suppressor gene. Therefore, we hypothesized that PID1 may sensitize brain tumor cells to therapy, and thus account for the longer survival in patients with higher PID1 mRNA. Indeed, while both cisplatin (10µg/ml) or transient PID1 overexpression increased apoptosis of glioma and medulloblastoma cell lines (indicated by increased AnnexinV and mitochondrial depolarization), combining cisplatin with PID1 caused a markedly higher apoptosis. Moreover, knockdown of PID1 by siRNA inhibited the cisplatin-induced mitochondrial membrane depolarization and apoptosis (AnnexinV), suggesting that PID1 may be required for cisplatin-induced apoptosis. Taken together, this suggests a sensitizing effect of PID1 to chemotherapy in both glioma and medulloblastoma cell lines. Intriguingly, PID1 mRNA and protein both increased in response to etoposide (5µg/ml), vincristine (50ng/ml) or cisplatin (5µg/ml) in a time- and dose-dependent manner in the brain tumor cell lines. This chemotherapy-induced increase in PID1 mRNA was blocked by inhibitors of NFKB, suggesting that regulation of PID1 may be an NFKB-dependent mechanism. Ongoing work seeks to decipher the mechanism of these effects and test them in vivo in mouse models.

CONCLUSIONS: Our data suggest that PID1 acts to sensitize glioma and medulloblastoma cells to chemotherapy, possibly explaining the correlation between higher PID1 mRNA and survival in patients.

Citation Format: Jingying Xu, Xiuhai Ren, Anthony Tran, Gregory M. Shackleford, Anat Erdreich-Epstein. PID1, a new growth-inhibitory gene, sensitizes brain tumor cell lines to chemotherapy. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3975. doi:10.1158/1538-7445.AM2014-3975
Abstract 3979: NSC305787 inhibits specific protein–protein interactions involving ezrin in osteosarcoma cells
Authors: Celik Haydar , Han Jenny , Hong Sung-Hyeok , Bulut Gulay , Toretsky Jeffrey , Uren Aykut
Published: May 2015

Osteosarcoma (OS) is the malignant tumor of bone that is most common in children and adolescents. OS is characterized by a high propensity for metastasis, especially to lungs, which is the main cause of death. The membrane-cytoskeleton cross-linker ezrin plays a key role in the metastatic progression of OS. In our earlier studies, we discovered that a small molecule, NSC305787, directly binds to ezrin and inhibits its function related to metastatic phenotype. In order to gain a better insight into the molecular mechanisms of NSC30787 action, an affinity pull-down coupled mass spectrometry was conducted to find the putative ezrin-interacting proteins whose interaction with ezrin is decreased or lost in the presence of NSC305787. Detailed mass spectrometry analysis identified 78 proteins after filtering for the potential background contaminants. Out of 78 proteins, 9 proteins were known ezrin-binding proteins including ezrin, radixin, moesin, polyadenylate-binding protein 1 (PABP-1), Na(+)/H(+) exchange regulatory cofactor (NHERF1/EPB-50), calpains, epidermal growth factor receptor kinase substrate 8 (EPS8), protein kinase C ioto (PKC-ioto) and focal adhesion kinase (FAK). For the remaining 69 proteins, we immunoprecipitated ezrin from K7M2 OS cells, and then western blotted for 8 selected functionally relevant proteins in order to test the validity of our approach. Of these 8 candidate proteins, Co-IP experiments verified binding of ezrin to 5 novel binding partners including DDX3 RNA helicase, caprin-1, AEG-1/MTDH/LYRIC, Rho guanine nucleotide exchange factor 2 (GEFH1) and catenin delta-1 (p120-catenin). NSC305787 effectively reduced the binding of DDX3 RNA helicase, AEG-1/MTDH/LYRIC and GEFH1 to ezrin in Co-IP experiments. Direct binding of ezrin to DDX3 RNA helicase was also demonstrated by surface plasmon resonance analysis using Biacore instrument. In conclusion, our results suggest that NSC305787 can modulate ezrin function through blocking its interaction with crucial proteins involved in adhesion, migration, invasion and survival. We also identified novel ezrin interactors that are involved in different levels of mRNA metabolism suggesting a completely new molecular function of ezrin that is independent from its classic role as a membrane cytoskeletal cross-linker at the plasma membrane. Thus, these results highlight novel molecular mechanisms for ezrin in cancer metastasis.

Citation Format: Haydar Celik, Jenny Han, Sung-Hyeok Hong, Gulay Bulut, Jeffrey Toretsky, Aykut Uren. NSC305787 inhibits specific protein–protein interactions involving ezrin in osteosarcoma cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3979. doi:10.1158/1538-7445.AM2014-3979
Abstract 4001: Interaction of CD200 overexpression on tumor cells with CD200R1 overexpression on stromal cells: An escape from the host immune response in rectal cancer patients
Authors: Bisgin Atil , Meng Wen Jian , Adell Gunnar , Sun Xiao Feng
Published: May 2015

Introduction: CD200 imparts an immunoregulatory signal through its receptor - CD200R1, leading to the suppression of tumor specific immunity. The mechanism of CD200:CD200R1 signaling pathway is still uncertain.

The aim of this study was to investigate the expression pattern and localization of CD200 and its receptor CD200R1 in normal mucosa and rectal cancer patients.

Material and Methods: The immunohistochemical expressions and localizations of CD200 and CD200R1 were examined in 140 rectal cancer patients. Seventy nine of 140 underwent the preoperative radiotherapy treatment and the others were untreated prior to the surgery. In addition, 121 matched normal rectal mucosa samples from the same patients were evaluated.

Results: CD200 was expressed in normal mucosal epithelium and rectal cancer. Expression of CD200R1 was seen in normal mucosal epithelium and stromal cells of tumor samples. We found significant up-regulation of CD200 in rectal cancer tissues compared to the normal mucosa (P = 0.001). Interestingly, CD200R1 was overexpressed in stromal cells of rectal cancer patients presenting metastases compared to patients without metastases (P = 0.002). However, there is no difference in the expression levels between stromal cells and tumor cells. More than that, the up-regulation of CD2OO in tumor cells and CD200R1 overexpression in stromal cells was observed in 87 % of metastatic rectal cancer patients. These high expression levels of tumoral CD200 and stromal CD200R1 were significantly correlated with lymph node metastasis.

Conclusions: Interaction of CD200 and CD200R1 and their expression profile might be useful to follow-up disease progression in terms of carcinogenesis and metastasis.

Citation Format: Atil Bisgin, Wen Jian Meng, Gunnar Adell, Xiao Feng Sun. Interaction of CD200 overexpression on tumor cells with CD200R1 overexpression on stromal cells: An escape from the host immune response in rectal cancer patients. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4001. doi:10.1158/1538-7445.AM2014-4001
Abstract 4009: N-ras codon 61 mutation is associated with distant metastasis in patients with follicular thyroid carcinoma
Authors: Jeon MinJi , Jang Eun Kyung , Song Dong Eun , Sim So Young , Kim Eui Young , Choi Yun M. , Han Ji Min , Kim Won G. , Kim Tae Yong , Shong Young Kee , Kim Won Bae
Published: May 2015

Distant metastasis is a main cause of death in follicular thyroid carcinoma (FTC) patients. Factors related to distant metastasis in FTC had been known as age, primary tumor size, and invasiveness. Ras mutations were also supposed to be associated with poor clinical outcomes.

We analyzed Ras mutations in FTC with a distant metastasis (FTC M1, n=28), size matched-FTC without a distant metastasis (FTC M0, n=28), follicular adenoma (FA, n=17), and nodular hyperplasia (NH, n=12) to figure out the roles of Ras mutations in follicular thyroid carcinogenesis and metastasis. In addition, we assess the relationship between Ras mutations and clinical outcomes in FTC patients. NRAS, HRAS, and KRAS mutations were assessed using direct sequencing method.

Among 85 patients, 39 patients (46%) had Ras mutations. NRAS codon 61 mutation (n=21; 25%) was the most common point mutation. HRAS codon 61, KRAS codon 12/13, and KRAS codon 61 mutations were found in 7, 6, and 4 patients, respectively. NRAS codon 12/13 mutation was found in only 1 patient, and HRAS codon 12/13 mutation was not found. Ras mutations were significantly more common in the FTC than FA or NH groups. Especially, NRAS codon 61 mutation was associated with distant metastasis in patients with FTC. However, there was no significant difference in survival between the Ras mutation positive-FTC and Ras mutation negative-FTC patients.

Ras mutation, especially NRAS codon 61 mutation, was significantly associated with the presence of distant metastases. NRAS codon 61 mutation status might be a potential prognostic factor in FTC patients.

Citation Format: MinJi Jeon, Eun Kyung Jang, Dong Eun Song, So Young Sim, Eui Young Kim, Yun Mi Choi, Ji Min Han, Won Gu Kim, Tae Yong Kim, Young Kee Shong, Won Bae Kim. N-ras codon 61 mutation is associated with distant metastasis in patients with follicular thyroid carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4009. doi:10.1158/1538-7445.AM2014-4009
Abstract 4025: Combination strategy of endoscopic resection and telomerase-targeting oncolytic virus for eradicating lymph node metastasis of submucosal invasive colorectal cancer
Authors: Hori Naoto , Kikuchi Satoru , Kishimoto Hiroyuki , Tazawa Hiroshi , Hashimoto Yuuri , Kuroda Shinji , Kagawa Shunsuke , Urata Yasuo , Hoffman Robert M. , Fujiwara Toshiyoshi
Published: May 2015

Endoscopic resection is now one of the options as minimally invasive therapy for early colorectal cancer, However, surgical resection has been still standard for submucosal invasive colorectal cancer, because about 10% of that have LN metastasis. To secure endoscopic resection for submucosal invasive colorectal cancer, an alternative, innovational strategy to purge potential LN metastasis is required. We previously reported that a telomerase-dependent oncolytic adenovirus (OBP-301) spreads to the regional LNs, selectively replicates, and eradicates metastatic cancer cells in LNs, when administered into the orthotopically xenotransplanted rectal tumor in mouse model. In this study we evaluated whether OBP-301 could inhibit LN metastasis after the local resection mimicking endoscopic resection in a submucosal invasive rectal tumor mouse model. Human colon cancer HCT-116 cells expressing green fluorescent protein (GFP) were orthotopically implanted into the submucosal layer of the rectum in nude mouse, to develop resultant rectal tumor mimicking submucosal invasive one. LN metastasis was observed in 78.5% of mice as early as 7 days after cancer cell inoculation. The rectal tumor was injected with OBP-301 peritumorally, and then got locally excised mimicking Endoscopic submucosal dissection (ESD) technique. Seven days after tumor resection, the treatment with OBP-301 showed a significant inhibition in LN metastasis, whereas the other control treatment did not. Moreover, LN metastatic recurrence was not detected in OBP-301-treated group even 4 weeks after tumor resection. Similar results were also observed in another model with human colon cancer Colo-205-GFP cells. These results suggest that a novel combination of ESD and OBP-301 might have potential to take place of the conventional surgical approach in treating submucosal invasive colorectal cancer.

Citation Format: Naoto Hori, Satoru Kikuchi, Hiroyuki Kishimoto, Hiroshi Tazawa, Yuuri Hashimoto, Shinji Kuroda, Shunsuke Kagawa, Yasuo Urata, Robert M. Hoffman, Toshiyoshi Fujiwara. Combination strategy of endoscopic resection and telomerase-targeting oncolytic virus for eradicating lymph node metastasis of submucosal invasive colorectal cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4025. doi:10.1158/1538-7445.AM2014-4025
Abstract 4041: Involvement of Notch1 signaling pathway in medulloblastoma metastasis
Authors: Kahn Suzana A. , Gholamin Sharareh , Zhang Michael , Nitta Ryan , Weissman Irving , Mitra Siddhartha , Cheshier Samuel
Published: May 2015

Medulloblastoma (MB) is a malignant brain tumor that originates in the cerebellum in children and spreads via the cerebrospinal fluid to the leptomeningeal spaces of the brain and of the spinal cord. MB is stratified into four distinct groups, according to genetic and clinical features, and patients from group 3 (also known as c-Myc-amplified group) have the highest risk of developing metastatic disease and, consequently, a poor prognosis. Treatment protocols involve surgery, craniospinal radiation, and high-dose chemotherapy, which frequently cause disabling neurotoxic effects in long-term survivors. We used MB cell lines and primary cells isolated from patients from the c-Myc-amplified group to develop a spontaneous spinal metastasis orthotopic xenograft model as a tool to understand the cellular determinants of leptomeningeal and spinal dissemination. Human cells isolated from the primary site tumors expressed 10 fold higher NICD1 (Notch1 Intracellular Domain), the active form of Notch1, than cells isolated from spinal metastatic sites (as quantified by western blot and imunohistochemistry), suggesting the downregulation of canonical Notch1 signaling pathway in MB metastasis. Moreover, flow cytometry analyses revealed that a higher percentage of cells isolated from metastatic sites expressed full-length surface Notch1 as compared to the primary site tumor. These differences cannot be explained by enrichment in the stem cell population at primary tumor sites, as we observed that MB cells isolated from primary tumors and metastatic sites exhibit equivalent self-renewal potential and express equivalent levels of CD133 and CD15. Our goal is to understand the role of notch in regulating medulloblastoma metastasis.

Citation Format: Suzana A. Kahn, Sharareh Gholamin, Michael Zhang, Ryan Nitta, Irving Weissman, Siddhartha Mitra, Samuel Cheshier. Involvement of Notch1 signaling pathway in medulloblastoma metastasis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4041. doi:10.1158/1538-7445.AM2014-4041
Abstract 4045: Pediatric osteosarcoma lung metastasis: variations in clinical management
Authors: Vo Andrea T. , Bhattasali Onita , Roth Michael , Gill Jonathan , Geller David , Randall Lor , Gorlick Richard
Published: May 2015

Background: Nearly 20% of patients with newly diagnosed osteosarcoma have detectable metastatic disease at presentation, most of which is located in the lungs. These patients have a poor prognosis with survival of generally less than 30%. NCCN guidelines as well as many osteosarcoma clinical trials endorse resection of metastases as part of standard treatment. There are limited recommendations as to the timing or method of resection.

Methods: An anonymous online questionnaire was distributed to establish practice patterns among members of the Connective Tissue Oncology Society, a multi-disciplinary sarcoma focused organization. Participants were asked about management of lung metastases in 3 clinical scenarios (2 cm unilateral, 0.5 cm unilateral, and multiple bilateral lung nodules). Questions evaluated timing of management, decision to resect, technique of resection, and decision to explore the contralateral lung.

Results: 183 individuals completed the survey. Most report initiating nodule management after neoadjuvant therapy. If the nodule disappeared after neoadjuvant therapy or definitive surgery, most report not performing surgical resection. Among nodules resected after neodjuvant therapy or definitive surgery that were unchanged in size, thoracotomy is the most commonly reported surgical technique. When only unilateral nodules were present on imaging, few choose to explore the contralateral lung (11% in 2.0cm and 10% in 0.5cm, compared to 70% in multiple bilateral nodules). Pediatric Oncologists were more likely to initiate management earlier than Medical Oncologists for 2.0cm nodule (90% vs 63%, p<0.001), 0.5 cm nodule (77% vs. 47%, p <0.005), and multiple bilateral nodules (79% vs. 51%, p =0.008).

Conclusion: There is marked heterogeneity among treating clinicians in their management of pulmonary metastases in osteosarcoma. Variation was also noted between medical specialties. Prospective trials are needed to evaluate if these differences have significant implications on outcomes for patients with newly diagnosed metastatic osteosarcoma.

Citation Format: Andrea T. Vo, Onita Bhattasali, Michael Roth, Jonathan Gill, David Geller, Lor Randall, Richard Gorlick. Pediatric osteosarcoma lung metastasis: variations in clinical management. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4045. doi:10.1158/1538-7445.AM2014-4045
Abstract 4046: N-myc interactor impacts autophagy via modulation of GSK3β/mTOR signaling
Authors: Metge Brandon J. , Mitra Aparna , Shevde Lalita A. , Samant Rajeev S.
Published: May 2015

Autophagy is a cellular mechanism for removal of dysfunctional cellular organelles, large protein complexes. It drives a specialized form of program cell death termed as programmed cell death II. Recent studies have demonstrated a prominent role of the process of autophagy in cancer progression and drug sensitivity. Under conditions of stress, cancer cells try to utilize autophagy to leverage a survival advantage; however, excessive autophagy renders cells vulnerable.

Our previous work has demonstrated a pivotal role of N-myc interactor(NMI) in impeding breast cancer progression and metastasis. Loss of NMI is one of the key events that precedes and contributes to invasive progression of breast cancer. Here we provide evidence that NMI plays a crucial role in regulation of mTOR signaling ultimately impacting the autophagic response in breast cancer cells. Our initial observations showed that breast cancer cells restored for NMI expression contained more acidic vacuoles as determined by staining with Lysotracker. Further indication of autophagy was evidenced by the formation of distinct punctae in NMI cells upon transfection with GFP-LC3 which was ablated upon treatment with an autophagy inhibitor. Cells restored for expression of NMI showed decreased levels of p62 along with an increased LC3II processing. The mTOR signaling pathway plays a pivotal role in driving the autophagy. We found that p70S6K phosphorylation dramatically decreases in NMI expressing cells indicating an inhibition of mTOR signaling in these cells. Our observations show that active GSK3β, a negative modulator of mTOR, is responsible for mediating the effects of NMI. Conversely, NMI silenced cells showed indication of inactive GSK3β concomitant with activated mTOR signaling.

Functionally, NMI mediated sensitization of cells to autophagy rendered breast cancer cells incapable of sustaining stress of cisplatin treatment. The sensitivity of these cells to cisplatin was notably reduced upon inhibition of the autophagic process. This suggests a possible role for autophagy in NMI induced drug sensitivity. Collectively, our results suggest that NMI plays an important role in modulation of autophagy via activation of GSK3β and consequent inhibition of mTOR signaling.

Citation Format: Brandon J. Metge, Aparna Mitra, Lalita A. Shevde, Rajeev S. Samant. N-myc interactor impacts autophagy via modulation of GSK3β/mTOR signaling. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4046. doi:10.1158/1538-7445.AM2014-4046
Abstract 4094: Identification of co-inhibitory receptor expression on T cells from gastric cancer patients
Authors: Zong Yunhui , Chu Fuliang , He Songbing , Jing Yu , Hunsucker Sally A , Yang Tina , Zhang Ming , Ye Sisi , Li Yafen , Liu Bin , Tang Jinle , Meng Huimin , An Gangli , Yang Lin
Published: May 2015

The critical role of tumour antigen-specific immune responses to restrain tumour growth in patients with gastric cancer furthers the need to dissect the co-inhibitory pathways involved in tumour infiltrating lymphocyte (TIL) exhaustion/dysfunction. It was previously reported that PD-1 expression was significantly increased on CD4+ and CD8+ T cells from patients with gastric cancer and in gastric cancer tissues, compared to normal donors. In this study, we observed up-regulation of TIM-3 and PD-1 expression on peripheral T cells and up-regulation of CTLA-4, TIGIT, TIM-3 and PD-1 expression on TILs from gastric cancer patients; however TIGIT was down-regulated on CD8+ TILs and TIM-3 up-regulation on CD8+ TILs was not statistically significant. Furthermore, the percentages of PD-1+/TIM-3+ and PD-1+/TIM-3- peripheral T cells was significantly higher in gastric cancer patients than normal donors, and the percentage of PD-1+/TIM-3+ TILs was significantly higher than the percentage of PD-1+/TIM-3+ peripheral T cells in gastric cancer patients. PD-1+/TIM-3+ cells represent the predominant fraction of TILs. PD-1 blockade enhanced cytokine production and the cytotoxicity of TILs and exhibited a synergistic effect with TIM-3 blockade. Collectively, our results suggest that the use of anti-PD-1 blockade in combination with anti-TIM-3 blockade could restore the cytotoxic activity of TILs in patients with gastric cancer.

Citation Format: Yunhui Zong, Fuliang Chu, Songbing He, Yu Jing, Sally A Hunsucker, Tina Yang, Ming Zhang, Sisi Ye, Yafen Li, Bin Liu, Jinle Tang, Huimin Meng, Gangli An, Lin Yang. Identification of co-inhibitory receptor expression on T cells from gastric cancer patients. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4094. doi:10.1158/1538-7445.AM2014-4094
Abstract 4097: Vacuolar ATPase subunit a2 is associated with immunosuppression in cancer
Authors: Katara Gajendra K. , Kulshrestha Arpita , Jaiswal Mukesh , Gilman-Sachs Alice , Beaman Kenneth D.
Published: May 2015

Macrophage polarization contributes to distinct human pathologies. In tumors, a polarized M2 phenotype called tumor-associated macrophages (TAMs), are associated with promotion of invasion and angiogenesis. In cancer cells, vacuolar ATPase (V-ATPase), a multi-subunit enzyme, is expressed on the plasma/vesicular membranes and critically influences the metastatic behavior. In addition, the soluble, cleaved N-terminal domain of a2 isoform of V-ATPase (a2NTD) is associated with in vitro induction of pro-tumorigenic properties in monocytes. Monocytes induced with recombinant a2NTD showed polarization towards M2 macrophage subtype by expressing high levels of mannose receptor-1, Arginase-1, IL-10, MMP-9, VEGF and TGF-β. Additionally, co-culture of these macrophages enhances invasion and angiogenesis of tumor cells and suppression of T cell activation. This activity of the a2 isoform of V-ATPase (a2V) caused us to further investigate its role in cancer progression by evaluating the effects of a2V knock down on tumor cells. In this study, we have successfully deleted a2V gene in mouse 4T1 breast cancer cells using shRNA mediated knock down. These knock down cell lines are co-cultured with macrophages to evaluate previously observed immuno-modulatory effect of a2NTD. In addition, a2V knock down cells are injected into the Balb/c mice to generate tumors. The effect of a2V gene deletion on tumor progression and immune responses are evaluated in mice. We show that in the absence of a2V (finally a2NTD) growth and progression of tumor are suppressed by dominated host immune responses.

Citation Format: Gajendra K. Katara, Arpita Kulshrestha, Mukesh Jaiswal, Alice Gilman-Sachs, Kenneth D. Beaman. Vacuolar ATPase subunit a2 is associated with immunosuppression in cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4097. doi:10.1158/1538-7445.AM2014-4097
Abstract 4102: p27 as a biomarker and determinant of risk for endometrial carcinoma arising in the setting of obesity
Authors: McCampbell Adrienne S. , Mittlestedt Megan L. , Dere Ruhee , Zhou Lijun , Djordjevic Bojana , Soliman Pamela T. , Zhang Qian , Wei Caimiao , Hursting Stephen D. , Lu Karen H. , Walker Cheryl L. , Broaddus Russell R.
Published: May 2015

Endometrial carcinoma exhibits the strongest association with obesity of any cancer. Tumor growth is driven by PI3K/AKT activation, and opposed by tumor suppressors, including the tuberous sclerosis complex (TSC) and p27, with inactivation of TSC2 and loss of cytoplasmic misloclization of p27 both being linked to PI3K/AKT signaling. However, little is known about the involvement of p27 in the development of endometrial carcinoma arising in the setting of obesity, expecially its role in early disease progression. We found that p27 was moderatley-to-severely reduced in both “normal” endometrial glands as well as endometrial hyperplasia of obese women. Obese Tsc-2-deficient Eker rats similarly developed endometrial hyperplasia with a high frequency and short latency, and like obese women, endometrial hyperplasia that developed in obese rats exhibited loss of p27. In Eker rats, even a short duration of caloric restriction (2-4 months of age) reduced weight, increased adiponectin and lowered leptin to produce favorable leptin:adiponectin ratio, and reduced circulating insulin levels. Caloric restriction also increased p27 levels, primarily in the nucleus, and significantly decreased endometrial hyperplasia incidence by up to 50%. These data demonstrate that in the setting of obesity, loss of p27 occurs very early in progression to carcinoma. Importantly, p27 can be modulated along with weight by diet, and perhaps other lifestyle interventions that decrease risk for development of carcinoma, suggesting it may be a useful biomarker, and possible determinant of risk for endometrial carcinoma in the setting of obesity.

Citation Format: Adrienne S. McCampbell, Megan L. Mittlestedt, Ruhee Dere, Lijun Zhou, Bojana Djordjevic, Pamela T. Soliman, Qian Zhang, Caimiao Wei, Stephen D. Hursting, Karen H. Lu, Cheryl L. Walker, Russell R. Broaddus. p27 as a biomarker and determinant of risk for endometrial carcinoma arising in the setting of obesity. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4102. doi:10.1158/1538-7445.AM2014-4102
Abstract 4106: Curcumin inhibit PhIP induced cytotoxicity by inhibiting ROS production, DNA strand breaks and DNA adducts formation in MCF 10A cells
Authors: Jain Ashok K. , Samykutty Abhilash , Jackson Carissa L. , Thangaraju Muthusamy , Browning Darren D.
Published: May 2015

PhIP is a known food carcinogen found in well done meat which causes several cancers including breast cancer. PhIP metabolites produce DNA adduct and DNA strand breaks. Curcumin, obtained from the rhizome of Curcuma longa, has potent anticancer activity. So far none of the study demonstrates the inhibition of PhIP induced cytotoxicity by the co-treatment of Curcumin in normal breast epithelial cells in vitro. Therefore, we developed a model system using the MCF 10A normal breast epithelial cells to study the PhIP cytotoxicity and if cells are co-cultured with Curcumin and PhIP how it affects. Consequently, the core signaling pathways have been explored to evaluate the efficacy of Curcumin. In this study, the effectiveness of Curcumin was investigated in MCF 10A cells along with PhIP. The cytotoxic ability was detected with MTT assay, ROS activation by DCF, the influence of the cell cycle was checked with flow cytometry, DNA damage by comet assay, DNA adduct formation by anti-PhIP DNA primary, and apoptosis by Annexin-V-FITC staining. The influence of the core signaling pathways was evaluated by RT PCR and/or Western blotting which included Nrf2 (GSR, GPX, NQO1), FOXO (Catalase, GADD45, PRDX3) targets; DNA repair genes/proteins BRCA1, H2AFX and PARP-1; and tumor suppressor P16 gene expression. PhIP cytotoxicity induced the expression of various antioxidant and DNA repair genes on MCF-10A cells but co-treatment of Curcumin retained its expression level similar to untreated groups. Additionally, Curcumin co- treatment increased the expression level of tumor suppressor expression gene P-53. Expression of antioxidants genes was induced by PhIP whereas Curcumin significantly suppress the PhIP induced ROS activation, DNA strand breaks and DNA adduct formation and consequently inhibited the cell death. In conclusion, Curcumin appears to be effective to inhibit P450 mediated ROS production and PhIP-DNA adducts which consequently reduces DNA damage.

Citation Format: Ashok K. Jain, Abhilash Samykutty, Carissa L. Jackson, Muthusamy Thangaraju, Darren D. Browning. Curcumin inhibit PhIP induced cytotoxicity by inhibiting ROS production, DNA strand breaks and DNA adducts formation in MCF 10A cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4106. doi:10.1158/1538-7445.AM2014-4106
Abstract 4115: GS-19, a novel GSK inhibitor suppresses the growth of pancreatic cancer cells by inhibiting EGFR/AKT/STAT-3 signaling
Authors: Ranjan Alok , Birkenfeld Ashlee , Lu Jianyu , Hua Duy , Srivastava Sanjay K.
Published: May 2015

Pancreatic cancer remains the fourth leading cause of cancer related deaths in the United States. Therefore, identification of novel strategies to control and treat pancreatic cancer is of great importance. GS-19 is a novel compound recently found to be a GSK inhibitor. In this study, we determined the anti-cancer effects of GS-19 in pancreatic cancer cells. GS-19 treatment inhibited the growth of Panc-1 and BxPC-3 cells in a concentration-dependent manner. Both the cell lines treated with GS-19 exhibited G-1 cell cycle arrest as well as apoptosis. Western blot analysis of both BxPC-3 and Panc-1 cells treated with GS-19 inhibited the activation (phosphorylation) and expression of key survival signaling proteins such as EGFR, STAT3 and AKT. The apoptosis induced by GS-19 was associated with cleavage of caspase-3 and PARP. Taken together, our results suggest that GS-19 suppress pancreatic cancer cell growth by inhibiting EGFR/AKT/STAT-3 signaling. However, further mechanistic studies are in progress to establish the role of EGFR/AKT/STAT-3 signaling in pancreatic cancer cells. [The studies in part were supported by R01 grant CA129038 awarded by NIH to S.K.S.]

Citation Format: Alok Ranjan, Ashlee Birkenfeld, Jianyu Lu, Duy Hua, Sanjay K. Srivastava. GS-19, a novel GSK inhibitor suppresses the growth of pancreatic cancer cells by inhibiting EGFR/AKT/STAT-3 signaling. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4115. doi:10.1158/1538-7445.AM2014-4115
Abstract 4120: Periodic fasting mimicking diet started late in life reduces and delays carcinogenesis
Authors: Brandhorst Sebastian , Wei Min , Navarre Gerrardo , Dubeau Louis , Conti Peter , Morgan Todd , Longo Valter D.
Published: May 2015

Nutrition can have profound effects on aging and age-related pathologies including cancer. As such, studies of fasting (the complete withdrawal of food) and dietary restriction (DR, generally described as a 20- 40% reduction in calories) have greatly contributed to the understanding of the relationship between nutrients, cellular aging, cancerogenesis and cancer cell survival. Our findings in yeast indicated that starvation decreases age-dependent mutations whereas our mouse xenograft tumor models indicate that fasting alone can be as effective as chemotherapy drugs in delaying the progression of certain tumor types, in part through the reduction of extracellular glucose and IGF-1 concentration/signaling. Here, we developed a fasting mimicking/enhancing diet (FMD) and tested its effects on cancerogenesis. In a cohort of female C57Bl6 mice, bi-monthly 4 day FMD feeding cycles followed by normal food intake, significantly improved survival in mice without causing excessive weight loss. Mice in the FMD cohort had a significantly reduced lifelong cancer incidence rate. Lymphoma was the most common cause of morbidity/mortality and affected ∼67% in the ad lib fed control group but only ∼40% of the FMD fed mice. In both cohorts lymphomas contributed to the great majority of all neoplasms found. Neoplasms were found in animals in the ad lib fed cohort starting 5 months after onset of our study, whereas no tumors occurred in the first 8 months in the FMD cohort. Furthermore, the FMD caused a shift from the presence of tumors in multiple organs (≥3) to tumors being either absent or present in 2 or less sites, thus indicating a potential shift to the development of benign tumors in the FMD group. In summary, the periodic FMD cycles had a significant impact on the development of cancer while allowing the preservation of lean body mass and animal weight until old age and extending longevity.

Citation Format: Sebastian Brandhorst, Min Wei, Gerrardo Navarre, Louis Dubeau, Peter Conti, Todd Morgan, Valter D. Longo. Periodic fasting mimicking diet started late in life reduces and delays carcinogenesis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4120. doi:10.1158/1538-7445.AM2014-4120
Abstract 4133: Disparities in colon cancer survival according to payer status: Findings from national cancer data bank
Authors: Devarakonda Srinivas S. , Shi Runhua , Neppalli Amarendra , Mills Glenn
Published: May 2015

BACKGROUND:

The objective of the current study was to examine the impact of payer status on relative survival outcomes in colon cancer patients.

METHODS:

A population-based analysis of National Cancer Data Bank (NCDB) records for invasive colon cancer was done using data from patients diagnosed between 1998 and 2006 and followed up to 2011. The cohort consisted of colon cancer patients of analytic stage I to stage IV. Information regarding age, race, gender, stage of cancer at the time of diagnosis, year of diagnosis, insurance status, income, treatment received and diagnostic facility, and type of treating facility were included in this study. Age, sex, race, calendar year specific US life table between 1970 and 2007 were used to compute expected survival. Multivariate Poisson modeling was used to estimate relative survival for payer status adjusting for other factors.

RESULTS:

436,235 colon cancer patients who met the inclusion criteria were included in the study. The mean age at diagnosis was 68.9 years. The stage distribution among the variables listed above is statistically significant. The 5-year cumulative relative survivals are 0.76, 0.84, 0.93, 1.74 and 1.10 for uninsured, Medicaid, private insurance, Medicare, and unknown payer status respectively. In multivariate analysis of relative survival, adjusting for other factors, the excess hazard ratios (EHR) were 1.74, 1.14, 0.89 and 1.29 for uninsured, Medicaid, Medicare, and unknown payer status versus private insurance respectively (P<0.05). The EHRs for blacks and other races versus whites were 1.07 and 1.53 respectively (p<0.05). The EHRs for follow-up year of 2, 3, 4, 5 versus year 1 were 1.17, 0.21, 0.18, and 0.54 respectively (P <0.05). Compared to academic/research program facility type, the EHRs for community cancer program, comprehensive community cancer program, and other specified types of cancer programs were 1.02,0.77 and 25.9 respectively (P <0.05). The relative survival rates for stage II, III, IV versus stage I were 0.95, 0.82, and 0.70 respectively (P<0.05). Diagnosis before or after 2002 and the type of treatment facility lead to a marginal difference in the relative survival.

CONCLUSIONS:

Our study shows the impact of payer status on the relative survival of patients with colon cancer and demonstrated increased mortality for uninsured patients in contrast to survival benefit for patients with Medicare and private insurance.

Citation Format: Srinivas S. Devarakonda, Runhua Shi, Amarendra Neppalli, Glenn Mills. Disparities in colon cancer survival according to payer status: Findings from national cancer data bank. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4133. doi:10.1158/1538-7445.AM2014-4133
Abstract 4157: Autosome instability and loss of chromosome Y in blood lymphocytes predict lung cancer risk
Authors: Sun Bing , Wang Ying , Kota Krishna , Shi Yaru , Makambi Kepher H. , Loffredo Christopher A. , Shields Peter G. , Harris Curtis C. , Zheng Yun-Ling
Published: May 2015

Background: Cancer cells are characterized by frequent chromosome aberrations (CA), which reflect defects of the underlying mechanisms that maintain genomic stability. Chromosome aberrations in peripheral blood lymphocytes has been shown to be associated with overall cancer risk in previous small studies. However, the relationship between types of chromosome aberrations in blood lymphocytes and lung cancer risk has not been well defined. In a case-control study of non-small cell lung cancer, we evaluated if CA in autosomes and sex chromosomes were predictive of lung cancer risk. Method: Chromosome aberrations were determined by G-banding analysis of metaphase chromosomes from short-term cultured blood lymphocytes. Logistic regression was used to estimate the strength of association between CAs and lung cancer risk. Results: Autosome instability (defined as CA in any of the 22 pairs of autosomes in > 5% of the cells) was significantly associated with lung cancer risk among individuals who were <= 67 years of age; adjusted odds ratio (OR) of 5.0 [95% confidence interval (CI) = 1.6 - 15.3]. There was no significant association between autosome instability and lung cancer risk in individuals who were older than 67 years of age. Loss of chromosome Y was significantly associated with an elevated risk of lung cancer among males; adjusted OR of 3.2 (95% CI = 1.2 - 8.8). This association appeared to be stronger in older males (> 67 years of age) and in African American males. We observed no significant association between loss of chromosome X and lung cancer risk in both males and females. Conclusion: Autosome instability and loss of chromosome Y in blood lymphocytes were significantly associated with lung cancer risk. Analysis of chromosome aberrations in blood lymphocytes may be a promising tool for the identification of sub-population who are at high risk of developing lung cancer.

Citation Format: Bing Sun, Ying Wang, Krishna Kota, Yaru Shi, Kepher H. Makambi, Christopher A. Loffredo, Peter G. Shields, Curtis C. Harris, Yun-Ling Zheng. Autosome instability and loss of chromosome Y in blood lymphocytes predict lung cancer risk. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4157. doi:10.1158/1538-7445.AM2014-4157