bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1691: SL-401 activity against AML blasts correlates with CD123 levels and does not down-regulate CD123 expression
Authors: Mani Rajeswaran , Gopalakrishnan Bhavani , Mo Xiaokui , Brooks Chris , Blum William , Lozanski Gerard , Vasu Sumithira , Muthusamy Natarajan
Published: August 2015

Despite the progress, the current therapies for acute myeloid leukemia (AML) are not curative partly due to failure to eradicate the leukemic stem cells (LSCs) that repopulate the leukemia. CD123 is the alpha-chain of interleukin-3 receptor (IL-3R) that is expressed on AML and AML LSCs, while it is undetectable in normal hematopoietic stem cells. We report here time and dose dependent cytotoxic activity of SL-401(Stemline Therapeutics, Inc), a diphtheria toxin interleukin-3 fusion protein which is designed to selectively bind, internalize and induce apoptosis of CD123+ primary AML cells isolated from patients.

AML blasts were obtained from leukapheresis of patients and cultured in RPMI1640 with 20% fetal bovine serum and antibiotics and supplemented with growth factors GM-CSF and SCF (10ng/ml) overnight before treatment with increasing concentrations of SL-401. CD123 expression of AML blasts was analyzed using multicolor flow cytometry before and 24 hours after treatment with SL-401. The viability of the cells before and after treatment with increasing concentrations of SL-401 was measured by annexinV-FITC and propidium iodide staining.

SL-401 induced dose-dependent (SL-401: 10, 100, 1000ng/ml) cytotoxicity in primary AML in nanomolar range (trend p = 0.0002 at 48hr), which correlated with surface expression levels of CD123. AML CD34+CD33+ blasts showed high CD123 expression levels and were susceptible to cytotoxicity with SL-401 (P<0.0001,1000ng/ml at 48hr). CD34−CD33+ committed leukemic cells were also CD123+ and responded to SL-401. To determine if SL-401 modulated CD123 levels after exposure, AML blasts pre-treated with SL-401 for 24hr were analyzed by flow cytometry for receptor expression. Importantly, SL-401 treatment after 24 hours did not down-regulate CD123 expression on primary AML blasts indicating a lack of an escape mechanism for malignant cells.

These results provide evidence that time- and dose-dependent activity of SL-401 against CD123+ AML blasts correlates with CD123 expression. Moreover, treatment does not down-regulate IL-3R expression. Clinical studies of SL-401 in patients with relapsed/refractory AML with evaluation of CD123 expression and blastic plasmacytoid dendritic cell neoplasm (BPDCN) are ongoing.

Citation Format: Rajeswaran Mani, Bhavani Gopalakrishnan, Xiaokui Mo, Chris Brooks, William Blum, Gerard Lozanski, Sumithira Vasu, Natarajan Muthusamy. SL-401 activity against AML blasts correlates with CD123 levels and does not down-regulate CD123 expression. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1691. doi:10.1158/1538-7445.AM2015-1691
Abstract 1707: PACE4-dependent cellular uptake and retention of the Multi-Leucine peptide inhibitor into cancer cells
Authors: Couture Frédéric , Ly Kevin , Levesque Christine , Kwiatkowska Anna , Desjardins Roxane , Guérin Brigitte , Day Robert
Published: August 2015

The proprotein convertase PACE4 is strongly overexpressed in prostate cancer and plays an important role in tumor progression by promoting cell proliferation and tumor angiogenesis. Because the high expression levels and the necessity of this enzyme for cancer progression, we have developed a peptide-based inhibitor known as the Multi-Leucine (ML) peptide, which is a PACE4-specific inhibitor. Just like PACE4-downregulation through stable shRNA transfections, this compound displays anti-proliferative properties when applied on cancer cells. We showed its target-specific uptake into prostate cancer xenografts by positron emission tomography, which allowed clear tumor visualization.

The ML-peptide has potent effects to block the further progression of prostate cancer cells, however, the uptake mechanism of this peptide is not clear and whether it is PACE4-dependent. We therefore tested the uptake and efflux rates of the ML-peptide peptide in prostate cancer cells and correlated these with PACE4 expression levels in wild type and stable PACE4-knockdown cell lines. In addition to prostate cancer cell lines LNCaP, DU145 and PC3, we tested HepG2, Huh7 and HT1080 cells, which also express PACE4. To measure peptide uptake, the ML peptide was modified through the addition a 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA) moiety to strongly chelate radiometal such as 64Cu. Our study shows that uptake of the ML-peptide is correlated with expression levels, but more importantly can be blocked in PACE4-knockdown cell lines, demonstrating a PACE4-dependant uptake mechanism. We also correlated uptake with the anti-proliferative effects of the ML-peptide and showed that entry into these cell lines predicted the effects of the ML-peptide on the growth of these cells.

These results confirm the notion that ML-peptide entry within cell is an important requirement to exert growth inhibition properties, and further provide a mechanism for that entry. This study demonstrates further support for the use of the ML-peptide or its analogs as potential drugs in prostate cancer, as well as any other PACE4-dependent tumors. These indications of peptide uptake within tumor could allows PACE4-status to be determined by positron emission tomography and may be of great theranostics uses in the context of pharmacological intervention with PACE4 inhibitors.

Citation Format: Frédéric Couture, Kevin Ly, Christine Levesque, Anna Kwiatkowska, Roxane Desjardins, Brigitte Guérin, Robert Day. PACE4-dependent cellular uptake and retention of the Multi-Leucine peptide inhibitor into cancer cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1707. doi:10.1158/1538-7445.AM2015-1707
Abstract 1710: Xanthohumol suppresses estrogen-signaling in endocrine resistant breast cancer through the specific inhibition of BIG3-PHB2 interactions
Authors: Katagiri Toyomasa
Published: August 2015

Estrogen-receptor-α (ERα) plays a pivotal role in the development and progression of breast cancer. The current endocrine therapies for breast cancer are mainly based on targeting ERα signaling using selective ERα modulators, ERα downregulators, and aromatase inhibitor (AI)1-3. However, tamoxifen treatment often fails, and patients succumb to recurrent, endocrine-resistant tumors. Moreover, some patients who have undergone AI treatment still relapse. The precise molecular events that determine alterations in the effectiveness of these endocrine therapies remain unknown.

We previously reported that the oncoprotein brefeldin A-inhibited guanine nucleotide-exchange protein 3 (BIG3) and tumor suppressor prohibitin 2 (PHB2) complex play a pivotal role in E2 signaling modulation in ERα-positive breast cancer. Namely, BIG3 binds PHB2, thereby inhibiting the E2-dependent suppressive ability of PHB2 and resulting in constitutive ERα activation. Considering these findings, strategies utilising the tumour suppressive activity of PHB2 upon its release from BIG3 by inhibitors of protein-protein interaction may represent novel therapies for breast cancer. Accordingly, we focus on whether XN potentially binds to the tumor suppressor protein PHB2, forming a novel natural anti-tumor compound targeting the BIG3-PHB2 complex and acting as a pivotal modulator of E2 /ERα signaling in breast cancer cells. XN treatment effectively prevented the BIG3-PHB2 interaction, thereby releasing PHB2 to directly bind to both nuclear- and cytoplasmic ERα. This event led to the complete suppression of the E2-signaling pathways and ERα-positive breast cancer cell growth both in vitro and in vivo, but did not suppress the growth of normal mammary epithelial cells. Our findings suggest that XN may be a promising natural compound to suppress the growth of luminal-type breast cancers, especially endocrine resistant breast cancers.

Citation Format: Toyomasa Katagiri. Xanthohumol suppresses estrogen-signaling in endocrine resistant breast cancer through the specific inhibition of BIG3-PHB2 interactions. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1710. doi:10.1158/1538-7445.AM2015-1710
Abstract 1711: SLC6A15, a novel potential metabolic target for non-small cell lung cancer
Authors: SRIVASTVA PALLEVI , Yoo Hyuntae
Published: August 2015

Current therapeutic targets for non-small cell lung cancer consist only of signaling pathways targeting VEGF, EGFR or ALK. Recent researches have put focus on understanding cancer metabolism (such as 2-hydroxyglutarate accumulation in gliomas with mutant IDH1) and have opened a new avenue in cancer therapeutics which can complement the current practices. Here we introduce a new potential metabolic target for non-small cell lung cancer, SLC6A15. It is a trans-membrane protein known to facilitate uptake of branched chain amino acids (BCAA) (leucine, isoleucine and valine) and proline into the cells. BCAA was recently reported to increase cellular proliferation by regulating m-TOR in normal cells like beta-pancreatic cells. Here we show that these BCAA are highly enriched in lung tumor cell lines (H1299 & H460) as well as in human lung tumor tissues as a result of overexpression of SLC6A15. We performed a transient knockdown of this transporter using siRNA for SLC6A15 and observed its effect on reduced clonogenicity of the two cancer cell lines by 3.3 and 1.6 fold, respectively. Further, gas chromatography mass spectrometry (GC-MS) analysis of the cells treated with si(SLC6A15) revealed selective reduction of BCAA levels by 1.3 to 1.5 fold but not in Proline, which can be synthesized in the cells unlike BCAA. These results support the role of these amino acids in tumor cell proliferation. Overexpression of SLC6A15 protein was confirmed by western blot analysis in 8 NSCLC cell lines and is indicative of its tumor specific nature. This finding is also supported by SLC6A15 gene expression data available on TCGA (The Cancer Genome Atlas), where 6 out of 10 patients show high expression of this gene. Therefore, we conclude that SLC6A15 is a new candidate of therapeutic target for NSCLC. Targeted inhibition may open new possibilities to specific reduction of tumor cell proliferation and possibly, increase life expectancy of patients suffering from NSCLC.

Citation Format: PALLEVI SRIVASTVA, Hyuntae Yoo. SLC6A15, a novel potential metabolic target for non-small cell lung cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1711. doi:10.1158/1538-7445.AM2015-1711
Abstract 1712: CAMKV as an immunotherapy target in MYCN-amplified neuroblastoma
Authors: Sussman Robyn T. , Huang Kevin , Raman Pichai , Maris John M.
Published: August 2015

Background: Neuroblastoma is an embryonal tumor of the developing sympathetic nervous system that accounts for 10-15% of pediatric cancer-related deaths. Although an intensive treatment regimen consisting of chemotherapy, surgery, radiation, immunotherapy and autologous stem cell transplant is used for patients with high-risk neuroblastoma, about half of these patients ultimately die. Reliable tumor-specific genomic biomarkers, such as MYCN amplification, have been identified for high-risk neuroblastoma and are used to stratify therapy. Recent clinical studies using immunotherapies to target cell surface molecules such as GD2 in neuroblastoma and CD19 in leukemia have shown impressive results, and have highlighted immunotherapy as a critical strategy in combating childhood cancer.

Methods: As part of the Stand Up 2 Cancer/St. Baldrick's Pediatric Cancer Dream Team's effort to streamline new immunotherapuetics into the clinic, we developed a computation approach to prioritize new targets for immunotherapy for high-risk neuroblastoma. Here we used MYCN as a biomarker for the most aggressive subset of neuroblastoma to define candidate cell-surface targets that are differentially expressed in MYCN-amplified neuroblastoma. We used RNAi to manipulate gene expression levels of MYCN to determine if these cell-surface targets are under the control of MYCN in neuroblastoma cells.

Results: The most significant differentially expressed cell surface molecule in high-risk neuroblastoma based on MYCN-amplification status was the calmodulin kinase-like vesicle-associated gene (CAMKV, p = 1×10-6), which encodes a protein that binds calmodulin in the presence of calcium, but lacks the kinase activity of other calmodulin kinase family members. We have confirmed that CAMKV protein is selectively expressed in MYCN-amplified neuroblastoma cell lines. From membrane fractionation and immunohistochemistry, we have verified that CAMKV is membranous in MYCN-amplified neuroblastoma cell lines and xeongraft tumors. Immunohistochemistry also revealed that CAMKV is not highly expressed in human pediatric normal tissues with the exception of neural tissues. Depletion of MYCN using RNAi resulted in a significant reduction in CAMKV RNA and protein expression (p = 2.3×10-6).

Conclusions: These results provide a template for the discovery and validation of immunotherapeutic targets in MYCN-amplified neuroblastoma and have the potential to lead to the development of a new immunotherapy targeting CAMKV.

Citation Format: Robyn T. Sussman, Kevin Huang, Pichai Raman, John M. Maris. CAMKV as an immunotherapy target in MYCN-amplified neuroblastoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1712. doi:10.1158/1538-7445.AM2015-1712
Abstract 1714: The role and mechanism of JAB1 as a therapeutic target in biliary tract cancer
Authors: Nam Ah-Rong , Kang Kyo Hwa , Park Ji Eun , Bang Ju-Hee , Jin Ling , Jin Mei Hua , Kim Tae Yong , Han Sae-Won , Song Sang-Hyun , Im Seock-Ah , Kim Tae-You , Oh Do-Youn , Bang Yung-Jue
Published: August 2015

Background: JAB1 (c-Jun activation domain binding protein-1) is a c-Jun coactivator, also known as COP9 signalosome subunit 5 (CSN5). Jab1 has an important role in cell proliferation and apoptosis. Overexpression of Jab1 is correlated with poor prognosis in various cancers. Biliary tract cancer (BTC) has a poor prognosis with a huge unmet medical need. The role of Jab1 has not been studied in BTC. We investigated the role and mechanism of Jab1 as a potential therapeutic target in BTC.

Methods: We used 8 kinds of BTC cells and designed Jab1 siRNA. MTT assay and colony formation assay were done to determine growth inhibitory effect of Jab1 knockdown. Cell cycle analysis was done by FACS Calibur flow cytometer and cell migration was evaluated by wound healing assay. We used cycloheximide chase assay for measuring of protein half-life.

Results: BTC cell lines showed high level of Jab1 expression. Among them, SNU478, SNU869 and SNU 1196 were indicated with especially higher level of Jab1 expression. Cell growth and proliferation of BTC cells were decreased by Jab1 knockdown. Depletion of Jab1 induced G1 arrest, as well as decreased CyclinD/CyclinA and increased p27. Cell migration was also inhibited by Jab1 knockdown. Inhibition of Jab1 showed the decrease of pSrc, pAkt. Interestingly, depletion of Jab1 led to the elevation of PTEN protein levels without change of PTEN mRNA levels. PTEN half-life was longer in Jab1 siRNA-transfected cells. Suppression of Jab1 increased the sensitivity of BTC cells to the cytotoxic chemotherapeutic agent.

Conclusion: Suppression of Jab1 shows antitumor effects in BTC cells by inhibiting cell proliferation, migration, cell cycle and increase of chemosensitivity. Taken together, Jab1 might be a potential therapeutic target in BTC.

Citation Format: Ah-Rong Nam, Kyo Hwa Kang, Ji Eun Park, Ju-Hee Bang, Ling Jin, Mei Hua Jin, Tae Yong Kim, Sae-Won Han, Sang-Hyun Song, Seock-Ah Im, Tae-You Kim, Do-Youn Oh, Yung-Jue Bang. The role and mechanism of JAB1 as a therapeutic target in biliary tract cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1714. doi:10.1158/1538-7445.AM2015-1714
Abstract 1715: IRES-transactivating factors (ITAFs) as therapeutic targets in breast and colorectal cancer
Authors: Ward Toby M. , Bockhorn Jessica , Harrell J C. , Liu Xiaofei , Pegram Mark D.
Published: August 2015

Background: The majority of cellular mRNAs are translated in a cap-dependent manner, via ribosome assembly at 5′ methylguanosine caps followed by initiation at a nearby favorable AUG codon. Alternatively, internal ribosome entry sites (IRESs) facilitate translation in a cap-independent manner, particularly during cell stress, and have been identified in mRNAs encoding many canonical cellular oncogenes, including c-MYC, c-SRC, XIAP, Aurora-A kinase, and IGF1R. IRES trans-activating factors (ITAFs) are accessory proteins that augment or inhibit translation of client IRES-containing mRNAs, and were found to be upregulated in many cancer types including breast and colorectal cancers.

Methods and Results: Analysis of microarray datasets from cancer and normal tissue specimens (The Cancer Genome Atlas, METABRIC, and University of North Carolina datasets) revealed that expression of several ITAFs is upregulated in malignant tissues compared to normal controls. mRNAs encoding ITAFs were significantly upregulated (p < 0.05) in cancer specimens, and ITAF protein levels were increased as measured by immunohistochemistry on tissue microarrays made up of malignant and healthy tissue specimens.

To investigate biological impact of ITAF silencing, gene knockdown was accomplished using pGIPZ and pTRIPZ lentiviral shRNA vectors. Silencing of ITAFs and concomitant reduction in protein levels (measured by western blot) led to significant proliferation impairment in MDA-MB-231, MCF7 and DLD1 cancer cells in 2-D culture (as measured by counting assays, p<0.05) as well as in soft agar assays (p< 0.05). In cancer cell lines, treatment with TNF-related apoptosis inducing ligand (TRAIL) led relocalization of ITAFs from cell nuclei to the endoplasmic reticulum (assayed by immunofluorescence), with increased translation of client IRES-harboring mRNAs encoding BAG-1 and c-MYC oncoproteins, despite impaired global cap-dependent translation.

Conclusions and future directions: Future experiments will include expanded investigation of protein-level ITAF expression in archival human specimens using immunohistochemistry, as well as investigation of the effects of ITAF silencing in cancer cell xenografts in immunocompromised mice. Based on their increased expression in malignant tissues and activity in translation of oncoproteins, cellular ITAF genes may represent novel therapeutic targets for intervention.

Citation Format: Toby M. Ward, Jessica Bockhorn, J C. Harrell, Xiaofei Liu, Mark D. Pegram. IRES-transactivating factors (ITAFs) as therapeutic targets in breast and colorectal cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1715. doi:10.1158/1538-7445.AM2015-1715
Abstract 1749: Bortezomib and JQ1 hold promise for human and canine osteosarcoma
Authors: Yang Yating , Yuzbasiyan-Gurkan Vilma
Published: August 2015

Osteosarcoma (OS) is the most common primary malignant bone tumor of both humans and dogs. Despite advances in treatment of other cancers, the clinical outcome of OS remains poor for the past decades. Therefore, there is a great need to identify novel and effective treatments for osteosarcoma. Several classes of novel small molecules with anti-tumor activities have become available in the last few years. Among these are tyrosine kinase inhibitors (TKIs) which are able to inhibit kinase activity of many receptor tyrosine kinases. Another class of molecules target bromodomains which modify chromatin and block transcription of key factors, another class of molecules target inhibition of the proteasome which induces cell death through various mechanisms. Previous findings from our lab identified two TKIs (sunitinib, sorafenib), a bromodomain inhibitor (JQ1), and a proteasome inhibitor (bortezomib) as having good growth inhibition effects in low concentrations on three canine OS cell lines (D17, Abrams, and Gracie) and two human OS cell lines (SAOS2 and U2OS) in a series of drug screening assay. Among these candidates, we hypothesized that bortezomib and JQ1 would be effective against canine and human OS cell lines. We then determined the exact IC50 values of these two drugs, the concentration capable of inhibiting proliferation by 50%. The IC50 values of bortezomib varied in cells from 0.01 to 0.22 μM and JQ1 from 0.5 μM to 12.5 μM, which all within achievable plasma concentration and tolerable levels. In addition, these two drugs shown to decrease cell migration ability in wound healing assay as well as invasion ability in matrigel assay. With the promising results from in vitro studies, we further exam JQ1 as therapeutic strategy in an orthotopic tumor xenograft mouse model. However, the evaluation of JQ1 efficiency need to be evaluated before human and canine OS clinical trials.

Citation Format: Yating Yang, Vilma Yuzbasiyan-Gurkan. Bortezomib and JQ1 hold promise for human and canine osteosarcoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1749. doi:10.1158/1538-7445.AM2015-1749
Abstract 1780: Targeted inactivation of CRL4-CDT2 E3 ubiquitin ligase as a novel therapeutic strategy for malignant melanoma
Authors: Benamar Mouadh , Guessous Fadila , Corbett Patrick , Slingluff Craig L. , Abbas Tarek
Published: August 2015

The CRL4CDT2 E3 ubiquitin ligase is emerging as a critical regulator cell cycle progression, primarily through the targeted ubiquitylation and degradation of critical cell cycle regulators including CDT1, p21 and SET8 via the 26S proteasome. CDT2, the substrate receptor of the CRL4CDT2 E3 ligase is overexpressed or amplified in several human malignancies, but its role in the development or progression of these cancers remains elusive. We show that CDT2 is overexpressed in primary and metastatic melanoma. Targeted depletion of CDT2 by si-RNA or its pharmacological inhibition by MLN4924, a selective NEDD8-activating enzyme (NAE) inhibitor, suppressed the proliferation of melanoma cancer cells in vitro and inhibited tumor growth of human melanoma xenografts in mice, irrespective of the BRAF mutation status. Inhibition of CRL4CDT2 in melanoma cells with various genetic backgrounds, but not in immortalized human melanocytes, resulted in robust p21- and SET8-dependent rereplication. These results identify the CRL4CDT2 E3 ubiquitin ligase as a promising new therapeutic target for cutaneous melanoma, and demonstrate the efficacy of MLN4924 as a promising anti-melanoma agent.

Citation Format: Mouadh Benamar, Fadila Guessous, Patrick Corbett, Craig L. Slingluff, Tarek Abbas. Targeted inactivation of CRL4-CDT2 E3 ubiquitin ligase as a novel therapeutic strategy for malignant melanoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1780. doi:10.1158/1538-7445.AM2015-1780
Abstract 1792: Radiation-induced downregulation of GLT-1 glutamate transporter mRNA expression is reversed by renin-angiotensin system inhibitors
Authors: Kooshki Mitra , Naczki Christine , Robbins Michael E. , Metheny-Barlow Linda J.
Published: August 2015

Fractionated partial or whole brain irradiation (fWBI) is often required to treat metastatic brain cancer. Unfortunately, fWBI can lead to the development of a progressive, irreversible cognitive impairment in patients who survive longer than six months following treatment. Prior studies suggest that neuroinflammation and alterations in neuronal plasticity may play a role in this treatment-related side effect. In the current study we determined that radiation-induced inflammatory activation of primary rat cortical astrocytes results in downregulation of mRNA and protein for the GLT-1 glutamate transporter, which functions to regulate glutamate signaling at excitatory synapses. Furthermore, we show that delivery of clinically-relevant fWBI (40Gy, 5Gy/fraction, 2x/week) to young adult male rats results in 51% decrease in Glt-1 mRNA in the cortex at 48 hours. Two months after completion of fWBI, cortical GLT-1 mRNA is further decreased to 25% the levels observed in sham controls, while at six months following fWBI GLT-1 levels remain 43% decreased compared to controls. We have previously identified two classes of compounds, the renin-angiotensin system inhibitors and PPAR agonists, which ameliorate the fWBI-induced cognitive impairment in this rodent model, although the full mechanisms by which these compounds protect cognition remain unknown. We found that dietary administration of the angiotensin receptor blocker L-158-809, but not the ACE inhibitor Ramipril, prevents fWBI-induced loss of cortical GLT-1 mRNA at 48 hours, while both cognition-sparing compounds restored GLT-1 expression at 2 months post-fWBI. Similarly, administration of the PPARδ agonist GW0742 prevented the fWBI-induced decrease of GLT-1 mRNA observed 6 months post-fWBI. Together these data suggest that radiation-induced downregulation of GLT-1 is a potential mechanism contributing to the development of radiation-induced brain injury, including cognitive impairment, and that GLT-1 may represent a druggable target to prevent this treatment-related cognitive decline. (Supported by NIH CA112593)

Citation Format: Mitra Kooshki, Christine Naczki, Michael E. Robbins, Linda J. Metheny-Barlow. Radiation-induced downregulation of GLT-1 glutamate transporter mRNA expression is reversed by renin-angiotensin system inhibitors. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1792. doi:10.1158/1538-7445.AM2015-1792
Abstract 1805: DUOX2: The key player for hyper-radiosensitivity in gastric cancer cells with low dose fractionation radio therapy (LDFRT)
Authors: Nguyan Duc , Parekh Palak R. , Chang Elizabeth T. , Sharma Navesh , Carrier France
Published: August 2015

The most conventional therapy for solid tumors is radiotherapy. Still there is a challenge when it comes to managing highly disseminated gastrointestinal cancers because of the toxicity issues. Earlier, low dose fractionated radiotherapy (LDFRT) was considered to be ineffective for tumor removal, rather believed to cause cancer. There are few improvements with the use of novel approach in few cases of solid malignancy. This paradigm exploits the advantage of LDFRT with the combination of chemotherapy. Because now it's known that LDFRT causes hyper-radiosensitivity (HRS). The aim of our study is to determine the efficacy of LDFRT in combination with modified regimen of docetaxel, cisplatin and 5′-fluorouracil (mDCF) and their mechanism of action in gastric cancer cells. Our data indicate that consecutive three days radiation with daily fraction of 0.15 Gy produced HRS in gastric cancer cells and potentiated the effect of chemotherapeutic drugs (mDCF). Colony survival assays showed only 10% of gastric cancer cell survival when LDFRT was combined with mDCF while ten times higher dose (1.35 Gy) of radiation alone was required to achieve the same results. RT2-PCR profiler array analysis showed marked upregulation of dual oxidase-2 (DUOX2), an enzyme of NADPH- oxidase family, without inducing the genes involved in DNA repair. This was evident from 3.5 fold increase in reactive oxygen species (ROS) in cells exposed to LDFRT and mDCF. Further, this was confirmed by increase in cancer cell survival hence radioresistance with downregulation of DUOX2 expression. In addition to this inhibition of DUOX2 activity also abrogated the efficacy of the combination therapy. Taken together these data suggest that chemopotentiation by LDFRT in gastric cancer cells may be partly due to increased ROS production by DUOX2 without inducing the DNA repair machinery. These data thus provide a rationale for further explorations of potential clinical applications of LDFRT as a chemopotentiator for advanced and metastatic gastric cancers. (* = Equally Contributed)

Citation Format: Duc Nguyan, Palak R. Parekh, Elizabeth T. Chang, Navesh Sharma, France Carrier. DUOX2: The key player for hyper-radiosensitivity in gastric cancer cells with low dose fractionation radio therapy (LDFRT). [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1805. doi:10.1158/1538-7445.AM2015-1805
Abstract 1823: A novel web interface to facilitate pathology directed mass spectrometry
Authors: Seeley Erin H. , Dunkerley R. Ryan , Moreira Andre L. , Downey Robert J. , Kilby Greg W.
Published: August 2015

Clinical proteomics studies often involve multiple investigators located at different physical locations necessitating a centralized platform for joint access to data and other information pertinent to the study. To help communicate pathology targets of interests from remote researchers to the on-site technical team, the Pathology Directed Mass Spectrometry (PDMS) web application was developed. PDMS was designed to be a secure, online digital pathology platform specifically configured for down-stream mass spectrometry imaging (MSI) analysis. Hosting ultra-high resolution digital microscopy images of stained serial tissue sections, PDMS allows researchers to zoom into their samples to review and annotate specific areas for analysis. Once finalized, annotated images are exported from PDMS for MSI targeting. Built for HTML5 browsers, PDMS was deployed in a secure LAMP (Linux, Apache, MySQL, PHP) environment, utilizing a Javascript visualization engine and server-side conversion tools.

In a pathology directed mass spectrometry profiling experiment, two sections of a tissue specimen are collected, one on a mass spectrometry target and one that is stained for histological evaluation. A pathologist annotates a digital microscopy image to indicate areas of interest for mass spectral analysis. The annotated image is then merged with an image of the unstained section and after appropriate sample preparation, mass spectra are collected from the annotated areas. A major advantage to this type of analysis is that it is high throughput and is conducive to downstream biostatistical analysis for biomarker discovery.

The PDMS software has been successfully used in the molecular differentiation of histological patterns in stage I pulmonary carcinomas as part of a joint collaboration with Memorial Sloan Kettering Cancer Center. Image annotation was carried out remotely prior to mass spectral analysis in house. Statistical analyses of the mass spectral data resulted in over 85% classification accuracies in differentiating histological tumor subtypes within individual pulmonary carcinoma patient samples.

Citation Format: Erin H. Seeley, R. Ryan Dunkerley, Andre L. Moreira, Robert J. Downey, Greg W. Kilby. A novel web interface to facilitate pathology directed mass spectrometry. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1823. doi:10.1158/1538-7445.AM2015-1823
Abstract 1832: Quantification of metabolic changes by magnetic resonance and mechanism of cell death in ovarian cancer
Authors: Belkacemi Louiza
Published: August 2015

Quantification of metabolic changes by magnetic resonance and mechanism of cell death in ovarian cancer Louiza Belkacemia*, Leila Maurmannb and Rathindra N Bosea

aDepartment of Biology and Biochemistry, University of Houston, Houston, TX 77204

bDepartment of Chemistry, Kansas State University, Manhattan, KS 6650

Ovarian cancer is the most frequent cause of death from gynecologic malignancy. Presently most ovarian cancers are diagnosed when the disease has progressed to stage III or IV. Therefore there is an urgent need for the identification of new biomarkers for early diagnosis and drug efficacy. Changes in metabolite concentrations have been associated with tumor regression or progression. Nuclear magnetic resonance (NMR) has been used to identify molecular markers during cell death (apoptosis), especially the onset of membrane disintegration. Cisplatin is the most commonly used drug in chemotherapy of ovarian cancer. To date, most research has been focused on the cisplatin-DNA reaction. Thus it is essential to evaluate the effect of cisplatin on binding to other biomolecules in the cell. Chinese hamster ovarian (CHO) cells, which, like resistant ovarian cancer cells, require high cisplatin concentrations to induce cell death, were treated with 50 μM cisplatin at different time intervals. Metabolite concentrations were determined by one and two dimensional 31P and 1H NMR. Moreover, the A2780 cisplatin-sensitive cell line and its resistant variant were evaluated for the mechanism of cell death. Analysis revealed that enhanced apoptosis was associated with significantly increased phosphocholine and glycerophosphocholine metabolites and marked elevation of cellular phosphocreatine and nucleoside triphosphate concentrations at 6 hrs and decreased thereafter. Plasma membrane alterations were paralleled by an upregulation of expression of Fas and related proapoptotic Bax and PUMA genes, and Fas and caspase-3 and -9 proteins. These findings suggest that NMR may be used as a tool to monitor tumors and drug efficacy in humans using for instance tissue biopsies. Further, NMR measurements of cell extracts may provide significant insight on pathways to apoptosis.

Citation Format: Louiza Belkacemi. Quantification of metabolic changes by magnetic resonance and mechanism of cell death in ovarian cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1832. doi:10.1158/1538-7445.AM2015-1832
Abstract 1872: Plasma carotenoids and breast cancer risk in the Cancer Prevention Study II Nutrition Cohort
Authors: Wang Ying , Gapstur Susan M. , Gaudet Mia M. , Furtado Jeremy D. , Campos Hannia , McCullough Marjorie L.
Published: August 2015

Background: Several plasma carotenoids have been inversely associated with postmenopausal breast cancer risk in large cohort studies and a pooled analysis. Whether associations differ by tumor or participant characteristics is unclear. We investigated the associations of plasma carotenoids with postmenopausal breast cancer risk overall and by estrogen receptor (ER) status, tumor stage, smoking status and body mass index in a case-control study nested in the Cancer Prevention Study II Nutrition Cohort.

Methods: A total of 561 breast cancer cases diagnosed between blood draw in 1998-2001 and June 30, 2007 and matched 1:1 with controls on race, birth date and blood draw date were included. Multivariable-adjusted conditional and unconditional logistic regression models were used to calculate odds ratios (ORs) and 95% confidence intervals (CIs).

Results: Plasma α-carotene was inversely associated with breast cancer risk even after adjustment for total fruit and vegetable intake (4th vs. 1st quartile OR = 0.58, 95% CI: 0.40, 0.85, P-trend = 0.03). This inverse association was stronger among ever smokers (P-interaction<0.05), and obese women (P-interaction = 0.02), but did not differ by tumor stage (P-heterogeneity = 0.61). Although there was an inverse trend between α-carotene and ER+ (P-trend = 0.01) but not ER- breast cancer (P-trend = 0.99), the test for heterogeneity by ER status was not statistically significant (P-heterogeneity = 0.28). There was some evidence of an inverse association with total carotenoids but this association was nonlinear.

Conclusions: These results suggest that higher plasma α-carotene is associated with lower risk of breast cancer, and this association was even stronger in smokers and obese women.

Citation Format: Ying Wang, Susan M. Gapstur, Mia M. Gaudet, Jeremy D. Furtado, Hannia Campos, Marjorie L. McCullough. Plasma carotenoids and breast cancer risk in the Cancer Prevention Study II Nutrition Cohort. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1872. doi:10.1158/1538-7445.AM2015-1872
Abstract 1876: Adolescent and mid-life diet: risk of pancreatic cancer in the NIH-AARP Diet and Health Study
Authors: Moy Kristin A. , Potischman Nancy , Thompson Frances E. , Subar Amy , Ruder Elizabeth H. , Thiebaut Anne C.M. , Stolzenberg-Solomon Rachael Z.
Published: August 2015

Background: To date, the data concerning diet and pancreatic cancer risk has focused on diet assessed during adulthood. Given the long latency period of this malignancy, examining diet during adolescence and midlife may shed light on the diet - pancreatic cancer association. Methods: We examined the associations between diet at ages 12-13 years, 10 years before baseline (ages 40-61 years), and the joint effects of both with risk of pancreatic cancer in the NIH-AARP Diet and Health Study. Six months after baseline, 303,094 individuals (175,991 men and 127,103 women) completed two 37-item food frequency questionnaires that assessed diet at ages 12-13 years and 10 years previously. Cox proportional hazards regression was used to estimate smoking, body mass index, and diabetes adjusted multivariable hazard ratios (HRs) and 95% confidence intervals (CIs) for pancreatic cancers (n = 1,322 by the end of 2006). Results: Comparing highest to the lowest tertile (T3 vs. T1), carbohydrate intake at age 12-13 (HR (95% CI): 0.86 (0.75, 0.98), P-trend = 0.06) and 10 years before baseline (HR (95% CI): 0.87 (0.76, 1.00), P-trend = 0.05) were inversely associated with pancreatic cancer. Intake of total fat at ages 12-13 (HR (95% CI): 1.15 (1.00, 1.33), P-trend = 0.07) and 10 years before baseline (HR (95% CI): 1.18 (1.03, 1.35), P-trend = 0.02) were positively associated with risk. High intake of fat in both adolescence and midlife was positively associated with risk (HR (95% CI): 1.32 (1.08, 1.62)). Calcium intake 10 years before baseline was associated with reduced risk (T3 vs T1 HR (95% CI): 0.88 (0.77, 1.00), P-trend = 0.05) as was changing from low intake in adolescence to high intake in midlife (HR (95% CI): 0.71 (0.54, 0.93)). Solid fat (butter and margarine) intake 10 years before baseline was associated with increased risk (T3 vs T1 HR (95% CI): 1.23 (1.06, 1.42), P-trend = 0.005) as was consumption of solid fats at both time points (HR (95% CI): 1.31 (1.08, 1.60)). Conclusion: Diet during adolescence and midlife may play a role in the etiology of pancreatic cancer.

Citation Format: Kristin A. Moy, Nancy Potischman, Frances E. Thompson, Amy Subar, Elizabeth H. Ruder, Anne C.M. Thiebaut, Rachael Z. Stolzenberg-Solomon. Adolescent and mid-life diet: risk of pancreatic cancer in the NIH-AARP Diet and Health Study. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1876. doi:10.1158/1538-7445.AM2015-1876
Abstract 1889: Paleolithic and Mediterranean diet pattern scores and their associations with biomarkers of inflammation and oxidative balance
Authors: Whalen Kristine , McCullough Marji , Flanders W. Dana , Hartman Terryl J. , Judd Suzanne , Bostick Roberd M.
Published: August 2015

Chronic inflammation is associated with poor diet quality as well as risk for cancer and other chronic diseases. A diet-inflammation association may relate to evolutionary discordance. We investigated associations of two diet pattern scores, the “Paleolithic” and the Mediterranean, with circulating levels of two inflammation-related biomarkers, C-reactive protein (CRP), an acute inflammatory protein, and F2-isoprostanes, a reliable marker of lipid peroxidation in vivo, in a colonoscopy-based, pooled case-control study of incident sporadic colorectal adenomas (N = 646). We created diet scores from responses on a modified Willett food-frequency questionnaire, and measured plasma levels of CRP and F2-isoprostanes by ELISA and GCMS, respectively. Both diet scores were calculated and categorized into quintiles, and their associations with the biomarker levels were estimated using unconditional logistic regression. The multivariable-adjusted odds ratios (OR) comparing those in the highest relative to those in the lowest quintiles of the Paleolithic and Mediterranean diet scores, were, respectively, 0.73 (95% confidence interval [CI] 0.39, 1.36; Ptrend = 0.35) and 0.71 (95% CI 0.36, 1.40; Ptrend = 0.12) for CRP levels, and 0.35 (95% CI 0.17, 0.74; Ptrend <0.01) and 0.64 (95% CI 0.29, 1.40; Ptrend = 0.03) for F2-isoprostanes levels. These findings, which are particularly strong for the inverse Paleolithic diet score-F2-isoprostanes association, suggest that diets that are more “Paleolithic-” or Mediterranean-like may be associated with lower levels of systemic inflammation and oxidative stress.

Citation Format: Kristine Whalen, Marji McCullough, W. Dana Flanders, Terryl J. Hartman, Suzanne Judd, Roberd M. Bostick. Paleolithic and Mediterranean diet pattern scores and their associations with biomarkers of inflammation and oxidative balance. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1889. doi:10.1158/1538-7445.AM2015-1889
Abstract 1925: Streamlined analysis and interpretation of RNA editing variants from melanoma cancer samples
Authors: Joecker Anika , Yip Rupert , Arens Anne , Oester Bodil , Basset Douglas , Macy Bryant
Published: August 2015

Of special interest in a clinical setting is the identification and interpretation of somatic variants from the DNA of a tumor/normal matched pair.

Studies have shown that a large proportion of variants present in DNA are not present in the RNA and vice versa due to RNA-Editing and other unknown molecular processes1. In addition, recent publications show that RNA-editing plays an important role in cancer2,3. These findings can have strong implications for clinical diagnostics where the correct identification and interpretation of somatic variants from tumor/normal matched samples is essential.

We took a published Whole Trancriptome/Whole Exome Sequencing dataset of tumor and peripheral blood from two patients with uveal melanomas5. We looked for somatic variants present in the RNA, but absent in the DNA using CLC Cancer Research Workbench. A strong mutation signature for C-T, A-G changes was found, which result from Adenosine to Inosine changes, a typical RNA-Editing event, which was recently published to be common in human and is mediated by ADAR proteins.

By running Ingenuity Variant Analysis directly from CLC Cancer Research Workbench we found RNA-Editing found in these two patients is highly associated with cancer and more specifically with Melanoma directly. Some of the variants directly associated with Melanoma were classified as likely pathogenic and were not described in COSMIC, but present in Ingenuity's large Knowledge Base.

1) Widespread RNA and DNA Sequence Differences in the Human Transcriptome. Mingyao Li, Isabel X. Wang, Yun Li, Alan Bruzel, Allison L. Richards, Jonathan M. Toung, and Vivian G. Cheung. Science 1 July 2011: 333 (6038), 53-58.

2) RNA editing in RHOQ promotes invasion potential in colorectal cancer. Sae-Won Han et al. J Exp Med 2014 211:613-621.

3) A disrupted RNA editing balance mediated by ADARs (Adenosine DeAminases that act on RNA) in human hepatocellular carcinoma. Tim Hon Man Chan et al.Gut 2014;63:5 832-843

4) A-to-I RNA editing occurs at over a hundred million genomic sites, located in a majority of human genes.Genome Res. March 2014 24: 365-376

5) Harbour JW et al., “Frequent mutation of BAP1 in metastasizing uveal melanomas.”, Science, 2010 Nov 4;330(6009)

Citation Format: Anika Joecker, Rupert Yip, Anne Arens, Bodil Oester, Douglas Basset, Bryant Macy. Streamlined analysis and interpretation of RNA editing variants from melanoma cancer samples. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1925. doi:10.1158/1538-7445.AM2015-1925
Abstract 1927: Evolutionary novel genes expressed in fish tumors determine progressive evolutionary characters
Authors: Matyunina Ekaterina , Emelyanov Alexander , Kozlov Andrei
Published: August 2015

The origin of evolutionary novel genes is connected with the origin and evolution of the novel organismal functions and the increase in morphological complexity of multicellular organisms. Novel gene acquire new or altered functions. Here we describe that evolutionary novel genes, expressed predominantly in transgenic fish tumors after their induced regression, determine progressive evolutionary characters and are conserved in human.

We used a sample of genes, which were activated in inducible krasV12 transgenic zebrafish tumors according to the results of RNASeq data. Genes from our sample were expressed after HCC regression upon kras inactivation. The search of orthologs by different tools, such as blastx and psiblast, with e-value cut off not less than 10-3 and query coverage more than 50%, discovered that considerable proportion of tumor-specifically expressed genes are evolutionary novel, i.e. their orthologs are not found in Lamprey. 63 of these novel genes have orthologs in humans. Gene ontology (GO) data were used to analyze their functions. According to the results of GO some of the genes have functions not characteristic to fish. Among known functions of the human orthologues of evolutionary novel genes of zebrafish (vs. Lamprey) there are some important morphogenetic functions. For example, genes ccdc40, lmx1ba and lepa acquired functions in lung, embryonic utera and placenta, i.e. organs originated in taxa higher than fish. A large proportion of genes from our sample are connected with retina type eye, heart and brain development.

Citation Format: Ekaterina Matyunina, Alexander Emelyanov, Andrei Kozlov. Evolutionary novel genes expressed in fish tumors determine progressive evolutionary characters. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1927. doi:10.1158/1538-7445.AM2015-1927