bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4222: Targeting HMGA2 suppresses GBM stemness, invasion and tumorigenicity
Authors: Kaur Harpreet , Hütt-Cabezas Marianne , Taylor Isabella , Asnaghi Laura , Rodriguez Fausto , Orr Brent A. , Eberhart Charles G. , Raabe Eric H.
Published: August 2015

Glioblastoma Multiforme (GBM) causes the majority of brain tumor-related deaths in the United States. The high invasiveness of GBM prevents complete surgical resection, allowing tumor recurrence. The lack of curative treatment highlights a dire need to develop specific targeted therapeutics. Identifying and targeting novel molecular markers in glioma stem-like cells (GSCs) thought to be critical for tumor invasion and recurrence could lead to development of better therapies. HMGA2, a non-histone transcriptional modulator, is a regulator of normal and cancer stem cells. The significance of HMGA2 as a potential therapeutic target in GBM has not been investigated. We found increased HMGA2 expression in a subset of primary human GBM tumors, in multiple patient-derived GBM neurosphere and adherent cell lines, and in the mesenchymal subgroup of GBM in The Cancer Genome Atlas (TCGA) database. Lentiviral short hairpin RNA (shRNA)-lowered HMGA2 protein levels and significantly reduced invasion and clonogenicity of GBM cell lines in transwell invasion and soft agar colony formation assays. Pharmacological inhibition of HMGA proteins using the DNA minor-groove binding drug Netropsin significantly inhibited growth of the U-87 MG GBM cell line (P<0.01) in MTS assays. Mice bearing orthotopic GBM neurosphere xenograft lines transduced with HMGA2 shRNA lived longer (median survival = 108 days) compared to control shRNA (median survival = 67.5 days) (P<0.0001 by log-rank analysis). Conversely, lentiviral-mediated overexpression of HMGA2 in GBM cell lines increased invasion, clonogenicity and resistance to radiation in vitro (P<0.01). Overexpression of HMGA2 also promoted GBM tumor formation in vivo in orthotopic xenografts. To elucidate the molecular mechanism by which HMGA2 promotes invasion and stemness in GBM, we performed mRNA expression profiling on GBM neurosphere lines lentivirally transduced to express HMGA2 or empty vector control. We found increased CD44 expression in HMGA2-overexpressing GBM neurosphere lines compared to control, and confirmed this result by western blotting. Together, our results suggest an oncogenic role of HMGA2 in promoting GBM tumorigenicity by increasing invasiveness and a stem cell-like state, and show that HMGA2 is a potential therapeutic target in GBM.

Citation Format: Harpreet Kaur, Marianne Hütt-Cabezas, Isabella Taylor, Laura Asnaghi, Fausto Rodriguez, Brent A. Orr, Charles G. Eberhart, Eric H. Raabe. Targeting HMGA2 suppresses GBM stemness, invasion and tumorigenicity. [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 4222. doi:10.1158/1538-7445.AM2015-4222
Abstract 4242: Metformin suppresses GRP78-dependent PI3-Kinase activity in clonogenic side population to enhance the anti-myeloma benefit of bortezomib
Authors: Driscoll James J. , Jagannathan Sajjeev , Abdel Malek Mohamed AY , Vad Nikhil , Malek Ehsan
Published: August 2015

Multiple myeloma (MM) patients invariably relapse through mechanisms that remain elusive. Disease recurrence after treatment suggests that rare, tumor-initiating subpopulations with chemoresistant phenotypes persist or emerge to propagate and mediate tumor regrowth. Stem cell-like side population (SP) cells were isolated from myeloma cell lines and patient samples and exhibited resistance to proteasome inhibitors relative to the main population (MP). SP cell-based high-throughput screening detected pharmacologics that enhanced the anti-proliferative effect of bortezomib and revealed the anti-diabetic agent metformin. Bortezomib treatment increased levels of the molecular chaperone glucose-regulated protein GRP78 but metformin co-treatment suppressed bortezomib-induced GRP78 upregulation. Bortezomib treatment also promoted GRP78 co-localization with the signal transducer phosphatidylinositol-3-kinase (PI3K) that activates cell growth and survival pathways. SP cells displayed greater levels of the PI3K product phosphatidylinositol (3,4,5)-trisphosphate (PIP3) and enhanced PI3K-mediated phosphorylation of protein kinase B/AKT compared to MP cells. Metformin treatment or knockdown of GRP78-encoding HSPA5 enhanced sensitivity to bortezomib preferentially in SP cells. Tumor growth was significantly delayed and overall survival prolonged after metformin co-treatment with bortezomib in mice injected with parental or SP cells. Taken together, the results support the pharmacologic repositioning of metformin to inhibit GRP78 induction in therapy-resistant cells and enhance the anti-myeloma benefit of bortezomib.

Note: This abstract was not presented at the meeting.

Citation Format: James J. Driscoll, Sajjeev Jagannathan, Mohamed AY Abdel Malek, Nikhil Vad, Ehsan Malek. Metformin suppresses GRP78-dependent PI3-Kinase activity in clonogenic side population to enhance the anti-myeloma benefit of bortezomib. [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 4242. doi:10.1158/1538-7445.AM2015-4242
Abstract 4249: High-Frequency of TMPRSS2-ERG fusion gene identifies a non-random androgen-driven event of chromosomal instability in BRCA mutated prostate cancer
Authors: Castro Elena , Van de Poll Floortje , Piñeiro Elena , Nombela Paz , Bancroft Elizabeth , Govindasami Koveela , Guy Michelle , Kovacs G , Ellis Steve , Antoniu Antonis , Easton Douglas , Kote-Jarai Zsofia , Al-Sharour Fatima , Olmos David , Eeles Rosalind
Published: August 2015

INTRODUCTION:

Germline BRCA mutations are an independent prognostic factor for PrCa prognosis and treatment outcomes. BRCA tumours are associated with chromosomal instability due to underlying DNA repair deficiency; however, recurrent patterns of genomic aberrations in this context have not been explored.

METHODS:

We collected PrCa formalin-fixed paraffin-embedded samples from diagnostic biopsies and/or prostatectomies from 5 BRCA1 and 24 BRCA2 germline mutation carriers, which were matched with at least 1 sporadic PrCa (ie PrCa with no germline mutation) by stage and treatment. All samples were reviewed by an expert pathologist, and the tumour relevant areas were selected for further analyses. Recurrent chromosomal abnormalities in PrCa were also analyzed using previously described fluorescence in situ hybridization (FISH) “break-apart” and/or “fusion” assays, including a novel FISH “combined break-fusion” assay for TMPRSS2-ERG designed with probes flanking 3´and 5´of ERG and 5´of TMPRSS2. Genomic abnormalities were confirmed by a second method (qPCR or IHC) whenever possible.

RESULTS:

No differences in Gleason score between mutation carriers and non-carriers were seen (p = 0.134). Neuroendocrine phenotype was not identified in any sample. The “break-apart” assay identified interstitial deletion of genomic sequences between TMPRSS2 and ERG in all BRCA mutated tumours in at least one focus (100%), compared with 45% of non-carriers (P<0.001). The novel “break-fusion” assay allowed us to identify TMPRSS2 as the fusion partner in all cases, excluding other potential partners previously described. ETV1 rearrangements were identified in 3.5% of carriers and in 4% of non-carriers (P = 0.662).

CONCLUSIONS:

The presence of TMPRRSS2-ERG fusion gene in all BRCA mutated tumours in our series identifies a non-random androgen-driven event on the background of high chromosomal instability. Our findings suggests that BRCA mutated PrCa is androgen-dependent which may have important implications for the management of these patients.

Citation Format: Elena Castro, Floortje Van de Poll, Elena Piñeiro, Paz Nombela, Elizabeth Bancroft, Koveela Govindasami, Michelle Guy, G Kovacs, Steve Ellis, Antonis Antoniu, Douglas Easton, Zsofia Kote-Jarai, Fatima Al-Sharour, David Olmos, Rosalind Eeles. High-Frequency of TMPRSS2-ERG fusion gene identifies a non-random androgen-driven event of chromosomal instability in BRCA mutated prostate cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4249. doi:10.1158/1538-7445.AM2015-4249
Abstract 4280: Utilizing a selective agonist of the intermediate-affinity IL-2 receptor with an improved pharmacokinetic profile leads to an enhanced immunostimulatory response With reduced toxicity in mice
Authors: Losey Heather C. , Lopes Jared E. , Dean Reginald L. , Flick Heather , Gomes Madison , Huff Michael R. , Moroso Rosemarie A. , Sun Lei , Wang Chunhua , Waters Julie F. , Alvarez Juan C.
Published: August 2015

RDB 1450 is an engineered fusion protein which selectively activates the intermediate-affinity IL-2 receptor present on NK cells and memory CD8 T cells. In rodents, RDB 1450 has a half-life of 6-8 hours, compared to a half-life of minutes for recombinant human IL-2 (rhIL-2). Repeat daily dosing (QD) of RDB 1450 in mice results in marked expansion of NK cells and CD8+CD44high T cells, with minimal expansion of CD4+ regulatory T cells (Tregs). Higher QD doses elicit more pronounced effector cell expansion with modest pulmonary edema. TID delivery of the same total daily dose of RDB 1450 enhances the immunological responses in the absence of pulmonary edema. These results suggest that prolonged delivery of reduced concentrations of RDB 1450 drive the desired immune responses with improved tolerability. In contrast, high circulating concentrations of rhIL-2 are required to stimulate effector cells due to its preferential activation of Tregs at lower concentrations. The selectivity profile and favorable pharmacokinetics of RDB 1450 highlight its potential as a novel, differentiated immunotherapeutic agent for the treatment of cancer.

Citation Format: Heather C. Losey, Jared E. Lopes, Reginald L. Dean, Heather Flick, Madison Gomes, Michael R. Huff, Rosemarie A. Moroso, Lei Sun, Chunhua Wang, Julie F. Waters, Juan C. Alvarez. Utilizing a selective agonist of the intermediate-affinity IL-2 receptor with an improved pharmacokinetic profile leads to an enhanced immunostimulatory response With reduced toxicity in mice. [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 4280. doi:10.1158/1538-7445.AM2015-4280
Abstract 4281: Determination of the Relative Potency of a Selective Agonist of the Intermediate-Affinity IL-2 Receptor on Lymphocytes from Human, Cynomolgus Monkey and Mouse
Authors: Rosentrater Emily E. , Flick Heather , Lopes Jared E. , Losey Heather C. , Wang Chunhua , Alvarez Juan C.
Published: August 2015

RDB 1450 is an engineered fusion protein designed to selectively activate the intermediate-affinity IL-2 receptor versus the high-affinity IL-2 receptor. The potencies of RDB 1450 and IL-2 for activation of distinct subsets of effector and regulatory lymphocytes from murine, non-human primate and human donors were determined. Splenocytes isolated from mice or leukocytes isolated from human or cyno blood were stimulated with either RDB 1450 or IL-2. Multicolor flow cytometry was used to identify distinct subpopulations and the extent of phosphorylation of STAT5 was measured. Whereas IL-2 is 2-3 orders of magnitude more potent on immunosuppressive Tregs relative to NK cells and memory CD8 T cells, RDB 1450 induces activation of NK cells, memory CD8 T cells and Tregs at comparable concentrations within each species. The non-differentiated potency of RDB 1450 on the lymphocyte subpopulations examined suggests that its effects are mediated through the intermediate affinity receptors even on CD25-expressing Tregs. The preferential activation of Tregs by IL-2 may lead to immunosuppression and limit its antitumor efficacy. In contrast, RDB 1450 does not exhibit the same preference for Treg activation and is expected to be more effective in driving antitumor immune responses. These results highlight the differentiated immunological profile of RDB 1450 and support its potential as a novel human immunotherapy.

Citation Format: Emily E. Rosentrater, Heather Flick, Jared E. Lopes, Heather C. Losey, Chunhua Wang, Juan C. Alvarez. Determination of the Relative Potency of a Selective Agonist of the Intermediate-Affinity IL-2 Receptor on Lymphocytes from Human, Cynomolgus Monkey and Mouse. [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 4281. doi:10.1158/1538-7445.AM2015-4281
Abstract 4290: Potent and selective next generation inhibitors of indoleamine-2,3-dioxygenase (IDO1) for the treatment of cancer
Authors: Powers Jay P. , Walters Matthew J. , Noubade Rajkumar , Young Stephen W. , Marshall Lisa , Melom Jan , Park Adam , Shah Nick , Bjork Pia , Fridman Jordan S. , Beck Hilary P. , Chian David , McKinnell Jenny V. , Osipov Maksim , Reilly Maureen K. , Shunatona Hunter P. , Walker James R. , Zibinsky Mikhail , Jaen Juan C.
Published: August 2015

The IDO1 pathway has been proposed to mediate immunosuppressive effects in the tumor microenvironment through its role in the catabolism of tryptophan, resulting in effects on the differentiation and proliferation of T cells. IDO1 inhibition has shown promising clinical benefit as well as exacerbated toxicity in the treatment of melanoma, when combined with the anti-CTLA-4 antibody ipilimumab.

We have discovered a novel class of highly selective small molecule inhibitors of IDO1 which surpass the potency of the compounds currently in clinical development. These compounds potently inhibit IDO1 activity in IFN-γ stimulated HeLa cells with single digit nM potency. Importantly, they also retain their potency in the presence of human serum, with IC50 values ranging between 5 and 15 nM in this more physiologically relevant media. Consistent with the role of IDO1+ dendritic cells in the suppression of T cell proliferation, this series of molecules is capable of restoring the proliferative capacity of human T cells (which is inhibited by allogeneic IDO1+ dendritic cells) with EC50 values of 2-3 nM. The molecules exhibit preclinical PK characteristics that are suitable for assessing the contribution of IDO1 to tumor growth in murine models, both alone and in combination with other therapeutic agents. The compounds have high metabolic stability against cultured human hepatocytes and exhibit preclinical PK and ADME characteristics consistent with once-daily dosing in humans. The full preclinical profile of one of these molecules, selected for clinical evaluation, will be the focus of this presentation.

In conclusion, we have discovered a novel class of small molecule inhibitors of IDO1, which provides a preclinical basis for the clinical evaluation of a next generation IDO1 inhibitor in combination with other therapeutic agents.

Citation Format: Jay P. Powers, Matthew J. Walters, Rajkumar Noubade, Stephen W. Young, Lisa Marshall, Jan Melom, Adam Park, Nick Shah, Pia Bjork, Jordan S. Fridman, Hilary P. Beck, David Chian, Jenny V. McKinnell, Maksim Osipov, Maureen K. Reilly, Hunter P. Shunatona, James R. Walker, Mikhail Zibinsky, Juan C. Jaen. Potent and selective next generation inhibitors of indoleamine-2,3-dioxygenase (IDO1) for the treatment of 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 4290. doi:10.1158/1538-7445.AM2015-4290
Abstract 4293: Histamine dihydrochloride as an immune modulator for solid tumors
Authors: Newick Kheng , O'Brien Shaun , Kapoor Veena , Wang Liang-chuan , Moon Edmund , Sterman Daniel , Albelda Steven
Published: August 2015

Our recent adoptive T cell transfer trial for mesothelioma revealed a striking anti-tumor response in a patient which was accompanied by anaphylaxis, suggesting endogenous histamine release. In animal models, histamine has direct and immune-based anti-tumor effects. In fact, histamine dihydrochloride is approved in Europe in combination with interleukin-2 (IL2) for leukemia treatment. We thus explored the anti-tumor role of histamine in mesothelioma and lung cancer. An in vitro MTT assay was used to measure cell proliferation. Daily doses of histamine (1 mg/kg) - alone, or in combination with other immunotherapies - was used to treat four subcutaneous flank tumor models in three different mouse strains. Flow cytometric analyses were subsequently performed on tumors. Histamine had in vitro anti-tumor activity against all five cell lines tested with an IC50 of 0.1 mg/ml. Histamine also significantly reduced tumor volume in all four in vivo models without toxic side effects. Immunodepletion studies revealed that the effect was partially due to T cells. When combined with two immunotherapy approaches (adenovirally-delivered interferon-α or a tumor antigen), anti-tumor efficacy was augmented. Histamine resulted in more interferon-γ and IL2 production by tumor-infiltrating lymphocytes, downregulation of CD206 expression on tumor-infiltrating macrophages, and reduced numbers of T regulatory cells (Tregs). This pre-clinical data suggests that histamine alone, or in combination with immunotherapy, should be explored in the treatment of mesothelioma, lung cancer, and perhaps other cancers.

Citation Format: Kheng Newick, Shaun O'Brien, Veena Kapoor, Liang-chuan Wang, Edmund Moon, Daniel Sterman, Steven Albelda. Histamine dihydrochloride as an immune modulator for solid tumors. [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 4293. doi:10.1158/1538-7445.AM2015-4293
Abstract 4299: MicroRNA 200c regulates cytochrome P450 1B1-mediated docetaxel resistance in renal cell cancers
Authors: Chang Inik , Mitsui Yozo , Fukuhara Shinichiro , Gill Ankurpreet , Wong Darryn K. , Yamamura Soichiro , Shahryari Varahram , Tabatabai Z. Laura , Dahiya Rajvir , Shin Dong Min , Tanaka Yuichiro
Published: August 2015

Cytochrome P450 1B1 (CYP1B1) is up-regulated in many types of cancer. It has been suggested that CYP1B1 may be a general tumor biomarker and have an important function in cancers. Despite its high protein expression and enzymatic activity in renal cell cancer (RCC), functional significance of CYP1B1 has not been elucidated. Here we explored the functional role and regulatory mechanism of CYP1B1 in RCC. Reduction of CYP1B1 levels fail to prevent in vitro tumorigenicity such as proliferation, apoptosis, and cell cycle progression of RCC cells. Moreover, the expression levels are not associated with tumor type, stage, Fuhrman grade and 5-year survival probability after surgery. Instead, alteration of CYP1B1 expression regulates the chemosensitivity of RCC cells to docetaxel suggesting its critical contribution to the RCC chemoresistance. Additionally, miR-200c, which is significantly down-regulated in RCC tissues and cell lines regulates CYP1B1 mRNA, protein and enzyme activity. An inverse association was also observed between the expression levels of miR-200c and CYP1B1 protein in RCC tissues. Thus, high expression of CYP1B1 could be a result of decreased miR-200c expression in RCC. Finally, alteration of miR-200c levels affects the chemosensitivity of RCC cells. Restoration of docetaxel resistance by exogenous expression of CYP1B1 in miR-200c-over-expressing cells indicates that CYP1B1 is a functional target of miR-200c. These results suggest that CYP1B1 up-regulation mediated by low miR-200c is one of the mechanisms underlying resistance of RCC cells to docetaxel. Therefore, expression of CYP1B1 and miR-200c in RCC may be useful as a prediction for docetaxel response.

Citation Format: Inik Chang, Yozo Mitsui, Shinichiro Fukuhara, Ankurpreet Gill, Darryn K. Wong, Soichiro Yamamura, Varahram Shahryari, Z. Laura Tabatabai, Rajvir Dahiya, Dong Min Shin, Yuichiro Tanaka. MicroRNA 200c regulates cytochrome P450 1B1-mediated docetaxel resistance in renal cell cancers. [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 4299. doi:10.1158/1538-7445.AM2015-4299
Abstract 4326: Co-amplification of FGF receptors and ligands in FGFR inhibitor-sensitive cell lines
Authors: Bell Katherine , Gaffney Dana , Martinez-Cardona Gabriela , Karkera Jayaprakash , Platero Suso
Published: August 2015

The fibroblast growth factor receptors (FGFR) play a key role during development and in adult function. FGFRs belong to a family of receptor tyrosine kinases which are single-pass transmembrane receptors with extracellular ligand binding domains and an intracellular tyrosine kinase domain. Upon binding of the ligand the kinase domain activates intracellular signaling networks that coordinate cellular processes such as proliferation, growth, differentiation, migration and survival. Fibroblast growth factors (FGFs) are a family of 18 ligands which are able to bind and activate distinct FGFRs. Deregulated FGFR activity, through mutations or translocations, is often associated with oncogenic events. Overexpression of FGFR or FGF may lead to increased cell proliferation, growth, differentiation, migration or survival, thus making it an interesting target.

In this study we optimized a TaqMan qRT-PCR-based approach to evaluate the copy number variation for several FGFRs (FGFR1, FGFR2, FGFR3 and FGFR4) and several FGF ligands (FGF1, FGF2, FGF3, FGF4, FGF10, FGF12, and FGF19) on a panel of 23 cell lines. The cell types covered a variety of diseases including bladder, breast, endometrial, gastric, kidney, liver, lymphoma, melanoma, sarcoma, small cell lung carcinoma and squamous cell carcinoma. These cell lines had been tested for sensitivity to the JNJ FGFR small molecule inhibitor JNJ42541707 which is a pan-FGFR inhibitor. We observed in the sensitive (IC50<100nm) and moderately sensitive (IC50 100 nM - 1000 nM) cell lines a co-amplification of FGF receptor and ligand. Two of the most sensitive cell lines to FGFR inhibitor treatments had the greatest amplification of FGFR2 with a copy number value greater than 100. This finding that co-amplification occurs in receptor and ligand offers a new potential biomarker for patients that may be sensitive to small molecule FGFR inhibitors.

Citation Format: Katherine Bell, Dana Gaffney, Gabriela Martinez-Cardona, Jayaprakash Karkera, Suso Platero. Co-amplification of FGF receptors and ligands in FGFR inhibitor-sensitive cell lines. [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 4326. doi:10.1158/1538-7445.AM2015-4326
Abstract 4329: Expression of serpin B2, neuroserpin and L1CAM in association with metastasis and survival in non-small cell lung cancer
Authors: Ramnefjell Maria , Helgeland Lars , Akslen Lars A.
Published: August 2015

Background: Lung cancer is a leading cause of cancer related deaths worldwide, and finding new prognostic and predictive biomarkers are of utmost importance.

Previous studies have shown that different serpins and adhesion molecules have an impact on disease progression and development of metastasis, especially to the brain. Here, we examined whether expression of the anti-plasminogen activators Serpin B2 and Neuroserpin, and the adhesion molecule L1CAM was associated with patient prognosis, brain metastasis or other clinicopathologic characteristics, in non-small cell lung cancer (NSCLC).

Study design: The material consists of surgical specimens from 100 NSCLC patients treated during 1993-1999 at Haukeland University Hospital, Bergen, Norway. Sections from tumor blocks were stained with antibodies to Serpin B2, Neuroserpin, and L1CAM.

Results: There were 66 men and 34 women, with a mean age at diagnosis of 65.7 (41 adenocarcinomas, 38 squamous cell carcinomas, and 21 carcinomas of other types).

Observation time ranged from 0 to 254 months (mean 74 months). 12 patients were fully recovered, 46 died of lung cancer, and 34 died from other causes. 11 of 89 patients developed brain metastasis.

Low expression of Serpin B2 was associated with reduced overall survival (p<0.05) in adenocarcinomas, and with a trend for all types combined (p = 0.10), whereas there was no significant difference in survival related to expression of Neuroserpin or L1CAM.

Expression of Neuroserpin was associated with adrenal metastasis (OR 13.3 (95%CI 1.5-119.7), p<0.05), and there were trends for associations between expression of Serpin B2 and metastases to skin and bone (p = 0.072 and p = 0.068, respectively). There were no significant associations between these markers and presence of brain metastasis.

Conclusions: Low expression of Serpin B2 in lung adenocarcinoma was associated with reduced overall survival in our cohort. In contrast to previous findings by others, we did not find a relationship between L1CAM expression and overall survival or metastasis. Neither was there any association between expression of Neuroserpin and survival, although we found an increase in adrenal metastasis in this group. Low expression of Serpin B2 could be a biomarker for prognosis in lung adenocarcinomas and should be studied in other cohorts.

Citation Format: Maria Ramnefjell, Lars Helgeland, Lars A. Akslen. Expression of serpin B2, neuroserpin and L1CAM in association with metastasis and survival in 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 4329. doi:10.1158/1538-7445.AM2015-4329
Abstract 4342: Differential expression of novel PKD1 related biomarker panel in African American men with prostate cancer
Authors: Nickkholgh Bita , Fang Xiaolan , Winters Shira M. , Fino Nora , Balaji KC.
Published: August 2015

Background: Prostate Cancer (PCa) exhibits striking racial disparity and is presented more aggressively in African Americans (AAs) than Caucasians (CAs). We have previously reported that the dysregulation of PKD1-centered signaling in primary AA prostate cancer cell line E006AA is akin to more aggressive Caucasian cell line C4-2. As PKD1 plays a central role in several cellular mechanisms that contribute in aggressive malignant phenotype, we defined a PKD1 related biomarker panel consisting of 11 biomarkers and explored whether PKD1 centered signaling would be differentially regulated in AA compared to CA PCa. Methods: PKD1 related biomarker panel consisted of PKD1, AR, HSP27, N-Cadherin, Snail, Vimentin, MMP-2, MMP-9, MT-1, E-cadherin, beta-Catenin. The validity of the PKD1 related biomarker panel in PCa was evaluated on available microarray databases (cbioportal.org). The differential expression of the biomarker panel was quantified on PCa tissue samples from15 AA and 15 CA men with matching Gleason score obtained from tumor tissue bank of Comprehensive Cancer Center of Wake Forest University. Results: The cbioportal consisted of six different PCa databases with approximately 662 human PCa samples in total. In average, PKD1 related biomarker panel was altered 47.7% in available cases. Quantification of the PKD1 related biomarker panel regarding gene expression demonstrated a trend toward higher expression of majority of biomarkers in AA samples in compare to CAs. However, only variances in expression of MMP2 (P<0.0001) and MT1A (P = 0.03) were significantly different between AAs and CAs. The results obtained from human PCa samples showed a moderate correlation with previously reported cell line data which showed significantly higher expression of EMT markers N-Cadherin, Vimentin and Snail in AA cell line in compare to CA cell lines. Conclusion: The differential expression of PKD1 related biomarker panel in AAs in compare to CAs suggests a biological basis for disparity in PCa. Besides, relatively high altered PKD1 related biomarker panel in available microarray databases suggests the potential of the panel as a tool in diagnosis of aggressive phenotype in PCa.

Citation Format: Bita Nickkholgh, Xiaolan Fang, Shira M. Winters, Nora Fino, KC. Balaji. Differential expression of novel PKD1 related biomarker panel in African American men with prostate cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4342. doi:10.1158/1538-7445.AM2015-4342
Abstract 4345: FGFR1, a new FISH biomarker, may predict recurrence of prostate cancer after prostatectomy
Authors: Zhang Ying , Pestova Katerina , Du Jing , Liu Ping , Buckingham Lela , Colpitts Tracey
Published: August 2015

Background: To identify prostate cancer (CaP) with high recurrence risk after Radical Prostatectomy (RP) is a challenge. FGFR1 (Fibroblast Growth Factor Receptor 1) plays a critical role in prostate development and tumorigenesis. In this study, we explored the possibility of using FGFR1 fluorescence in situ hybridization (FISH) to predict recurrence after RP in the Formalin fixed paraffin embedded (FFPE) tissue from CaP patients.

Methods: FFPE histological specimens from 52 RP cases were evaluated with FGFR1 (8p12) / CEP 8 probe mix. While 32 of the 52 patients recurred within 5 years (PSA progression or death of disease), and 20 remained disease-free with 8 to 15 years. Median age of patients was 62 with Gleason Score range from 4 to 9. Percent of FGFR1 gain and FGFR1 loss per specimen, as well as mean number of FGFR1 signals per specimen were calculated. CEP8 was used as a control to assess hybridization quality. Statistical analysis (Cox Proportional Hazards model) was conducted using SAS 9.2.

Results: Survival analysis demonstrated that percent FGFR1 loss in a specimen could identify recurrent CaP with the Harzard Ratio (HR) of 3.34 (p = 0.0011), which indicated that the risk of recurrence for FGFR1 loss-positive group is 3.34 times than that for FGFR1 loss-negative group. On the other hand, survival analysis of the percent FGFR1 gain showed that FGFR1 gain (amplification) is favorable for survival with HR of 0.36 (p = 0.0043), which indicated that the risk of recurrence for FGFR1 gain-positive group is 0.36 times than that for FGFR1 gain-negative group. By using mean FGFR1 signals per specimen in the Kaplan-Meier analysis, the patient specimens stratified into three distinct groups (overall logrank p = 0.0011): normal for FGFR1, FGFR1 gain (more favorable outcome), and FGFR1 loss (highest risk of recurrence).

Conclusions: This study shows the utility of FGFR1 copy number abnormalities in CaP. Our results indicate that gain and loss of FGFR1 may have a different effect on disease recurrence. Therefore, FGFR1 FISH may provide predictive value for CaP recurrence in post RP patients, either by itself, or as an adjunct to the currently clinical prognostic indicators.

Citation Format: Ying Zhang, Katerina Pestova, Jing Du, Ping Liu, Lela Buckingham, Tracey Colpitts. FGFR1, a new FISH biomarker, may predict recurrence of prostate cancer after prostatectomy. [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 4345. doi:10.1158/1538-7445.AM2015-4345
Abstract 4350: Prognostic potential of epithelial to mesenchymal transition in prostate cancer at first presentation
Authors: Al-Sukaini Ahmad , Hart Claire , Robinson Richard , Brown Mick , Clarke Noel
Published: August 2015

Introduction and Objective: Metastatic progression of prostate cancer requires a change of state, epithelial to mesenchymal transition (EMT), reducing cell to cell adherence and contact, with associated cell motility and invasion. EMT is one of the hallmarks of cancer and characterisation of the EMT process has defined a panel of EMT biomarkers. Here we investigate the potential of an EMT panel of biomarkers in an unscreened population at first presentation with prostate cancer.

Methods: A prostate Tissue Micro Array (TMA) comprising 2,450 cores containing normal adjacent, prostatic intraepithelial neoplasia and malignant prostate tissue from 524 diagnostic biopsies, linked to an extensive clinical database with long-term outcome, was studied. Expression of a panel of EMT biomarkers, including E-cadherin and vimentin, was assessed using an automated, high throughput fluorescent staining (Leica BOND-MAX), image capture (Carl Zeiss Microimaging Mirax scanner) and quantitative image analysis (Definiens Tissue Studio) protocol. Definiens H-Score was converted to pathological scores prior to univariate and multivariate Cox proportional hazards regression analysis in SPSS.

Results: Expression of an EMT profile predicted a significant reduction in both overall and cancer specific survival. In univariate analysis, low or negative E-cadherin staining was associated with a reduction in median overall survival by 45 months (95% CI: 10.4-79.6; P<0.0001) and a cancer specific mean survival of 80.1 months (95% CI: 45.7-114.5; P<0.0001) when compared to high and intermediate E-cadherin staining. Similarly, high EMT expression, defined as loss of E-cadherin and presence of vimentin, was compared to low EMT expression, defined as preserved E-cadherin and the absence of vimentin. There was a greater reduction in overall median survival in high EMT tumours of 54 months (95% CI: 12.5-95.5; p<0.0001) and cancer specific survival of 93.2 months (95% CI: 62.8-123.7; p<0.0001) when compared to low EMT tumours. Stage, grade, PSA at diagnosis and EMT status was included in the multivariate analysis for cancer specific survival. Both Gleason grade (p = 0.017) and EMT status (p = 0.030) were found to be significant prognostic factors in predicting cancer specific survival.

Conclusions: Loss of E-cadherin and concomitant expression of the EMT marker vimentin are powerful predictors of prostate cancer outcome at first presentation.

Citation Format: Ahmad Al-Sukaini, Claire Hart, Richard Robinson, Mick Brown, Noel Clarke. Prognostic potential of epithelial to mesenchymal transition in prostate cancer at first presentation. [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 4350. doi:10.1158/1538-7445.AM2015-4350
Abstract 4353: Improved prediction of prostate cancer prognosis by using a multiparametric molecular classifier
Authors: Tsourlakis Maria Christina , Kluth Martina , Schlomm Thorsten , Simon Ronald , Sauter Guido , Minner Sarah
Published: August 2015

Background: Because no single molecular marker have proven to be sufficiently powerful for routine estimation of prostate cancer prognosis as to yet, it is hoped that the combined analysis of multiple markers will allow for distinguishing between indolent and aggressive forms of prostate cancer. In fact, first generation commercial multiparametric test assays show promising results. We hypothesized that such tests may be further optimized.

Design: A tissue microarray (TMA) containing 0.6 mm tissue cores from more than 12,000 prostate cancer patients with long-term follow-up data was analyzed with more than 100 molecular markers by immunohistochemistry or FISH. Markers with strong and independent prognostic value in uni- and multivariate analyses were combined to obtain panels of markers that yield additional prognostic information beyond the standard parameters (including pre-operative PSA, Gleason grade, tumor stage, nodal stage) in ROC analysis. For this purpose, a simple score (0-10 points) was build based on the number and type of adverse features per tumor.

Results: Markers with strong independent prognostic relevance included deletions of 6q15 and PTEN, overexpression of EZH2, TUBB3, and nuclear accumulation of p53, indicating dominant-negative TP53 gene mutation. A total of 5,129 cancers harbored none of these alterations (score 0) and showed the best prognosis in a Kaplan-Meier plot using biochemical recurrence as the study endpoint. The worst prognosis was found for 610 tumors harboring ≥8 (score 8-10) of these alterations, while an intermediate prognosis with decreasing time to recurrence was found for cancers with scores 1-2 (n = 2,418), 3-4 (n = 1,367) and 5-7 (n = 1038). The differences between the groups were high significant (p<0.0001). The 5 genes classifier provided prognostic information independent from PSA, Gleason, stage, and nodal stage (p<0.0001).

Conclusion: Multiparametric classifiers have a high potential to improve the predictive accuracy of current models using established clinic-pathological models. Analyses of very large sets of patient tissues are instrumental to identify the best candidates for such molecular marker panels. It is likely, that continued analysis of our TMA will identify even better marker panels than the current 5 genes classifier.

Citation Format: Maria Christina Tsourlakis, Martina Kluth, Thorsten Schlomm, Ronald Simon, Guido Sauter, Sarah Minner. Improved prediction of prostate cancer prognosis by using a multiparametric molecular classifier. [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 4353. doi:10.1158/1538-7445.AM2015-4353
Abstract 4360: Validation of phosphodiesterase 10A as a cancer target
Authors: Lee Kevin , Li Nan , Chen Xi , Zhu Bing , Yet Larry , Madeira da Silva Luciana , Russo Suzanne , Keeton Adam B. , Boyd Michael R. , Piazza Gary A.
Published: August 2015

Phosphodiesterase 10A (PDE10) is a cAMP and cGMP degrading PDE isozyme that is highly expressed in the brain striatum where it plays an important role in cognition and psychomotor activity. PDE10 inhibitors are being developed for the treatment of schizophrenia and Huntington's disease and are generally well tolerated, likely because of low expression levels in peripheral tissues. We recently reported high levels of PDE10 in tumors and that genetic silencing by siRNA inhibits tumor cell growth with a high degree of selectivity over normal cells (Li et al., Oncogene 2014). These observations suggest that PDE10 may have an unrecognized role in tumorigenesis and represents a novel cancer target. To further test this possibility, we studied the effects of a highly specific PDE10 inhibitor, Pf-2545920 (MP-10) on colon tumor cell growth. Here we show that Pf-2545920 selectively inhibits tumor cell growth, causes G1 cell cycle arrest, and induces apoptosis. The concentration range by which Pf-2545920 inhibits tumor cell growth parallels the concentration range required to increase intracellular cyclic nucleotide levels and activate PKA and PKG. Moreover, PDE10 knockdown by siRNA reduces the sensitivity of tumor cells to the growth inhibitory activity of Pf-2545920. Using the crystal structure of PDE10 to design novel inhibitors, a series of compounds were synthesized and screened for tumor cell growth inhibitory activity and PDE10 isozyme specificity. A lead compound, ADT-030 was found to inhibit tumor cell growth and PDE10 enzymatic activity with IC50 values in the nanomolar range, but did not significantly affect the growth of normal cells. Unlike Pf-2545920, ADT-030 exhibits high selectivity for activating PKG signaling without affecting PKA signaling. Inhibitors of PKA and PKG were used to confirm that the tumor cell growth inhibitory activity associated with PDE10 inhibition involves PKG activation, while PKA activation appears to be ancillary. These findings serve to validate PDE10 as a cancer target, whereby novel inhibitors can be designed to specifically activate cGMP/PKG signaling with a high degree of tumor cell selectivity. Supported by NIH grants 1R01CA155638 and 1R01CA131378 (Piazza).

Citation Format: Kevin Lee, Nan Li, Xi Chen, Bing Zhu, Larry Yet, Luciana Madeira da Silva, Suzanne Russo, Adam B. Keeton, Michael R. Boyd, Gary A. Piazza. Validation of phosphodiesterase 10A as a cancer target. [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 4360. doi:10.1158/1538-7445.AM2015-4360
Abstract 4378: Targeting the embryonic morphogen Nodal reduces viability of doxorubicin-treated breast cancer cells in vitro
Authors: Bodenstine Thomas M. , Chandler Grace S. , Margaryan Naira V. , Strizzi Luigi , Gilgur Alina , Seftor Elisabeth A. , Seftor Richard E B , Khalkhali-Ellis Zhila , Ugolkov Andrey , Mazar Andrew P. , Hendrix Mary J C
Published: August 2015

Triple negative breast cancer represents an aggressive malignancy for which few targeted therapies exist. The embryonic morphogen Nodal is re-expressed in highly aggressive breast cancers and reduction of Nodal signaling decreases tumor cell metastatic characteristics. This represents a potential avenue for the development of therapeutics aimed at inhibiting the Nodal pathway. To explore this further, we evaluated Nodal expression in breast cancer cells following chemotherapy. In vitro treatment of MDA-MB-231 and MDA-MB-468 human breast cancer cell lines with Doxorubicin, a chemotherapy commonly used for the treatment of malignant breast cancer, resulted in the generation of surviving subpopulations that retained Nodal expression (>80%) and remained viable. To validate these findings in vivo, we analyzed Doxorubicin treated metastatic patient derived breast cancer xenografts (PDX) which displayed a residual population of Nodal positive cells when analyzed by immunohistochemistry. Based on these findings, we hypothesize that targeting Nodal in a combinatorial approach with chemotherapy will lead to a reduction in remaining viable populations of cancer cells. When we utilized an anti-Nodal neutralizing antibody to inhibit Nodal signaling, antibody treatment reduced cell proliferation, attachment and increased rates of apoptosis in Doxorubicin treated breast cancer cell lines. Decreases in phosphorylation of Histone 3, a marker of cellular proliferation, and increases in the cleavage of PARP, correlating to induction of apoptosis, were noted in response to antibody treatment. These data suggest that Nodal signaling plays a critical role in the growth of metastatic breast cancer cells and targeting Nodal may improve therapeutic outcome in malignancies such as triple negative breast cancer. The initial results highlight translational potential for the development of humanized antibodies capable of inhibiting Nodal function.

Citation Format: Thomas M. Bodenstine, Grace S. Chandler, Naira V. Margaryan, Luigi Strizzi, Alina Gilgur, Elisabeth A. Seftor, Richard E B Seftor, Zhila Khalkhali-Ellis, Andrey Ugolkov, Andrew P. Mazar, Mary J C Hendrix. Targeting the embryonic morphogen Nodal reduces viability of doxorubicin-treated breast cancer cells in vitro. [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 4378. doi:10.1158/1538-7445.AM2015-4378
Abstract 4386: Docetaxel-trastuzumab stealth immunoliposome: design and non-clinical study in HER2+ and HER2- breast cancer models
Authors: Fanciullino Raphaelle R. , Brunel Jean-Michel , Correard Florian , Giacometti Sarah , Ciccolini Joseph
Published: August 2015

Targeted liposomal drug delivery is expected to enhance the therapeutic effects of anti-cancer agents through both active and passive targeting of tumor cells. We have developed a stealth liposomal formulation of docetaxel anchored with trastuzumab (HER2-ANC) to target HER2+ breast cancers. Pegylated SUV liposomes with 95% docetaxel encapsulation yield were obtained using the thin-film method. Homogenous distribution in liposome size of 100-150 nm diameter was achieved by extrusion, prior to anchoring trastuzumab through maleimide coupling. Anchoring was evidenced by electronic microscopy with anti-mouse gold antibody. HER2-ANC antiproliferative activity was assessed on a representative panel of human breast cancer cells expressing (SKBR3, MDA-MB451) or not expressing HER2 (MDA-MB231). Free docetaxel and trastuzumab combination was used as control. Cytotoxicity was evaluated in vitro on 2D culture cells by MMT assay, and in 3D spheroids co-cultured with HMEC endothelial cells. HER2-ANC proved to have a similar efficacy than the free combination, including in triple negative MDA-MB231 cells in 2D models. In 3D models, the immunoliposome displayed higher efficacy as compared with the free combination, possibly because of a better and deeper diffusion of the antibody inside the spheroids. Surprisingly, and although less marked, this difference in favor of HER2-ANC was observed as well with the triple negative MDA-MB231 cells, thus suggesting that a better delivery of docetaxel was achieved with the liposomal form. Fluorescence imaging showed that HER2-ANC was internalized inside tumor cells. Measurement of HER2 expression and immunofluorescence staining of alpha-microtubulline supported this increase in efficacy observed in vitro. After tagging HER2-ANC with DIR fluorescent probe, in vivo preliminary distribution studies in nude mice bearing both the HER2+ SKBR3 and HER2- MDA-MB231 tumors showed that a more specific tumor uptake is achieved in HER2+ tumors, thus suggesting that higher antiproliferative effect and lower toxicities can be expected in further efficacy studies. Overall, this proof of concept study strongly suggest that redesigning existing combinations of biologics and cytotoxics as immunoliposomes could help to optimize the efficacy/toxicity balance of anticancer treatments.

Citation Format: Raphaelle R. Fanciullino, Jean-Michel Brunel, Florian Correard, Sarah Giacometti, Joseph Ciccolini. Docetaxel-trastuzumab stealth immunoliposome: design and non-clinical study in HER2+ and HER2- breast cancer models. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4386. doi:10.1158/1538-7445.AM2015-4386