bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract A221: A phase II study of SB939 in patients with recurrent or metastatic castration resistant prostate cancer (CRPC).
Authors: Eigl Bernhard J. , North Scott , Murray Nevin , Heng Daniel YC , Winquist Eric , Powers Jean , Walsh Wendy R. , Eisenhauer Elizabeth , Squire Jeremy , Cox Michael , Chi Kim N.
Published: June 2012

Background: SB939 is a potent oral inhibitor of class I, II and IV histone deacetylases (HDACs) with high tumor tissue selectivity. These 3 classes of HDAC are highly expressed in prostate cancer and associated with poor clinical outcomes. HDAC inhibition has been shown to abrogate androgen receptor signaling via inactivation of HSP90, and may have relevance particularly in TMPRSS2-ERG fusion positive prostate cancer.

Methods: In this multicenter phase II study, pts with CRPC received SB939 60mg every other day 3 times per week × 3 weeks on a 28-day cycle. Primary endpoints were PSA response rate (RR) (>50% decline from baseline for > 4 weeks) and progression free survival (PFS). Secondary endpoints included objective response rate; response duration, overall survival; circulating tumor cell (CTC) enumeration at baseline, 6 and 12 weeks; TMPRSS2-ERG fusion analysis; correlative analysis of archival tissue; and safety. Planned sample size was 29 pts, the hypotheses (PSA RR <2% vs. ≥15%) could be tested at 10% and levels.

Results: Twenty-nine pts have been accrued. Median age is 69, 25 pts had received no prior chemotherapy, ECOG performance status was 0/1 in 48%/52% of pts. Bone/lymph node/visceral metastases were present in 86%/69%/10% of pts. The median number of cycles of SB939 administered is 2 (range 1–6) with 7 pts still continuing treatment. Adverse events were generally grade 1–2, with 5 patients experiencing grade 3 events (fatigue (5 pts), vomiting (1 pt) and dyspnea (1pt)). To date, a confirmed PSA response has been noted in 2 pts (7%). In 8 RECIST-evaluable pts, there were no objective responses, and 3 pts had stable disease lasting from 3.4 to 5.6 months. CTC favorable conversion (from ≥5 at baseline to <5 at 6 or 12 weeks) was observed in 9/14 evaluable patients (64%) and unfavorable CTC levels (>5) at 6 weeks correlated with PSA progression. RT-PCR for presence of TMPRSS2-ERG transcripts from whole blood and FISH analyzes of CTC and archival tissue for TMPRSS2-ERG fusion and PTEN deletion status are ongoing.

Conclusion: SB939 is well tolerated and has shown early evidence of activity in CRPC. Analysis of more mature data will be required to determine whether this agent warrants further study, and whether TMPRSS2-ERG status might predict for response to SB939.

Supported by grants from the Canadian Cancer Society and the Ontario Institute for Cancer Research.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr A221.
Abstract A230: Discovery and characterization of ARQ 092, an ATP-independent, potent and selective inhibitor of AKT kinases.
Authors: Chan Thomas C.K. , Lapierre Jean-Marc , Ashwell Mark A. , France Dennis S. , Chen Chang-Rung , Cornell-Kennon Susan , Bull Cathy , Eathiraj Sudhi , Palma Rocio , Liu Yanbin , Makhija Sapna , Waghorne Carol , Savage Ron , Fujiwara Kosaku
Published: June 2012

The AKT family of serine-threonine kinases is a critical signal transduction node serving a variety of cellular functions including survival, proliferation, protein synthesis, and glucose metabolism. Small molecule inhibitors of AKT are emerging as potential targeted agents to treat cancers that exhibit aberrant AKT pathway signaling. A screening strategy was employed to identify inhibitors which utilize the intrinsic negative regulatory function of hydrophobic clusters in the ATP-binding cleft to inhibit the kinase activity of AKT1. ARQ 092 was identified through this screening paradigm following in vitro and in vivo optimization and was selected for further development. Crystallographic studies provided evidence that this mechanism of inhibition of AKT had been achieved. ARQ 092 binds to inactive, unphosphorylated AKT1 with sub-nanomolar affinity (Kd = 0.28 nM measured by surface plasma resonance) and biochemically inhibits all three isoforms (IC50 values of 3 nM, 4.5 nM, and 5.5 nM respectively for AKT1, AKT2, and AKT3). ARQ 092 displayed inhibition kinetics against AKT1 which were not affected by ATP concentrations up to 1000 M. When screened against a panel of over 300 kinases, ARQ 092 inhibited only three kinases within 100-fold of its IC50 against AKT1. ARQ 092 potently inhibited AKT phosphorylation (Ser473 & Thr308) in AN3CA human endometrial carcinoma cells (EC50 values of 39 nM and 61 nM, respectively for p-Ser473 & p-Thr308) and demonstrated concentration-dependent inhibition of phosphorylation of the downstream AKT substrate PRAS40. ARQ 092 inhibited growth of AN3CA cells (GI50 = 555 nM), A2780 cells (GI50 = 400 nM), and IGROV-1 cells (GI50 = 66 nM), in addition to LNCaP, ZR-75–1, and BT-474 human cancer cell lines (GI50 values ranging from 1 to 4 μM). Following a single oral dose of ARQ 092, inhibition of AKT and PRAS40 phosphorylation was documented in vivo in both AN3CA human endometrial tumor and BT474 human breast tumor xenograft models. The growth of AN3CA tumor xenografts was markedly suppressed after daily oral administration of ARQ 092 for 10 days. Finally, ARQ 092 was shown to have good pharmaceutical properties and was advanced into preclinical development.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr A230.
Abstract A66: Antiproliferative effects of CDK4/6 inhibition alone and combination with chemotherapy in KRAS-mutated NSCLC.
Authors: Hong Jihyung , Zhang Xiang-Hua , Shin Jung-Young , Kim Jeong-Oh , Oh Ji-Eun , Kang Jin-Hyoung
Published: June 2012

Background: Deregulation of the p16INK4A-cyclin D-cyclin-dependent kinases (CDK) 4/6-retinoblastoma (RB) pathway is a common paradigm in various solid tumors and represents one of the attractive targets in cancer treatment. KRAS mutation confers intrinsic resistance to cytotoxic chemotherapeutic agents, and to EGFR TKIs leading to treatment failure. An effective systemic treatment for adenocarcinomas harboring KRAS mutations are urgently needed to improve clinical outcomes.

Purpose: CINK4, a chemical inhibitor of cyclin-dependent kinase 4, has demonstrated effective anti-proliferative activity in vivo in the presence of pRb. Herein, we investigated that CINK4 may increase its anti-proliferative activity in the NSCLC cells harboring KRAS mutation

Methods: We measured the anti-proliferative activities of CINK4 in A549 (KRASG12S), H358 (KRASG12C), PC14 (KRASwild), and Calu-3 (KRASwild) cell lines using SRB assay, cell cycle distribution quantified by flow cytometry, and the expression of cell cycle regulators and apoptosis-related proteins was evaluated by Western blotting. Multiple drug effect was analyzed to study an interaction between two drugs. The nature of the interaction between two agents was calculated for the combination index (CI).

Results: Concentration- and time-dependent anti-proliferative effects of CINK4 (0.1∼40 μM concentration range) and pacitaxel (0.1∼300 nM) alone were seen in four NSCLC cell lines, and were not significantly different between each cell line (48 h IC50: 10.4 ± 0.2 μM and 5.5 ± 0.9, 10.1± 1.9 μM and 5.2 ± 0.9, 6.6 ± 0.8 μM and 3.6 ± 1.0, and 8.4 ± 0.2 μM and > 10 nM, in A549, H358, PC14, and Calu-3, respectively). After 48 h CINK4 treatment alone, the G0/G1phase cell population increased with increment of subG1 faction in concentration-dependent manner. CINK4 reduced cyclinD1 and Rb phosphorylation and increase the expression of cleaved PARP in concentration-dependent manner in A549, H358, and PC14 cell lines. And CINK4 combined with paclitaxel in two KRAS mutated cell lines showed enhanced anti-proliferative activity of paclitaxel. The CI values of CINK4 and palitaxel combination in A549 and H358 were 0.51 ± 0.16 and 0.63± 0.12, respectively.

Conclusions: CINK4 showed promising anti-tumor activity in NSCLC cell lines. NSCLC harboring KRAS mutation may have clinical benefit from CDK4/6 inhibitor in combined with chemotherapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr A66.
Abstract C202: Discovery and initial characterization of the clinical compound ABT-348, a potent inhibitor of the VEGF, PDGF, and Aurora kinase families.
Authors: Curtin Michael L. , Heyman Robin , Frey Robin R. , Soni Niru B. , Marcotte Patick A. , Pease Lori J. , Glaser Keith B. , Magoc Terrance J. , Tapang Paul , Albert Daniel H. , Osterling Donald J. , Olson Amanda M. , Bouska Jennifer J. , Guan Zhiwen , Martin Ruth L. , Stewart Kent D. , Tse Chris , Davidsen Steven K. , Michaelides Michael R.
Published: June 2012

The Aurora kinases are a family of serine/threonine kinases that mediate multiple events in cell division. Humans have three Aurora kinases, A, B and C, that are differentially localized and mediate distinct functions during mitosis. Because the Aurora kinases play a key role in mitosis and are overexpressed in multiple human tumor types, there has been considerable interest in developing Aurora kinase inhibitors as antitumor agents. A number of small-molecule Aurora kinase inhibitors have been reported and there are several compounds currently in Phase I/II clinical trials for cancer. ABT-348 is a novel, potent and orally bioavailable inhibitor of the Aurora kinases as well as the VEGF and PDGF families of receptor tyrosine kinases and is currently in Phase I clinical trials. Herein we describe the discovery and preparation of this molecule including the SAR work that provided improved Aurora kinase potency, aqueous solubility and oral bioavailability with reduced hERG and CYP liabilities relative to earlier analogs.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr C202.
Abstract 1009: Leptin induces Src/Gbr2/Gab2/STAT3 activation and Rac-1 crosstalk to regulate VEGF/VEGFR2 in breast cancer
Authors: Fuller Miles , Byrd Manzy , Guo Shanchun , Gonzalez-Perez Ruben Rene
Published: June 2012

Leptin and its receptor, OB-R, are overexpressed in cancer cells and their levels correlate to worse prognosis of cancer patients. Leptin is an important inducer of VEGF/VEGFR2 in breast cancer under normoxia through the activation of several signaling pathways and HIF-1α/NFκB/SP1. However, the specific mechanisms of leptin-mediated regulation of VEGF/VEGFR2 genes are still partially unknown. VEGF/VEGFR2 directly regulates tumor angiogenesis and also works as an essential autocrine/paracrine process for cancer cell proliferation and survival. We have previously shown that disruption of leptin signaling with our novel antagonists, PEG-LPrAs, significantly impaired the growth of breast tumors and reduced the levels of VEGF/VEGFR2. We hypothesize that leptin regulation of VEGF/VEGFR2 in breast cancer involves the activation of Src and Gbr2/Gab2/STAT3 and crosstalk to Rho-GTPases. Methods: To test this hypothesis, VEGF/VEGFR2 expression and activation of signaling intermediaries were determined in mouse and human breast cancer cells challenged with leptin as well as genetic and pharmacologic kinase inhibitors. Results: Inhibition of JAK2 partially reduced leptin-induced pSTAT3 levels but increased VEGF protein and mRNA. Moreover, leptin-induced pSTAT3 was related to Src and Gab2. Inhibition of Src increased leptin-induced VEGF and dramatically reduced VEGFR2. Rac1 inhibition also abrogated leptin-induced pSTAT3 and decreased pGab2 and VEGF/VEGFR2 levels. Conclusions: Leptin activation of Rac1 upregulates VEGF/VEGFR2 and promotes a negative feed-back, which downregulates leptin activation of Src/Gbr2/Gab2/STAT3. In addition, leptin activation of Src/Gbr2/Gab2 and Rac1 are needed for the induction of VEGFR2. These data suggest a high complexity of signaling crosstalk is involved in leptin regulation of pro-angiogenic factors and breast cancer growth. Therefore, PEG-LPrA combined with inhibitors for leptin-sensitive regulatory signaling partners could provide novel multiple target therapies for breast cancer. This could be of paramount importance for the higher risk patients: obese and postmenopausal women who show highest leptin levels. Our proposed studies could also be relevant to patients having the very aggressive and difficult to treat triple negative breast cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1009. doi:10.1158/1538-7445.AM2011-1009
Abstract 1287: Profile-related evidence to determine individualized cancer therapy (PREDICT): Preliminary results of the Personalized Phase I Clinical Trials program at MD Anderson Cancer Center
Authors: Tsimberidou Apostolia Maria , Hong David S. , Wheler Jennifer J. , Fu Siqing , Piha-Paul Sarina , Naing Aung , Falchook Gerald S. , Luthra Raja , Iskander Nancy G. , Wen Sijin , Kurzrock Razelle
Published: June 2012

Introduction: To assess the outcomes of molecularly targeted therapy, we analyzed the results of mutational analyses in patients seen in our Phase I clinic and assessed patient survival by the number of mutations in patients treated with and without matched targeted therapy. Methods: Mutational analysis was mostly performed in the CLIA-certified pathology laboratory. DNA was extracted from microdissected paraffin-embedded tumor samples. The following genes were analyzed: PIK3CA (exon 9: codons 532-554; exon 20: codons 1011-1062); BRAF (exon 15: codons 595 to 600); KRAS and NRAS (codons 12, 13, 61); EGFR (exons 18 to 21, kinase domain); KIT (exons 9, 11, 13, 17); and RET (exon 10: codons 609, 611, 618, 620; exon 11: codon 634; exon 16: codon 918). The loss of the tumor suppressor nuclear protein (PTEN) was determined by immunohistochemistry. The allocation of patients to clinical trials varied according to protocol availability; eligibility; patient's prior response to therapy, toxicity and preference; and physician's choice. Patients whose tumor had a mutation were preferably treated with a matched targeted agent. Survival was compared between patients treated with and without matched targeted therapy. Results. Of 849 patients who had tissue available, 354 (41.7%) had ≥ 1 mutation (1, n=313; 2, n=38; 3, n=3) (Table 1). The median survival of patients with 0 vs. 1 vs. 2-3 mutations was 9.4 vs. 11.7 vs. 7.3 months, respectively (p =0.02; Table 2). In 278 evaluable patients with 1 mutation, the 2-yr survival rates were 42% and 17% (p = 0.02) for patients treated with and without matched targeted therapy. Conclusion: In patients with 1 mutation, treatment with matched targeted therapy was associated with superior survival compared to treatment without matching. Patients with 1 mutation had longer survival than those with no or >1 mutation, perhaps because most of them were treated with a matched targeted therapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1287. doi:10.1158/1538-7445.AM2011-1287
Abstract 1401: SYK promotes tumor progression in ovarian cancer cell lines
Authors: Sultan Aya , Wang Cindy Y. , Duran George E. , Francisco E. Brian , Berek Jonathan S. , Sikic Branimir I.
Published: June 2012

Identification of novel drug targets in ovarian cancers has focused on expression of genes in the tyrosine kinase family. We have previously shown that the spleen tyrosine kinase (SYK) gene was expressed in 73% of 55 ovarian cancer specimens, compared to 26% of 60 breast cancer specimens. Previous studies established the role of SYK in tumor progression. Among its many targets, SYK can bind microtubules and other molecules associated with the cytoskeleton. We found SYK to be expressed in 6 of 10 ovarian carcinoma cell lines. Because of the ability of SYK to bind to microtubules, we studied the relationship of SYK expression to cellular migration. We examined several ovarian cancer cell lines that variably express SYK to elucidate the potential effects of SYK on cell migration. Our results demonstrate that knockdown of SYK expression by siRNA resulted in a decreased capability of cells to migrate using a traditional wound scratch assay. The duration of this effect was dependant upon cell line used suggesting that additional protein targets may be involved in the migratory process. The potential effects of SYK expression on invasion are currently being investigated. We will evaluate potential upstream and downstream targets to elucidate the pathway(s) responsible for SYK effects and compare within the cell lines examined. Understanding the growth signals and proteins potentially responsible for invasion and metastasis in ovarian cancer cell lines and tumor samples may eventually lead to new therapeutic approaches for ovarian cancers.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1401. doi:10.1158/1538-7445.AM2011-1401
Abstract 1416: Development and initial evaluation of the antitumor activity of a functionalized benzophenone thiosemicarbazone inhibitor of cathepsin L
Authors: Trawick Mary Lynn , Pinney Kevin G. , Chaplin David J. , Horsman Michael R. , Chavarria Gustavo E. , Arispe Wara M. , Kishore Kumar G. D.
Published: June 2012

The discovery of new chemotherapeutic strategies for inhibiting the growth and spread of cancer represents an important goal in the effort to enhance both cancer patient quality of life and survival. Cysteine cathepsins are a family of lysosomal proteases that are often upregulated in a variety of cancers and have been implicated in key processes in cancer progression. Our focus has centered on the development of novel non-peptidic inhibitors of cathepsin L. Small molecule inhibitors of the cysteine protease cathepsin L have the potential to limit degradation of extracellular matrix in the normal tissue surrounding the tumor thus retarding cancer metastasis. It has also been established that cathepsin L is involved in blood vessel development and thus inhibitors have the potential to suppress tumor growth and dissemination through an antiangiogenic effect. We have recently reported the synthesis of a series of functionalized benzophenone, thiophene, pyridine, and fluorene thiosemicarbazone derivatives several of which are potent inhibitors of cathepsin L with activity in the nM range while exhibiting IC50 values greater than 10 micromolar for cathepsin B. In addition, selected compounds were found to inhibit the migration, and invasion of DU-145 prostate cancer cells through Matrigel. Initial evaluation of the in vivo activity of the lead compound, designated KGP94, has been undertaken in a C3H mammary carcinoma. We have evaluated the effects of five daily doses of 20 mg/kg given to mice with treatment initiated either on the day of tumor inoculation or once the tumors reached 200 mm3 in size. Results to date indicate significant growth retardation is achieved against recently implanted and established tumors. Based on these encouraging results further work with this compound is ongoing.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1416. doi:10.1158/1538-7445.AM2011-1416
Abstract 1492: FoxM1 expression and its association with matrix metalloproteinases in diffuse large B-cell lymphoma
Authors: Siraj Abdul K. , Hussain Azhar R. , Ahmed Maqbool , Ahmed Saeeda O. , Bu Rong , Al-Dayel Fouad , Bavi Prashant , Uddin Shahab , Al-Kuraya Khawla S.
Published: June 2012

Forkhead Box M1 (FoxM1) has been shown to play a critical role in pathogenesis of various malignancies however its role in lymphoid malignancies is not fully elucidated. Therefore, in this study, we first investigated the role of FoxM1 in a large series of DLBCL tissues in a tissue micro array (TMA) format by Immunohistochemistry (IHC). We then followed this with in vitro studies on DLBCL cell lines using either pharmacological inhibitors of FoxM1 or siRNA knock down strategy. Using IHC, FoxM1 was detected in 84.6% of DLBCL tumors. FoxM1 expression was found to be significantly associated with early stage (p=0.0149), germinal centre phenotype (p=0.0126) and high proliferative tumor marker Ki67 (p<0.0001). In addition, FoxM1 expression was also significantly associated with expression of SKP2 (p<0.0001) and inversely associated with p27 expression (0.0215). Metalloproteinases, such as MMP-2 and MMP-9 play an important role in invasion and migration of cancer cells were also found to be significantly associated with expression of FoxM1 (p=0.0008 and p=0.0002) respectively. Our in vitro studies showed that treatment of DLBCL cell lines with thiostrepton, a specific inhibitor of FoxM1 inhibited cell proliferation, invasion and migration of DLBCL cells and induced apoptosis in a dose dependent manner. Inhibition of FoxM1 led to down-regulation of MMP-2, MMP-9 and SKP2 expression while the expression of p27 was up-regulated in DLBCL cells. Similar results were obtained using siRNA targeted against FoxM1. Furthermore, treatment of DLBCL cells with thiostrepton caused a dose dependent apoptosis in all DLBCL cell lines used in the study. On dissecting the apoptotic pathway, we found that thiostrepton induced its apoptotic effect via the mitochondrial pathway and activation of caspases. Finally, treatment of DLBCL cells with thiostrepton down-regulated the expression of XIAP, cIAP1 and Survivin. Altogether, these results suggest that FoxM1 signaling contribute in pathogenesis of germinal centre subtype of DLBCL and may serve as useful molecular biomarker and potential therapeutic target.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1492. doi:10.1158/1538-7445.AM2011-1492
Abstract 1561: The white adipose tissue is a rich reservoir of two distinct CD34+ mesenchymal and endothelial progenitor populations with a catalytic role in cancer progression
Authors: Martin-Padura Ines , Mancuso Patrizia , Gregato Giuliana , Marighetti Paola , Calleri Angelica , Corsini Chiara , Lohsiriwat Visnu , Petit Jean-Yves , Bertolini Francesco
Published: June 2012

A catalytic role has been proposed in neoplastic angiogenesis and cancer progression for bone marrow-derived circulating endothelial progenitors (CEPs). However, in preclinical and clinical studies the quantitative role of marrow-derived CEPs in cancer vascularization was found to be extremely variable. We have found that the human and murine white adipose tissue (WAT) is a very rich reservoir (containing a mean of 320 times more CD45-CD34+ progenitors/mL than the marrow) of two distinct CD45-CD34+ perivascular cell populations with mesenchymal and endothelial differentiation potential. When compared to marrow-derived CD34+ cells in gene expression array studies, purified WAT-CD34+ cells expressed similar levels of stemness-related genes such as Nanog, Sox2, Lin28 and others, and significantly increased levels of angiogenesis-related genes such as CD144, VEGFR2, ALK1 and others. In vitro the two WAT-CD34+ human populations generated fibroblasts, mature mesenchymal cells and mature endothelial cells and endothelial tubes. In aging mice, the number of WATCD34+ cells significantly declined in some WAT compartments but not in the mammary fat pad. When compared to marrow-derived progenitor cells, WAT-CD34+ cells expressed significantly increased (8 fold) levels of FAP-alpha, a crucial suppressor of anti-tumor immunity (Kraman et al, Science 2010). The co-injection of human WATCD34+ cells from lipofilling procedures significantly increased tumor growth and metastases in orthotopic models of human breast cancer in NSG mice, where human WATCD34+ cells contributed to tumor stroma and vascularization. A novel brain/adipocyte-BDNF/leptin axis has been recently proposed to play a potentially relevant role in cancer progression (Cao et al, Cell 2010). WATCD34+ progenitors express high levels of the receptors involved in this axis and our studies suggest a catalytic role in cancer progression and metastases.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1561. doi:10.1158/1538-7445.AM2011-1561
Abstract 160: Altered levels of circulating microRNAs in the plasma of non-small cell lung cancer patients: Potential as biomarker
Authors: Qian Xuemin , Fujioka Naomi , Kratzke Robert , Xu Wayne W. , Melkamu Tamene , Kassie Fekadu
Published: June 2012

The development of molecular markers for lung cancer could offset the deficiencies of currently available early detection and staging approaches. To this end, blood markers are particularly attractive since samples are obtained in a minimally invasive way at any stage of lung carcinogenesis. In this study we compared the levels of 10 microRNAs (miR-21, miR-31, miR-155, miR-147, miR-218, miR-377, miR-551b*, miR-569, miR-1283, and miR-1468) in the blood plasma of 27 non-small-cell lung cancer (NSCLC) patients and 38 healthy controls, using quantitative reverse transcription- polymerase chain reaction (qRT-PCR), to determine if these miRNAs might have a diagnostic value for NSCLC. The results of the area under the receiver operating characteristic curve (ROC), which measures the ability of the test to correctly classify those with and without the disease, were 0.98, 1.00, 0.8, and 0.98 for miR-21, miR-155, miR-377 and miR-1468, respectively, indicating that plasma levels of miR-21, miR-155, miR-377 and miR-1468 are powerful discriminators between patients and controls. At the optimum cutoff point for the cycle threshold (CT) value of the four miRNAs, the sensitivities of miR-21, miR-155, miR-377 and miR-1468 were 92.5%, 100%, 84%, and 100%, respectively; the corresponding values for the test specificity were 100%, 100%, 88%, and 100%, respectively. These results indicate the great potential of miR-21, miR-155, miR-377 and miR-1468 as diagnostic markers for NSCLC but need to be proved in prospective studies with larger samples.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 160. doi:10.1158/1538-7445.AM2011-160
Abstract 1624: SIRT1 overexpression in pancreatic adenocarcinomas correlates with shortened patient survival and small molecule inhibition as well as target knockdown of SIRT1 induces growth arrest and apoptosis in pancreatic cancer cells
Authors: Stenzinger Albrecht , Göppert Benjamin , Kamphues Carsten , Sinn Bruno , Bahra Markus , Neuhaus Peter , Weichert Wilko
Published: June 2012

Aims: Several lines of evidence indicate that the nuclear enzyme SIRT1 belonging to the class III histone deacetylases is implicated in the initiation and progression of various malignancies. SIRT1 exerts its function by regulating deacetylation of histone and non-histone substrates leading to transcriptional repression of tumor suppressor genes. Recently, SIRT1 gained attraction as druggable target. Since chemotherapeutic treatment options are still very limited in pancreatic cancer we aimed at investigating the potential of SIRT1 in this regard by evaluating the effects of SIRT1 inhibition in vitro and SIRT1 expression in vivo.

Methods: Employing immunohistochemistry SIRT1 expression was analyzed in a large cohort of pancreatic ductal adenocarcinomas and subsequently correlated with clinicopathological data and patient survival. Furthermore, we investigated the impact of SIRT1-specific small molecule inhibition and target knock down as well as combinatorial regimens including conventional chemotherapy and small molecule inhibitors directed against the EGFR in pancreatic cancer cell culture models. Using the xCELLigence system, cellular events were measured quantitatively in real time and corroborated by secondary conventional readouts including FACS analysis.

Results: We found nuclear SIRT1 expression in 36 (27.9%) of 129 pancreatic carcinomas. SIRT1 expression was significantly higher in poorly differentiated carcinomas (p=0.002). SIRT1 expression was positively correlated with the expression of conventional HDACs suggesting a shared regulation. Strong SIRT1 expression was a significant predictor of poor survival both in univariate (p=0.002) and multivariate (HR 1.65, p=0.045) analysis under inclusion of WHO stage and grade.

Small molecule inhibition and target knockdown of SIRT1 lead to a rapid growth arrest and influenced cell viability. This effect was even more pronounced in combinatorial regimens with Gefitinib, but not in combination with Gemcitabine.

Conclusions: SIRT1 is shown to be an independent prognostic biomarker for pancreatic adenocarcinomas. Moreover our data suggest that SIRT1 plays an important functional role in pancreatic cancer cell growth, which can be levered out by combinatory small molecule inhibition.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1624. doi:10.1158/1538-7445.AM2011-1624
Abstract 1626: Polyribosome-associated proteins targeted by HDACi promote the selective degradation of oncogenic transcripts
Authors: Scott Gary K. , Gabriel Bianca , Held Jason , Lee Elbert , Benz Christopher C.
Published: June 2012

We previously demonstrated that histone deacetylase inhibitors (HDACi), including several HDAC6-selective inhibitors, rapidly induce the accelerated decay of ERBB2 mRNA as well as ∼ 300 other oncogenic transcripts possessing U-rich sequences in their 3’UTR. While the mechanism underlying this oncogene transcript decay switch remains unknown, more recent evidence indicates that HDACi induced ERBB2 mRNA decay occurs on actively translating polyribosomes; and current studies are attempting to define the polyribosome proteins mediating this switch as they might represent novel therapeutic target candidates. A screen for proteins rapidly modified, recruited to, or released from polyribosomes following HDACi exposure (trichostatin-A, TSA) was undertaken in the ERBB2-positive human breast cancer cell line, SKBR3. Monosome and polyribosome enriched cytosols from control or TSA treated cells were either sucrose gradient fractionated for Northern and Western blot analysis of fractions, or the proteins were resolved by 2-dimensional (2-D) gel electrophoresis using three different isoelectric focusing gradients (ranging from pH 12 to pH 4). Western blots confirmed that monosome and polyribosome fractions contained the 40S ribosome scaffold protein, RACK1. Northern blots confirmed that HDACi (TSA × 6h) induced decay of ERBB2 mRNA, which could be completely prevented by co-treatment with the translational inhibitor cycloheximide, was confined to the polyribosome fractions. Comparison of the protein stained 2-D gels from control and treated samples indicated that 2-4 h of TSA exposure changes the isoelectric migration of >80 proteins, suggesting that HDACi induces post-translational modifications (PTMs) in various polyribosome proteins. Altered 2-D protein spots were excised, trypsin digested, and identified by nano-liquid chromatography (LC)-tandem mass spectrometry (MS/MS) using a 4000 QTRAP (Applied Biosystems, with Mascot v. 2.2 search database); in addition to RACK1, other TSA modified proteins include YB-1 (Y-box binding protein), DBPA (DNA-binding protein A), PRKRA (interferon-inducible double stranded RNA-dependent protein kinase activator A), NPM (nucleophosmin), and HNRPC (heterogeneous nuclear ribonucleoproteins C1/C2). Western blots looking for differential lysine acetylation (Ac-K) revealed that TSA induces Ac-K protein isoforms in the sucrose gradient polyribosome fractions containing decaying ERBB2 mRNA; induced Ac-K isoforms were also observed on the 2-D gels. MS/MS analysis of these TSA induced Ac-K isoforms is expected to reveal novel polyribosome HDAC substrates and candidate effectors of oncogene transcript decay.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1626. doi:10.1158/1538-7445.AM2011-1626
Abstract 1653: Vasodilator-stimulated phosphoprotein ser157 and ser239 are antimetastatic targets in colon cancer
Authors: Ali Mehboob , Pitari Giavanni Mario
Published: June 2012

Colorectal cancer is the third deadly cancer worldwide. Patient mortality is largely the result of metastatic disease progression, in which tumor cells undergo dramatic morphological changes characterized by the formation of membrane protrusive structures mediating invasion (lamellipodia, filopodia, invadopodia). These events are driven by the activity of actin-regulatory proteins, including the vasodilator-stimulated phosphoprotein (VASP), which reorganize the actin cytoskeleton to promote cell migration and invasion. In colon cancer, cAMP- and cGMP-dependent VASP phosphorylation at Ser157 (pVASP-Ser157) and Ser239 (pVASP-Ser239) represent opposing molecular switches, which regulate the invasive cell shape. Here, pVASP-Ser157 and pVASP-Ser239 differentially affected the metastatic potential of colon cancer cells, including tumor proliferation, colony formation, migration, adhesion and metastasis in vivo. Investigations employed intestinal cancer cells, human tumor cells expressing serine phosphoresistant VASP mutants, cGMP or cAMP analogs, agonists selectively elevating intracellular cGMP or cAMP levels, and orthotopic mouse models of colorectal cancer. Collectively, results demonstrated pVASP-Ser239 suppresses, while pVASP-Ser157 promoted, the metastatic phenotype of colon cancer cells. Thus, differential VASP serine phosphorylation is a novel targeted strategy to prevent colon cancer metastasis. Clinical translation of these findings may provide original therapeutic interventions to reduce mortality in patients with colon cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1653. doi:10.1158/1538-7445.AM2011-1653
Abstract 166: MicroRNA profiling analysis identify new signature of tumor-promoting M2-type macrophages
Authors: Ng Yee Seng , Roca Hernan , Sud Sudha , Pienta Kenneth J. , Chinnaiyan Arul M. , Keller Evan T. , Varsos Zachary S. , Maher Christopher A. , Cao Xuhong
Published: June 2012

M2-type macrophages are a major inflammatory component in prostate cancer and are associated with cancer cell growth, survival, and metastasis. Since recent findings have demonstrated the importance of microRNAs in the differentiation of human cells, we hypothesize that microRNAs are involved in regulating monocytic differentiation into M2-type macrophages and we aim to identify the microRNAs involved. Methods: Human CD14+ PBMCs were isolated from healthy donors and stimulated with cytokines known to promote differentiation into the M2-type macrophages (IL4, IL13, IL6, and MCSF). As controls, we also treated cells with media only and with INFγ, a cytokine known to promote M1-type polarization. Cells were collected at 48 hours post-stimulation and sent for microRNA profiling by microarray. Microarray findings were then confirmed by qRT-PCR. Results: Microarray profiling of cytokine-stimulated M2-type macrophages demonstrated an altered miRNA expression compared to M1-type macrophages and untreated monocytic cells. Specifically, of the 858 microRNAs that were profiled, miR-135a, miR-193b, miR-203, miR-302, miR-375, miR-452, miR-511, miR-603, miR-622, miR-654, miR-938 and miR-1238 were found to be up-regulated by greater than 2 folds. Furthermore, qRT-PCR analysis confirmed the microarray results. When the qRT-PCR analysis was repeated for cells treated with cytokines for 6 hours, 18 hours and 24 hours, we noticed that the overexpression in these microRNAs could already be observed at 6 hours post-treatment, although the expression levels were noticeably greater at later timepoints. Flow cytometric analysis showed the surface expression of CD206 (mannose receptor: the well characterized surface marker of M2-type macrophages) in both MCSF and IL-4-treated cells but only after at least 18 hours of stimulation. Conclusions: MicroRNA expression in PBMCs is altered when they are stimulated by M2-type promoting cytokines and the change in microRNA expression preceded the expression of the M2-specific CD206 surface marker. This change in microRNA expression can therefore be used as a marker for M2-polarized macrophages and may play critical roles in the regulation of macrophage differentiation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 166. doi:10.1158/1538-7445.AM2011-166
Abstract 1666: Targeting tumor-initiating cells for the treatment of hepatocellular carcinoma
Authors: Zhang Zhiqian , Zhao Wei , Wang Limin , Han Haibo , Xing Baocai , Li Jiyou , Wang Yu
Published: June 2012

Tumor-initiating cells (TICs) or so-called cancer stem cells (CSCs) are a minor subpopulation of poor differentiated cells that are responsible for tumor initiation, maintenance, and spreading. Hence, eliminating these cells is supposed to be an effective way of the treatment of cancer. By immunizing mice with TIC-enriched hepatocellular carcinoma (HCC) cells (Xu XL, et al. The properties of tumor-initiating cells from a hepatocellular carcinoma patient's primary and recurrent tumor. Carcinogensis. 2010; 31:167-74.) and by subsequent hybridoma technique, we have obtained an antibody 1B50-1, which recognizes the antigen of a 150 KD ion channel-composing protein. Despite the percentage of 1B50-1 positive cells varies from <1% to ∼90% in established hepatocellular carcinoma cell lines, the administration of this antibody by i.p. injection at a dose of 40mg/kg could suppress the growth of HCC cell lines’ engraftments in NOD/SCID mice with an inhibition rate that could be as high as 87%, comparing with control groups. Furthermore, 100 of 1B50-1 positive cells, but not the negative compartments, from several HCC cell lines were sufficient to initiate tumor growth in NOD/SCID mice, and displayed self-renewal ability as demonstrated by serial transplantation assay in mice. The 1B50-1 positive cells also expressed higher levels of Nanog, Sox-2, and Oct 4 than the negative ones. Immunohistochemical staining of 29 HCC specimen with 1B50-1 has revealed that only a small population of positive cells were scattered in 16 HCC tissues, and that these cells with similar distribution pattern were only found in 4 out of 12 paracancerous tissues. These data suggest that the antigen for 1B50-1 is a new marker for HCC tumor-initiating cells, and that it is a promising target for HCC molecular therapy. 1B50-1 is worthy of further developing into drug for the treatment of HCC.

Funding: This study was supported by the National Basic Research Program of China (973 Program, No 2010CB529402), National Natural Science Foundation of China, and “863” Program (2008AA02Z108).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1666. doi:10.1158/1538-7445.AM2011-1666
Abstract 1691: Niche inflammation strengthens the IGF-1R-Akt signaling and results in poor prognosis of HBV-HCCs
Authors: Lin Hsin-Yi , Chang Te-Sheng , Jen Hsin-Wei , Yang Ya-Ting , Lin Ya-Ping , Lee Kam-Fai , Tung Shui-Yi , Kuo Liang-Mou , Huang Yen-Hua
Published: June 2012

Hepatocellular carcinoma (HCC) is the fifth most common cancer in the world. Inflammation of various etiologies has been documented to highly associate with the development of HCC. However, the role of inflammation in virus-induced hepatocarcinogenesis still remains largely unknown. Here we demonstrate that the potential synergistic effect of HBV and proinflammatory cytokines might associate with the poor prognosis of HBV-related HCC (HBV-HCC) via IGF-1R signaling activation. In this study, human HCC frozen tissues and tissue blocks were collected for clinical analysis under IRB approval. Real-time quantitative PCR analysis of HCC frozen tissues revealed a reciprocal correlation between the transcriptional levels of IGF-1R and disease-free interval (DFI) in HBV-HCC tissues (N=117). Immunohistochemical staining of HCC tissue blocks also showed a strong correlation between IGF-1R signaling activation (IGF-1R/phospho-IGF-1R) and short DFI. To test the collaborative effect of HBV and inflammation, human HCC cell lines including HBV-related cells (HepG2.2.15, PLC5, Hep3B, and HA22T) and non-HBV-related cells (HepG2 and Huh 7) were cultivated with inflammation condition medium (inflam-CM). We found that inflam-CM treatment significantly increased the levels of pluripotent regulator IGF-1/IGF-1R and IGF-1R signaling (phospho-IGF-1R and phospho-Akt) in HBV- HCC cells (HepG2.2.15 and Hep3B). Additionally, inflam-CM also markedly increased the drug resistance capacity of HepG2.2.15 and Hep3B cells. Blockage of IGF-1R phosphorylation by picropodophyllin (PPP) or down-regulation of IGF-1R by siRNA dramatically increased the drug sensitivity of HBV-HCC cells. Further analysis using neutralizing antibodies against potential proinflamatory cytokines is ongoing to identify the cross-talk between inflammation and IGF-1R signaling in HBV- HCCs. In summary, our works demonstrate that niche inflammation may activate the IGF-1R-Akt signaling and result in poor prognosis of HBV-HCC. This finding may provide a potential therapeutic target for HBV-HCC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1691. doi:10.1158/1538-7445.AM2011-1691
Abstract 1697: Gene signature, histopathological classification, and cisplatin-activated expression profiles in human epithelial ovarian cancer
Authors: Guo Yi , Zhu Honglan , Liang Xiaobing , Mueller Michael D. , Yu Jing Jie
Published: June 2012

PURPOSES: To identify biomarkers in epithelial ovarian cancer (EOC) and investigate mechanism of cisplatin-induced activation and expression, 50 genes were screened from 131 ovarian cancer patient samples. Twenty expressed genes were assessed in 30 samples by qPCR. Ovarian cancer cells were treated with cisplatin and studied for expression of cisplatin-activated genes.

RESULTS: Studies of gene signature of histopathological classification TAL2, EGF, ILF3 and UBE21 shown unique expression patterns in EOC histological types. The expression of ILF3 in tumor tissues was significantly higher than normal samples, and especially high in serous carcinomas, compared to mucinous, endometrial and clear cell carcinomas. ILF3 and UBE21 showed overexpression in advanced stage and poorly differentiated patient samples. Investigation of cisplatin-activated signaling indicated increased mRNA expression of ILF3, UBE21 and other kinases involved in cell cycle checkpoint, DNA repair, and apoptosis such as p53, Chk2, c-jun, and hMLH1.

CONCLUSIONS: Overexpression of ILF3 and UBE21 in advanced stage and advanced grade of EOC specimens suggests that these two genes may play important roles in tumorgenesis, tumor progression and pathological differentiation of the disease. Results from in vitro studies demonstrate that activated genes p53, Chk2, c-jun and hMLH1 play a role in chemo-resistance to cisplatin therapy. Further investigation of the molecular mechanisms of the identified genes is warranted.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1697. doi:10.1158/1538-7445.AM2011-1697
Abstract 1699: Thymidylate synthase and thymidine phosphorylase expression contributes to cisplatin resistance
Authors: Lin Yun-Wei , Tsai Min-Shao , Chiu Yu-Fan , Weng Shao-Hsing , Kuo Ya-Hsun , Chuang Show-Mei
Published: June 2012

Chemotherapy for advanced human non-small cell lung cancer (NSCLC) includes platinum-containing compound such as cisplatin in combination with a second- or third-generation cytotoxic agents. 5-Fluorouracil (5-FU) belongs to anti-metabolite chemotherapeutics, and its mechanism of cytotoxicity is involved in inhibition of thymidylate synthase (TS). TS and thymidine phosphorylase (TP) were key enzymes of the pyrimidine salvage pathway. In this study, we have examined the molecular mechanism of TS and TP in regulating the drug sensitivity to cisplatin in NSCLC cell lines. Cisplatin could increase the phosphorylation of MKK1/2-ERK1/2 and the protein levels of TS and TP through enhancing the protein stability in A549 and H1975 cells. Blocking ERK1/2 activation by MKK1/2 inhibitor (U0126) decreases TS and TP protein levels in both cell lines treated with cisplatin. Depletion of endogenous TS or TP expression by specific small interfering RNA transfection significantly increases cisplatin-induced cell death and growth inhibition. Moreover, pemetrexed (TS inhibitor) enhances cisplatin-induced cytotoxic effects. Combined treatment with 5-FU can decrease cisplatin-induced ERK1/2 activation and the induction of TS and TP, which can subsequently result in synergistic cytotoxic effects. Enforced expression of constitutive active MKK1/2 vectors rescue the protein levels of phospho-ERK1/2, TS and TP, and the cell viability which were decreased by cisplatin and 5-FU combination. In contrast, U0126 enhances the drug sensitivity to cisplatin and/or 5-FU in lung cancer cells. We conclude that the up-regulation of ERK1/2-dependent TS and TP can protect human lung cancer cells from cisplatin-induced cytotoxicity.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1699. doi:10.1158/1538-7445.AM2011-1699