bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract A187: Matrix metalloproteinase-9 as a novel therapeutic target in B-cell chronic lymphocytic leukemia.
Authors: Bailón Elvira , Ugarte-Berzal Estefanía , Amigo-Jiménez Irene , Van den Steen Philippe , Opdenakker Ghislain , García-Marco José A. , García-Pardo Angeles
Published: August 2014

B-cell chronic lymphocytic leukemia (B-CLL) cells secrete matrix metalloproteinase-9 (MMP-9) at higher levels than normal B-cells. We previously showed that MMP-9 is also constantly present on the surface of B-CLL cells, where it binds to a docking complex containing alpha4beta1 integrin and CD44v. Interaction of MMP-9 with these receptors contributes to B-CLL progression since: 1) elicits a signaling pathway that leads to cell survival, and 2) regulates in vitro cell migration (Redondo-Muñoz et al, Blood 2008, Cancer Cell 2010). We have now addressed the effect of MMP-9 overexpression at the cell surface, a situation likely occurring at B-CLL niches. We have used primary B-CLL cells as well as stable MEC-1 cell transfectants, containing either empty (Mock) or MMP-9-encoding lentiviral vectors. In vivo experiments using NOD/SCID mice showed that MMP-9-MEC-1 transfectants had significantly impaired homing to bone marrow and spleen, compared to Mock-cells. Importantly, incubation of primary B-CLL cells with MMP-9 inhibited their homing to these organs. Inhibition of migration was dose-dependent and only partially involved the catalytic MMP-9 activity, as the inactive mutant MMP-9MutE had a partial effect. MMP-9-MEC-1 cells or primary B-CLL cells incubated with MMP-9 showed reduced activation of key signaling pathways involved in cell migration, such as Akt, ERK, FAK and RhoAGTPase. In contrast, PTEN, a phosphatase that antagonizes Akt, ERK and FAK signaling and inhibits migration, and the RhoA inhibitor p190RhoGAP were upregulated in MMP-9-cells and B-CLL cells overexpressing MMP-9. Moreover, MMP-9-cells were more resistant to fludarabine or arsenic trioxide cytotoxic treatments than Mock-cells, indicating that MMP-9 also protected B-CLL cells from drug-induced apoptosis. Altogether our results reveal novel functions for MMP-9 likely contributing to malignant cell retention in lymphoid organs and drug resistance. MMP-9 thus contributes to B-CLL pathology and may represent a therapeutic target in this disease.

Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A187.

Citation Format: Elvira Bailón, Estefanía Ugarte-Berzal, Irene Amigo-Jiménez, Philippe Van den Steen, Ghislain Opdenakker, José A. García-Marco, Angeles García-Pardo. Matrix metalloproteinase-9 as a novel therapeutic target in B-cell chronic lymphocytic leukemia. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A187.
Abstract A191: DHMEQ: A novel NF-kappaB inhibitor with good results in pediatric medulloblastoma cell lines.
Authors: Ramos Priscila Maria Manzini , Castro-Gamero Angel Mauricio , Pezuk Julia Alejandra , Scrideli Carlos Alberto , Umezawa Kazuo , Tone Luiz Gonzaga
Published: August 2014

Medulloblastoma (MB) is the most common malignant central nervous system tumor of childhood representing 15-30% of pediatric brain tumors. The standard treatment involves surgery and chemotherapy and radiotherapy is used only for patients older than 3 years old. The risk of neurocognitive sequel, growth and endocrine systems leading the search for new therapeutic approaches. Great efforts have been made to improve treatments with high success rates and minimal toxicity, among them DHMEQ is a drug that has shown a low toxicity and high effectivity in blocking the NFkappaB, a key transcription factor that control the expression of a large number of genes related to cell proliferation, growth and apoptosis, showing antineoplasic effects in several cancer cell lines. There are no reports showing the effects of DHMEQ in MB. Cell proliferation, clonogenic capacity, wound healing assay, apoptosis (Annexin V), and combination with chemotherapeutic drugs (Cisplatin (CDDP), Temozolomide (TMZ) and Etoposide) or radiation were performed on three pediatric MB cell lines: UW402, UW473 e ONS76. Functional tests were performed using 0-20 µg/mL DHMEQ at 24, 48 and 72h. IC50 values of each drug and drug combination analyses were based in Chou-Talalay method, and simultaneous treatments of DHMEQ with CDDP (2:1 ratio), TMZ (1:200 ratio) or Etoposide (1:0,8 ratio) were performed for 48h or 72h. For gamma radiation clonogenic survival assays were used the doses of 2, 4 and 6 Gy. Statistical analysis was made by ANOVA and Pairwise Multiple Comparison de Holm-Sidak tests (SigmaStat 3.5). All experiments showed DHMEQ caused a decrease on cell proliferation, viability, migration ratio and clonogenic capacity, while increase apoptosis (P<0.05). In the drug combination assay, DHMEQ acted additively with CDDP and synergistically with TMZ and Etoposide in all cell lines. Moreover, DHMEQ enhanced significantly the radiation effects on the three cell lines studied. These results support the biological importance of the NFkappaB as a promising therapeutic target for medulloblastoma treatment, and the potencial effect radio- and chemo- sensitizing of DHMEQ on MB cells.

Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A191.

Citation Format: Priscila Maria Manzini Ramos, Angel Mauricio Castro-Gamero, Julia Alejandra Pezuk, Carlos Alberto Scrideli, Kazuo Umezawa, Luiz Gonzaga Tone. DHMEQ: A novel NF-kappaB inhibitor with good results in pediatric medulloblastoma cell lines. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A191.
Abstract A205: Development and application of a platform for analysis of protein biomarkers in clinical biopsies of limited quantity.
Authors: Du Jinyan , Lahdenranta Johanna , Zhang Bo , Kudla Art , McDonagh Charlotte
Published: August 2014

MM-111 is a bispecific antibody that inhibits HER3 activation in HER2-expressing tumor cells. Using preclinical models, we have generated hypotheses on both pharmacodynamic and predictive biomarkers of MM-111’s antitumor activity. To evaluate these hypotheses in patients receiving MM-111, we are collecting tumor tissue samples from patients in clinical trials both prior to and following exposure to MM-111. Because of the limited quantity of tissue available, it is challenging to measure a broad spectrum of potential protein biomarkers in these specimens. Here we have optimized tissue processing procedures and developed multiplex Luminex protein assays in order to explore a more extensive biomarker panel from the frozen tumor biopsies collected from patients. To develop a method suitable for use with small tissue quantities, we simulated the clinical biopsy procedures in tumor-bearing mice. We collected 18G core needle biopsies from tumors in mouse xenograft models and flash-froze the samples in the same manner as is being executed in the clinic for patient biopsies. We then prepared total protein lysates from these biopsy samples and measured MM-111 biomarkers with multiplex Luminex protein assays. We found that a sufficient amount of protein could be isolated from 18G core needle biopsies for replicate Luminex assays. Signals were well correlated between two independent biopsies from the same xenograft. We also used this preclinical model system to define the parameters for sample collection in the clinic. We found that the phospho and total protein levels of key MM-111 biomarkers were maintained in tumor biopsy tissue stored for up to 60 minutes at ambient temperature before freezing. We are now using this system to further explore the mechanisms of MM-111 activity compared to other therapeutics that target HER2-expressing tumors. We have analyzed the signaling effects of MM-111 and pertuzumab in biopsies obtained from an esophageal xenograft model derived from a human cancer cell line and observed that MM-111 but not pertuzumab could effectively down-regulate EGFR and HER2 phosphorylation, which is consistent with in vitro data. In summary, we have developed a method for accurately measuring multiple biomarkers in core needle biopsy samples using multiplex Luminex protein assays. We plan to use this approach to test our biomarker hypotheses in MM-111 clinical trials.

Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A205.

Citation Format: Jinyan Du, Johanna Lahdenranta, Bo Zhang, Art Kudla, Charlotte McDonagh. Development and application of a platform for analysis of protein biomarkers in clinical biopsies of limited quantity. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A205.
Abstract A241: Differential roles of STAT3 depending on the mechanism of STAT3 activation in gastric cancer cells.
Authors: Okamoto Wataru , Okamoto Isamu , Tuchihara Katsuya , Yanagihara Kazuyoshi , Nishio Kazuto , Nakagawa Kazuhiko
Published: August 2014

Background: Signal transducer and activator of transcription 3 (STAT3) is a transcription factor that is activated in response to growth factors and cytokines, and which contributes to the regulation of cell proliferation, apoptosis, and motility in many human tumor types.

Methods: We investigated the mechanisms of STAT3 activation and the function of STAT3 depending on its mechanism of activation in gastric cancer cells.

Results: All cell lines positive for constitutive MET activation manifested STAT3 phosphorylation. The MET-tyrosine kinase inhibitor (TKI) and cell transfection with a small interfering RNA (siRNA) specific for MET mRNA inhibited STAT3 phosphorylation in MET-activated cells, indicating that STAT3 activation is linked to MET signalling. Forced expression of a constitutively active form of STAT3 also attenuated MET-TKI-induced apoptosis, suggesting that inhibition of STAT3 activity contributes to MET-TKI-induced apoptosis in MET-activated gastric cancer cells. On the other hand, MKN1 and MKN7 cells, both of which are negative for MET activation, produced interleukin-6 (IL-6) that activated STAT3 through the Janus kinase (JAK) pathway but which did not appear to be required for cell survival. Depletion of STAT3 by transfection with a specific siRNA inhibited both the migration and invasion activities of MKN1 and MKN7 cells, suggesting that STAT3 activated by IL-6 contributes to regulation of cell motility.

Conclusion: Our data thus show that activated STAT3 contributes to either cell survival or cell motility in gastric cancer cells, and that these actions are related to different mechanisms of STAT3 activation.

Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A241.

Citation Format: Wataru Okamoto, Isamu Okamoto, Katsuya Tuchihara, Kazuyoshi Yanagihara, Kazuto Nishio, Kazuhiko Nakagawa. Differential roles of STAT3 depending on the mechanism of STAT3 activation in gastric cancer cells. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A241.
Abstract A280: Small molecule inhibition of atypical protein kinase C iota reveals a functional basis for oncogenesis.
Authors: Linch Mark , Sanz-Garcia Marta , Kjaer Svend , Soriano Erika , Riou Philippe , Knowles Philip , Dillon Christian , Roffey Jon , McDonald Neil , Parker Peter
Published: August 2014

Background: Atypical protein kinase C iota (PKCι) is a serine/threonine kinase that has been implicated in a variety of cellular processes including polarity, migration, proliferation and survival. PKCι is amplified in many cancers correlating with poor prognosis, however the upstream activating inputs and downstream effectors are not well defined.

Experimental Procedures: 3D MDCK epithelial cell spheroids were grown in Matrigel and used to assess polarized morphogenesis and growth. PKCι was manipulated by inhibition with CRT0066854, or transfection with siRNA or cDNA variants. A PKCι binding interface was identified by X-ray crystallography and interrogated by mass spectrometry of wildtype or binding mutant immunocomplexes.

Results: To identify potential inputs into PKCι we assessed a panel of transformed MDCK cells; p110α, Src, Ras, ErbB2 and Raf. Parental MDCK cells form polarized cysts in Matrigel, however Ras, ErbB2 and p110α MDCK cells formed dysplastic spheroids. In the case of Ras and ErbB2, but not p110α MDCK spheroids, normal polarized morphogenesis could be rescued by siRNA-PKCι or partial PKCι inhibition with decreased spheroid size seen for all transformed cells lines. An effector binding site (PKCι RIPR motif) identified from the crystal structure of the kinase and characterized as a recruitment site for select targets including LLGL2 and Myosin X, was found to be critical for PKCι to confer polarized morphogenesis. Interestingly this binding motif shows a low frequency of mutation in a subset of human tumors.

Conclusions: Downstream of Ras and ErbB2, PKCι activity is enhanced with a resulting dysplasia and loss of polarization, that can be rescued by PKCι inhibition. Mutation of the PKCι-RIPR effector binding motif that is found in cancer, prevents interaction with LLGL2 and its expression also elicits growth as non-polarized, dysplastic epithelial spheroids. This indicates that PKCι may promote cancer, not only by amplification, but by upstream oncogenic inputs and somatic mutation. The oncogene dependent PKCι hyperactivation is pro-proliferative in 3D culture, providing compelling evidence that PKCι is a good target for Ras mutant tumors.

Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A280.

Citation Format: Mark Linch, Marta Sanz-Garcia, Svend Kjaer, Erika Soriano, Philippe Riou, Philip Knowles, Christian Dillon, Jon Roffey, Neil McDonald, Peter Parker. Small molecule inhibition of atypical protein kinase C iota reveals a functional basis for oncogenesis. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A280.
Abstract A285: Inhibition of histone deacetylase enhances radiosensitivity in hepatocellular carcinoma by inhibiting nuclear translocation of HDAC4 and DNA repair.
Authors: Liu Wei-Lin , Cheng Jason Chia-Hsien , Gao Ming , Cheng Ann-Lii
Published: August 2014

Purpose: Tumor control of hepatocellular carcinoma (HCC) by radiotherapy (RT) remains unsatisfactory. The use of an effective molecularly targeted agent in combination with RT is a potential therapeutic approach. Histone deacetylase (HDAC) inhibitors are undergoing clinical evaluation for cancer therapy. HDAC modulates chromatin structure, involves the inhibition of DNA repair, and is considered to influence radio-response. The study aim was to assess the effect of combining a hydroxamic acid derivative HDAC inhibitor, LBH589, with RT on human HCC cells.

Methods and Materials: Human HCC cell lines (Huh-7 and PLC-5) were used to evaluate the in vitro synergism of combining LBH589 with irradiation. Western blot and immunofluorescence investigated the protein expressions after the combined treatment. Western blot analysis of proteins from nuclear and cytoplasmic fractions was done by using anti-HDAC4 antibody. Annexin-V immunofluorescence was used to measure apoptotic cells after the combined treatment.

Results: Clonogenic survival showed a greater than additive effect with LBH589 administered before RT as compared to RT alone. LBH589 significantly enhanced the accumulation of acetyl-histone H3. Notably, radiation itself induced translocation of HDAC4 to the nucleus. In contrast, treatment with LBH589 followed by irradiation resulted in HDAC4 confinement to the cytoplasm. The findings indicated that HDAC inhibition affects the nuclear localization of HDAC4. We studied the combined effect of LBH589 and RT on DNA repair in both cell lines. γ-H2AX foci representing DNA double-strand breaks (DSBs) were detected in the nuclei at 24 hours following 10-Gy irradiation. LBH589 administered before irradiation increased the persistence of ϒ-H2AX foci beyond 24 hours. Akt phosphorylation, active caspase-3, cleaved PARP in Western blot analysis, and annexin V immunofluorescence assay revealed a synergistic effect of combining LBH589 with RT on inducing apoptosis.

Conclusion: This study identifies the inhibitory nuclear translocation of HDAC4 by adding LBH589 to RT, and recognizes the effect of LBH589 on sensitizing HCC cells to radiation-induced DNA DSBs and apoptosis. Taken together, LBH589 is a potent, orally bioavailable inhibitor of HDAC with therapeutic value as a radiosensitizer of HCC.

Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A285.

Citation Format: Wei-Lin Liu, Jason Chia-Hsien Cheng, Ming Gao, Ann-Lii Cheng. Inhibition of histone deacetylase enhances radiosensitivity in hepatocellular carcinoma by inhibiting nuclear translocation of HDAC4 and DNA repair. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A285.
Abstract A39: Analysis of endoscopic ultrasound (EUS) - guided fine needle aspiration (FNA) samples from patients with suspected pancreatic ductal adenocarcinomas (PDACS) using plectin-1 (PLEC1) and KRAS oncogene mutation analysis.
Authors: Park Joo Kyung , Song Byung Jun , Ryu Ji Kon , Kim Yong-Tae , Lee Se-Hoon
Published: August 2014

Introduction: Pancreatic cancer remains an incurable and rapidly lethal cancer, with a 5-year survival rate of less than 10%. Little to nothing is known about advanced pancreatic cancer because of problems with tissue availability and usually insufficient quantity of tissue available for diagnosis and study. One of the key features of genetic basis of PDAC is the point mutation of KRAS oncogene occurred in 95 % of its pathogenesis. Plec1 was also recently identified as PDAC specific biomarker.

Aims and Methods: The aim of this study was to find out the techniques using minimal specimens in order to achieve accurate diagnosis of patients with suspected PDACs. DNA was extracted from EUS-guided FNA samples in 80 patients with suspected PDACs. Seven different somatic mutations of KRAS oncogene were evaluated using ARMS and Scorpions technology, and immunohistochemical staining of Plec1 antibody was done.

Results: The total of 80 study patients with suspected of PDACs underwent EUS-guided FNA and the final diagnosis were as follows; PDAC: 63 patients, pancreas neuroendocrine tumor: 4, autoimmune pancreatitis: 3, chronic pancreatitis: 1, metastasis to pancreas: 6, others: 3. The sensitivity and specificity of pathologic diagnosis in EUS-guided FNA samples were 87% and 88% respectively. When we combine KRAS oncogene mutational analysis and pathologic reports of FNA samples, we could get the following sensitivities and specificities; 95% vs. 97%. Also, adding the results of plectin-1 antibodies of immunohistochemical staining could increase the sensitivity of PDAC diagnosis.

Conclusion: Triple combinations of the techniques (pathologic reports, KRAS oncogene mutational analysis, Plec1 staining) could increase accuracy, however, cost-effectiveness and laborious work should be taken into consideration in clinical practice. Further investigation of using minimal specimens to study PDACs may enable us to understand biological behavior and to identify better tools for the diagnosis and treatment of this deadly cancer.

Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A39.

Citation Format: Joo Kyung Park, Byung Jun Song, Ji Kon Ryu, Yong-Tae Kim, Se-Hoon Lee. Analysis of endoscopic ultrasound (EUS) - guided fine needle aspiration (FNA) samples from patients with suspected pancreatic ductal adenocarcinomas (PDACS) using plectin-1 (PLEC1) and KRAS oncogene mutation analysis. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A39.
Abstract A5: Effects of rifampin on the pharmacokinetics (PK) of lenvatinib (L) in healthy participants.
Authors: Shumaker Robert , Aluri Jagadeesh , Fan Jean , Martinez Gresel , Thompson Gary A. , Ren Min
Published: August 2014

Background: L is an orally administered multitargeted tyrosine kinase inhibitor of VEGF receptors 1, 2, 3; FGF receptors 1, 2, 3, 4; PDGFR-β; RET; and KIT under clinical investigation in solid tumors at doses of 24 mg/d. This study in volunteers assessed the influence of P-glycoprotein (Pgp) inhibition (single-dose R) and simultaneous CYP3A4 and Pgp induction (multiple-dose R) on L PK parameters.

Methods: This was a single-center, single-dose, open-label, sequential, 3-period study. Subjects were administered 3 regimens with a 14-day washout between each treatment. Subjects received a single oral dose of L 24 mg on Day 1 (Period 1), Day 15 (Period 2), and Day 43 (Period 3). In Period 2 on Day 15, subjects received a single oral dose of rifampin 600 mg coadministered with L. In Period 3, subjects received oral rifampin 600 mg/d for 21 days (Days 29 to 49).

Results: 15 subjects were enrolled and completed the study. Single-dose rifampin increased free (unbound) L AUC0-∞ (32%) and Cmax (30%). Multidose rifampin decreased free L AUC0-∞ (9%) without a corresponding decrease in Cmax; mean t1/2(free L) decreased by ∼2.5 h (to ∼17 h) following single-dose rifampin and by ∼5 h following multidose R. The 90% confidence intervals (CIs) of the geometric least squares (GLS) mean ratios for Cmax and AUCs were above the upper CI boundary of the prespecified bioequivalence interval of 80%-125% following a single rifampin dose but within the prespecified bioequivalence interval following multidose R. Headache (n=3, 20%) was the most frequently reported treatment-emergent adverse event (TEAE); 5 subjects reported mild or moderate treatment-related TEAEs (n=5, 33%) most frequently nausea (n=3, 20%) and diarrhea (n=2, 13%).

Conclusions: L exposure was increased by single-dose rifampin but decreased by multiple rifampin doses. The magnitude of these changes was relatively small and likely not clinically meaningful.

Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A5.

Citation Format: Robert Shumaker, Jagadeesh Aluri, Jean Fan, Gresel Martinez, Gary A. Thompson, Min Ren. Effects of rifampin on the pharmacokinetics (PK) of lenvatinib (L) in healthy participants. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A5.
Abstract A79: Delivery of multiple genes to neuroblastoma using peptide-conjugated polymeric nanoparticles for cancer therapy.
Authors: Lee Jangwook , Choi Chang Seon , Lee Sang Kyung , Lee Kuen-Yong
Published: August 2014

Tissue-specific delivery of functionalized polymeric nanoparticles has found potential for the treatment of various diseases including cancer. Theragnosis is an emerging approach that can perform diagnosis and therapy simultaneously, and has made great progress in cancer diagnosis and therapy. In this presentation, we report the preparation and characterization of tumor-targeting polymeric nanoparticles delivering both imaging probe and multiple anticancer genes. Poly(D,L-lactide-co-glycolide) (PLGA) nanoparticles containing 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI) and siRNAs (c-Myc, VEGF, and Bcl-2) were fabricated by a W/O/W double emulsion method, followed by chemical modification with rabies virus glycoprotein (RVG) peptide using a carbodiimide chemistry. The RVG peptide, derived from rabies virus, binds specifically to acetylcholine receptors substantially found on the membrane of neuroblastoma. We evaluated in vitro cellular uptake and gene silencing efficiency of the nanoparticles using N2a cells. The cellular uptake and gene silencing efficiency of RVG-conjugated nanoparticles significantly increased compared with non-conjugated nanoparticles. We also monitored biodistribution and tumor-targeting ability of the nanoparticles after intravenous injection into a neuroblastoma-bearing mouse model. At 24 h post-injection, the whole body fluorescence was clearly detected at the tumor site when RVG-conjugated nanoparticles were injected into the mice. From ex vivo quantitative analysis, prominent fluorescence intensity of RVG-conjugated nanoparticles was found in tumor, compared with non-conjugated nanoparticles. Finally, in vivo anticancer efficacy of the nanoparticles was evaluated. The target specificity of the nanoparticles led to remarkable anticancer efficacy in the mouse model. This approach suggests that tumor-targeting polymeric nanoparticles are useful for delivery of both imaging probe and multiple anticancer genes specifically to neuroblastoma, which may be critical in the design and tailoring of theragnostic systems for various diseases including cancer.

Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A79.

Citation Format: Jangwook Lee, Chang Seon Choi, Sang Kyung Lee, Kuen-Yong Lee. Delivery of multiple genes to neuroblastoma using peptide-conjugated polymeric nanoparticles for cancer therapy. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A79.
Abstract A82: Localized tumor delivery of radiosensitizers and chemotherapeutics using ‘INCeRT’ implants.
Authors: Kumar Rajiv , Belz Jodi E. , Markovic Stacey , Korideck Houari , Ngwa Wilfred F. , Niedre Mark , Berbeco Ross I. , Cormack Robert , Makrigiorgos Mike G. , Sridhar Srinivas
Published: August 2014

In cancer therapy, effective delivery of the therapeutic plays a critical role in determining the success of the treatment planning system. In last decade, there is a major thrust in developing novel nanoparticles based delivery systems to improve the therapeutic benefits in cancer. An efficient delivery system should be able to target the diseased site with minimal systemic toxicity and a slow sustained release of the drug. Here, we have fabricated a nanoparticles based smart ‘INCeRT’ (Implantable Nanoplatform for Chemo-Radiation Therapy) implant for localized delivery of radiosensitizing nanoparticles/ drugs in prostate cancer in conjunction with brachytherapy. These implants are physically similar to the clinically used brachytherapy spacers but have the added capability of imaging and local drug delivery. We have fabricated INCeRT spacers with biocompatible and biodegradable polymer, PLGA (poly-(lactide-co-glycolide)) impregnated with nanoparticles encapsulating imaging probe (Cyanine 7.5) and chemotherapeutic drug, docetaxel (DTX). Using a similar approach, we have also fabricated PLGA spacers impregnated with high Z (atomic number) gold nanoparticles (Hi-Z-CuRE: High Z-Customizable Radiotherapy Enhancement) for effectively boosting the radiation dose locally. The morphology, composition and nanoparticle's distribution inside the spacers was studied by SEM (scanning electron microscopy) and EDS (Energy-dispersive X-ray spectroscopy). Further, preliminary in vivo imaging experiments with subcutaneous prostate cancer tumored mice implanted with INCeRT spacers showed a size dependent diffusion of nanoparticles from the spacers in the tumor matrix. Also, from in vivo therapeutic studies with these spacers showed a sustained and slow release of the DTX from the spacers and showed a better response in suppressing the tumor as opposed control mice with saline injections. Further experiments for studying the combined chemo-radiation therapy are underway. This new treatment approach would be of crucial benefit for patients with local relapse who require salvage radiotherapy but have reached their radiotherapy normal tissue dose limits. This work was supported partially by NSF-DGE-0965843, HHS/1U54CA151881 CORE1, 1R03 CA164645-01 and a seed grant from the BWH Biomedical Research Institute.

Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A82.

Citation Format: Rajiv Kumar, Jodi E. Belz, Stacey Markovic, Houari Korideck, Wilfred F. Ngwa, Mark Niedre, Ross I. Berbeco, Robert Cormack, Mike G. Makrigiorgos, Srinivas Sridhar. Localized tumor delivery of radiosensitizers and chemotherapeutics using ‘INCeRT’ implants. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A82.
Abstract A85: Peptides specific for chronic lymphocytic leukemic cells: Characterization and use in targeted drug delivery.
Authors: Firer Michael A. , Lubin Bat-Chen , Cohen Sharon , Luboshits Galia
Published: August 2014

The ability to selectively target drugs to Chronic Lymphocytic Leukemia (CLL) cells may significantly improve the efficacy of current chemotherapy protocols for this disease. Our studies in model leukemic systems have already demonstrated that a strategy involving Peptide-Drug Conjugates (PDCs) may achieve this goal. In a previous preliminary study, we used a phage display peptide library to isolate a series of peptides that bind CLL cells. The current research has expanded on this work and began by absorbing a 7-mer linear and 7-mer cyclic phage display peptide libraries on a series of normal human or murine primary cells. Rigorous pre-absorption reduces the original vast phage library population (1011 clones) and enhances the ability to select peptides specific for the target CLL cells. The cells used included endothelial, heart, kidney, lung, liver, skin, fibroblasts and several subpopulations of blood cells. These absorptions resulted in a 7-log reduction in the size of the phage libraries. The stock libraries were also injected into mice. Recovery of blood circulating phage 24hrs later showed that in vivo absorption reduced the pool size of viable phage by approximately 8-logs. These "absorbed" libraries now form the basis for isolating CLL cell-bound and cell-internalizing phage. Target cells include murine cell lines derived from spontaneously-developed aggressive CLL (BCL-1 and A20) and human patient CLL cells. Analysis of peptide sequences of A-20 binding or internalizing phage clones revealed overlap in amino acid usage but not one consistent consensus sequence, suggesting that these peptides target different CLL-specific membrane components, some of which can induce phage internalization and should be applicable for PDCs. Using a similar approach, we are isolating peptides for BCL-1 cells as well as peptides to cells from an initial cohort of CLL patients. The bioinformatic and biochemical results for all these peptides will be presented. Peptides selected on the basis of uptake kinetics will be conjugated to cytotoxic drugs using our dendrimer technology. This technology has already allowed us to demonstrate improved efficacy of known CLL drugs. Progress on this stage of the project will also be presented.

Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A85.

Citation Format: Michael A. Firer, Bat-Chen Lubin, Sharon Cohen, Galia Luboshits. Peptides specific for chronic lymphocytic leukemic cells: Characterization and use in targeted drug delivery. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A85.
Abstract A95: Intermittent, but not chronic, hypoxia increased chromosomal copy number variations of prostate cancer cells.
Authors: Zhang Jingsong , Wang Xue
Published: August 2014

Background: Intra-tumoral hypoxia as a potential mechanism for therapy resistance has been well documented in prostate cancer. Screening drugs under hypoxic conditions, however, has not been adopted for drug discovery. Part of this is due to the difficulty to recapitulate the heterogeneity of intra-tumoral hypoxia in vitro. Indeed, a spatio-temporal irregularity of the microvascular network of a growing tumor creates regions that are either chronically (diffusion-limited) or intermittently (perfusion-limited) hypoxic. If genomic instability is the fundamental driving force for development of therapy resistance, the hypoxic culture condition with the most genomic instability would serve as an alternative in vitro model for drug testing. We therefore accessed chromosomal copy number variations (CCNV) in prostate cancer cells grew under intermittent as well as chronic hypoxia.

Method: Metastatic prostate cancer cell lines, PC3 and DU145, were grown under normoxia (21% oxygen), chronic hypoxia (0.2% oxygen), or intermittent hypoxia (16 hours of 0.2% oxygen alternating with 8 hours of 21% oxygen) for total of 3 weeks. PC3 cells were also treated with the 1 µM poly(ADP-ribose) polymerase (PARP) inhibitor, veliparib under intermittent or chronic hypoxia conditions for 3 weeks. Genomic DNA was extracted for molecular karyotyping with the nanoString nCounter human karotyping platform, which covers 24 human chromosomes with 338 probes. DNA from human foreskin fibroblast (HFF) grew under normal culture condition (21% oxygen) was used as a reference.

Results: Both PC3 and DU145 cells continue to proliferate under intermittent as well as chronic hypoxia. Compared to untreated PC3 cells, daily treatment with PARP inhibitor under chronic or intermitent hypoxia for 3 weeks did not affect the proliferation or survival of the PC3 cells. Compared to HFF, frequent CCNVs were observed in PC3 and DU145 cells. This is consistent with the known complicated karyotype of these 2 cell lines. When compared to PC3 or DU145 cells cultured under normoxia, CCNVs were enhanced under intermittent hypoxia, but not chronic hypoxia. Daily treatment with PARP inhibitor did not alter CCNVs under either chronic hypoxia or intermittent hypoxia.

Conclusions: Intermittent hypoxia is probably a more relevant model to study genomic instability and drug resistance than chronic hypoxia. Neither chronic hypoxia nor intermittent hypoxia sensitizes prostate cancer cells to PARP inhibitor, veliparib.

Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A95.

Citation Format: Jingsong Zhang, Xue Wang. Intermittent, but not chronic, hypoxia increased chromosomal copy number variations of prostate cancer cells. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A95.
Abstract B101: Design and optimization of a new series of HSP70 inhibitors for use in the treatment of pancreatic cancer.
Authors: Maxwell David S. , Sun Duoli , Peng Zhenghong , Qi Wei , McConkey David , Abbruzzese James L. , Bornmann William G.
Published: August 2014

Through a combination of structure-based design and similarity methods, we have identified a novel series of HSP70 inhibitors that are directed towards the ATPase domain. The structural core was identified through a multi-stage docking and similarity search of screening compounds available from multiple commercial sources. Upon selection of the core, a virtual library was constructed, pre-filtered, and screened in two docking packages. By combining the resulting scores with those from a docking decoy library, workflow software was used to calculate a Z-score, which has recently been shown to assist with separating actives from in-actives. ADME predicted values were also calculated for each structure in the virtual library. Filters were applied in the workflow at several points to focus on the best compounds, thirty of which were ranked and considered for synthesis. The compounds were synthesized in five steps using commercially available starting materials. At present, six compounds from the series have been synthesized and four of those were tested alone and in combination with a known proteasome inhibitor, bortezomib. The third highest ranked compound showed the most potent effect on pancreatic cancer cell death in multiple cell lines. We will report on the design, synthesis, and biological characterization of this new class of HSP70 inhibitors.

Citation Information: Mol Cancer Ther 2013;12(11 Suppl):B101.

Citation Format: David S. Maxwell, Duoli Sun, Zhenghong Peng, Wei Qi, David McConkey, James L. Abbruzzese, William G. Bornmann. Design and optimization of a new series of HSP70 inhibitors for use in the treatment of pancreatic cancer. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr B101.
Abstract B103: Inhibitors of S100B (SBiXs) in malignant melanoma.
Authors: Cavalier Michael , Wilder Paul T. , Coop Andrew , MacKerell Alexander , Weber David J.
Published: August 2014

Summary: In malignant melanoma (MM), elevated levels of the tumor marker, S100B, binds directly to wild-type (wt) p53, dissociates the p53 tetramer, and down-regulates p53-dependent tumor suppression functions; therefore, it is the goal to develop S100B inhibitors (SBiXs; X=compound number) to restore active p53 in this deadly cancer. As a proof of principle for a drug design project, inhibiting S100B with small interfering antisense RNA (siRNAS100B) and several SBiXs was shown to restore wild-type p53 levels and its downstream gene products, as necessary to induce cell growth arrest and apoptosis in malignant melanoma.

Purpose: It is our purpose to discover/synthesize and/or to improve existing inhibitors of S100B to restore p53 activity in human malignant melanoma. Such compounds can then be examined for in vivo efficacy.

Results and conclusions: Development of S100B inhibitors (SBiXs) is ongoing at the University of Maryland Schools of Pharmacy and Medicine with the goal of obtaining higher affinity and better specificity towards inhibiting S100B. Computer aided drug design (CADD) followed by synthesis and experimental assays (i.e. NMR, binding, and cellular assays) have identified compounds that bind S100B and inhibit melanoma cell growth. Available binding data, IC50 data, and 3D structures of S100B-SBiX complexes will be presented that will act as the basis for the design of improved SBiXs. SBiXs may also have therapeutic value for treating other cancers with elevated S100B and wt p53 such as astrocytoma, renal tumors, and some forms of leukemia.

Citation Information: Mol Cancer Ther 2013;12(11 Suppl):B103.

Citation Format: Michael Cavalier, Paul T. Wilder, Andrew Coop, Alexander MacKerell, David J. Weber. Inhibitors of S100B (SBiXs) in malignant melanoma. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr B103.
Abstract B118: Histone demethylase inhibitor, trans-2-phenylcyclopropylamine hydrochloride inhibits KDM3A and KDM5A activities, resulting in overcoming resistance against bortezomib.
Authors: Terui Yasuhito , Kuniyoshi Ryoko , Tomida Akihiro , Hatake Kiyohiko
Published: August 2014

Purpose: The proteasome inhibitor bortezomib has shown impressive clinical activity alone and in combination with conventional and other novel agents for the myeloma treatment. Although bortezomib is known to be a selective proteasome inhibitor, the downstream mechanisms of cytotoxicity and drug resistance remain to be understood yet. However, resistance to bortezomib as a single agent develops in the majority of patients, and activity in other malignancies has been less impressive. To overcome bortezomib resistance, we compared differential gene expression profiles of bortezomib-resistant and bortezomib-sensitive IM-9 myeloma cell lines in response to bortezomib, and expressions of KDM3A and 5A genes were significantly incresed.

Methods: The differential gene expression profiles of bortezomib-resistant IM-9 and bortezomib-sensitive IM-9 multiple myeloma cell lines in response to bortezomib was performed using Affymetrix GeneChip. To confirm the results, real-time PCR and Western blot analysis were performed. bortezomib-resistant IM-9 cells were treated with or without bortezomib and/or histone demethylase inhibitor, trans-2-phenylcyclopropylamine hydrochloride (2-PCPA). Moreover, level of histone methylation was examined by western blot analysis. Results: At concentrations that effectively inhibited proteasome activity (maximum dose with 100nM), bortezomib induced cell death in bortezomib-sensitive IM-9 cells, but not in bortezomib-resistant IM-9. In comparison of differential gene expression profiles between bortezomib-resistant IM-9 and bortezomib-sensitive IM-9 cells, we showed overexpression of KDM3A and KDM5A, which are associated with chromatin-mediated reversible drug-tolerant state, Moreover, 2-PCPA overcomes bortezomib resistance in a dose-dependent manner. The histone demethylation at H3K4, H3K9 and H3K27 was suppressed with 2-PCPA.

Conclusion: Histone lysine-specific demethylase inhibitor, 2-PCPA with bortezomib may be useful for overcoming bortezomib-resistance in myeloma cells by suppression of KDM3A and KDM5A activities.

Citation Information: Mol Cancer Ther 2013;12(11 Suppl):B118.

Citation Format: Yasuhito Terui, Ryoko Kuniyoshi, Akihiro Tomida, Kiyohiko Hatake. Histone demethylase inhibitor, trans-2-phenylcyclopropylamine hydrochloride inhibits KDM3A and KDM5A activities, resulting in overcoming resistance against bortezomib. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr B118.
Abstract B138: The landscape of targetable master regulators and genomic drivers of glioblastoma.
Authors: Iavarone Antonio
Published: August 2014

We are experiencing the exciting time of the genome era. For our work this means being able to contextualize the alterations of each genetic network in the natural environment of a specific tumor and identify the key driving modules on which specific tumor subgroups rely for growth, survival and progression. With this information in hand, we can target the critical alterations with specific drugs, often already available for other types of diseases. By focusing on one of the most lethal forms of human tumors, the glioblastoma multiforme (GBM), we have been able to make incredible progress along this line in the last two years. Our recent work led to efficient targeting in preclinical mouse models of the most aggressive gene expression signature of high-grade glioma, the mesenchymal signature. We have identified the first examples of highly oncogenic and recurrent gene fusions in GBM, target their dependency in a particular tumor subtype, and observe dramatic anti-tumor effects. Recurrent gene fusions in GBM result in the constitutive activation of receptor tyrosine kinase genes (FGFR, EGFR and NTRK1) that render tumors addicted to the driver events. Both lines of investigation are maturing towards clinical trials. The integrated computational-experimental pipeline that we developed plus our ability to functionalize any genetic brain tumor module was recently applied to the entire landscape of CNVs, somatic mutations and gene fusions of human GBM. This information will advance our ability to quickly translate each new genetic finding into the personalized context of the clinical setting.

Citation Information: Mol Cancer Ther 2013;12(11 Suppl):B138.

Citation Format: Antonio Iavarone. The landscape of targetable master regulators and genomic drivers of glioblastoma. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr B138.
Abstract B177: Non-invasive tumor mutation detection of cell-free urinary DNA using massively parallel deep DNA sequencing.
Authors: Poole Jason C. , Janku Filip , Holley Veronica R. , Wheler Jennifer J. , Meric-Bernstam Funda , Luthra Rayjalakshmi , Leppin Lorieta , Hassaine Lafifa , Kosco Karena , Erlander Mark G.
Published: August 2014

Background: Previous studies have demonstrated the ability to detect tumor DNA mutations by PCR methodologies within cell-free urinary DNA in metastatic cancer patients. This has opened up the possibility to use massively parallel deep sequencing approaches for more global profiling of tumor mutations using cell free urinary DNA from these patients.

Methods: In order to achieve clinical utility the sequencing of cell-free urinary DNA must overcome two technical hurdles: 1) the need for extremely high sensitivity, between 0.01-0.05% and 2) an ultra-short DNA footprint (∼30 bp). Here we report the development of a method using cell free DNA extracted from urine that enriches for extremely low levels of mutated tumor DNA and thereby provides a high detection sensitivity. Detection of the KRAS gene mutations were used as a model system; the developed assay utilizes a 31bp footprint, contains a pre-amplification step that specifically enriches mutated DNA fragments and detects 8 KRAS mutations at the codon 12 and 13 sites. Spiking experiments with DNA derived from cell lines harboring the KRAS G12D mutation were completed to determine the limits of sensitivity of our method.

Results: Mutant DNA input at amounts of 0.2%, 0.05%, 0.01% and 0.0% of the total DNA returned observed detection levels of 18.25%, 4.45%, 1.84% and 0.54% respectively as a percentage of the total sequence reads with the detection of mutant DNA of 0.01% within the theoretical limit of the assay. To test our assay, we sequenced 30 ng of extracted urinary DNA from a stage IV colorectal carcinoma patient with a known mutation at the G12D site. Enrichment of the mutation presented at a level of 13.06% of our total sequence, corresponding to an input amount of approximately 0.14% mutational load in the patient's urine. Ongoing analysis of a larger number of patients will be presented to further evaluate the clinical sensitivity and specificity of KRAS mutations.

Conclusion: Further development of massively parallel DNA sequencing to detect mutations from cell-free urinary DNA has the potential to monitor metastatic patients for response, non-response and the emergence of resistance mechanisms to molecularly targeted therapies.

Citation Information: Mol Cancer Ther 2013;12(11 Suppl):B177.

Citation Format: Jason C. Poole, Filip Janku, Veronica R. Holley, Jennifer J. Wheler, Funda Meric-Bernstam, Rayjalakshmi Luthra, Lorieta Leppin, Lafifa Hassaine, Karena Kosco, Mark G. Erlander. Non-invasive tumor mutation detection of cell-free urinary DNA using massively parallel deep DNA sequencing. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr B177.