bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2601: EphB4/EphrinB2 targeting increases the efficacy of cisplatin in triple negative breast cancer
Authors: Yang-Kolodji Gloria W. , Liu Ren , Mehta Arjun , Gill Parkash , Tripathy Debu
Published: May 2015

Triple negative breast cancer (TNBC) represents a heterogeneous group of breast cancers that do not respond to either HER2 or hormonally targeted therapy and for which, chemotherapy is the only proven therapeutic option. However, certain biological pathways are known to be activated to a greater extent in TNBC.

The EphB4 receptor and its ligand EphrinB2 are expressed aberrantly in several types of cancer to promote angiogenesis and cell survival. In breast cancer, including TNBC, EphB4 expression is associated with a worse prognosis. An EphB4/Ephrin B2 inhibitor, sEphB4-HAS or SB4 (soluble protein EphB4 peptide stabilized with human serum albumin), developed at our institution, has shown inhibitory effects on cancer cell growth in both cell and animal models. Our goal here is to test the feasibility of EphB4 targeting in triple negative breast cancer cells, study the synergistic effects of SB4 and cisplatin, a common chemotherapy drug used for triple negative breast cancers, and to determine optimal combination doses of SB4 and cisplatin for eventual clinical trials.

Using two representative triple negative breast cancer cell lines MDA-MB-231 and HCC 1937, we show that SB4 inhibits cell growth in a dose dependent manner (IC50: 216.5μg/ml and 207.1 μg/ml, respectively). When SB4 is used in combination with cisplatin, synergy is observed. HCC 1937 cells treated with 200 μg/ml of sEphB4-HSA or 50 nM cisplatin for 7days, viability is decreased by 57 % and 8 %, and by 71% when given in combination. Similar results are seen in MDA-MB-231 cells, at 55%, 11%, and 63 % viability reduction. Mouse MDA-MB-231 xenografts are inhibited by SB4 treatment, with 20% reduction compared to control.

In addition, cell proliferation marker Ki67 assessed by immunohistochemical staining and pS6 (phosphorylated ribosomal protein S6) expression studied by immunoblotting demonstrate that SB4 in combination with cisplatin generates greater growth inhibitory effects on cell proliferation and pS6 expression than either alone.

In conclusion, EphB4 signaling in TNBC provides a niche to apply targeted therapy in combination with chemotherapy. The growth inhibitory effect of SB4 on TNBC cells and mouse xenografts supports the feasibility of EphB4 targeting. The effects of SB4 and cisplatin using xenograft models will be analyzed for synergy to provide a rationale to combine SB4 and chemotherapy in the clinic.

Citation Format: Gloria W. Yang-Kolodji, Ren Liu, Arjun Mehta, Parkash Gill, Debu Tripathy. EphB4/EphrinB2 targeting increases the efficacy of cisplatin in triple negative breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2601. doi:10.1158/1538-7445.AM2014-2601
Abstract 2602: KRAS G12D and G12V specific alkylating agent (KR12) inhibits growth of colon cancer with those KRAS mutations in vitro as well as in vivo
Authors: Nagase Hiroki , Hiraoka Kiriko , Inoue Takahiro , Watanabe Takayoshi , Shinohara Ken-Ichi , Koshikawa Nobuko , Toshinori Ozaki
Published: May 2015

Despite extensive efforts to develop chemotherapeutic drug(s) targeting mutated RAS, the successful drug discovery, especially targeting KRAS codon 12 mutation, has never been made. We have found that Pyrrole-Imidazole polyamide seco-CBI conjugate (KR12) selectively recognized mutated KRAS sequence (ACGCCT-A/T-CA) at codon 12 and significantly suppressed tumor growth specifically in human colon cancer cells with G12D or G12V mutation. KRAS expression suppressed preferentially at mutated allele and active KRAS were markedly reduced. G2/M arrest, senescence and subsequent apoptosis by activating the p53 pathway were observed in KR12-exposed LS180 cells with G12D heterozygous mutation. In LS180 and SW480 (G12V homozygous mutation) xenograft colon cancer models, KR12 treatment induced massive tumor growth suppression with low host toxicity. Collectively, our current results strongly suggest that KR12 is a specific alkylating agent against KRAS codon 12 mutations, and could become a powerful candidate compound for the unmet need of KRAS-mutant tumor treatment.

Citation Format: Hiroki Nagase, Kiriko Hiraoka, Takahiro Inoue, Takayoshi Watanabe, Ken-Ichi Shinohara, Nobuko Koshikawa, Ozaki Toshinori. KRAS G12D and G12V specific alkylating agent (KR12) inhibits growth of colon cancer with those KRAS mutations in vitro as well as in vivo. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2602. doi:10.1158/1538-7445.AM2014-2602
Abstract 2606: Cetuximab-mediated ADCC activity is correlated with the cell surface expression level of EGF-R but not the K-RAS/B-RAF mutational status in colorectal cancer
Authors: Ishii Yoshiyuki , Seo Yuki , Akimoto Shingo , Fukuda Kazumasa , Hayashida Tetsu , Okabayashi Koji , Tsuruta Masashi , Hasegawa Hirotoshi , Kitagawa Yuko
Published: May 2015

Cetuximab, an IgG1 monoclonal antibody against epidermal growth factor receptor (EGF-R), is widely used for the treatment of metastatic colorectal cancer (mCRC). One of the working mechanisms is considered to be antibody-dependent cell-mediated cytotoxicity (ADCC) triggered by Fcγ−R on natural killer cells. However, whether ADCC is associated with EGF-R expression and/or the mutational status of EGF downstream effectors (K-RAS and B-RAF etc.) in colorectal cancer (CRC) remains unclear. The aim of this study was to verify whether ADCC activities are associated with the cell surface expression levels of EGF-R and/or the mutational status of K-RAS and B-RAF. Five human CRC cell lines with different cell surface expression levels of EGF-R and different K-RAS and B-RAF mutational statuses were selected to evaluate ADCC activity using peripheral blood mononuclear cells (PBMC) from healthy human donors. Furthermore, tumor cells from resected specimens of CRC patients were used to evaluate the cell surface expression level of EGF-R using immunohistochemistry and the K-RAS and B-RAF mutational statuses using direct sequencing, while the ADCC activity was examined using PBMCs from the same CRC patients. A strong correlation was observed between the expression levels of EGF-R and the ADCC activities in the cell lines (correlation coefficient: 0.949; P = 0.003). Of the 13 resected specimens, a high ADCC activity level was significantly observed in tumor cells with high expression levels, compared with those with low expression levels, of cell surface EGF-R (P = 0.027). In both CRC cell lines and tumor cells from CRC patients, the ADCC activities were significantly associated with the cell surface expression levels of EGF-R (standard partial regression coefficients: 0.911 [P = 0.017] and 0.660 [P = 0.018], respectively), but not the mutational status of K-RAS and B-RAF (standard partial regression coefficient: -0.101 [P = 0.631] and 0.160 [P =0.510], respectively). Cetuximab-mediated ADCC activity may be correlated with the cell surface expression level of EGF-R, regardless of the mutational statuses of K-RAS and B-RAF, in CRC.

Citation Format: Yoshiyuki Ishii, Yuki Seo, Shingo Akimoto, Kazumasa Fukuda, Tetsu Hayashida, Koji Okabayashi, Masashi Tsuruta, Hirotoshi Hasegawa, Yuko Kitagawa. Cetuximab-mediated ADCC activity is correlated with the cell surface expression level of EGF-R but not the K-RAS/B-RAF mutational status in colorectal cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2606. doi:10.1158/1538-7445.AM2014-2606
Abstract 2616: RFX1-dependent activation of SHP-1 inhibits STAT3 signaling in hepatocellular carcinoma cells
Authors: Su Jung-Chen
Published: May 2015

Obatoclax is under clinical trial currently. Previously, we have demonstrated that SC-2001, an obatoclax analogue, has potent anti-tumor effect in HCC cells through the SHP-1/p-STAT3 cascade. However, the molecular mechanism by which SC-2001 activates SHP-1 is still elusive. Regulatory factor X (RFX) 1 is a transcription factor that has been linked to negative regulation of tumor progression. Nevertheless, its biological function in HCC cells is far from clear. In this study, by modulating its expression, we demonstrated that RFX1 is important in regulating SHP-1 in HCC cells. Ectopic expression of RFX1 can lead to activation of SHP-1 and inhibition of colony forming in HCC cells. Importantly, in a xenograft mouse model, RFX-1 overexpression obviously inhibited the growth rate of HCC tumors, indicating its tumor suppressor role. In addition, we found that SC-2001-induced apoptosis effect is regulated by RFX1. We showed that SC-2001 enhanced RFX1 nuclear translocation and binded to SHP-1 promoter to activate SHP-1 transcription. Moreover, knockdown of RFX1 reversed the anti-tumor effect of SC-2001 in vivo. Most important, we proved that SC-2001 displayed more potent anti-tumor effect than sorafenib, a clinically approved drug for HCC, in in vitro and in vivo assays. Furthermore, combination treatment of SC-2001 and sorafenib showed synergistic effect in HCC cells. Collectively, our study confirmed that RFX1 acts as a tumor suppressor in HCC and might be a new target for HCC therapy. The findings of this study also provide a new lead compound for target therapy via the activation of the RFX1/SHP-1 pathway. (Supported by NSC-102-2325-B-010-007)

Citation Format: Jung-Chen Su. RFX1-dependent activation of SHP-1 inhibits STAT3 signaling in hepatocellular carcinoma cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2616. doi:10.1158/1538-7445.AM2014-2616
Abstract 2659: The MedImmune ADC platform: Building highly potent and specific cancer drugs
Authors: Hollingsworth Robert E. , Kamal Adeela , Howard Philip W. , Hartley John A. , Tice David , Gao Changshou , Dimasi Nazzareno , Zhong Haihong , Harper Jay , Xiao Zhan , Toader Dorin , Martin Chris , Wu Herren , Greenberg Norman , Jallal Bahija
Published: May 2015

MedImmune has made a major commitment to developing antibody-drug conjugates for cancer. Our ADC initiative builds on our previous experience with this area and with other armed antibody technologies, which have generated several drugs currently undergoing clinical development. We have leveraged our expertise in antibody engineering to develop new antibody constructs for ADC development, including variants that facilitate site-specific conjugation of the payload to the antibody. This technology circumvents problems associated with random payload conjugation, resulting in a more homogeneous drug product as well as improving the stability and potency of the ADC. We have developed new, potent payloads, teaming up with Spirogen to apply their expertise in the pyrrolobenzodiazapine (PBD) dimer payload technology. The PBD payloads are versatile and potent, allowing use of multiple types of linkers and adjustment of potency to very high levels (picomolar IC50 range). The mechanism of action of the PBDs is different from other commonly used ADC payloads, inducing DNA damage that may evade DNA repair mechanisms and killing both bulk cancer cells and cancer stem cells. We have also developed other, novel ADC payloads. Our ADC target discovery approach permits rapid identification and validation of targets specifically suited for this technology. This includes the early generation and use of tool ADCs for target evaluation. We are applying this state-of-the-art ADC platform to advancing multiple projects as a major component of our oncology drug development strategy.

Citation Format: Robert E. Hollingsworth, Adeela Kamal, Philip W. Howard, John A. Hartley, David Tice, Changshou Gao, Nazzareno Dimasi, Haihong Zhong, Jay Harper, Zhan Xiao, Dorin Toader, Chris Martin, Herren Wu, Norman Greenberg, Bahija Jallal. The MedImmune ADC platform: Building highly potent and specific cancer drugs. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2659. doi:10.1158/1538-7445.AM2014-2659
Abstract 2662: Site specific ADC generation using SMARTag technology with programmable payload placement
Authors: Rabuka David
Published: May 2015

Antibody-drug conjugates (ADCs) are an exciting, new therapeutic option for cancer treatment.  ADCs offer the promise of targeted drug delivery with increased efficacy and safety.  Currently approved ADCs are heterogeneous mixtures resulting from nonselective ligation of drug to either cysteine or lysine residues.  This heterogeneity complicates the pharmacokinetics and pharmacodynamics of these drugs. 

Redwood Bioscience, Inc. has developed the SMARTagTM technology platform that enables precise, programmable, site-selective chemical protein modification and stable bioconjugate generation. Leveraging the target sequence of Formylglycine Generating Enzyme (FGE), proteins are chemoenzymatically modified to generate a precisely placed aldehyde functionality that can be chemically elaborated.   Subsequently, novel ligation chemistry is employed that exploits this “aldehyde tag” site.  This new reaction, the hydrazino-iso-Pictet-Spengler (HIPS) ligation, possesses two distinct advantages over current carbonyl ligations. First, the HIPS ligation proceeds quickly at near neutral pH in the absence of catalysts, allowing for one-step labeling of aldehyde-functionalized proteins under mild conditions. Second, HIPS ligation products are very stable in human plasma relative to an oxime-linked conjugates. Thus, the HIPS ligation exhibits a combination of stability and speed at near neutral pH that is unmatched by current carbonyl bioconjugation chemistries.  We will present our novel protein modification platform and its application to generating ADCs, including our new conjugation chemistries and linker libraries.  These technologies have been applied to the generation of a panel of site-specifically modified bioconjugates. Data from preclinical tumor models and PK studies highlighting the efficacy and safety of these ADCs and how these results are impacted by payload placement to the antibody will be presented.

Citation Format: David Rabuka. Site specific ADC generation using SMARTag technology with programmable payload placement. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2662. doi:10.1158/1538-7445.AM2014-2662
Abstract 2689: Breast cancer inhibition by a novel and potent biguanide, N1-hexyl-N5-benzyl-biguanide
Authors: Guo Zhijun , Chavez Kathryn J. , Alvarez Juan , Zhang Xia , Norris Beverly , Maher Michael , Morgan Monique , Schumacher Robert J. , Cuellar Rebecca , Sevrioukova Irina F. , Poulos Thomas L. , Denisov Ilia , Sligar Stephen G. , Gupta Kalpna , Blair Ian A. , Capdevila Jorge , Kelekar Ameeta , Amin Elizabeth , Georg Gunda , Potter David A.
Published: May 2015

Metformin is a widely used biguanide diabetes drug that is associated with decreased breast cancer risk and is currently being studied for treatment and prevention of breast cancer. While metformin and biguanides buformin and phenformin exhibit inhibitory activity against breast cancer in vitro and in vivo, they lack potency (IC50=5-20 mM) and their mechanisms of action remain unclear. More potent biguanides may provide insights into biguanide anti-cancer activity and we therefore studied the novel biguanide N1-hexyl-N5-benzyl-biguanide mesylate (HBB), which potently inhibits the MCF-7 and MDA-MB-231 breast cancer lines (IC50=20 uM for both lines). HBB induces AMPK phosphorylation in both lines at 10 uM concentration, whereas similarly dosed metformin, buformin or phenformin exhibits no activity. HBB also inhibits STAT3 phosphorylation at 10 uM concentration, whereas metformin dosed at 10 uM exhibits no activity. HBB reduced the mitochondrial membrane potential of both lines, but the effect was more prominent in the MDA-MB-231 line. HBB also induced ROS within 2.5 hours of exposure in the MCF-7 and MDA-MB-231 lines and caused rapid necrosis, but not apoptosis. N-acetylcysteine provides partial protection from HBB for MDA-231 line, but not the MCF-7 line. HBB provides proof of principle that highly potent biguanides can be synthesized with at least 250-fold greater potency than metformin, which can provide insights into the cancer inhibitory mechanisms of biguanide drugs. R01 CA113570, Randy Shaver Foundation, CTSI University of Minnesota

Citation Format: Zhijun Guo, Kathryn J. Chavez, Juan Alvarez, Xia Zhang, Beverly Norris, Michael Maher, Monique Morgan, Robert J. Schumacher, Rebecca Cuellar, Irina F. Sevrioukova, Thomas L. Poulos, Ilia Denisov, Stephen G. Sligar, Kalpna Gupta, Ian A. Blair, Jorge Capdevila, Ameeta Kelekar, Elizabeth Amin, Gunda Georg, David A. Potter. Breast cancer inhibition by a novel and potent biguanide, N1-hexyl-N5-benzyl-biguanide. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2689. doi:10.1158/1538-7445.AM2014-2689
Abstract 2709: Identification of Aurintricarboxylic Acid (ATA) as an inhibitor of TWEAK-Fn14 signaling in glioblastoma cells
Authors: Dhruv Harshil D. , Holiday Ethan , Chow Donald , Yin Holly , Berens Michael E. , Tran Nhan L.
Published: May 2015

The long-term survival of patients with glioblastoma (GB) is compromised by the proclivity for local invasion into the surrounding normal brain, escaping surgical resection and contributing to therapeutic resistance. Tumor necrosis factor-like weak inducer of apoptosis (TWEAK), a member of the tumor necrosis factor superfamily, can stimulate glioma cell invasion via binding to fibroblast growth factor-inducible 14 (Fn14) and subsequent activation of the transcription factor NF-κB. In order to discover a small molecule inhibitors that disrupt Fn14-TWEAK signaling axis and subsequently glioma cell invasion, we developed a cell-based drug screening assay using HEK293 cells that overexpresses Fn14 and harbors NF-κB driven Firefly luciferase protein expression. In preliminary drug screening assay using LOPAC1280 library of 1280 pharmacologically active compound, Aurintricarboxylic Acid (ATA) showed suppression of NF-κB driven Firefly Luciferase activity downstreatm of TWEAK-Fn14 signaling. Conversely, ATA did not show any suppression in NF-κB driven Firefly Luciferase activity downstream of TNFα-TNFR1 signaling, suggesting that ATA is a specific inhibitor of TWEAK-Fn14-NF-κB signaling. In vitro, we demonstrated that ATA suppresses TWEAK induced chemotactic migration of glioma cells (T98G and A172) and display no general cytotoxicity. In summary, this work reports a new small molecule inhibitor of TWEAK-Fn14 signaling that could be useful in enhancing the therapeutic targeting of this deadly disease.

Citation Format: Harshil D. Dhruv, Ethan Holiday, Donald Chow, Holly Yin, Michael E. Berens, Nhan L. Tran. Identification of Aurintricarboxylic Acid (ATA) as an inhibitor of TWEAK-Fn14 signaling in glioblastoma cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2709. doi:10.1158/1538-7445.AM2014-2709
Abstract 2710: Lead based development and evaluation of selective estrogen mimics in tamoxifen resistant breast cancer
Authors: Patel Hitisha , Xiong Rui , Zhao Jiong , Molloy Mary Ellen , Tonetti Debra , Thatcher Gregory R.J
Published: May 2015

Tamoxifen is the standard of care for many patients with ER+ breast cancer and works by antagonizing the actions of estrogen at ER. However it poses an increased risk of endometrial cancer and thrombotic events, and most significantly, 30-50% women develop resistance to tamoxifen therapy, underlining the need for superior therapeutic options. Paradoxically, prior to tamoxifen therapy, estradiol (E2) and the ER agonist, diethylstilbestrol, had been used in breast cancer therapy, though with serious side effects. Development of Selective Estrogen Mimics (SEMs) that cause regression of tamoxifen-resistant breast cancer, but without the side effects of E2, represents a rational therapeutic strategy. Novel SEMs were developed, which in vitro in 3D cultures and in vivo caused complete regression of tamoxifen-resistant xenografts, with characteristics similar to those of E2. These SEMs did not fuel growth of estrogen-dependent T47D xenografts and did not cause uterine growth. SEM mediation of classical ER-signaling was profiled in MCF7 and MDA-MB231:β41 cells. A tamoxifen-resistant cell line, MCF-7: 5C, was used to assay induction of cell death by both E2 and the SEMs and to probe the mechanism of action. Several SEMs were observed to act as partial agonists at ERα , suggesting ideal characteristics as therapeutics. Structure-activity relationships were explored, suggesting that partial ERα agonist have the capacity to cause regression of tamoxifen-resistant tumors, without adverse effects associated with estrogenic actions in normal gynecological tissues of the breast and uterus, which might contribute to carcinogenesis. The ER-mediated agonist/antagonist activity, regression efficacy, and pharmacokinetic profiles of these SEMs were examined to obtain lead compounds that are effective with minimal estrogenic side effects. In MCF-7:5C cells, SEM-induced cell death was mediated by activation of classical nuclear ERα signaling, via induction of apoptosis. This research paves the way for use of SEMs with partial agonist activity at ER for use in tamoxifen-resistant breast cancer with enhanced safety profiles.

Citation Format: Hitisha Patel, Rui Xiong, Jiong Zhao, Mary Ellen Molloy, Debra Tonetti, Gregory R.J Thatcher. Lead based development and evaluation of selective estrogen mimics in tamoxifen resistant breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2710. doi:10.1158/1538-7445.AM2014-2710
Abstract 2728: Thermal ablative therapy with novel gold nanorods in an orthotopic model of urinary bladder cancer
Authors: Yang Xiaoping , Su Lih-Jen , Rosa Francisco G. La , Smith Elizabeth Erin , Cho Suehyun Katherine , Kavanagh Brian , Park Won , Flaig Thomas W.
Published: May 2015

Background

The use of nanoparticles has shown great therapeutic potential, but delivery and clearance concerns have limited their use systemically. Intravesical instillation of nanoparticles in the urinary bladder provides a unique administration route to treat superficial bladder cancer. We examined the anti-tumor effect of epidermal growth factor receptor (EGFR)-directed gold nanorods combined with photothermal treatment in a superficial bladder cancer model.

Methods

A novel orthotopic nu/nu murine xenograft model was established through intravesical instillation of T24 human bladder cancer cells labeled with Luciferase. After confirmation of positive tumor cell implantation via bioluminescence, gold nanorods linked to the anti-EGFR antibody C225 were instilled into the bladder cavity. The nanorods were specifically fashioned to respond to near infrared (NIR) light, which was externally administered via an 808 nm diode laser to the bladder. A safe and effective regimen of in vivo photothermal ablation was determined by optimizing the duration, power and distance from the tumor to the laser source. The anti-cancer effect was monitored by an in vivo imaging system in a non-invasive manner.

Results

The tumor implantation rate with the T24 human cancer cell was approximately 90%. The optimized regimen of photothermal treatment in this setting was laser power of 2.1 W/cm2 for 30 seconds at 2.5 centimeters from the skin. Using this in vivo tumor model, and near-infrared energy combined with C225-conjugated gold nanorods, we demonstrate significant therapeutic efficacy against the implanted orthotopic bladder tumor (Table) with minimal systemic toxicity using this regimen.

Conclusions

Photothermal ablation with EGFR-directed gold nanorods is effective and well tolerated in an in vivo model of urinary bladder cancer. Future clinical development would include NIR light delivery via a cystoscope in a human application

Summary of luciferase results of in vivo photothermal treatmentTotal miceLuciferase DecreaseLuciferase IncreaseLuciferase No Significant ChangeConjugated Nano Rods+Laser161303Laser Only14275

Citation Format: Xiaoping Yang, Lih-Jen Su, Francisco G. La Rosa, Elizabeth Erin Smith, Suehyun Katherine Cho, Brian Kavanagh, Won Park, Thomas W. Flaig. Thermal ablative therapy with novel gold nanorods in an orthotopic model of urinary bladder cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2728. doi:10.1158/1538-7445.AM2014-2728
Abstract 2764: The depletion of LRP5, unlike that of LRP6, promotes apoptosis in triple-negative breast cancer cells, making it an interesting therapeutic target
Authors: Maubant Sylvie , Maire Virginie , Tesson Bruno , Némati Fariba , Gentien David , Marty-Prouvost Bérengère , Depil Stéphane , Cruzalegui Francisco , Tucker Gordon , Roman-Roman Sergio , Dubois Thierry
Published: May 2015

Treatment of patients with triple-negative breast cancers (TNBCs) remains a major challenge for oncologists and alternative treatments to conventional chemotherapies are needed to improve their survival. The Wnt/beta-catenin signaling, recently reported to be activated in TNBCs, may represent an interesting pathway to target. We report that both LRP5 and LRP6 Wnt coreceptors are more strongly expressed in TNBCs than in other breast tumor subtypes. As very few studies have explored potential differences between LRP5 and LRP6, we investigated the effects of modulating specifically LRP5 or LRP6 expression on Wnt signaling, cell viability and tumorigenesis in HCC38 and MDA-MB-468 TNBC cells. We found that these two cell lines are more similar to TNBC biopsy specimens in terms of Wnt pathway gene expression profiles than any other tested cell line. Unlike LRP5, LRP6 was involved in activating the canonical Wnt pathway in response to Wnt3a. LRP5 knockdown induced caspase-dependent apoptosis, whereas LRP6 knockdown had no such effect. Importantly, LRP5-depleted cells were more sensitive to conventional chemotherapy than cells depleted of LRP6. The knockdown of LRP5 or LRP6 decreased tumorigenesis both in vitro and in vivo. Overall, these data suggest that the LRP5 and LRP6 coreceptors have different functions in TNBCs, with LRP5 playing a preponderant role in survival control. Our data suggest that both LRP5 and LRP6 are potential treatment targets in TNBCs, but that LRP5 may be the most useful target, given the impact of its depletion on cell survival as well as on the response to anti-cancer drugs.

Citation Format: Sylvie Maubant, Virginie Maire, Bruno Tesson, Fariba Némati, David Gentien, Bérengère Marty-Prouvost, Stéphane Depil, Francisco Cruzalegui, Gordon Tucker, Sergio Roman-Roman, Thierry Dubois. The depletion of LRP5, unlike that of LRP6, promotes apoptosis in triple-negative breast cancer cells, making it an interesting therapeutic target. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2764. doi:10.1158/1538-7445.AM2014-2764
Abstract 2776: Overexpression of HGF is an effective indicator for Met-targeting therapy in hepatocellular carcinoma
Authors: Xie Qian
Published: May 2015

The success of molecular targeted therapy against cancer depends on discovering the essential pathways that contribute to carcinogenesis and cancer progression and the molecular targets that control the pathway activity. By using genetically engineered mouse models, we show that overexpression of Hepatocyte growth factor (HGF) stimulated HBV surface antigen (HBsAg) production and accelerated the HCC progression. HGF transgenic mouse model produced spontaneous HCC with genomic profiles similar to that HBV-positive HCC patients with poor prognosis. Mechanistically, MET receptor expression is required for repair of virus-induced liver damage and HCC development. Importantly, HGF overexpression induces an autocrine activation which determines the sensitivity to small-molecule Met inhibitors. MET inhibitors are entering clinical trials against cancer, and our data provide a molecular basis for targeting the Met pathway in hepatitis B-induced HCC. Our results demonstrate that HGF is a driver of HBV-induced HCC progression and may serve as an effective biomarker for Met-targeted therapy, and that hgf-transgenic mouse model serves as a good tool for studying human HCC biology and preclinical evaluating therapeutics.

Citation Format: Qian Xie. Overexpression of HGF is an effective indicator for Met-targeting therapy in hepatocellular carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2776. doi:10.1158/1538-7445.AM2014-2776
Abstract 2777: Lycopene synergistically enhances quinacrine action to inhibit Wnt-TCF signaling in breast cancer cells through APC
Authors: Preet Ranjan , Shanmugam Rajasubramaniam , Mohapatra Purusottam , Das Dipon , Satapathy Shakti R. , Wyatt Michael D. , Kundu Chanakya N.
Published: May 2015

We previously reported that quinacrine (QC) has anticancer activity against breast cancer cells. Here, we report the mechanism of action of QC and its ability to inhibit Wnt-TCF signaling in two independent breast cancer cell lines. QC altered Wnt-TCF signaling components by increasing the levels of adenomatous polyposis coli (APC), DAB2, GSK-3β and axin and decreasing the levels of β-catenin, p-GSK3β (ser 9) and CK1. QC also reduced the activity of the Wnt transcription factor TCF/LEF and its downstream targets cyclin D1 and c-MYC. Using a luciferase-based Wnt-TCF transcription factor assay, it was shown that APC levels were inversely associated with TCF/LEF activity. Induction of apoptosis and DNA damage was observed after treatment with QC, which was associated with increased expression of APC. The effects induced by QC depend on APC because the inhibition of Wnt-TCF signalling by QC is lost in APC-knockdown cells, and consequently, the extent of apoptosis and DNA damage caused by QC was reduced compared with parental cells. Further, previously we have shown that QC inhibits topoisomerase, therefore we tested the effect of another topoisomerase inhibitor, etoposide, on Wnt signaling. Interestingly, etoposide treatment also reduced TCF/LEF activity, β-catenin and cyclin D1 levels commensurate with induction of DNA damage and apoptosis. Lycopene, a plant-derived antioxidant, synergistically increased QC activity and inhibited Wnt-TCF signaling in cancer cells without affecting the MCF-10A normal breast cell line. Collectively, the data suggests that QC-mediated Wnt-TCF signal inhibition depends on APC and that the addition of lycopene synergistically increases QC anticancer activity.

Note: This abstract was not presented at the meeting.

Citation Format: Ranjan Preet, Rajasubramaniam Shanmugam, Purusottam Mohapatra, Dipon Das, Shakti R. Satapathy, Michael D. Wyatt, Chanakya N. Kundu. Lycopene synergistically enhances quinacrine action to inhibit Wnt-TCF signaling in breast cancer cells through APC. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2777. doi:10.1158/1538-7445.AM2014-2777
Abstract 2786: Downregulated expression of hepatoma-derived growth factor (HDGF) reduces glioma cancer cell tumor growth and angiogenesis
Authors: Liu Li-Feng , Hsu Shu-Shong , Chen Chih-Hao
Published: May 2015

Hepatoma derived growth factor (HDGF) is a mitogenic growth factor in the progression of human glioma. In this paper, we study the gliomagenesis role of HDGF in syngenic glioma rat model by Lenti-virus sh-HDGF (LV-shHDGF) infection. The tumor inhibition of HDGF depletion in rat glioma C6 cells was measured by MRI scanning. We found all glioma tumors growth after implanting but only HDGF knock down tumors had a significant reduction in the tumor size at the 4th-week MRI-scanning. After tumor volume was quantified on serial brain section, we found a 10-folds of reduction in the tumor size was observed in HDGF knock down tumors than scrambles (p<0.005). By survival analysis, rats with gliomas had a less median survival of 25 days and the HDGF knock down group was still alive from the time point of implanting. According to the pathological analysis, the HDGF knock down tumors presented a significant cell apoptosis and reduction in the Ki67 mitotic index, CD31 angiogenesis index, and ANG-4 that exhibited markedly reduced tumor growth and blood vessel formation than controls. In addition, Western blot analysis showed exogenous HDGF increased the expression of PI3K, phospho-Akt, phospho-FAK, phospho-MDM2, PCNA and transcription factors of Iκ-B, NF-κB, cFos, ANG-4, and HIF-α in a dose-dependent manner; however, these tumorigenesis moleculars were significantly decreased in HDGF knock down cells. We concluded that HDGF is a mitogenic growth factor in the progression of glioma and it can be a useful therapeutic target for clinical management of glioma in the future.

Note: This abstract was not presented at the meeting.

Citation Format: Li-Feng Liu, Shu-Shong Hsu, Chih-Hao Chen. Downregulated expression of hepatoma-derived growth factor (HDGF) reduces glioma cancer cell tumor growth and angiogenesis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2786. doi:10.1158/1538-7445.AM2014-2786
Abstract 2800: Immunotherapy of malignant pleural mesothelioma using MET specific t – cells
Authors: Thayaparan Thivyan , Stegen Sjoukje v.d , Pereira Ana C. Parente , Petrovic Roseanna , Spicer James , Maher John
Published: May 2015

Background: Malignant pleural mesothelioma (MPM) is an incurable cancer predominantly induced by asbestos exposure. This aggressive tumor commonly presents at an advanced stage of disease. Although surgery, combination chemotherapy and radiotherapy may be used in disease management, median survival from diagnosis is less than 12 months. New therapeutic approaches are required to improve outcome in this disease.

Chimeric antigen receptors (CAR) are fusion molecules that couple the HLA-independent binding of a selected cell surface target antigen to the delivery of a tailored T-cell activating signal. Using CAR technology, we aim to develop a novel immunotherapy for MPM that is both safe and effective. These molecules are delivered to patient T-cells using retroviral gene transfer, thereby stably re-directing their specificity for antigen. The receptor tyrosine kinase MET is overexpressed in >80% of MPM patients making it an attractive candidate for CAR directed immunotherapy.

Methods: Three MET-targeted peptide-binding motifs have been incorporated into a second generation CAR framework containing a fused CD28/CD3ζ endodomain. Functional assessment of MET-targeted CARs has been assessed by co-cultivation of genetically enhanced test and control T-cells with MPM cell lines that express MET at various levels.

Results: In preliminary studies we have shown that human T-cells engineered to express MET CARs can destroy mesothelioma tumor cell monolayers, accompanied by T-cell proliferation and cytokine production. Xenograft models are currently under development using bioluminescence to further evaluate functionality and toxicity in vivo.

Conclusions: These findings demonstrate proof of concept for the utility of CAR engineered T-cells to recognise and destroy MET-expressing MPM tumor cells. We envisage that regional i.e. intrapleural delivery of human CAR positive T-cells could be used to maximise therapeutic index of this approach.

Citation Format: Thivyan Thayaparan, Sjoukje v.d Stegen, Ana C. Parente Pereira, Roseanna Petrovic, James Spicer, John Maher. Immunotherapy of malignant pleural mesothelioma using MET specific t – cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2800. doi:10.1158/1538-7445.AM2014-2800
Abstract 2801: The use of induced pluripotent stem (iPS) cells for the safety testing of enhanced affinity TCR-transduced T cells
Authors: Adams Katherine J. , Gerry Andrew B. , Brewer Joanna E. , Pumphrey Nicholas J. , Bennett Alan D. , Jakobsen Bent K.
Published: May 2015

Engineering T cells with enhanced affinity T cell receptors (TCRs) is a promising therapeutic strategy for treating cancer. Engineered TCRs overcome the limitations of naturally occurring cancer-antigen specific TCRs which are typically of extremely low affinity and fail to efficiently recognize tumor cells. However, the use TCRs with enhanced affinity for adoptive therapy carries with it the risk that they might recognize normal tissue, either as on-target or off-target reactivity. Two fatalities in a clinical trial of an affinity-enhanced MAGE-A3-specific TCR have highlighted the need for more biologically relevant cellular testing systems to minimize cross-reactivity-related toxicity in future adoptive immunotherapy trials.

Cellular Dynamics International Inc. supply a range of iCell products that are highly pure induced pluripotent stem (iPS) cell-derived cells that reproduce normal human biology in an in vitro setting. The advantage of iCell cultures over standard cultures of primary cells is that they better represent human organs with respect to their antigen presentation by HLA.

In this study, a panel of iCell sub-types, representing major organs of the human body, were used to test the reactivity of T cells transduced with an enhanced affinity TCR for HLA-A2-restricted peptide antigen NY-ESO-1/LAGE-1157-165. IncuCyte FLR technology, which enables direct visualization of caspase-3/7-dependent apoptosis at 37°C in real time, was used to assess the killing of the iCells by NY-ESO TCR-transduced T cells over a 4 day time course.

While potent killing by the NY-ESO TCR-transduced T cells was observed against an HLA-A2+, NY-ESO-1+/LAGE-1+ tumor cell line, A375, no reactivity was observed against any of the tested iCells sub-types unless the iCells were pulsed with 1 µM NY-ESO peptide.

The screening of TCR-transduced T cells against biologically appropriate cell cultures, such as iCells, further advances preclinical safety testing of new TCRs for use in adoptive immunotherapy.

Citation Format: Katherine J. Adams, Andrew B. Gerry, Joanna E. Brewer, Nicholas J. Pumphrey, Alan D. Bennett, Bent K. Jakobsen. The use of induced pluripotent stem (iPS) cells for the safety testing of enhanced affinity TCR-transduced T cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2801. doi:10.1158/1538-7445.AM2014-2801
Abstract 2813: Survival benefit of an add-on dendritic cell vaccine for patients with inoperable pancreatic cancer receiving chemotherapy: a multicenter study
Authors: Kobayashi Masanori , Shigetaka Shimodaira , Nagai Kazuhiro , Ogasawara Masahiro , Takahashi Hidenori , Abe Hirofumi , Tanii Mitsugu , Okamoto Masato , Tsujitani Shun-ichi , Nagaya Masaki , Yonemitsu Yoshikazu ,   The DC vaccine study group of Japan Society of Innovative Cell Therapy (J-SICT)
Published: May 2015

Purpose: Dendritic cell (DC)-based cancer vaccine may have a significant benefit to patients with advanced pancreatic cancer. However, variations among clinical studies make it hard to detect its clinical outcome. Here we statistically identified factors determining the clinical benefit using data obtained at seven Japanese institutions that employed the same DC preparation and treatment regimen.

Patients and Methods: Among the 354 patients who met the inclusion criteria, 255 patients who received standard chemotherapy combined with peptide-pulsed DC vaccines were analyzed.

Results: The mean survival time from diagnosis was 16.5 months (95%CI=14.4-18.5), and that from the first vaccination was 9.9 months (95%CI=8.0-12.9). Known prognostic baseline factors related to advanced pancreatic cancer, namely ECOG-PS, peritoneal metastasis, liver metastasis, and prognostic nutrition index (PNI) were also representative. Importantly, erythema reaction after vaccination was an independent and treatment-related predictive factor for better survival, and that OK432 might be a good adjuvant enhancing the antitumor immunity during DC vaccination.

Conclusion: This is the first report of a multicenter clinical study suggesting the feasibility and possible clinical benefit of an add-on DC vaccine in patients with advanced pancreatic cancer who are undergoing chemotherapy. These findings need to be addressed in well-controlled prospective randomized trials.

Citation Format: Masanori Kobayashi, Shimodaira Shigetaka, Kazuhiro Nagai, Masahiro Ogasawara, Hidenori Takahashi, Hirofumi Abe, Mitsugu Tanii, Masato Okamoto, Shun-ichi Tsujitani, Masaki Nagaya, Yoshikazu Yonemitsu, The DC vaccine study group of Japan Society of Innovative Cell Therapy (J-SICT). Survival benefit of an add-on dendritic cell vaccine for patients with inoperable pancreatic cancer receiving chemotherapy: a multicenter study. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2813. doi:10.1158/1538-7445.AM2014-2813
Abstract 2816: Immune-tolerant elastin-like polypeptide (iTEP) particles promote peptide vaccine presentation by dendritic cells
Authors: Chen Mingnan , Dong Shuyun , Cho Scott
Published: May 2015

T cell epitope-based peptide vaccines are promising experimental therapeutics for cancer and infection due to their unparalleled specificity, yet their presentations by dendritic cells (DCs) are hindered by their intrinsic limitations: short half-lives, non-typical antigen sizes for DCs to take in and process, lack of intrinsic adjuvants, and poor cross-presentation efficiencies. We aimed to boost the presentation by fabricating an immune-tolerant elastin-like polypeptide (iTEP) nanoparticle (NP) to resolve the limitations. The properties of iTEPs that render them as ideal vaccine carriers include their non-immunogenic and non-inflammatory natures, their simple purification method through phase transition, and their ability to fuse the vaccines and peptide adjuvants together.

We first fused a cytotoxic T lymphocyte (CTL) epitope vaccine, SIINFEKL, from ovalbumin to an iTEP amphiphilie and found the fusion is able to self-assemble into NPs. The NP was turned into a size similar to virus-like particles by modulating the iTEP sequences. The NP enhanced the presentation of SIINFEKL with H2b on the surface of DC2.4 cells in comparison with free peptide, ovalbumin, or soluble vaccine-iTEP fusions. Interestingly, another iTEP-vaccine fusion which forms larger, micrometer-sized aggregates led to even stronger vaccine presentation, an observation still under investigation. Moreover, our preliminary data showed that co-delivering the vaccines and an adjuvant peptide from flagellin by iTEPs also augmented the peptide vaccine presentation. Lastly, the iTEP NPs that deliver tumor antigen Kras- and hTERT-derived peptide vaccines were also generated and are currently under study with a hope to improve the efficacy of these vaccines.

In summary, iTEP particles enhanced peptide vaccine presentation. The underlying mechanisms of the enhancing effects of iTEP particles on the vaccine presentation will be studied next.

Supported by the University of Utah Start-up fund and the University of Utah SEED grant

Citation Format: Mingnan Chen, Shuyun Dong, Scott Cho. Immune-tolerant elastin-like polypeptide (iTEP) particles promote peptide vaccine presentation by dendritic cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2816. doi:10.1158/1538-7445.AM2014-2816
Abstract 2845: Expression of FGFR2b in gastric cancer as measured by immunohistochemistry with a highly specific monoclonal antibody
Authors: Deshpande Amit M. , Palencia Servando , Bellovin David I. , Gemo Abigael T. , Giese Tina , Stohr Bradley , Pierce Kristen L. , Los Gerrit
Published: May 2015

Gastric cancer is the second leading cause of death from cancer worldwide. Amplification of the FGFR2 gene has been reported in ∼ 5 to 7% of gastric cancers, and several studies have also reported overexpression of the FGFR2 protein using immunohistochemistry (IHC). FGFR2 gene amplification and protein overexpression have also been shown to correlate with an aggressive disease phenotype, making FGFR2 an attractive therapeutic target. We have reported elsewhere at this meeting that the b-isoform of FGFR2 is predominantly expressed in these tumors and have developed a therapeutic antibody, FPA144, which specifically targets this isoform. Here we report initial development of a separate monoclonal antibody, FPT mAb, for use as a companion IHC diagnostic (CDx) for selecting patients with high levels of FGFR2b protein.

In specificity studies using cells transfected with various FGFRs we show that FPT mAb specifically detects the b-isoform of FGFR2. In contrast, 7 commercially available anti-FGFR2 antibodies showed cross-reactivity with other FGF receptors. In an assay using cells with inducible FGFR2 gene expression, FPT mAb also was more sensitive for detection of FGFR2b than any of the commercially available antibodies. Furthermore, FPT mAb was able to detect FGFR2b protein expression in gastric cancer cases without FGFR2 gene amplification. We examined FGFR2b protein expression by IHC using the FPT mAb and FGFR2 gene amplification using FISH in 186 gastric cancer samples. 6% of samples showed amplification of the FGFR2 gene as measured by FISH (FGFR2:CEN10 >3), compared to 16.7% of samples positive for FGFR2b expression as measured by IHC (2-3+ staining in >50% of the tumor cells).

In conclusion, we show that the FPT mAb is sensitive and specific for detecting by IHC the b-isoform of FGFR2 expressed in gastric cancers. Further, we demonstrate that more gastric tumors overexpress FGFR2b protein than bear amplifications of the FGFR2 gene. These results suggest that a potentially larger percentage of patients with gastric cancer may be appropriately identified as candidates for FGFR2b-specific therapies.

Citation Format: Amit M. Deshpande, Servando Palencia, David I. Bellovin, Abigael T. Gemo, Tina Giese, Bradley Stohr, Kristen L. Pierce, Gerrit Los. Expression of FGFR2b in gastric cancer as measured by immunohistochemistry with a highly specific monoclonal antibody. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2845. doi:10.1158/1538-7445.AM2014-2845