bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4182: Statins induce apoptosis in osteosarcoma cells by activation of Ampk and p38-MAPK via suppression of mevalonate pathway
Authors: Kamel Walied Abd-Elghani , Sugihara Eiji , Yamaguchi Sayaka Iwai , Nobusue Hiroyuki , Maki Kenta , Muto Akihiro , Saya Hideyuki , Shimizu Takatsune
Published: July 2016

Osteosarcoma (OS) is the most common, non-hematopoietic, primary malignant bone tumor. OS is characterized by its aggressive local growth and systemic dissemination. Although combination of surgical operation and adjuvant chemotherapy greatly improved the prognosis, more than 20% of patients still cannot get long-term survival. Therefore, novel therapeutic approaches should be expected to be developed. Previously, we developed an OS mouse model by overexpressing c-MYC in bone marrow stromal cells derived from Ink4a/Arf knockout mice. We isolated highly tumorigenic cells (designated AXT cells) from tumors after serial transplantation. Inoculation of AXT cells into syngeneic C57BL/6 mice results in the development of lethal OS with metastatic lesions including lung, which pathologically and clinically mimics human osteoblastic osteosarcoma. To obtain the novel therapeutic agents for OS, we performed drug screening using existing drug collections and found that statins strongly suppressed AXT cell growth and migration in vitro. Simvastatin induces caspase dependent apoptosis, which can be completely rescued by the supplement of mevalonate and geranylgeranyl pyrophosphate but not by farnesyl pyrophosphate, suggesting that OS cells are susceptible to the inhibition of the mevalonate pathway. As the responsible effectors, we found treatment of simvastatin drives RhoA-GTP translocation from cell membrane to cytosol as well as the disruption of interaction between RhoA and Rho-GDI, indicating that, in spite of the accumulation of RhoA-GTP in OS cells, RhoA cannot work properly, which appears to affect cell motility and anchorage dependent survival. As downstream signaling of RhoA, p38-MAPK and AMPK are strongly activated by simvastatin treatment. Activation of p38-MAPK is not accompanied by the ROS production. The specific inhibitor for p38-MAPK or AMPK can rescue this activation as well as the simvastatin-induced cell death, respectively, suggesting that activation of both kinases is responsible for the anti-tumor effect of simvastatin. A single treatment of statins attenuated OS tumor growth in vivo. These findings suggest that the activation of p38-MAPK and AMPK by statins become a potential therapeutic option for OS.

Citation Format: Walied Abd-Elghani Kamel, Eiji Sugihara, Sayaka Iwai Yamaguchi, Hiroyuki Nobusue, Kenta Maki, Akihiro Muto, Hideyuki Saya, Takatsune Shimizu. Statins induce apoptosis in osteosarcoma cells by activation of Ampk and p38-MAPK via suppression of mevalonate pathway. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4182.
Abstract 4184: Pre-clinical study using KRAS mutant mouse models for lung cancer immunotherapy together with MEK inhibition
Authors: Deng Jiehui , Li Shuai , Herter-Sprie Grit , Smith Paul D. , Freeman Gordon J. , Engelman Jeffrey A. , Hammerman Peter , Wong Kwok-Kin
Published: July 2016

Activating KRAS mutations are identified as the most common driver oncogene for lung adenocarcinomas, accounts for 20∼30% of lung cancer patients. The development of therapeutic strategy for KRAS mutant lung cancer patients remains to be an unmet need, due to poor prognosis and therapeutic resistance to conventional therapy. The purpose of our study is through utilizing different types of genetically engineered mouse models (GEMMs), to understand the molecular heterogeneity of KRAS mutant lung cancer, to define the immune microenvironment, and to determine the feasibility and strategy of using MEK inhibition in combination with immune checkpoint inhibitors for the treatment of KRAS mutant lung cancer.

We generated two different types of GEMMs with KRAS mutations that develop tumor in the lung: Kras with G12D mutation or KRAS with G12C mutation. Continuous treatment of either KrasG12D or KRASG12C mice with MEK inhibitor Selumetinib (AZD6244, ARRY-142886) shows short-term response followed by drug resistance to single regimen treatment. The immuno-profiling shows that Selumetinib can increase cytotoxic T cells percentage and decrease PD-L1 expression on both myeloid cells and lymphocytes. Prolonged treatment of Selumetinib on KRAS mutant mice will lead to down-regulation of the expression of immune-checkpoint inhibitory factors Ctla-4 and Pd-1 on both CD4+ and CD8+ T cells. When KRAS mutant mice are treated with Selumetinib intermittently, they show improved response comparing with continuous treatment. When combining MEK inhibition with immune checkpoint blockade by using Selumetinib together with Pd-1 antibody treatment, Tim3 and Ctla-4 expression on T cells is increased potentially leading to T cell exhaustion and immune suppression in lung cancer. These pre-clinical results provide molecular insight for the immune response to MEK inhibition in KRAS driven lung cancer. Furthermore, these data support our hypothesis that MEK inhibition combination with immune checkpoint blockade treatment will have better outcome for lung cancer patients that have KRAS mutation through activating T cell response.

Citation Format: Jiehui Deng, Shuai Li, Grit Herter-Sprie, Paul D. Smith, Gordon J. Freeman, Jeffrey A. Engelman, Peter Hammerman, Kwok-Kin Wong. Pre-clinical study using KRAS mutant mouse models for lung cancer immunotherapy together with MEK inhibition. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4184.
Abstract 4186: Syngenomic fingerprint: the biomic characterization of the mouse syngeneic tumor models
Authors: Prime John E. , Mosely Suzanne , Koopmann Jens-Oliver , Wang Dennis YQ , Greenawalt Danielle , Harper James , Ahdesmaki Miika J. , Leyland Rebecca , Harris Olivia , Stewart Ross , Brohawn Philip , Higgs Brandon , Langford Bryony , Herath Athula , Kozarski Robert , Coates-Ulrichsen Jane , Anderton Judith , Morrow Michelle , Sainson Richard C. A , Wilkinson Robert W.
Published: July 2016

The pre-clinical assessment of immuno-oncology (IO) therapies can be enabled by the use of murine syngeneic tumors established in immuno-competent mice. With the aims of selecting relevant models and of minimizing animal experimentation by reducing the number of models tested, the full characterisation of syngeneic models at the transcriptomic and genomic level is a key objective for pre-clinical scientists.

Model characterisation includes global aCGH, exon array analysis and FACS profiling alongside exome sequencing. The model data is undergoing hypothesis free and driven analyses which are already generating valuable insights. Comparison of in vivo tumor samples with their in vitro equivalents has highlighted enrichment for a number of immune pathways; as has the comparison of different tumor lines. The genomic, transcriptomic and ‘proteomic’ model data are being integrated to give a functional output which will act as a ‘Syngenomic Fingerprint’ for each model.

The resulting Syngenomic fingerprints will help pre-clinical scientists to refine their in vivo plans through an improved understanding of the limits and advantages as well as the clinical relevance of some of our preclinical models. It is also supporting the targeted modification of models to better match specific human cancer types.

Citation Format: John E. Prime, Suzanne Mosely, Jens-Oliver Koopmann, Dennis YQ Wang, Danielle Greenawalt, James Harper, Miika J. Ahdesmaki, Rebecca Leyland, Olivia Harris, Ross Stewart, Philip Brohawn, Brandon Higgs, Bryony Langford, Athula Herath, Robert Kozarski, Jane Coates-Ulrichsen, Judith Anderton, Michelle Morrow, Richard C. A Sainson, Robert W. Wilkinson. Syngenomic fingerprint: the biomic characterization of the mouse syngeneic tumor models. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4186.
Abstract 4196: Real-time single-walled nanotube (SWNT)-based imaging system improves tumor detection and survival in ovarian cancer animal model
Authors: Ceppi Lorenzo , Na YoungJeong , Bardhan Neelkanth M. , Siegel Andrew , Rajan Nandini , Belcher Angela M. , Birrer Michael J.
Published: July 2016

Residual tumor detection in advanced stage ovarian cancer (OC) represents a critical challenge in the treatment of this disease. Optimal abdominal cytoreduction to reduce tumor to no evidence of macroscopic disease is the main endpoint of tumor debulking surgery, with demonstrated survival advantages, but the current clinically available imaging modalities have shown low tumor detection sensitivity of tumors smaller than 10 mm diameter. Instead, targeted molecule-based imaging systems offer higher detection rate and are likely to extend the aim of complete resection to the microscopic scale. We applied a custom designed imaging system to a preclinical ovarian cancer tumor model and observed higher microscopic tumor detection accuracy and survival advantage compared to non-image guided debulking surgery.

A two component imaging system was developed. The contrast agent is an intra-peritoneal injectable nanomolecular probe, composed of a single walled carbon nanotube (SWNT) coupled with an engineered M13-bacteriophage carrying a modified peptide targeting secreted protein, acidic and rich in cysteine (SPARC), extracellular protein overexpressed in OC. The imaging device system is capable of detecting SWNT inherent fluorescence in near-infrared second window (NIR2) and of displaying real-time images to guide the intraoperative tumor debulking.

In an orthotopic ovarian cancer animal model, the imaging system detected microscopic tumors in the millimeter and sub-millimeter scale resolution with a mean tumor diameter of 2.1 vs 3.6 mm (p< 0.01) compared to eye detected tumors. The imaging system demonstrated sensitivity of 97.2% and specificity of 69.7%, and a Receiver Operator Area under the Curve of 0.85 + 0.05. In a survival surgery animal model, the imaging guided resection surgery compared to non guided surgery showed a survival advantage for the experimental arm, with median survival time of 41.0 days vs 28.5 days, respectively (HR 0.22, 95% CI of ratio 0.04 to 1.08 Log Rank test: p: 0.063).

The accuracy of the imaging system and its survival advantage are promising. The survival outcome improvement in this preclinical model supports the translation to first-in-human investigation. Moreover, the imaging system accuracy suggests further exploitation in targeted delivery therapies.

Citation Format: Lorenzo Ceppi, YoungJeong Na, Neelkanth M. Bardhan, Andrew Siegel, Nandini Rajan, Angela M. Belcher, Michael J. Birrer. Real-time single-walled nanotube (SWNT)-based imaging system improves tumor detection and survival in ovarian cancer animal model. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4196.
Abstract 4199: Acidic pH-targeted nanorods facilitate detection of orthotopic pancreatic tumors via multispectral optoacoustic tomography
Authors: Zeiderman Matthew , McMasters Kelly M. , Chuong Phillip , McNally Lacey R.
Published: July 2016

We present a precision cancer nanomedicine based on acidic pH targeted chitosan coated mesoporous silica coated gold nanorods (CMGs) designed for multispectral optoacoustic tomography (MSOT). We have conjugated pH-sensitive variant 7 pHLIP peptide to the CMGs (V7-CMG) to provide targeting specificity to the acidic tumor microenvironment. For treatment and detection of resistant malignancies such as pancreatic cancer, such targeting modalities may help to improve treatment and enhance tumor detection. The V7-CMGs provide the ideal contrast and targeting specificity necessary for MSOT imaging of retroperitoneal orthotopic pancreatic tumors. The pH-sensitivity of the targeting pHLIP peptide and chitosan coating makes the particles suitable for simultaneous in vivo tumor imaging and drug delivery. In vitro studies confirm the pH-sensitivity of CMG drug release and tumor cell specific nano-cargo delivery.

Citation Format: Matthew Zeiderman, Kelly M. McMasters, Phillip Chuong, Lacey R. McNally. Acidic pH-targeted nanorods facilitate detection of orthotopic pancreatic tumors via multispectral optoacoustic tomography. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4199.
Abstract 420: Translational strategy for targeting MCL1 amplified tumors with CDK9 inhibitor
Authors: Yang Ziping , Bellin Ricky , Hessler Paul , Lu Xin , Uziel Tamar , Lam Lloyd T.
Published: July 2016

BCL-2, BCL-XL, and MCL-1 are members of the prosurvival family of proteins that regulate the mitochondrial apoptotic pathway. These proteins are often amplified or overexpressed in multiple tumor types. Previously, we showed that non-small cell lung cancer (NSCLC) cell lines with low BCL-XL expression (high MCL-1 / BCL-XL ratio) are MCL-1-dependent. TCGA data shows MCL1 amplification is one of the frequent genetic events in NSCLC adenocarcinoma (20%) and breast cancer (15%), while BCL2 is frequently amplified in activated B-cell like diffuse large B cell lymphoma (11%) and BCL2L1 in colorectal cancer (15%). We hypothesize that cancer cell lines with MCL1 amplification and/or high MCL-1 / BCL-XL ratio depend on MCL-1 for survival and are sensitive to inhibition of CDK9, a component of the transcriptional elongation complex that regulates MCL-1 expression. In this study, we demonstrate that NSCLC, triple negative breast cancer (TNBC) and ovarian cell lines with amplified MCL1 or low BCL-XL expression (high MCL-1 / BCL-XL expression ratio) are MCL-1-dependent, and are sensitive to dinaciclib, a CDK9 inhibitor. Cell lines with high BCL-XL expression could be re-sensitized to dinaciclib when co-treated with BCL2/BCL-XL inhibitor navitoclax (ABT-263) or BCL-XL-selective inhibitor A-1155463, suggesting BCL-XL as a resistance factor. Indeed, exogenous expression of BCL-XL rescues sensitive cell lines from dinaciclib. As reported in TCGA, ovarian, NSCLC and TNBC patients with MCL1 amplification also have high MCL-1 expression, and we hypothesized that these patients to be sensitive to CDK9 inhibitors. We developed a fluorescence in situ hybridization (FISH) assay to detect MCL1 amplification which can potentially select patients that may benefit from dinaciclib treatment. We show that fifteen percent of NSCLC patients have high MCL1 amplification, similar to published data. In addition, we have identified other predictive biomarkers associated with sensitivity in TNBC cell lines from gene expression analysis. In conclusion, we have developed a translational strategy for identifying MCL-1-dependent cancers that may be sensitive to CDK9 inhibitors in the clinic.

Disclosures:

All authors are employees of AbbVie. The design, study conduct, and financial support for this research were provided by AbbVie. AbbVie participated in the interpretation of data, review, and approval of the publication

Citation Format: Ziping Yang, Ricky Bellin, Paul Hessler, Xin Lu, Tamar Uziel, Lloyd T. Lam. Translational strategy for targeting MCL1 amplified tumors with CDK9 inhibitor. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 420.
Abstract 4202: Peptide probes binding to radiation-inducible cell surface GRP78 as a novel targeting strategy for various tumors
Authors: Kapoor Vaishali , Dadey David , Nguyen Kim , Li Hua , Rogers Buck , Thotala Dinesh , Hallahan Dennis
Published: July 2016

Traditional radiation therapy is often associated with significant toxicity to normal cells that could limit the success of cancer therapy. The greater understanding in the molecular differences between cancer cells and normal cells has lead to targeted therapies in cancer treatment. We have discovered novel radiation-inducible antigens in cancer using phage-display peptide libraries. One such inducible-antigen is Glucose regulated protein 78kDa (GRP78) that was shown to bind the hexapeptide GIRLRG. GRP78 is known to regulate cellular stresses, including hypoglycemia, hypoxia and the ER stress response. It is over-expressed in different cancer subtypes and has been correlated to their poor prognosis.

In this study, we evaluated the cell surface induction of GRP78 post irradiation (IR) on lung cancer (A549 and LLC), glioma (D54 and GL261) and endothelial (HUVEC and 3B11) cell lines using flow cytometry. Significantly higher surface expression of GRP78 was observed in all cell lines. We determined the specificity of GIRLRG to bind tumors in vivo using nano SPECT technology. GIRLRG was conjugated to a 40KDa PEG (for longer circulation time) and radiolabelled with 111Indium (111In) using diethylene triamine pentaacetic acid (DTPA) as the chelator. The A549 tumor bearing nude mice were irradiated (3Gy x 3 times) or sham irradiated prior to tail vein injections of 111In labeled GIRLRG. The mice were imaged 48h and 96h post injection using nano CT-SPECT imager. The SPECT images revealed that GIRLRG specifically bound to the irradiated A549 tumors while little or no binding was seen in the sham irradiated tumors. The post-SPECT imaging bio-distrubution also revealed maximum uptake in irradiated tumors. We further evaluated tumor binding of GIRLRG in Glioma (D54), esophageal cancer (OE33), cervical cancer (HT3), and pancreatic cancer (BxPC3). The nano-SPECT imaging showed that GIRLRG specifically bound to all the irradiated tumors tested. Phosphor images of the tumor sections showed that GIRLRG specifically bound to the tumors and not to normal tissues.

In conclusion, GIRLRG peptide has high affinity to GRP78 in vitro and in vivo. Radiolabeled GIRLRG is a novel tool for imaging tumors and may be developed further as a therapeutic agent to deliver cancer specific drugs or therapeutic radioisotopes.

Citation Format: Vaishali Kapoor, David Dadey, Kim Nguyen, Hua Li, Buck Rogers, Dinesh Thotala, Dennis Hallahan. Peptide probes binding to radiation-inducible cell surface GRP78 as a novel targeting strategy for various tumors. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4202.
Abstract 4204: Intraarterial delivery of doxorubicin-loaded hollow gold nanospheres for photothermal ablation chemotherapy of hepatocellular carcinoma in the liver of rats
Authors: Li Junjie , Zhou Min , Xiong Chiyi , Wang Wanqin , Cao Qizhen , Wen Xiaoxia , Ji Xin , Wang Andrew , Gupta Sanjay , Li Chun
Published: July 2016

Purpose: We sought to use combretastatin A-4 phosphate disodium (CA4P) to enhance the tumor uptake of doxorubicin (Dox)-loaded, polyethylene glycol (PEG)-coated hollow gold nanospheres (HAuNS; Dox@PEG-HAuNS) for improved photothermal ablation (PTA)-chemotherapy of hepatocellular carcinoma (HCC) in rats under image guidance. Materials and Methods: Two groups of 10 HCC-bearing rats each received intrahepatic arterial (IA) injection of either PEG-HAuNS/Lipiodol alone or CA4P followed by PEG-HAuNS/Lipiodol 5 min later. Each group was subdivided, and the Au content of tumors and tissues excised at 1 h or at 24 h was quantified using neutron activation analysis (n = 5/time point). Five rats received CA4P plus PEG-[64Cu]-HAuNS/Lipiodol and underwent μPET/CT. Therapeutic effects were compared among 3 groups of 6 rats each that received IA injection of saline (control group), CA4P plus Dox@PEG-HAuNS/Lipiodol (chemotherapy group), or CA4P plus Dox@PEG-HAuNS/Lipiodol plus near-infrared irradiation (PTA-chemotherapy group). Findings were verified with postmortem histopathology and/or autoradiography. Results: PEG-HAuNS uptake in CA4P-pretreated tumors was significantly higher than that in non-CA4P-pretreated tumors at both 1 h (P<0.05) and 24 h (P<0.01). The tumor-to-liver PEG-HAuNS uptake ratios at 1 h and 24 h were 5.63±3.09 and 1.68±0.77, respectively, in the CA4P-treated group and 1.29±2.40 and 0.14±0.11, respectively, in the non-CA4P-treated group. μPET/CT clearly delineated the tumors, enabling quantitative imaging guidance. Tumor volumes 10 d after therapy were 1.68±1.01, 3.96±1.75, and 6.13±2.27 cm3 in the PTA-chemotherapy, chemotherapy, and control groups, respectively, with significant differences between the PTA-chemotherapy group and other groups (P<0.05). Conclusion: CA4P pretreatment causes a higher concentration of Dox@PEG-HAuNS to be trapped inside the tumor, thereby enhancing the anti-HCC efficacy of μPET/CT-guided PTA-chemotherapy in rats. (Supported in part by the John S. Dunn Foundation)

Citation Format: Junjie Li, Min Zhou, Chiyi Xiong, Wanqin Wang, Qizhen Cao, Xiaoxia Wen, Xin Ji, Andrew Wang, Sanjay Gupta, Chun Li. Intraarterial delivery of doxorubicin-loaded hollow gold nanospheres for photothermal ablation chemotherapy of hepatocellular carcinoma in the liver of rats. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4204.
Abstract 4205: Targeting a variety of cancers with NOTA-derivatized pH (low) insertion peptide (pHLIP) complexes with 64Cu and 18F: What cancers are targetable
Authors: Demoin Dustin W. , Edwards Kimberly J. , Wyatt Linden C. , Sarparanta Mirkka , Pourat Jacob , Andreev Oleg A. , Reshetnyak Yana K. , Viola-Villegas Nerissa , Lewis Jason S.
Published: July 2016

Introduction: Although heterogeneous, the extracellular tumor environment is generally marked by decreased pH relative to nontumor tissues due to cancer cell metabolism. Using a pH-sensitive peptide to deliver diagnostic PET isotopes can provide clinicians with improved diagnostic tools for cancer-specific imaging and an alternative to receptor-based cancer targeting. pH (low) Insertion Peptides (pHLIPs) have previously been shown to target cancerous tissues and are able to deliver PET metals to the cancer site utilizing a NOTA-based chelator.

Methods: The thiol-containing cysteine moiety on the pHLIP peptide sequence was conjugated to the NOTA chelator. The NOTA-derivatized peptides were labeled with 64CuCl2 in 100 mM NH4OAc (pH 5) solution; the [18F]-fluoride was reacted with AlCl3 before forming the [18F]-AlF-NOTA-pHLIP complexes in NH4OAc buffered reaction mixtures (pH ∼ 4.1). After purification and formulation, the radiolabeled peptides were injected intravenously into 4T1 breast, PC-3 prostate, LNCaP prostate, or B16-F10 melanoma tumor-bearing mice.

Results: The 4T1 tumor-bearing mice showed significant uptake > 10%ID/g by 4 h p.i. and continued to show increased uptake of the 64Cu-NOTA-cysVar3 over 20 h. Tumors were visible in the 4T1 tumor-bearing mice as early as 2 h, but had the greatest tumor-to-background ratios at 24 h. Initial studies with B16-F10 tumor-bearing mice showed > 9%ID/g uptake in the tumor by 4 h p.i. with slight increase of 64Cu-NOTA-cysVar3 uptake over 24 h. Due to the placement of the melanoma tumors away from internal organs, tumors were visible by 1 h p.i. Additionally, the size of the melanoma tumors did not affect the%ID/g of radiotracer uptake.

Conclusions: Our studies indicate that the NOTA-cys(pHLIP), specifically NOTA-cysVar3, radiolabeled with 64Cu or 18F is a viable imaging tool for detecting highly metabolic tumors in preclinical mouse models.

Funding: NIH F32 CA186721 (D.W.D.), NIH R01 CA138468 (J.S.L.), NIH MSKCC Center Grant (P30-CA08748).

Citation Format: Dustin W. Demoin, Kimberly J. Edwards, Linden C. Wyatt, Mirkka Sarparanta, Jacob Pourat, Oleg A. Andreev, Yana K. Reshetnyak, Nerissa Viola-Villegas, Jason S. Lewis. Targeting a variety of cancers with NOTA-derivatized pH (low) insertion peptide (pHLIP) complexes with 64Cu and 18F: What cancers are targetable. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4205.
Abstract 422: Functional analysis of family with sequence similarity 83, member B in lung adenocarcinoma
Authors: Yamaura Takumi , Okabe Naoyuki , Takagi Hironori , Owada Yuki , Inoue Takuya , Watanabe Yuzuru , Fukuhara Mitsuro , Muto Satoshi , Matsumura Yuki , Hasegawa Takeo , Yonechi Atsushi , Hoshino Mika , Osugi Jun , Shio Yutaka , Higuchi Mitsunori , Ezaki Junji , Waguri Satoshi , Gotoh Mitsukazu , Suzuki Hiroyuki
Published: July 2016

Background:

We have reported the usefulness of Family with sequence similarity 83, member B (FAM83B) as a novel biomarker for diagnosis and prognosis of lung squamous cell carcinoma. FAM83B expresses not only in lung cancer but also in several types of solid cancer such as breast, esophageal, and so on. More recently, novel finding showing FAM83B involve to the resistance of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) in breast cancer cell lines has been shown. EGFR signaling pathway also plays an important role in lung adenocarcinoma, therefore we hypothesize that FAM83B associates to the tumor proliferation in lung adenocarcinoma, and its results in the resistance to EGFR TKI and in a novel therapeutic target in lung adenocarcinoma.

Materials and methods:

Sixty-four paired tumor and corresponding normal tissue samples obtained from patients with lung adenocarcinoma who underwent surgical resection since 2008 to 2011 were subjected. FAM83B mRNA expression were examined and analyzed relationship between the level of FAM83B and clinicopathological variables. Then, FAM83B knock down study was performed in several types of cancer cell lines (PC-14, H820, H1650, H1975, HLC-1, H2347) to understand whether FAM83B involve to cell proliferation.

Results:

Multivariate analysis show that smaller cancer(≦3cm vs >3cm, p = 0.044), and wild type EGFR(wild type vs. mutant, p = 0.004) tend to express higher FAM83B mRNA. There is no significant difference between FAM83B expression and progression-free survival. FAM83B knock down results in suppression of proliferation in HLC-1, H1650, which express high level of FAM83B protein.

Conclusions:

FAM83B could involve to tumor proliferation of lung adenocarcinoma in part, and could be the novel therapeutic target.

Citation Format: Takumi Yamaura, Naoyuki Okabe, Hironori Takagi, Yuki Owada, Takuya Inoue, Yuzuru Watanabe, Mitsuro Fukuhara, Satoshi Muto, Yuki Matsumura, Takeo Hasegawa, Atsushi Yonechi, Mika Hoshino, Jun Osugi, Yutaka Shio, Mitsunori Higuchi, Junji Ezaki, Satoshi Waguri, Mitsukazu Gotoh, Hiroyuki Suzuki. Functional analysis of family with sequence similarity 83, member B in lung adenocarcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 422.
Abstract 4222: Usefulness of VE1 immunohistochemical detection of BRAFV600E in aggressive thyroid cancers (PDCs and UCs)
Authors: Feás-Rodríguez Noa , Costa Angela M. , Alvarez Gago Tomás , Mateos Otero Juan José , Cameselle Teijeiro Jose Manuel , Muñoz Raquel , Garcia-Rostan Ginesa Maria
Published: July 2016

Activating BRAF mutations are frequent in thyroid follicular cell carcinogenesis. The BRAFV600E point mutation represents the most common oncogenic event in sporadic papillary thyroid cancer (PTC). Although at a much lower frequency the BRAFV600E mutation is also present among less differentiated, more aggressive, I131 resistant forms of thyroid cancer as poorly differentiated carcinomas (PDCs) and undifferentiated carcinomas (UCs). Small molecule inhibitors targeting either the BRAF V600E protein or upstream or downstream kinases involved in MAPK signalling are currently under pre-clinical or clinical investigation in advanced, metastatic, I131 resistant thyroid cancers. Recently, immunohistochemical (IHC) studies on PTCs, using the VE1 mouse anti-human BRAF V600E antibody have shown to be a reliable means for detecting the BRAFV600E mutation in a clinical setting without molecular genotyping units. In this study we sought to determine the usefulness of the VE1 antibody for detecting the BRAF V600E mutant protein in a series of 103 aggressive thyroid cancers (59 PDCs and 44 UCs) previously characterized by PCR-SSCP for the presence of BRAF mutations in exons 11 and 15. The BRAFV600E mutation was present in 10/59 PDCs (17%) and 11/44 UCs (25%). Immunohistochemistry revealed 9 mutated PDCs (sensitivity 90%) and 9 mutated UCs (82% sensitivity). The staining intensity in BRAF V600E mutated samples ranged from weak to strong. Non valuable results were found in 3 PDCs and 4 UCs. Overall the results indicate that immunohistochemistry with VE1 antibody may be an alternative to molecular biology approaches for the routine detection of BRAFV600E point mutations in clinical settings without molecular genotyping facilities. It may help clinicians in targeted therapy decision making.

Citation Format: Noa Feás-Rodríguez, Angela M. Costa, Tomás Alvarez Gago, Juan José Mateos Otero, Jose Manuel Cameselle Teijeiro, Raquel Muñoz, Ginesa Maria Garcia-Rostan. Usefulness of VE1 immunohistochemical detection of BRAFV600E in aggressive thyroid cancers (PDCs and UCs). [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4222.
Abstract 4223: Novel strategy for diagnosis of solid tumors by visualization of fusion transcripts
Authors: Badiane Markey Fatu , Batish Mona
Published: July 2016

The discovery of chromosomal translocations as one major cause of cancers has led to the introduction of numerous diagnostic techniques to detect these chromosomal abnormalities. Chromosomal banding, DNA fluorescence in situ hybridization (DNA FISH), and RT PCR are all techniques used for the diagnosis of cancers. Despite the advancement these techniques have brought to the field, they are primarily optimized for the detection of blood cancers and are not very efficient for the detection of solid tumors which are very heterogeneous. One such cancer, Ewing's sarcoma (ES), is a prevalent pediatric, bone and soft tissue tumor that is caused by a chromosomal translocation between Ewing's sarcoma breakpoint region 1 (EWSR1) on chromosome 22 and various members of the ETS protein family. The most common fusion partner, in 85% of cases, is Friend leukemia integration 1 (FLI1) on chromosome 11 which leads to the formation of the fused EWS/FLI1 gene that codes for an aberrant transcription factor. However, even within this single translocation there are multiple breakpoint-fusion scenarios that can occur which results in heterogeneity and increases the chances of a faulty diagnosis with standard techniques. Considering the diversity that exists within the molecular abnormalities that result in Ewing's sarcoma and other solid tumors, there is a need for an alternative and more robust method of detection. The technique of single molecule fluorescence in situ hybridization (smFISH) has been modified by our group to target the EWS/FLI1 transcript that results from the chromosomal fusion; this new technique is referred to as “Fusion FISH”. In Fusion FISH, two differently labeled probe sets are designed to target each half of the EWS/FLI1 fusion transcript. A co-localization of the two probe sets indicates the presence of the EWS/FLI1 translocation and a positive test for Ewing's sarcoma. Preliminary data has established the success of Fusion FISH in detecting both type 1 and type 2 translocations in several Ewing's sarcoma cell lines. The fusion transcript is expressed at similar levels in all cell lines and clearly distinct from the signal of wild type EWS and FLI1 mRNA. Additional studies will examine this new technique in other solid cancers, patient tumor samples, and for validation of additional relevant mRNA biomarkers.

Citation Format: Fatu Badiane Markey, Mona Batish. Novel strategy for diagnosis of solid tumors by visualization of fusion transcripts. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4223.
Abstract 4226: The rodent eye as a non-invasive window for understanding cancer nanotherapeutics
Authors: Wang Xinlei , Xiao Wenwu , Li Yuanpei , Zhang Pengfei , Goswami Mayank , Zawadzki Robert J. , Pugh Edward N. , Lam Kit S.
Published: July 2016

We have successfully used the mouse eye as a non-surgical window for highly efficient, optical investigation of xenograft models, using a state-of-the-art ocular imaging facility, the UC Davis “EyePod”. The EyePod employs single-cell resolution intravital confocal microscopy and optical coherence tomography, performed completely non-invasively through the natural optics of the eye (1). This technology enables repeatable in vivo imaging over days, weeks and even months, quantitative tracking of tumor development and delivery of theranostic nanoparticles, and the measurement of tumor and tumor microenvironment responses. Moreover, the retina is a highly “approachable part of the brain”, so that non-invasive study of ocular tumors provides a platform for examining such critical issues as drug delivery across the blood retinal barrier (BRB) and blood brain barrier (BBB). Clinical metastasis of solid tumor to the uvea is not uncommon. Therefore, uvea/subretinal xenograft implant, even though not orthotopic, is clinically relevant.

To visualize and photo-manipulate nanoparticle delivery to intraocular xenograft, we have used our recently reported nanoporphyrin (2) as the model system. This novel multifunctional porphyrin-based micellar nanoplatform allows (i) efficient encapsulation of hydrophobic chemotherapeutic drugs or fluorescent dyes, (ii) near-infra red fluorescent (NIRF) detection of the tumor via the intrinsic fluorescence of porphyrins, (iii) photodynamic therapy (PDT) and photothermal therapy (PTT) via efficient free radical and heat generation at the tumor site, respectively. Thorough understanding of how this nanocarrier distributes within the tumor microenvironment, and how it responds to controlled optical stimulation will enable us to maximize its therapeutic potential as a nano-theranostic agent. Preliminary FRET study with doxorubicin-loaded nanoporphyrin has allowed us to visualize in vivo and in real time the release of doxorubicin at the tumor site upon illumination with 680nm laser. In addition, the EyePod has enabled us to follow the tumor response over a period of 35 days. Work is currently underway to (i) develop patient-derived xenograft (PDX) model in the eye of NSG mice, and (ii) use EyePod to study the effect of tumor targeting and brain endothelial cell targeting peptides, recently developed in our laboratory, for nanodelivery into the tumor cells and across the BRB.

Citation Format: Xinlei Wang, Wenwu Xiao, Yuanpei Li, Pengfei Zhang, Mayank Goswami, Robert J. Zawadzki, Edward N. Pugh, Kit S. Lam. The rodent eye as a non-invasive window for understanding cancer nanotherapeutics. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4226.
Abstract 4233: Early detection of pemetrexed-induced inhibition of thymidylate synthase in non-small lung cancer with FLT-PET imaging
Authors: Chen Xiao , Berger Ian , Khalid Urooj , Cai Jenny , Patel Akash , Katz Sharyn I.
Published: July 2016

Introduction: Successful inhibition of thymidylate synthase (TS) can be imaged with FLT ([18F]thymidine)-positron emission tomography (PET) thereby providing a measure of therapy response on day 1 of therapy. Here we characterize this imaging strategy for implementation into 1st line therapy with pemetrexed and cisplatin for NSCLC.

Materials and Methods: The kinetics of pemetrexed-mediated TS inhibition mediated DNA salvage pathway “flare” in DNA was first characterized in vitro using 3H-thymidine DNA incorporation assays, analysis of TK1 protein expression and mobilization of the equilibrative nucleoside transporter 1 to the cell surface membrane. Kinetics of the pemetrexed-mediated “flare” in the salvage pathway was then confirmed in an in vivo mouse model of NSCLC. Finally, a proof-of-principle clinical trial was performed to demonstrate feasibility of imaging of pemetrexed-mediated TS inhibition in human subjects with NSLC.

Results: In vitro examination of pemetrexed-induced changes in the salvage pathway revealed a burst in DNA activity that peaked at 2 hrs following the administration of pemetrexed. The addition of cisplatin did not impact the amplitude or timing of the pemetrexed-induced “flare” in the salvage pathway. In vivo imaging of TS inhibition with FLT-PET confirmed a peak in salvage pathway activity at 2 hrs. Imaging of pemetrexed-induced TS inhibition in a human clinical trial demonstrated feasibility with imaging at the 2 hr time point.

Conclusion: FLT-PET measured efficacy of pemetrexed-induced TS inhibition is optimal at 2 hrs from the start of therapy. This timing of FLT-PET imaging is feasible in human clinical trials.

Citation Format: Xiao Chen, Ian Berger, Urooj Khalid, Jenny Cai, Akash Patel, Sharyn I. Katz. Early detection of pemetrexed-induced inhibition of thymidylate synthase in non-small lung cancer with FLT-PET imaging. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4233.
Abstract 423A: A tumor-associated mRNA localizing in circulating exosomes as a novel serological marker for pancreatic cancer: the retrospective clinical study
Authors: Taniuchi Keisuke
Published: July 2016

Introduction: Pancreatic ductal adenocarcinoma (PDAC) is the fourth most common cause of cancer death in the Western world. Early diagnosis of PDAC is difficult, and no biomarkers in blood can identify patients with pancreatic cancer at an early stage. Serum cancer antigen 19-9 (CA19-9) remains the gold standard serum marker for patients with PDAC; however, inadequate sensitivity and specificity limit the use of CA19-9 in the early diagnosis of PDAC. Therefore, the discovery of biomarkers derived from blood that facilitate the distinction of PDAC would greatly affect patient management. Exosomes are extracellular lipid microvesicles, secreted by nearly all cells in body fluids such as peripheral blood. The exosome-associated RNA is called exosomal shuttle RNA that includes microRNAs and messenger RNAs (mRNAs).

Methods: The aims of the present study are to assess if the mRNA that we found in the intracellular exosomes of PDAC cells is secreted from PDAC cells, and it is useful as a serum diagnostic marker for differentiating PDAC from individuals without pancreatic disorders compared with CA19-9. 20 PDAC patients (stage III: n = 5, stage IVa: n = 7 and stage IVb: n = 8 according to the classification of pancreatic carcinoma of the Japan Pancreas Society) and 30 control individuals without pancreatic diseases were analyzed in a retrospective cohort study. Circulating exosomes were isolated from serum, and then the mRNA localizing in exosomes was measured by real-time quantitative reverse transcriptase polymerase chain reaction (qRT-PCR).

Results: We confirmed that the mRNA was localized in secreted exosomes from cultured PDAC cells: a moderately differentiated PDAC cell line (S2-013) by the use of condition media real-time RT-PCR. Furthermore, the retrospective clinical study showed that the area under the curve (AUC) of reciever operating characteristic curves (ROC curves) was 0.805 (95%CI 0.691-0.9199) for this RNA and 0.8 (95%CI 0.675-0.926) for CA19-9. If the mRNA was combined with serum CA19-9, the AUC increased (0.921 [95%CI 0.851-0.991]).

Conclusions: Our data suggest that measuring the level of this exosome-associated mRNA has the potential to improve detection of PDAC. Further research is necessary to understand whether this mRNA has clinical implications for early detection of PDAC (stage I and II) and how much this information adds to serum CA19-9.

Citation Format: Keisuke Taniuchi. A tumor-associated mRNA localizing in circulating exosomes as a novel serological marker for pancreatic cancer: the retrospective clinical study. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 423A.
Abstract 4245: BLZ-100 provides optical contrast in animal models of breast cancer
Authors: Hansen Stacey , Fidel Janean , Kennedy Katie , Dernell William , Fransson Boel , Ramsay Josh , Wiss Valorie , Stroud Mark , Parrish-Novak Julia E.
Published: July 2016

Over 230,000 new breast cancer cases occur in the US annually. Complete surgical removal of tumor tissue is critical to reduce the risk of local recurrence. About 24% of patients who undergo breast conservation surgery require a second surgery due to positive margins. Second surgeries can be psychologically and financially costly to patients. BLZ-100, a peptide-fluorophore conjugate, is a Tumor Paint product candidate being developed to provide real-time, high resolution, intraoperative optical contrast between tumor and normal tissue, potentially helping surgeons accomplish more complete and accurate tumor resections. Intraoperative imaging with BLZ-100 may potentially improve the percentage of breast cancer patients achieving a complete resection and negative margins at their first surgical procedure. To better define the relationship between tumor pathology and BLZ-100 fluorescence, data from mouse models and canine mammary cancer cases were evaluated. Human breast cancer was modeled in mice using cell lines and xenograft models using patient-derived tumors. Review of histopathology was also conducted for each dog case. Animals received a single IV dose of BLZ-100, and tumors were excised one day after dosing. Excised tumor and normal tissues were imaged using the Odyssey Clx (LI-COR) to determine the ratio of signal in tumor compared to normal (background) tissue (TBR). Detailed histopathology reports for the dog cases were also analyzed to determine factors potentially associated with fluorescence intensity and TBR. A significant BLZ-100 TBR was observed. Histopathologic features potentially associated with TBR in dogs included cellularity and the degree of infiltration and demarcation. The studies supported the concept that BLZ-100 might potentially serve as an intraoperative optical imaging agent for breast cancer surgery.

Citation Format: Stacey Hansen, Janean Fidel, Katie Kennedy, William Dernell, Boel Fransson, Josh Ramsay, Valorie Wiss, Mark Stroud, Julia E. Parrish-Novak. BLZ-100 provides optical contrast in animal models of breast cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4245.
Abstract 4254: Dynamic change of differentiation in spheroids derived from mixed small cell carcinoma/adenocarcinoma of the uterine cervix
Authors: Kubota Satoshi
Published: July 2016

Small cell carcinoma of the uterine cervix (SCCC) occurs mainly in young women with poor prognosis. Clear treatment algorithm remains difficult to develop for its rarity. Herein, we report a case of SCCC with adenocarcinoma component, and analyzed the mechanism of differentiation using a spheroid culture method.

CTOS (cancer tissue-originated spheroid) is a primary culture system by which highly purified and viable cancer cells are cultivated. The principle of this method is maintaining cell-cell contact throughout the preparation and culture process. CTOS preserves characters of original tumors, especially their differentiation status.

We generated a panel of CTOSs from 9 SCCCs. The success rate of CTOS preparation in SCCC was 100%, indicating SCCC is one of the most suitable cancers for CTOS method.

We focused on SCCCs with adenocarcinoma component. These admixed tumors have been reported in other organs, such as lung, ovary, stomach, and so on. However, the mechanism underlying mixed tumor remains unclear. One of the CTOSs retained the characteristics of mixed tumor in original tumor.

Immunostaining revealed that CD99 was positive in the region of small cell carcinoma region, and CK7 in that of adenocarcinoma. CTOSs were sorted by CD99 and EpCAM (a possible stem cell marker) expression levels, and transplanted into NOD-Scid mice. Xenotumors derived from both CD99lowEpCAMlow cells and CD99lowEpCAMhigh cells showed mixed cell type in H&E staining. Immunohistochemistry revealed that CD99 was positive in the region of small cell carcinoma, and CK7 in that of adenocarcinoma. On the other hand, CD99highEpCAMhigh cells-derived xenotumor consisted of only small cell carcinoma. The tumors were positive for CD99, but negative for CK7.

Next, we searched for the key factor of differentiation. It has been reported that hypoxia induced adenocarcinoma in lung adenosquamous cell carcinoma. When CTOSs were incubated in hypoxia (3%O2), FACS analysis revealed that proportion of CD99low cells increased (20.6% in normoxia, 40.2% in hypoxia). Immunostaining of CTOSs showed similar results. Thus differentiation status of a mixed type SCCC showed dynamic change of differentiation in spheroids depending on culture conditions.

The research can provide insight into the origin of mixed tumor of SCCC, and may be useful for developing treatment strategy.

Citation Format: Satoshi Kubota. Dynamic change of differentiation in spheroids derived from mixed small cell carcinoma/adenocarcinoma of the uterine cervix. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4254.