bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4209: Targeting GPNMB with 89Zr-CR011 for PET imaging of triple negative breast cancer
Authors: Marquez Bernadette V. , Lee Supum , Keler Tibor , Hawthorne Thomas , Hoog Jeremy , Li Shunqiang , Ma Cynthia , Lapi Suzanne E.
Published: July 2016

Glycoprotein non-metastatic melanoma B (GPNMB) is a transmembrane protein overexpressed in 30 - 40% of triple negative breast cancer (TNBC) and has shown to be associated with metastasis and disease recurrence. An anti-GPNMB antibody drug conjugate, Glembatumumab Vedotin (CDX-011), is currently in Phase II clinical trials for the treatment of metastatic TNBC patients, with promising outcomes. Positron Emission Tomography using radiolabeled antibodies could be advantageous in stratifying patients who may benefit from CDX-011, tracking the biodistribution of CDX-011, and assessing GPNMB expression in vivo. To this end, we radiolabeled the naked antibody, Glembatumumab (CR011), with the positron-emitting 89Zr (half life = 3.3 days). We characterized the stability, affinity, rate of cellular internalization, and specificity of 89Zr-CR011 using various cell-binding assays in human TNBC cell lines. We determined that 89Zr-CR011 is stable in serum solution for up to 5 days, binds specifically to GPNMB+ TNBC cells with high affinity (KD = 16 nM), and internalizes rapidly (50% within 30 - 60 min). We conducted a biodistribution study from 1 - 12 days post administration via tail vein in GPNMB+ MDA-MB-468 xenografts to determine the time point at which we achieve the optimal tumor-to-nontarget ratios. A subset of mice was administered a blocking dose of unlabeled CR011 (100-fold excess, 1 mg/mouse), where we observed a 2.5-fold reduction in 89Zr-CR011 tumor uptake, confirming the specificity of 89Zr-CR011 for GPNMB+ TNBC tumors. PET imaging studies and dosimetry calculations are currently in progress. This preliminary study demonstrates that 89Zr-CR011 may be an excellent companion diagnostic agent for CDX-011 therapy and an essential tool to assess the function of GPNMB in vivo.

Citation Format: Bernadette V. Marquez, Supum Lee, Tibor Keler, Thomas Hawthorne, Jeremy Hoog, Shunqiang Li, Cynthia Ma, Suzanne E. Lapi. Targeting GPNMB with 89Zr-CR011 for PET imaging of triple negative 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 4209.
Abstract 421: Non reportable results - maternal neoplasm can alter results from non-invasive prenatal testing (NIPT)
Authors: Grosu Daniel , Dharajiya Nilesh G. , McCullough Ron M. , Sun Youting , Saldivar Juan-Sebastian , van den Boom Dirk , Ehrich Mathias
Published: July 2016

Cell free DNA is a powerful new analyte for the detection of chromosomal abnormalities of fetuses from the plasma of pregnant women. In non-invasive prenatal testing (NIPT), cfDNA is contributed into the plasma by the placenta. In rare cases, these results are distorted by additional sources of cfDNA in the maternal bloodstream. For example, organ transplantation can severely affect the cfDNA levels and lead to uninterpretable results. Another source of cfDNA that can lead to similar difficulties and ultimately to a non-reportable NIPT results, can be a maternal neoplasm. In our clinical laboratory, we have processed samples from over 450,000 pregnant patients. In cases of uninterpretable findings due to aberrant genomic profiles, potential reasons for non-reportable results were discussed with the caregiving physician. When feedback about these cases was available, it was collected in a database. In summary, we observed non-reportable NIPT results with aberrant genomic profile for 55 cases. A set of 43 had sufficient information available to allow inclusion in this summary. In 40 cases, a maternal neoplasm was confirmed (18 malignant, 20 benign, and two with imaging but without pathological confirmation). These observational results support further investigation by prospective controlled trials.

Citation Format: Daniel Grosu, Nilesh G. Dharajiya, Ron M. McCullough, Youting Sun, Juan-Sebastian Saldivar, Dirk van den Boom, Mathias Ehrich. Non reportable results - maternal neoplasm can alter results from non-invasive prenatal testing (NIPT). [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 421.
Abstract 4211: Discovery of a novel peptide for improvement of radiotherapeutic efficacy and development of prognotic diagnosis
Authors: Lee Kyoung Jin , Lee Jae Hee , Song Si Yeol , Jeong Seong-Yun , Choi Eun Kyung
Published: July 2016

Cancer can be treated by the various modalities including surgery, chemotherapy, radiotherapy, among which the radiotherapy is the most important therapeutics in the cancer treatment. Despite the progression of modalities, cancer therapy has been constrained by impasse of prediction of treatment outcome and direction of cancer therapeutics because of diagnosis date, metastasis and genetic diversity of the individual patients.

In this study, we discovered specific tumor targeting peptides and radiotherapy-responding peptides suitable to apply to cancer therapy and monitoring of radiotherapy response. For a non-invasive imaging for potential cancer diagnosis system, we screened peptides using the phage display technique. Phage display is used in various pharmaceutical biotechnologies, and the one of the most powerful technologies to identify novel peptides regulating carcinogenesis. To emphasize the function of peptide ligands in a living body and consider the tumoral microenvironment, we used the patient-derived xenograft (PDX) tumor models for in vivo peptide displayed phage screening. It is warranted that the peptides specifically targeting in vivo human cancer xenografts with radiation treatment or not in colorectal cancer PDXs. We performed in vivo peptide screening in the two different cases of colorectal cancer PDX mouse models, and selected peptides were confirmed to radiation responding peptides and radiation resistant peptides. The results suggested that selected peptides were a promising diagnostic lead molecule for rapid and accurate detection for the radiotherapy response.

Taken together, we described a strategy to identify the peptides by in vivo phage display, and peptides possessed the specific targeting ability which would be potentially feasible for the development of personalized precision treatment. Its might be potential IR-guided probe and prognostic diagnosis in radiotherapy for colorectal cancer.

Keywords: Peptide, Colorectal cancer, Patient-derived xenograft (PDX), Radiation, In vivo peptide screening

Citation Format: Kyoung Jin Lee, Jae Hee Lee, Si Yeol Song, Seong-Yun Jeong, Eun Kyung Choi. Discovery of a novel peptide for improvement of radiotherapeutic efficacy and development of prognotic diagnosis. [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 4211.
Abstract 4217: First pre-clinical validation of radiogenomics in glioblastoma
Authors: Zinn Pascal , Singh Sanjay , Luedi Markus M. , Zandi Faramak , Kotrotsou Aikaterini , Hatami Masumeh , Thomas Ginu , Elakkad Ahmed , Gumin Joy , Sulman Erik P. , Lang Frederick , Piwnica-Worms David , Colen Rivka R.
Published: July 2016

A plethora of Magnetic Resonance Imaging (MRI) features have been correlated to cancer genomics to date, however, none have established causality. Here, we present an in vivo xenograft RNA interference validated, potentially clinically applicable test method termed “Magnetic Resonance Radiomic Sequencing” (MRRS) for the noninvasive detection of cancer genomics in Glioblastoma. MRRS comprehensively assesses the entire tumor mass using imaging texture-based algorithms that generate thousands of variables (features) inherent to the tumor. Two independent glioblastoma stem cells (GSC1 and GSC3) harboring doxycycline inducible short hairpin RNA against Periostin (POSTN), a gene previously identified in our radiogenomic screen, were implanted at orthotopic location in nude mouse brain. In vivo knockdown of >90% and ∼40% POSTN gene was achieved in GSC3 and GSC1 respectively. The T2 and T1 post MRI texture features, in edema and contrast enhancement phenotype features were compared between doxycycline (POSTN knockdown) and sucrose (control) group of mice using T test statistics. The significant features were included in a Stepwise Forward Logistic Regression analysis to build the final predictive model. The accuracy of the model was tested using ROC cure analysis. Among 3600 features in GSC3 mice cohort, 117 features were significantly (p value<0.05) different between the two groups. The significant features were included in a Stepwise Forward Logistic Regression analysis, 2 textures features (feature 234 of edema T1 and feature 251 of edema T2) were selected to be included in the final predictive model. The AUC of the model with leave one out cross validation method was 100%. The similar analyses were done in the GSC1 mice. The final predictive model in the GSC1 group was statistically insignificant (p value = 0.15) with AUC (95% CI) = 73% (46%-98%), suggesting that MRRS is reflective of underlying gene expression levels. Our results therefore describe the ‘first mouse model derived MRRS signature to describe a causal link of gene alteration to MRRS. This novel test method may open an avenue for human-mouse matched co-clinical trials and noninvasive Radiogenomic diagnostics.

Citation Format: Pascal Zinn, Sanjay Singh, Markus M. Luedi, Faramak Zandi, Aikaterini Kotrotsou, Masumeh Hatami, Ginu Thomas, Ahmed Elakkad, Joy Gumin, Erik P. Sulman, Frederick Lang, David Piwnica-Worms, Rivka R. Colen. First pre-clinical validation of radiogenomics in glioblastoma. [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 4217.
Abstract 4219: Surface-displayed RGD enhanced the targeting and therapeutic efficacy of attenuated Salmonella typhimurium
Authors: Park Seung-Hwan , Zheng Jin Hai , Yun Hee-Seung , Park In-Kyu , Hong Yeongjin , Choy Hyun E. , Min Jung-Joon
Published: July 2016

Anticancer therapy using engineered bacteria aims to overcome limitations of current cancer therapy by actively targeting and efficiently removing cancer. In order to achieve this goal, new approaches are essential to target therapeutically resistant regions of tumors, to maintain enough number of bacteria in tumor and to deliver drugs at sufficient concentrations during the period of therapeutic process. Therefore, it is necessary to enhance the bacterial targeting efficiency through bacterial surface engineering. Here, we demonstrate that Salmonella tumor tropism can be strengthened significantly via surface displaying of RGD peptide sequence (ACDCRGDCFCG; RGD sequence) in the external loop of outer membrane protein A (OmpA) of attenuated Salmonella typhimurium. RGD-displaying Salmonella strongly bound to αvβ3 over-expressing cancer cells while showed weak binding to non-expressing cancer cells, indicating the feasibility of surface display with preferential homing peptide. In vivo bioluminescence imaging showed strong targeting efficiency of RGD-displayed Salmonella in αvβ3 over-expressing cancer xenografts (MDA-MB-231, MDA-MB-435, M21, and U87MG). The surface engineered bacteria significantly suppressed both human breast tumor (MDA-MB-231) and human melanoma (MDA-MB-435), and prolonged survival in mice. In conclusion, engineered bacteria displaying RGD peptides on the surface could advance both targeting efficiency and therapeutic effects.

Citation Format: Seung-Hwan Park, Jin Hai Zheng, Hee-Seung Yun, In-Kyu Park, Yeongjin Hong, Hyun E. Choy, Jung-Joon Min. Surface-displayed RGD enhanced the targeting and therapeutic efficacy of attenuated Salmonella typhimurium. [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 4219.
Abstract 4220: Tumor-targeting Salmonella typhimurium A1-R efficacy on colon cancer liver metastasis alone and in combination with fluorescence-guided surgery
Authors: Murakami Takashi , Hiroshima Yukihiko , Zhang Yong , Zhao Ming , Matsuyama Ryusei , Chishima Takashi , Tanaka Kuniya , Bouvet MIchael , Endo Itaru , Hoffman Robert M.
Published: July 2016

Liver metastasis is the most frequent cause of death from colon and other cancers. Surgical treatment of liver metastasis has had limited success. The aim of this study is to determine the efficacy of tumor-targeting Salmonella typhimurium A1-R alone and in combination with fluorescence-guided surgery on liver metastasis in orthotopic mouse models. HT-29 human colon cancer cells expressing red fluorescent protein (RFP) developed liver metastasis after orthotopic implantation. In the orthotopic nude-mouse models, S. typhimurium A1-R targeted liver metastases and significantly reduced their growth (p < 0.01) after i.v. administration. Adjuvant treatment with S. typhimurium A1-R was highly effective to increase overall survival (p = 0.01) and disease-free survival (p < 0.01) after bright-light surgery (BLS) of liver metastasis. The results of this study demonstrate the future clinical potential of S. typhimurium A1-R treatment of liver metastasis.

Citation Format: Takashi Murakami, Yukihiko Hiroshima, Yong Zhang, Ming Zhao, Ryusei Matsuyama, Takashi Chishima, Kuniya Tanaka, MIchael Bouvet, Itaru Endo, Robert M. Hoffman. Tumor-targeting Salmonella typhimurium A1-R efficacy on colon cancer liver metastasis alone and in combination with fluorescence-guided surgery. [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 4220.
Abstract 4221: Imaging formation of the tumor microenvironment in orthotopic liver tumors in red fluorescent protein (RFP) transgenic nude mice
Authors: Suetsugu Atsushi , Hiroshima Yukihiko , Matsumoto Takuro , Hasegawa Kosuke , Nakamura Miki , Shimizu Masahito , Saji Shigetoyo , Moriwaki Hisataka , Bouvet Michael , Hoffman Robert M.
Published: July 2016

We report here dual-color imaging of the formation of the TME in orthotopic liver cancer in transgenic red fluorescent protein (RFP) nude mice, which express RFP in all organs. Non-colored HUH-7 human hepatoma cells were injected in the spleen of RFP nude mice to establish an orthotopic liver cancer model. TME formation in the liver tumor was observed using the Olympus OV100 small animal fluorescence imaging system. Non-colored liver cancer cells formed colonies in the liver 28 days after cell transplantation to the spleen. In the liver, RFP host-mouse cells accumulated around and in the liver tumor as visualized by fluorescence imaging. A desmin and sirus-red positive area increased around and within the liver tumor over time. These results indicate cancer-associated fibroblasts (CAFs) were recruited by the liver metastasis suggesting that CAFs, along with the angiogenic tumor blood vessels, could serve as visible therapeutic targets.

Citation Format: Atsushi Suetsugu, Yukihiko Hiroshima, Takuro Matsumoto, Kosuke Hasegawa, Miki Nakamura, Masahito Shimizu, Shigetoyo Saji, Hisataka Moriwaki, Michael Bouvet, Robert M. Hoffman. Imaging formation of the tumor microenvironment in orthotopic liver tumors in red fluorescent protein (RFP) transgenic nude mice. [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 4221.
Abstract 4225: Dynamic control of tumor vasculature with direct observation using intravital microscopy
Authors: Gabriel Emmanuel M. , Fisher Daniel , Kim Minhyung , Powers Colin , Visioni Anthony , Muhitch Jason , Skitzki Joseph
Published: July 2016

Introduction

Intravital microscopy (IVM) provides in vivo real-time imaging of tumor-associated vasculature. Our group was the first to use IVM successfully in human subjects to image melanoma associated vessels. In this preclinical study, we hypothesized that intravenous (IV) nicardipine increases observable blood flow to tumor through vasodilatation and therefore may facilitate drug delivery.

Methods

Standard window chambers for IVM were implanted into female BALB/c mice. The murine colon cancer cell line CT26HA was inoculated in the skin within the chamber. Fluorescein isothiocyanate dextran (FITC-dex) was injected IV to enhance vessel observation prior to nicardipine (8 μg/mouse). Observations were performed at 100x magnification using a modified Olympus microscope for 10 minutes. Naïve mice were used for control observations. Mice bearing CT26HA were observed on post-inoculation day 5, 12 and 20. Vessel architecture and vessel diameter in response to nicardipine were compared.

Results

In non-CT26HA bearing mice, nicardipine resulted in a greater than 10% increase in skin vessel diameters directly measured using IVM (pre-nicardipine diameter 82.4 ± 3.6 μm versus post-nicardipine diameter 93.0 ± 3.9 μm. In CT26HA bearing mice, abnormal vessels were observed as early as day 7 even in the absence of a solid tumor mass. Vessel abnormalities included haphazard areas of increased vessel density, aberrant vessel branching patterns, nonfunctional vessels, and altered flow rates. Following nicardipine injection, the diameter of larger tumor-associated vessels did not increase and remained constant (pre: 111.2 ± 2.7 μm vs post: 109.1 ± 3.3 μm). Paradoxically, for smaller vessels (< 20 μm), tumor-associated vessels vasoconstricted following nicardipine injection and returned to baseline diameter after approximately five minutes. Blood flow in these small vessels decreased or stopped during this observation.

Conclusions

Observation of dynamic vessel changes from vasoactive agents is feasible using IVM. For tumor-associated vessels, nicardipine was not shown to increase vessel diameter and counterintuitively decreased diameters of smaller vessels. Implication on drug delivery from vasoactive agents will lead to further pre-clinical study of applications for human IVM.

Citation Format: Emmanuel M. Gabriel, Daniel Fisher, Minhyung Kim, Colin Powers, Anthony Visioni, Jason Muhitch, Joseph Skitzki. Dynamic control of tumor vasculature with direct observation using intravital microscopy. [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 4225.
Abstract 4237: Decreased hypoxia in a HER2+ breast cancer model following trastuzumab therapy
Authors: Sorace Anna G. , Quarles C. Chad , Sanchez Violeta , Yankeelov Thomas E.
Published: July 2016

Introduction: Hypoxic tumors demonstrate treatment resistance with an increased risk of metastasis. Therefore, alleviating hypoxia has potential to improve efficiency of combination therapies. The primary goal of this study is to quantify alterations in hypoxia in response to trastuzumab in a murine model of HER2+ breast cancer through imaging and histology.

Experimental Design: Mice were implanted subcutaneously with BT474 breast cancer cells (107) and randomly assigned into treated (10 mg/kg trastuzumab) or control (saline) groups. After tumors reached ∼250 mm3, animals (n = 32) were utilized to identify longitudinal changes in functional vascular through an intravenous injection of Hoechst 33342 nuclear stain (immediately prior to sacrifice) and hypoxia through pimonidazole intravenous injection (one hour prior to sacrifice). Tumors were extracted for immunofluorescence between days 0 through 7. Tumor sections were flash frozen and stained with anti-pimonidazole and propidium iodide (nuclear counterstain). Additionally, a set of tumors (n = 36) were sacrificed for immunohistochemistry (days 0 through 7), formalin fixed and stained for CA-IX. All stained sections were scanned in high resolution (20×) and quantitatively analyzed with Leica SCN400 software. Another cohort of animals (n = 10) were imaged with 18F-FMISO (fluoromisonidazole) PET between days 0 through 7 and quantified via mean tracer concentration (%ID/g).

Results: Immunohistochemistry revealed significantly increased hypoxia in the control group compared to treated on days 3 (p = 0.03) and 7 (p = 0.002), as measured through CA-IX staining. Additionally, on day 4, functional vascular delivery was increased while hypoxia (pimonidazole) decreased in treated tumors compared to control. 18F-FMISO PET imaging corroborated histology findings with significantly decreased hypoxia in treated tumors compared to control tumors on day 7 (-47%; p<0.0001).

Conclusion: Trastuzumab has been shown to decrease hypoxia, as measured through gold-standard immunofluorescence. Additionally, this trastuzumab-induced improvement in tumor hypoxia can be measured via clinically relevant, noninvasive imaging (18F-FMISO PET). Temporarily improving the tumor's functional vasculature and decreasing hypoxia during trastuzumab treatment has potential to enhance the effectiveness of combination therapies. Identifying windows of improved vascular and cellular normalization with noninvasive medical imaging provide opportunity to decrease resistance to standard-of-care treatments and optimize therapeutic timing and regimens.

Citation Format: Anna G. Sorace, C. Chad Quarles, Violeta Sanchez, Thomas E. Yankeelov. Decreased hypoxia in a HER2+ breast cancer model following trastuzumab therapy. [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 4237.
Abstract 4238: Multispectral optoacoustic tomography detects orthotopic pancreatic tumors in vivo using a nano-contrast agent
Authors: Chuong Phillip , Zeiderman Matthew , Grizzle William E. , McNally Lacey R.
Published: July 2016

There have been very modest developments in pancreatic cancer detection and treatment with mortality essentially remaining unchanged in the last four decades. The major challenge for both early detection and treatment of pancreatic tumors lies in overcoming various biological barriers, especially poor perfusion, insufficient contrast agents/drugs uptake by the tumor, and limitations of traditional imaging modalities. The combination of accurate real-time imaging with tumor-specific nano-contrast agents could mitigate these impediments. We constructed highly stable nano-contrast agents by encapsulating GNRs having aspect ratio 3:1 in PAA (1.5 ± 0.5 nm), and MS (6.25 ± 0.25 nm) shell, respectively and targeted to insulin growth factor 1 receptor (IGF1-R) positive pancreatic tumor cells via Syndecan-1 ligand. IGF1-R specific binding of Syndecan-MS GNRs and Syndecan-PAA-GNR was determined using in an IGF1-R negative cell line (MiaPaca-2) or a blocking antibody to IGF1-R in S2VP10L pancreatic cancer cells determined via flow cytometry. In vivo, mice bearing orthotopic pancreatic tumors were iv injected with 100 nM of Syndecan-MS GNRs and accumulation was determined four hours post injection via Multispectral Optoacoustic Tomography. Biodistribution was determined using a Region of Interest method. The cellular uptake of Syndecan-MS-GNRs significantly enhanced in the S2VP10L cells (503.4 counts) compared to MiaPaca-2 cells (64.1 counts), blocking agent (323.7counts) and untargeted MS GNRs (average 47 counts) revealing that Syndecan-MS-GNRs enhanced both tumor specificity and OA signals by binding with IGF1-R in S2VP10L cells. In vivo, Syndecan-MS-GNRs significantly accumulated in S2VP10L orthotopic pancreatic tumors (233.1 MSOT a.u.) with minimal accumulation in kidney(29.7 MSOT a.u.) and liver (31.4 MSOT a.u.). The Syndecan-MS-GNRs did not accumulate within the orthotopic MiaPaca-2 (IGF1-R negative) mouse model with the biodistribution of pancreas tumor (12.4 MSOT a.u.), kidney (114.3 MSOT a.u.), and liver (35.2 MSOT a.u.).

Citation Format: Phillip Chuong, Matthew Zeiderman, William E. Grizzle, Lacey R. McNally. Multispectral optoacoustic tomography detects orthotopic pancreatic tumors in vivo using a nano-contrast agent. [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 4238.
Abstract 4242: Rhodamine esters as fluorescent tumor painting agents for glioblastoma
Authors: Lin Yu-Hsi , Millward Steven , Gammon Seth , Satani Nikunj , Hammoudi Naima , Gray Joshua Philip , Kelderhouse Lindsay E. , Ornelas Argentina , Schroeder Haley , Muller Florian L.
Published: July 2016

GBM remains one of the most difficult cancers to treat. Despite surgery, radiation and chemotherapy, tumors invariably recur. There is a strong correlation between extend of resection and subsequent time to recurrence and ultimate patient survival. A key challenge for the neurosurgeon is to minimize removal normal brain tissue whilst being as aggressive as possible with regards of removal of tumor tissue. Defining tumor boundary during surgery remains challenging. Fluorescent dyes such as 5- Aminolevulinic acid, have been studied as aids for tumor delineation but have failed to gain widespread acceptance in clinical practice. It has long been known that a substantial number of cancers, including Glioblastoma, exhibit a hyperpolarization of the plasma and mitochondrial membrane potential. This hyperpolarization is manifest by increased retention of lipophilic cationic dyes (Nernstian probes), such as the SPECT-CT probes 99Tc-Sestamibi and 99Tc-Tetrofosmin in both glioma cells in culture and primary tumors. We synthesized a series of ester derivatives of Rhodamine B with high fluorescence quantum yield in the red spectrum and a very favorable biological safety profile. Utilizing a series of intracranial xenografted glioblastoma pre-clinical mouse models, we show that a single I.V. injection of 1 mg/kg fluoroethylrhodamine ester (RhoFe) results in dramatic selective fluorescence in tumor but not surrounding normal brain tissue. This was observed even in tumors with minimal breakdown of the blood brain barrier, as determined by T1-contrast enhancing agents. RhoFe-tumor-selective fluorescence can remain visible up to 72 hours after injection. Similar results were obtained with Tetramethylrhodamine ester (TMRE) and rhodamine 123 (Rho123), but unlike RhoFe, TMRE proved highly toxic, while Rho123 fluorescence was both weaker and in the green spectrum, accompanied by higher endogenous auto-fluorescence. While well tolerated in vivo, RhoFe shows strong photoxicity in cell culture, suggesting potential as a photodynamic therapy agent. Taken together, these data suggest that RhoFe may be a promising tumor painting agent, with potential utility in fluorescence guided surgery as well as photodynamic therapy.

Citation Format: Yu-Hsi Lin, Steven Millward, Seth Gammon, Nikunj Satani, Naima Hammoudi, Joshua Philip Gray, Lindsay E. Kelderhouse, Argentina Ornelas, Haley Schroeder, Florian L. Muller. Rhodamine esters as fluorescent tumor painting agents for glioblastoma. [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 4242.
Abstract 4244: Optical imaging of ovarian cancer metastasis in mouse models of obesity
Authors: Leevy W. Matthew , Liu Yueying , Metzinger Matthew , Lewellen Kyle , Stack. M. Sharon
Published: July 2016

Epithelial ovarian cancer (EOC) is a deadly gynecologic malignancy, owing to its propensity for intraperitoneal (IP) metastasis. Indeed, almost 75% of women with EOC are diagnosed during the later stages of the disease in which metastasis is already underway. Pathogenesis of EOC is marked by diffuse and widely disseminated metastatic sites within the IP compartment, often with peritoneal ascites. Mouse models of EOC have been developed to facilitate the pre-clinical research of this deadly disease, however the quantification of metastatic tumor burden in these animals has remained challenging due to the broad distribution of lesions in the IP space. Here we describe a combined In Vivo/Post-Mortem method to monitor and measure EOC metastasis in mouse models of obesity. RFP-labeled ovarian cancer cells were injected IP in obese and control cohorts of mice, followed by weekly in vivo imaging until time of sacrifice. Subsequent post-mortem imaging was conducted after surgical exposure of the peritoneal cavity to reveal organs that were then imaged in situ. Ex vivo imaging was performed last on dissected organs from each animal. Imaging signal was enhanced via multispectral imaging methods to remove tissue auto-fluorescence. Semi-quantitative methods to measure tumor burden are demonstrated in their application to the study of metastasis in both obese and control animals. Ultimately these results demonstrated that obesity dramatically increased ovarian cancer metastatic success, and likely contributes to the reduced EOC survival rates noted in clinical reports of obese patients.

Citation Format: W. Matthew Leevy, Yueying Liu, Matthew Metzinger, Kyle Lewellen, M. Sharon Stack. Optical imaging of ovarian cancer metastasis in mouse models of obesity. [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 4244.
Abstract 4255: EpCAM positive lung cells are involved in organoid formation
Authors: Semba Takashi , Sato Ryo , Suzuki Makoto , Saya Hideyuki , Arima Yoshimi
Published: July 2016

A combination of fluorescence activated cell sorting (FACS) and 3D culture techniques enable us to isolate specific lung cells and to form organoids, which recapitulate organ architectures in vitro. To clarify the cells of origin in the formation of lung organoids and to apply for oncogene-mediated transformation, we conducted spheroid culture and 3D culture experiments using mice lung cells.

We isolated the CD31-/CD45- cells from the lungs of wild-type (wt) or Ink4a/Arf-knockout (Ink4a/Arf-/-) C57BL/6 mice. The single cells were then cultured in serum-free medium in a low-attachment dish for 7 days to form spheres. The spheres were dissociated into single cells again, and were then infected with retroviruses expressing KRASG12V, EML4-ALK, EZR-ROS1 (Arai et al., PLoS ONE 2013), or KIF5B-RET (Saito et al., Carcinogenesis 2014) genes, which were identified as oncogenes in human lung adenocarcinomas. Those oncogene-expressing sphere-derived cells from wt and Ink4a/Arf-/- lung cells neither expressed EpCAM (the epithelial marker) nor TTF-1 (the lung lineage marker). We found that those cells formed undifferentiated invasive lethal tumors in lung lobes when the cells were inoculated into the lungs of mice via the trachea, after bleomycin treatment.

We then isolated the EpCAM+/CD45-/CD31- lung cells from wt or the Ink4a/Arf-/- C57BL/6 mice and these cells were subjected to a 3D culture. Those cells were suspended in a serum-free medium with keratinocyte growth factor, and the 1×104 cells were seeded on a 24 well 0.4μm insert pre-coated with growth factor-reduced Matrigel. After 2 weeks, the cells formed morphologically distinct phenotypes of colonies, such as cystic colonies, dense sphere colonies, and branched colonies. A histopathological study revealed that the EpCAM-positive colonies were composed of TTF-1-positive cells, and included differentiated lung cell markers, SP-C, Aquaporin 5, and CC-10. Interestingly, the EpCAM-/CD45-/CD31- cells from the lungs of the Ink4a/Arf-/- mice did not form organoids under 3D culture conditions.

Our data indicates that the EpCAM-positive normal mice lung cells, but not the EpCAM-negative cells, formed lung organoids including TTF-1-positive cells in a 3D culture. Inoculation of EpCAM+/CD45-/CD31- lung cells expressed oncogenes into the lungs of mice might allow us to develop a well-differentiated lung adenocarcinoma model.

Citation Format: Takashi Semba, Ryo Sato, Makoto Suzuki, Hideyuki Saya, Yoshimi Arima. EpCAM positive lung cells are involved in organoid formation. [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 4255.
Abstract 4259: Long term cell culture models to identify biomarkers of response
Authors: Warrior Usha , Crane Jonathan , Bernards Karen , Nelson Brian , Snead Katie , Angione Alison , Sherman Amber , DeVault Shawn , Hnilo Jim , Shah. jameel
Published: July 2016

In order to test the efficacy of many cancer therapeutics, culturing cells in the presence of compounds for longer than the conventional 72-hr cell growth assay may be necessary. It is shown that the inhibition of many epigenetic targets results in a significantly delayed drug response not revealed by conventional shorter term cell growth assays. OncoPanel™ from Eurofins Pharma Discovery Services is a collection of 300 cell lines with broad cancer type and subtype representation and extensive genomic characterization. We have developed a 10-day assay to test inhibitors of epigenetic targets. Using the EZH2-selective inhibitor, GSK343, we have demonstrated the advantages of cell line profiling under long-term assay conditions when drug response reflects a protracted mechanism of action. We also tested Olaparib, a PARP inhibitor, in this system and showed substancial increased sensitivity against several cell lines versus the 72 hr assay. The significant biomarkers identified from the genomic analysis of the data from both GSK343 and Olaparib substantiate the relevance of long term cell profiling.

Citation Format: Usha Warrior, Jonathan Crane, Karen Bernards, Brian Nelson, Katie Snead, Alison Angione, Amber Sherman, Shawn DeVault, Jim Hnilo, jameel Shah. Long term cell culture models to identify biomarkers of response. [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 4259.
Abstract 4261: Novel tumor microenvironment (TME) restored 3D cell culture model and humanized patient-derived xenograft model for cancer research and drug evaluation
Authors: Zhang Jing , Yang Dabing , Wang Huilin , Song Qiong , Li Hui , Shaw JP , Deng Tianjing
Published: July 2016

New anticancer drug development is an expensive and time consuming process, which typically can take more than 10 years and cost over a billion dollars. Although a majority of late discovery drug candidates demonstrate potency in preclinical models, most will ultimately fail to achieve efficacy in clinical trials. While recent strides have been made to improve preclinical models, the continued widespread use of traditional models creates translational challenges during clinical development. An intrinsic challenge for drug developers is to close the gap between preclinical modeling and translational efficiency for patients. Accurately modeling core attributes of the human tumor microenvironment (TME) in preclinical models is key for identifying novel targets, lowering toxicity and improving translational success. Even for advanced patient-derived xenograft (PDX) in vivo models, the cytokines generated by murine stromal cells fail to replicate many important paracrine networks, which can mislead preclinical pharmacology outcome data. To address this gap, we have begun to establish and characterize nearly 1,000 novel patient tumor samples, spanning over a dozen cancer indications, into robust human TME-restored 3D cell culture models and humanized patient derived xenograft models. Our goal is to improve overall patient-derived tumor model initiation success rates and subsequent translational pharmacology.

Citation Format: Jing Zhang, Dabing Yang, Huilin Wang, Qiong Song, Hui Li, JP Shaw, Tianjing Deng. Novel tumor microenvironment (TME) restored 3D cell culture model and humanized patient-derived xenograft model for cancer research and drug evaluation. [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 4261.
Abstract 4262: Combined 3D quantitative imaging and 3D cell culture for cancer drug discovery
Authors: Vidal Guillaume , Lobjois Valérie , Souguir Zied , Pannetier Pauline , Demange Elise , Lagarde Jean-Michel
Published: July 2016

Monitoring drug activity in multicellular tumor spheroid in vitro models is an important challenge as these 3D models have demonstrated their major relevance in pharmacological screening and anticancer drugs evaluation (1). Optical microscopy coupled to the selective plane illumination microscopy (SPIM)3D microscopy, and, and automatic automated images processing and quantitative analysis are promising approaches to process optimizes the evaluation of drug response in a 3D cell culture models with, in a high/mediumhigh throughput screening waycapabilities.

BIOMIMESYS® used as a 3D cell model is based on hyaluronic to provide an in vivo like environment to the cells.

The In this study the effects of the topoisomerase I inhibitor Topotecan were investigated on colon adenocarcinoma HCT-116 3D cell culture. Spheroids were prepared and grown in BIOMIMESYS® used as a 3D cell model isa matrix based on hyaluronic acid tothat provides an in vivo like environment to the cells.

was monitored and coupled with mMorphometric parameters and with nuclei fluorescence markers were used to monitor analysis for drug activity. Data provideand to characterization characterize of both cytostatic and cytotoxic effects of this drug withon the basis of both quantitative 3D and imaging of the mechanisms.

These data highlight the relevance of combining a biomimetic microenvironment for 3D cell culture recreating the the in vivo features of a tumor cell population biology, together with and high resolution content microscopy-based quantification for drug screening.

This biomimetic model provides an innovative platform for the in-depth analysis of tumor development and for the discovery or the characterization of novel therapeutic targets.

Ref:

1) Breslin S, O’Driscoll L. Drug Discov Today.18: 240-249 (2013)

Citation Format: Guillaume Vidal, Valérie Lobjois, Zied Souguir, Pauline Pannetier, Elise Demange, Jean-Michel Lagarde. Combined 3D quantitative imaging and 3D cell culture for cancer drug discovery. [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 4262.
Abstract 4265: Automation-enabled 3D tumor chemosensitivity screening by imaging and flow cytometry analysis
Authors: Kowalski Michael , Raksakulthai Vipat , Prasauckas Kristin , Jones-Roe Tara
Published: July 2016

Three-dimensional (3D) cancer cell cultures are being utilized for drug screening with increasing frequency since they provide a more physiologically relevant solid tumor model than two-dimensional cell cultures. Spheroids are a well characterized, in vitro tumor model system that has been utilized with multiple cancer cell lines. Growth and manipulation of these cultures in hanging drop plates (HDPs) can be challenging to manipulate and these challenges are amplified as sample throughput increases. We automated the culture and compound treatment of cancer spheroids in HDPs, as well as the preparation of these spheroids for analysis by high content imaging and flow cytometry. Automated cancer spheroid cultures showed excellent consistency in size and shape, providing higher confidence in the screening results. Creating single cell suspensions from the cancer spheroids for flow cytometry analysis was another significant challenge overcome through automation. Imaging provided information such as the location of drug-responding cells within the cancer spheroid while flow cytometry removed the potential for spheroid exclusion of staining reagents leading to false negatives. Automation enabled the use of these complementary analyses, thereby achieving a more complete understanding of compound sensitivity in solid tumors and illustrating how spheroids can be utilized to screen for novel cancer therapeutics.

Citation Format: Michael Kowalski, Vipat Raksakulthai, Kristin Prasauckas, Tara Jones-Roe. Automation-enabled 3D tumor chemosensitivity screening by imaging and flow cytometry analysis. [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 4265.
Abstract 4270: UMR957: a new osteogenic osteosarcoma cell-line derived from an osteoprotegerin null mutant mouse
Authors: Navet Benjamin , Amiaud Jérome , Charrier Céline , Quillard Thibaut , Brounais-Le Royer Bénédicte , Heymann Dominique , Lezot. Frederic P.R.
Published: July 2016

Background: Mouse osteosarcoma cell-lines enabling in vivo studies in syngeneic mouse strains are rare but important for preclinical studies. Existing cell-lines when injected are mainly osteolytic with a moderate tumor osteoid tissue formation. There is a crucial need for new cell-lines with higher osteogenic characteristic.

Methods: A female osteoprotegerin null mutant mouse, breed in the animal house of the Faculty of Medicine from the University of Nantes (France), spontaneously developed an osteosarcoma on the left tibia. The mouse was sacrificed, a biopsy was place in culture for further in vitro and in vivo (injections) studies and the tumor was analyzed using μ-CT, μ-radiography, classical histology and immunohistochemistry.

Results: μ-CT and μ-radiography analyses evidenced that the tumor developed in the Opg-/- mouse was highly osteogenic with an important tumor osteoid tissue that colonized most of the surrounding soft tissues. A total destruction of the tibia was observed. Histology and immunohistochemistry confirmed the presence of an osteosarcoma with an important expression of most bone markers as Osterix. An important vascularization was evidenced in the tumor as the presence of numerous TRAP positive cells. Monocyte/macrophage and T-lymphocyte cells were also present in this tumor. From the biopsy a cell-line was obtained and named UMR957. This cell-line evidenced all markers of the osteoblast cell. When injected in the tibia vicinity of either naïve C57BL/6J, Nude or Opg-/- mouse a highly osteogenic osteosarcoma was observed with similar features to the initial tumor.

Conclusions: A new mouse osteosarcoma cell-line was obtained from a spontaneous tumor developed in an Opg-/- mouse. This cell-line when injected in syngeneic mouse recapitulates all features of the initial tumor more particularly its massive osteoid tissue formation.

Citation Format: Benjamin Navet, Jérome Amiaud, Céline Charrier, Thibaut Quillard, Bénédicte Brounais-Le Royer, Dominique Heymann, Frederic P.R. Lezot. UMR957: a new osteogenic osteosarcoma cell-line derived from an osteoprotegerin null mutant mouse. [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 4270.
Abstract 4281: Mitochondrial DNA copy number, urinary isoprostanes and colorectal cancer risk
Authors: Thyagarajan Bharat , Wang Renwei , Koh Woon-Puay , Barcelo Helene , Adams-Haduch Jennifer , Guan Weihua , Bostick Roberd M. , Yuan Jian-Min , Gross Myron D.
Published: July 2016

Previous epidemiological studies on mitochondrial DNA (mtDNA) copy number and risk of colorectal cancer have yielded mixed results. One study reported a positive association, another reported an inverse association, and a third, a U-shaped association of mtDNA copy number with colorectal cancer risk. Since oxidative stress is one of the proposed underlying mechanisms that may link mtDNA copy number variation to colorectal cancer risk, we conducted a matched case-control study of incident colorectal cancer nested within the Singapore Chinese Health Study. Pre-diagnostic blood and urine samples were obtained from 448 colorectal cancer cases and 869 age- and sex-matched controls. The mtDNA copy number in peripheral white cells was measured using real time PCR and urinary F2-isoprostanes were quantified using a liquid chromatography mass spectrometry based method. Conditional logistic regression was used to estimate associations of mtDNA copy number and urinary F2-isoprostanes with incident colorectal cancer. The odds ratios (OR) and 95% confidence intervals (95% CI) for those in the highest relative to those in the lowest quartiles of relative mtDNA copy numbers and urinary F2-isoprostanes were 1.02 (95% CI 0.72 - 1.43; p for trend = 0.56) and 1.11 (95% CI 0.78-1.59; p for trend = 0.35) respectively. Urinary F2-isoprostanes levels were not associated with the mtDNA copy number among the controls (r = 0.03; p = 0.33). The results of this prospective study suggest that white blood cell mtDNA copy number or urinary levels of F2-isoprostanes in biospecimens collected several years prior to the diagnosis of colorectal cancer may not be associated with risk of developing colorectal cancer.

Citation Format: Bharat Thyagarajan, Renwei Wang, Woon-Puay Koh, Helene Barcelo, Jennifer Adams-Haduch, Weihua Guan, Roberd M. Bostick, Jian-Min Yuan, Myron D. Gross. Mitochondrial DNA copy number, urinary isoprostanes and colorectal cancer risk. [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 4281.