bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5493: A new strategy to assess cytotoxicity using Raman spectroscopy of single isolated nuclei
Authors: Lin Hsin-Hung , Li Yen-Chang , Liu Chun , Chen Chung-Hsuan , Yu Alice L. 
Published: February 2011

Abstract
Single-cell biodetection systems have potential applications in biomedicine and pharmacotherapy, and Raman spectroscopy is an ideal tool for single-cell analysis. Raman spectroscopy biosensors can provide an information-rich “fingerprint” of the overall biochemical composition of single cell undergoing time-dependent changes of cellular biochemistry without reacting with many biologically active compounds. To minimize the interfering signals form cytoplasm, nuclei were isolated from cells for analysis by Raman spectrum to compare the cellar responses and biochemical changes that occur in the nuclei of the HL60 promyelocytic leukemia cell line and drug-resistant cells HL60/ADR after treatment with the antineoplastic drug, doxorubicin. With increasing concentrations of doxorubicin up to 80 nM, the intensities of individual peaks representing nucleic acids and proteins displayed a concentration dependent decrease in the nuclei of HL60 but not HL60/ADR. At 80 nM doxorubicin, there was a 55.6±2.2% (p&lt;0.001) of control in the intensities of Raman peaks representing nuclei acid, but no change of Raman peak intensities of the nuclei of HL60/ADR (96.4±3.1% of control). A significant drop in the intensities of Raman peaks representing nucleic acid was evident around 8 to 10 hours after doxorubicin exposure. Upon incubation with 80nM doxorubicin for 24 hours, thymidine incorporation was 74.2±1.9% and 94±3.8% of control in HL60 and HL60/ADR, respectively and Alamar Blue assay was even less sensitive with the value of 102.0±1.8% and 101.7±2.5% of control in HL60 and HL60/ADR, respectively. To verify the sensitivity and reproducibility of single nuclei Raman spectroscopy, treatment of CCRF-CEM, a T lymphoblastoid cell line, and drug-resistant subline CEM/VBL with anti-microtubule agent, vinblastine was also examined. At 0.4 nM, the intensities of Raman spectrum for nucleic acid dropped to 77.2±1.5% (p&lt;0.001) of control while thymidine incorporation was not significantly inhibited (94.8±4.1% of control). In comparison, CEM/VBL was resistant to vinblastine up to 50 nM with negligible changes in Raman peak intensities of CEM/VBL nuclei (96.8±2.7% of control), and thymidine incorporation was 102.5±1.2% of control. These findings demonstrate that this single nuclei Raman spectroscopy has the ability to detect and identify nuclear changes related to cytotoxicity at lower concentrations and in shorter times than conventional cell based assays. Thus, this new cytotoxicity detection strategy using Raman spectroscopy of single, isolated nuclei may be useful for rapid and sensitive detection of cellular changes in response to chemotherapeutic agents.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5493.
Abstract 5496: Determination of drug-kinase residence time in a homogenous, low-volume format
Authors: Marks Bryan , Lebakken Connie , Riddle Steve , Somberg Richard , Hereley Sara 
Published: February 2011

Abstract
In addition to drug-target affinity, drug-target residence time is becoming recognized as a critical parameter influencing drug efficacy. Desired compound action can only take place when a drug is interacting with its target protein, which is governed both by the affinity of the compound for the target, as well as the dissociative off-rate of the compound from the drug-target complex. A compound with a slower dissociative off-rate may require reduced dosing schedule relative to a compound with a rapid off-rate, and effective target specificity may be increased if the off-rate for the desired drug-target complex is greater than that for off-target complexes. With respect to kinase inhibitors, a slow off-rate for Tykerb relative to the related EGFR inhibitors Tarceva and Iressa has been proposed to explained longer lasting effects in cell culture. Currently, off rates for kinases can be measured by surface plasmon resonance (SPR)-based instruments or by detailed enzyme kinetics experiments. While SPR yields highly quantitative data, it is low-throughput and requires costly instrumentation. Furthermore, it often involves covalent conjugation of protein targets to surfaces, which can lead to loss of protein function. Off-rate measurements by classical enzyme kinetics methods are established, but fairly labor-intensive. We have developed and will present data on a fully homogenous (in-solution) method for determining kinase-compound off-rates that is simple to perform in a standard fluorescence-based plate reader. In this method, a drug-kinase complex is first formed at supra-Kd concentrations of drug, and then rapidly diluted into a solution containing a fluorescent, ATP-site directed small molecule probe. As the compound dissociates from the kinase the fluorescent probe associates with the kinase, causing an increase in the fluorescence signal that can be monitored in real time.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5496.
Abstract 5502: Therapeutic efficacy prediction by quantitative protein analysis in response to anti-cancer agents
Authors: Nishizuka Satoshi S. , Ishida Kazushige , Matsuo Teppei , Noda Hironobu , Iwaya Takeshi , Wakabayashi Go 
Published: February 2011

Abstract
Background: The conventional chemosensitivity test employs a growth suppression assay. However, surgically removed specimens often have low viability which makes the assay unreliable, since the poor cell growth may be associated with the low viability instead of the chemosensitivity of the cell. On the other hand, we do know that the level of proteins involved in signal transduction is changed prior to cellular responses to the anti-cancer agents. In the present study, we investigated if protein signaling within a 24-hour exposure of the anti-cancer agents can predict the cellular fate at 72-hour time points.
Materials and Methods: The human colon cancer cell line, HCT116, was exposed to three different DNA-damaging anti-cancer agents including gamma irradiation, ultraviolet, and doxorubicin. The anti-cancer agent-exposed cell pellets from different conditions (e.g., doses, and time-points) were collected to produce a lysate array, which finally accommodated more than 500 samples on a single glass slide. Array image and quantitative protein analyses were performed with the P-SCAN and ProteinScan programs.
Results: A total of 19 protein species were tested on a lysate array, providing kinetic information of the protein. In response to all three agents, p53 protein was stabilized and the level increased over time in a dose-dependent manner. Upon detailed time-course analysis, however, an oscillation was seen in response to gamma irradiation, indicating the presence of a feedback loop. Comparing the rate of increasing levels of p21 and CyclinD3 within 24 hours in response to doxorubicin administration, the rates are well associated with the status of cell fate. In other words, when the rate of p21 is higher than CyclinD3, it is associated with a G2 arrest at 72 hours, whereas a higher CyclinD3 rate is associated with apoptosis at 72 hours after drug administration.
Conclusion: Some events occurring at 72 hours can be predicted by seeing the quantitative level of protein kinetics within 24 hours. Most of the primary culture cells can survive for only a short period of time. The protein kinetics may be a useful clinical chemosensitivity predictor even when using low viability specimens.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5502.
Abstract 5508: Screening Of Novel Thymidylate Synthase Inhibitors
Authors: Repalli Jayanthi , Robinson Matthew R. , Gumina Giuseppe , Berger Sondra H. 
Published: February 2011

Abstract
The nucleotide stress response (NSR) is defined as activation of nucleoside salvage pathways to rescue cells from the detrimental consequences caused by a deficiency of or increased demand for nucleotides. Thymidylate depletion due to loss of function of the enzyme, thymidylate synthase (TS) or exposure to TS inhibitors disrupts the balance in nucleotide pools and damages DNA leading to stress-induced cell death. In order to replete nucleotide pools, cells activate nucleoside (ENT 1) transporters to increase the influx of extracellular nucleosides. TS catalyzes the de-novo synthesis of thymidylate for DNA replication. The enzyme in humans undergoes conformational switching between active and inactive forms. We are testing the hypothesis that stabilization of the inactive conformation will not induce the resistance mechanisms that are observed after exposure to active site-targeted TS inhibitors. To identify lead compounds that bind to the inactive conformation, a NSR assay has been developed, which measures a stress response to TMP depletion. TS inhibitors that bind to either the active or inactive conformations will induce TMP stress which is measured as induction of uptake of a fluorescent probe, N2, N3-etheno-6-thiomethylpurine riboside (ETMPR). In order to differentiate TS inhibitors that are selective for active and inactive conformations, we conducted an experiment on paired isogenic cell lines that express either wild-type hTS (undergoes conformational switching) or a mutant hTS (R163K - stabilized in the active conformation). Cells exposed to the potent active site TS inhibitor, raltitrexed (RTX) exhibited a similar level of uptake of ETMPR. When cells are exposed to glutarate (inactive site inhibitor predicted by structure-based drug design), fluorescent nucleoside uptake was preferentially induced in cells expressing wild-type hTS, consistent with the prediction that glutarate is an inactive site targeted drug. Protein turnover studies revealed that glutarate decreases TS expression, which is opposite to the effect of active TS inhibitors, which stabilize TS to degradation. Collectively, the data indicate that the NSR assay will be useful in identifying lead inactive site-targeted drugs and that these drugs will not stabilize TS, a mechanism thought to cause resistance to active state TS inhibitors.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5508.
Abstract 551: Protein biomarkers of human prostate cancer stromal cells
Authors: Khamis Zahraa I. , Iczkowski Kenneth A. , Sang Qing-Xiang Amy 
Published: February 2011

Abstract
In situ human prostate tumors are not lethal unless they become invasive and metastatic. For cells to become invasive, the prostate gland must undergo disruption of the basal cell layer and degradation of the basement membrane underneath the luminal epithelial cell layer. Although the roles of proteinases in breaking down the basement membrane have been well-studied, little is known about the factors that induce basal cell layer disruption, degeneration, and its eventual disappearance in invasive cancer. It is hypothesized that microenvironmental factors may affect the degradation of the basal cell layer which if protected may prevent tumor progression and invasion. In this study, we have revealed differential protein expression patterns between cells separated from different prostate pathologies and identified several important stromal proteins that may contribute to inflammation and malignant transformation of benign prostate tissues to cancerous tissues using mass spectrometry and proteomics methods. The identifications of these proteins may generate new strategies for the prevention of basal cell layer disruption leading to the eventual blockade of invasive prostate carcinomas. Furthermore, human matrix metalloproteinase-26/endometase/matrilysin-2 was also investigated and its expression in preinvasive human prostate cancer cells may be associated with the expression of some apoptotic markers. This work may contribute to our understanding of epithelial-stromal interaction in the progression of human prostate cancer. (This work was supported in part by grants DAMD17-02-1-0238 and W81XWH-07-1-0225 from DOD, US Congressionally Directed Medical Research Programs, a grant from the Elsa U. Pardee Foundation, and grants from Florida State University to Dr. Q.-X. Sang)
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 551.
Abstract 5513: Synergistic antitumor activity of NK012, a micelle-forming macromolecular prodrug of SN-38, combined with platinum compounds in human cancer xenograft models
Authors: Masuda Kuniko , Onda Takeshi , Akatsu Yuichi , Ichimura Eiji , Nakanishi Takeshi , Okamoto Kazuya , Morino Tomio 
Published: February 2011

Abstract
Irinotecan (CPT-11) is clinically used to treat various tumors, but low enzymatic conversion to its active metabolite, SN-38, limits its therapeutic effect. NK012, a micelle-forming macromolecular prodrug of SN-38, accumulates in tumor tissue and gradually releases SN-38 in enzyme-independent manner. NK012 showed much stronger growth inhibitory effects than CPT-11 at maximum tolerated dose (MTD) against all tested xenografted human tumors. Furthermore, NK012 showed much stronger antitumor activities than other cytotoxic agents clinically used for cancer treatment (taxanes, doxorubicin, carboplatin etc.).
Here, we report combination antitumor effects of NK012 with platinum compounds in human breast cancer, non-small cell lung cancer, colorectal cancer and hepatocellular cancer xenografts in nude mice. The dose of NK012 was set at 1.9-3.8 mg/kg/day to be sub-optimum for complete regression depending on the cell line. NK012 and carboplatin were administered by q7dx3. Although both of NK012 and carboplatin were ineffective as single agent (minimum T/C values &gt;50%), their combination was effective (minimum T/C value = 8%) to cause tumor regression in MC-05-JCK breast cancer. In MC-05-JCK breast cancer, the TGDs (Tumor Growth Delay; tumor tripling time over control group delayed by treatment / tumor tripling time of control group × 100) by NK012 1.9 mg/kg/day, carboplatin 50 mg/kg/day (67% of MTD), and their combination were 41%, 29%, and 286%, respectively. In NCI-H460 non-small cell lung cancer, the TGDs by NK012 3.8 mg/kg/day, carboplatin 75 mg/kg/day (MTD), and their combination were 90%, 71%, and 403%, respectively. In Co-3 colorectal cancer, the TGDs by NK012 3.8 mg/kg/day, carboplatin 75 mg/kg/day, and their combination were 358%, 97%, and &gt;489%, respectively. Thus, combination of NK012 with carboplatin showed remarkably synergistic effects, and it didn't accompany toxicity exacerbation. Similarly, combination of NK012 with cisplatin exhibited remarkable synergistic effects in Li-07-JCK hepatocellular cancer. The TGDs by NK012 3.8 mg/kg, one shot, cisplatin 10 mg/kg, one shot, and their combination were 58%, 136%, and 455%, respectively.
These results indicate that NK012 and platinum compounds combination regimen would be effective for treatment of patients with solid tumors including breast, non-small cell lung, colorectal and hepatocellular carcinomas.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5513.
Abstract 5518: Bladder tumor-targeting peptide-mediated anti-tumor activity of a pro-apoptotic peptide
Authors: Jung Hyun-Kyung , Kwak Mi-Kyung , Lee Eun-Ju , Son Ji Woong , Park Rang-Woon , Kim In-San , Lee Byung-Heon 
Published: February 2011

Abstract
Bladder carcinoma is the most common urothelial malignancy in developed countries and the rate of recurrence is pretty high. Using phage display, we have previously identified a bladder tumor-targeting peptide with the amino acid sequence of CSNRDARRC (namely, Bld-1). Bld-1 was shown to target bladder tumor in carcinogen-induced tumor-bearing rats. A synthetic pro-apoptotic peptide KLAKLAKKLAKLAK, called (KLAKLAK)2, has been demonstrated to induce apoptotic cell death of tumor cells. In this study, we used Bld-1 as a homing domain to help the pro-apoptotic peptide selectively bind to bladder tumor cells and allow its internalization. Bld-1 preferentially bound to HT-1376 bladder tumor cells, while little binding to A549 lung tumor cells was observed. Confocal microscopic analysis demonstrated that Bld-1 was efficiently internalized into HT-1376 cells. Bld-1-conjugated D-enantiomer D(KLAKLAK)2 was synthesized and incubated with HT-1376 cells for 24, 48, and 72 h. Bld-1-D(KLAKLAK)2 exerted its cytotoxic activity on HT-1376 cells in a dose- and time-dependent manner. In addition, D(KLAKLAK)2 alone also showed cytotoxic activity after 48 h treatment but this was in a lesser extent as compared to Bld-1-D(KLAKLAK)2. The cell death induced by Bld-1-D(KLAKLAK)2 was due to apoptosis as shown by annexin V and propidium iodide staining. Bld-1-D(KLAKLAK)2 more efficiently inhibited tumor growth in HT-1376 bladder tumor-bearing mice than D(KLAKLAK)2. These results suggest that Bld-1-D(KLAKLAK)2 might be a useful therapeutic agent for targeted therapy of bladder tumor.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5518.
Abstract 552: Microglia-stimulation of glioma cell invasion is dependent on EGFR and CSF1R signaling
Authors: Coniglio Salvatore J. , Kedrin Dmitriy , Dobrenis Kostantin , Symons Marc , Segall Jeffrey E. 
Published: February 2011

Abstract
The ability of glioma tumor cells to invade surrounding brain parenchyma is what makes complete removal of tumor nearly impossible resulting in a median survival of only 12 months for patients afflicted by this disease. Targeting the motile properties of this tumor therefore, is an attractive strategy for therapeutic intervention. Many recent studies have demonstrated a key role for the tumor microenvironment in promoting tumor cell invasion and metastasis. Our laboratory and collaborators have demonstrated that tumor associated macrophages (TAMs) play a critical role in breast carcinoma invasion and metastasis. Here I propose a similar mechanism may occur between glioma cells and microglia within the brain. In this study we demonstrate that murine microglia are able to enhance GL261 glioma cell invasion into Matrigel in co-culture assays by up to 3 fold. Furthermore, we have found using pharmacological and/or antibody mediated inhibition of EGFR or CSF1R signaling abrogates this effect. GL261 cells constituitively secrete CSF-1, the levels of which are unaffected by EGF stimulation, EGFR inhibition or coculture with microglia. Disruption of EGF binding to EGFR using an EGF-specific function-blocking antibody also inhibits microglia-stimulated invasion, showing that EGF is responsible for mediating invasion. This data strongly suggests a paracrine loop exists between microglia and glioma similar to that seen in the breast cancer model. It is our hope that the results of this study may offer novel therapeutic approaches against glioblastoma by targeting functions of the tumor associated microglia as well as the glioma cells.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 552.
Abstract 5520: Thermally targeted proapoptotic peptide for cancer therapy
Authors: Moktan Shama , Raucher Drazen 
Published: February 2011

Abstract
Peptide based therapy is a promising field of anti-cancer therapy. The biologically active therapeutic peptides, derived from an endogenous protein or designed synthetically, can inhibit or activate important cellular pathways to promote cancer cell death. The major challenges of peptide based therapy are stability and tumor-specificity of the therapeutic peptides. One of the methods to increase stability and target the therapeutic peptide to the tumor site is to utilize stimulus-responsive macromolecular carriers.
We have designed a therapeutic peptide, SynB1-ELP-(KLAKLAK)2 based on the thermally responsive biopolymer, elastin-like polypeptide (ELP) and the mitochondrial membrane disrupting proapoptotic peptide, (KLAKLAK)2. Additionally, a cell penetrating peptide, SynB1 has been included in the N-terminus of this polypeptide in order to increase the polypeptide's membrane permeability. SynB1 is an 18-mer peptide derived from protegrins, which are antimicrobial peptides found in porcine leukocytes. The utilities of ELP are two-fold - 1) it is a macromolecule and therefore can provide stability to the overall polypeptide system, and 2) it is thermally responsive and therefore can be targeted to the tumor site by external application of local hyperthermia. The unique phase transition property of ELP allows it to remain soluble at physiological temperature but forces it to aggregate in response to mild hyperthermia (40 - 42 °C). Therefore, SynB1-ELP-(KLAKLAK)2 can be targeted to solid tumors by the application of local hyperthermia without being degraded before reaching the tumor.
The anti-proliferative effect of SynB1-ELP-(KLAKLAK)2 was examined in MCF-7 breast cancer cells. While the polypeptide was not cytotoxic to cells at 37 °C, it killed about 40% of cells at 42 °C after 6 days with two one hour treatments. Similar treatment schedule resulted in approximately 4-fold increase in apoptosis induction at 42 °C compared to 37 °C. Confocal microscopy showed that SynB1-ELP-(KLAKLAK)2 localizes in the mitochondria which supports the mitochondrial membrane disrupting feature of the (KLAKLAK)2 peptide. In conclusion, SynB1-ELP-(KLAKLAK)2 localizes in the mitochondria and inhibits the proliferation of MCF-7 cells in combination with hyperthermia by inducing apoptosis. Further studies include determining the proapoptotic effects of SynB1-ELP-(KLAKLAK)2 in vivo.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5520.