bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 824: Targeting epidermal growth factor receptors and urokinase-type plasminogen activator receptors for sarcoma therapy
Authors: Graef Ashley J. , Kim Jong-Hyuk , Vallera Daniel A. , Modiano Jaime F. 
Published: August 2018

Abstract
eBAT is a bispecific angiotoxin consisting of truncated, deimmunized Pseudomonas exotoxin fused to epidermal growth factor (EGF) and the amino terminal fragment of urokinase. This drug was designed to use epidermal growth factor receptors (EGFR) and urokinase-type plasminogen receptors (uPAR) as baits to deliver the lethal toxin into the cells, rather than as a means to interrupt signaling by the receptors. We previously showed that eBAT is remarkably safe, and that it reduced tumor burden in mice with established human rhabdomyosarcoma xenografts. It also was safe and showed a strong efficacy signal when used in the adjuvant setting in a clinical trial for dogs with splenic hemangiosarcoma. Nevertheless, the therapeutic mechanism of action of eBAT remains to be elucidated. Our goal is to identify patterns of uPAR expression in sarcomas that promote sarcoma growth and that mediate sensitivity for therapeutic activity by eBAT. We generated uPAR-knockout (KO) cells from MC1A-C1 mouse fibrosarcoma cell line using the CRISPR-Cas system to target exon 2 of the uPAR gene. The resulting cell lines D10 and H6 are clonal uPAR knockouts, whereas K02 and K06 represent mixtures where approximately 80% and 30% of the cells retain uPAR expression. uPAR expression in K02 and K06 has remained stable over numerous passages, indicating that loss of uPAR does not create selective pressure to enhance or inhibit cell growth in vitro. Furthermore, the growth kinetics of D10 and H6 are comparable to parental MC1A-C1 cells. eBAT treatment significantly inhibited proliferation of parental cells, but not of uPAR KO cells. In contrast, all of the cells were resistant to treatment with EGF- and uPA-monospecific toxins. The engraftment efficiency and tumor growth of uPAR KO cells in syngeneic hosts was substantially diminished when compared to K02 and K06 uPAR+ cells, suggesting that, even while uPAR-KO cells might be more resistant to eBAT therapy, uPAR expression appears to be necessary to create the tumor niche and/or to promote tumor growth in vivo. In our ongoing work, we are investigating how uPAR expression in stromal cells and in cells of the innate immune system influence sarcoma growth and the response to eBAT.
Citation Format: Ashley J. Graef, Jong-Hyuk Kim, Daniel A. Vallera, Jaime F. Modiano. Targeting epidermal growth factor receptors and urokinase-type plasminogen activator receptors for sarcoma therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 824.
Abstract 829: SAS1B protein: Determining whether ASTL or SAS1B has a role in tumor progression
Authors: Khokhar Zunair S. , Pires Eusebio S. , Kim Alexander J. , D'Souza Ryan 
Published: August 2018

Abstract
The matrix metalloproteinase Sperm Acrosomal SLLP1 Binding (SAS1B) is an ideal molecular and cellular candidate for both female contraception and for cancer therapies. SAS1B translation is restricted among adult tissues to the ovary and oocytes, and SAS1B first appears in oocytes of follicles at the primary-secondary transition and also in tumors of varied types and stages. Research conducted using SAS1B antibodies suggested that the protein is localized at the cell membrane of cancer cells and can get internalized in a classical endocytosis assay and can be killed using an immunotoxin linked onto antibodies. The current study aims at deciphering whether the ASTL gene or the SAS1B protein is important for tumor progression, invasion and metastasis through cell biology and molecular biology tools and ensuring tumor cell-killing properties via an ADC-mediated killing using biochemistry and molecular biology tools. PCR, immunohistochemistry, Immunofluorescence, and Western blots revealed that a noncancerous breast cell line, MCF10A5E, is SAS1B null and can be used to study cancer-like properties once stably transfected with the SAS1B construct. Previous work in the lab indicated that an SAS1B-ADC strategy is a viable immunotherapy that could benefit patients who have failed to respond to chemotherapy, if indeed SAS1B plays a role in cancer development and progression. The outcomes of an SAS1B-ADC in a breast cancer model will be presented.
Citation Format: Zunair S. Khokhar, Eusebio S. Pires, Alexander J. Kim, Ryan D'Souza. SAS1B protein: Determining whether ASTL or SAS1B has a role in tumor progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 829.
Abstract 852: Specificity of PSMA-617 radiotherapy for prostate cancer
Authors: Reddy Joseph A. , Nelson Melissa , Xu LeCun , Westrick Elaine , Vetzel Marilynn , Wheeler Leroy , Felten Albert , Santhapuram Hari , Vlahov Iontcho , Leamon Christopher P. 
Published: August 2018

Abstract
Prostate-specific membrane antigen (PSMA) is a biomarker that is overexpressed on prostate cancer as well as on the neovasculature of many non-prostate solid tumors. Lately, several small molecular PSMA binding ligands chelated to various radionuclides have been studied for imaging and treatment of prostate cancer lesions. One such clinically tested agent is PSMA-617, a DOTA derivative of a PSMA-specific targeting ligand. In our hands, radiochemical yields of PSMA-617 labeled with either 177Lu (β emitter) or 225Ac (α emitter) were greater than 97%. Binding experiments with 177Lu and 225Ac chelates of PSMA-617 and D-Glu PSMA-617 (a predicted non-PSMA binding, D-glutamic acid isomer of PSMA-617) were performed using the PSMA-positive, 22RV1 cell line. PSMA-617, was found to bind in a concentration dependent manner with very high affinity, whereas the cellular association of D-Glu PSMA-617 was found to be &gt; 60-fold lower. The in vivo biodistribution of these two agents were investigated in BALB/c nu/nu mice bearing subcutaneous 22RV1 xenografts. Organ distribution revealed specific PSMA-617 uptake in the 22RV1 tumors and the PSMA+ kidneys, while the uptake of D-Glu PSMA-617 remained near background in both of these tissues. Different from tumor, PSMA-617 exhibited a rapid clearance from the kidneys to yield a high tumor-to-background contrast (tumor:blood = 300; tumor:muscle = 290; and tumor:kidney = 15) at 24 h post dose. Complementary antitumor activity studies were performed with 225Ac-labeled PSMA-617. Here, significant tumor growth inhibition was observed without detectable hematologic or renal toxicity to the mice. These data provide additional support for a phase 3 prospective trial currently being planned to evaluate 177Lu-PSMA-617 in patients with metastatic PSMA+ prostate cancers, along with the continued clinical investigation of 225Ac-PSMA-617 in a similar patient population.
Citation Format: Joseph A. Reddy, Melissa Nelson, LeCun Xu, Elaine Westrick, Marilynn Vetzel, Leroy Wheeler, Albert Felten, Hari Santhapuram, Iontcho Vlahov, Christopher P. Leamon. Specificity of PSMA-617 radiotherapy for prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 852.
Abstract 861: Angiopoietin-2 blocking antibodies improve tumor growth inhibition and survival in mice treated with low doses of radiation
Authors: Kallio Pauliina , Jokinen Elina , Das Suvendu , Högström Jenny , Heino Sarika , Lähde Marianne , Alitalo Kari 
Published: August 2018

Abstract
Almost 50 % of all cancer patients receive radiation therapy during their treatment, yet several tumor types show radiation therapy resistance that compromises treatment efficacy. Many radiation sensitizers have been tested for improvement of the outcome of radiation therapy. For example, inhibitors of VEGF and VEGF receptors were tested in attempt to normalize the tumor vasculature and to improve the outcome of radiation therapy. Here, we have analysed if antibodies blocking the angiogenic factor angiopoietin-2 could improve radiation therapy of mouse tumor isografts and human tumor xenografts in mice. To test if the anti-tumor effect of small doses of radiation can be increased by Ang2 blocking antibodies, we analysed the radiation sensitivity of subcutaneous tumor isografts in C57bl/6JRj mice, and human xenografts in NOD scid gamma mice. In several models, the combination treatment with anti-Ang2 plus radiation was superior to the monotherapies in decreasing tumor growth and increasing the lifespan of the tumor-bearing mice both in immunocompetent and immunocompromised mice. Based on our results, the anti-Ang2 blocking antibody treatment could be tested in the clinics to improve the effect of radiation therapy, especially in those cases in which an optimal radiation dose cannot be given because of the radiation sensitivity of the surrounding tissues.
Citation Format: Pauliina Kallio, Elina Jokinen, Suvendu Das, Jenny Högström, Sarika Heino, Marianne Lähde, Kari Alitalo. Angiopoietin-2 blocking antibodies improve tumor growth inhibition and survival in mice treated with low doses of radiation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 861.
Abstract 862: Effects of fixation and antigen retrieval on the RNA integrity number
Authors: Otali Dennis , Mullen Maitlyn , Qinghua He , Oelschlager Denise , Grizzle William E. 
Published: August 2018

Abstract
The effect of fixation and antigen retrieval (AR) on RNA as measured by the RNA integrity number (RIN) has not been adequately explored. RIN is an index (pseudo-measure) used to measure stability of mRNA; however, RIN is an actual measure of the stability of ribosomal RNA and not an actual measure of mRNA. RIN values range from 1 for highly degraded to 10 for intact RNA.
Most surgical specimens are fixed in 10% neutral buffered formalin (3.7% formaldehyde) because of its ability to preserve morphological detail and prevent degradation. However, formaldehyde fixation causes changes in proteins through formation of crosslinks, which masks the immunorecognition of some antigens. In immunohistochemical evaluations, antigen retrieval (AR) approaches e.g., boiling of tissue sections under pressure are used to reverse these effects of fixation.
It is believed that fresh or snap frozen specimens serve as most the suitable source of intact mRNA, DNA and other macromolecules for molecular analysis. However, more recent studies using rt-Q PCR indicate that RNA and DNA can be measured in formalin fixed paraffin embedded (FFPE) tissue samples which usually have RIN values of 3 or less. The question arises as to what is the usefulness of RIN in FFPE tissues.
This study used three cancer cell lines DU145 (prostate), MDA231 (breast) and S2VP10 (pancreas) to examine the relationship between 10% NBF fixation, RIN and AR. The first objective was to analyze whether the duration of fixation of cells in 10% NBF correlated with decreases in RIN. Three fixation times i.e., 5 minutes, 20 minutes and 120 minutes were studied. The second objective was to investigate the effects of two AR approaches on RIN. Two AR conditions at pH 6 and pH 9 were used on the same cell lines and fixation times.
For each cell line and treatment i.e., no fixation, or fixation at different times (with and without AR), RNA was extracted and RIN analyzed using the Agilent 2100 Bioanalyzer. RIN values obtained from the Agilent 2100 Bioanalyzer were statistically analyzed. T-tests show that the large decreases to about 3 in RIN values after fixation (5, 20 and 120 minutes) are statistically significant for all cell lines. All p-values are less than 0.0001. This study confirms that fixation of cells in 10% NBF lowers RIN to about 3 the value observed for fixed tissues. RIN did not vary significantly with time of fixation in 10% NBF and RIN did not change from the fixed values after either condition of AR. Thus RIN is not a useful measure of RNA stability in 10% NBF fixed tissue.
Citation Format: Dennis Otali, Maitlyn Mullen, He Qinghua, Denise Oelschlager, William E. Grizzle. Effects of fixation and antigen retrieval on the RNA integrity number [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 862.
Abstract 877: Sigma 2 ligands kill pancreatic cancer cells via disruption of lysosomal cholesterol transport
Authors: Jin Linda X. , Vangveravong Suwanna , Sankpal Narendra V. , Cullinan Darren R. , Goedegebuure Peter , Spitzer Dirk , Hawkins William G. 
Published: August 2018

Abstract
Introduction: The sigma 2 receptor is overexpressed in pancreatic cancer and has been an attractive target for development of cancer selective therapies. However, the mechanism of sigma 2 mediated cancer cell death remains elusive. The sigma 2 receptor has been recently identified as TMEM97, a lysosomal/endoplasmic reticulum protein involved in regulation of cholesterol export in conjunction with NPC1.
Methods: Uptake of SW43 in pancreatic cancer cells was visualized using a NBD fluorophore conjugated ligand. Intracellular cholesterol staining was visualized using filipin staining. Protein expression was quantified by both Western blot and RT qPCR. Cell death was assayed in triplicate using Titer-Glo viability assay.
Results: We hypothesized that sigma 2 ligands may kill pancreatic cancer cells through disruption of lysosomal cholesterol export. Using ASPC1 pancreatic cancer cells, we first demonstrate that sigma 2 ligand SW43 is rapidly taken up and targets to endo-lysosomal compartments, and that treatment with SW43 causes a rapid and dose dependent sequestration of cholesterol in lysosomal compartments as visualized by filipin cholesterol staining. This correlated to an increase in TMEM97 and corresponding decrease in NPC1 expression following SW43 treatment. We next demonstrate that this cholesterol sequestration phenotype is required for cell death, and might be related to the free amine side chain of SW43, as its N-phthalimide derivative fails to induce both lysosomal cholesterol sequestration and cell death despite high doses of ligand used. Lysosomal cholesterol sequestration from sigma 2 ligand treatment sensitizes cells to further targeting of cholesterol metabolism. Using low doses of simvastatin and SW43, we demonstrate that targeting both cholesterol export and cholesterol synthesis displays significant synergism in killing ASPC1 pancreatic cancer cells.
Conclusions: These findings provide further insight into how the sigma 2 receptor/ligand interaction mediates cancer cell death and provides the basis for rationally combined targeted therapy regimens to be tested in vivo.
This work was supported by NIH R01CA163764 and NIH T32CA009621-26A1 and the SUS Foundation Resident Research Award.
Citation Format: Linda X. Jin, Suwanna Vangveravong, Narendra V. Sankpal, Darren R. Cullinan, Peter Goedegebuure, Dirk Spitzer, William G. Hawkins. Sigma 2 ligands kill pancreatic cancer cells via disruption of lysosomal cholesterol transport [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 877.
Abstract 915: AKT inhibition sensitizes TNBC cells with acquired drug resistance to UM-164
Authors: Merrill Nathan M. , Lachacz Eric J. , Gilani Rabia A. , Bao LiWei , Wu Zhi Fen , Merajver Sofia D. , Soellner Matthew B. 
Published: August 2018

Abstract
One of the major clinical challenges in breast cancer treatment is the rapid development of resistance to targeted therapies. In light of this, there is significant interest in identifying markers of drug resistance. In particular, markers of drug resistance are sought that can be targeted to overcome acquired drug resistance. To this end, we have used reverse phase protein array (RPPA) analysis to identify differences in protein expression levels between cell lines that are sensitive or have acquired resistance to UM-164, a mechanistically novel, dual c-Src/p38 kinase inhibitor with potent anti-TNBC activity. UM-164 resistant cells acquire increased resistance also to many diverse chemotherapeutics and targeted agents relative to their drug-sensitive counterparts. We analyzed RPPA data for both UM0164-sensitive and UM-164-resistant cells and identified an increase in phospho-AKT, phospho-ERK, and phospho-FAK levels in the resistant cells. Using inhibitors of each upregulated pathway, we found that combination of UM-164 with MK2206, an inhibitor of AKT, resulted in an increased sensitization of cells to treatment with UM-164. Furthermore, formal Chou-Talalay synergy analyses demonstrate that AKT inhibitors + UM-164 are effectively synergistic in reducing the proliferation of drug-resistant cells. We further evaluated this combination treatment in mice, combining UM-164 and MK2206 in a xenograft of UM-164-resistant cells, showing significant anti-tumor efficacy. Finally, we show that the combination treatment resulted in increased cell death in ex vivo cultures of patient derived TNBC materials. Together, we have demonstrated that hyperactivation of AKT leads to UM-164 resistance and AKT inhibitors can be used to re-sensitize TNBC cells to UM-164. These results demonstrate that RPPA analyses are a valuable tool for the systematic and unbiased identification of truly actionable markers of therapeutic resistance.
Citation Format: Nathan M. Merrill, Eric J. Lachacz, Rabia A. Gilani, LiWei Bao, Zhi Fen Wu, Sofia D. Merajver, Matthew B. Soellner. AKT inhibition sensitizes TNBC cells with acquired drug resistance to UM-164 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 915.
Abstract 916: A screen to target EGFR (C797S) in osimertinib-resistant lung cancer identifies brigatinib-based combinations that synergistically inhibit cell growth
Authors: Martin Matthew J. , Floc'h Nicolas , Michaloglou Chrysiis , Finlay M Raymond , Ward Richard A. , Smith Paul D. , Cross Darren A. 
Published: August 2018

Abstract
Up to 20% of non small-cell lung cancers are driven by activating mutations in EGFR (EGFRm). While EGFR tyrosine kinase inhibitors (TKIs) have proven highly effective in the clinic (e.g. gefitinib in first line EGFRm and osimertinib in T790M post-TKI resistance), acquired resistance mechanisms present an obstacle to curing EGFR-driven disease. To-date, approximately 20% of osimertinib-resistant cases in the 2nd line setting are EGFRm/T790M/C797S, and there is currently no approved therapy for these “triple-mutant” (TM) patients. We have obsrerved that the ALK inhibitor brigatinib shows moderate, equipotent activity against both EGFR-TM and the single activating mutant. We used this information to screen a panel of compounds to identify brigatinib-based combinations that could effectively target EGFR-TM-expressing cells. Among the most potent combination partners were selumetinib and other inhibitors of the MAP kinase pathway. These agents were able to synergistically impair growth of TM cells when combined with brigatinib in vitro, and accordingly show prolonged suppression of downstream pathway biomarkers in combination- vs. brigatinib monotherapy-treated cells. Further, we have shown effective combinations can dramatically enhance apoptotic cell death compared to brigatinib treatment alone. Critically, neither monotherapy treatment with effective combination partners, nor combinations based on osimertinib showed growth inhibition in the EGFR triple-mutant setting. Taken together, these data suggest novel therapeutic strategies for patients in this critical area of unmet need that warrant further preclinical testing.
Citation Format: Matthew J. Martin, Nicolas Floc'h, Chrysiis Michaloglou, M Raymond Finlay, Richard A. Ward, Paul D. Smith, Darren A. Cross. A screen to target EGFR (C797S) in osimertinib-resistant lung cancer identifies brigatinib-based combinations that synergistically inhibit cell growth [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 916.
Abstract 96: Novel inverse functioning molecules, GSN as metastatic promoter and OAS2 as metastatic repressor in colorectal adenocarcinoma
Authors: Kim Jin Cheon , Ha Je Jin , Tak Ka Hee , Roh Seon Ae , Kim Chan Wook , Yoon Yong Sik , Lee Jong Lyul , Kim Seon-Kyu , Kwon Ee Hong , Cho Dong-Hyung , Kim Seon-Young , Kim Yong Sung 
Published: August 2018

Abstract
Background LVI and PNI, recognized as major histological traces of metastatic CRC, have been recognized as a category I prognostic factor representing aggressive CRC. The primary aim of our study was to find out molecules associated with LVI and PNI and to examine their biological behavior with their prognostic significance.
Methods LVI- and PNI-associated molecules were identified and verified by utilizing (1) RNA-seq and selecting 117 genes by differential expression values in primary cancer samples of 130 patients with and without systemic recurrence (n = 72 and 58) (2) association analysis to extract six genes associated with LVI or PNI (3) assessment of biological property measuring proliferation, invasion/migration, epithelial-mesenchymal transition, and autophagy flux (4) verification of disease-free survival using public data sets of gene
Results GSN-overexpressing LoVo cells showed more than 2-fold greater invasion compared with control cells 5 (p &lt; 0.001 - 0.005), whereas OAS2-overexpressing RKO cells showed a significantly reduced invasion compared with control cells (p &lt; 0.001 - 0.005). GSN repressed expressions of E-cadherin, β-catenin, claudin-1, and snail, whereas it enhanced N-cadherin and ZEB1 expressions. Reverse findings were noticeably identified in OAS2-overexpression. Interestingly, expressions of multiple autophagy-related proteins including ATG5-12, ATG6/BECN1, ATG7, and ATG101 were decreased in GSN-overexpressing Lovo cells. In addition, accumulation of p62, a well-known substrate of autophagic degradation, suggested that an autophagic flux was also reduced in the GSN-overexpressing Lovo cells. In contrast to GSN overexpression, reverse findings of autophagy flux was identified in OAS2-overexpressing RKO cells. The 5-year RFS rates were significantly greater in the GSN underexpression group than in the overexpression group (73.6% and 64.7%, p = 0.038), whereas patients with OAS2 overexpression presented a greater RFS rate than those with underexpression (73.4% and 63.7%, p = 0.01).
Conclusion GSN and OAS2, as a recurrence promoter and suppressor, respectively, suggest their potential value as recurrence predictor or therapeutic targets in future clinical validation studies.
Citation Format: Jin Cheon Kim, Je Jin Ha, Ka Hee Tak, Seon Ae Roh, Chan Wook Kim, Yong Sik Yoon, Jong Lyul Lee, Seon-Kyu Kim, Ee Hong Kwon, Dong-Hyung Cho, Seon-Young Kim, Yong Sung Kim. Novel inverse functioning molecules, GSN as metastatic promoter and OAS2 as metastatic repressor in colorectal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 96.