bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract LB-006: Intracellular protein therapy with SOCS3 utilizing aMTD-enabled macromolecule intracellular transduction technology (MITT)
Authors: Choi Youngsil , Shin Seulmee , Lee Kuysook , Jo Daewoong 
Published: August 2018

Abstract
Background Macromolecule intracellular transduction technology (MITT), a platform for the discovery/development of new medicinal drugs, is enabled with a new generation of hydrophobic cell-penetrating peptides (CPPs), termed advanced macromolecule transduction domains (aMTDs). Originally derived from the hydrophobic signal peptides of secreted proteins, earlier generations of MTDs were hampered by poor solubility and less efficient cell-permeability. The MITT platform addresses these limitations, first, by the design of synthetic aMTD sequences that incorporate 6 critical factors found to enhance the intracellular delivery of proteins into cells and tissues and second, by the use of humanized solubilization domains to enhance the solubility of aMTD-containing recombinant proteins. Object The goal of this study is to develop and evaluate an aMTD-SOCS3 (suppressor of cytokine signaling 3) protein (iCP-SOCS3) as cell-permeable biotherapeutic to selectively target tumors dependent on JAK/STAT signaling and to compare the cell-/tissue-penetration efficiencies of iCP-SOCS3 with a previous generation of cell-permeable SOCS3 (MTM-SOCS3: CP-SOCS3) enabled by a hydrophobic CPP, termed membrane translocating motif (MTM) derived from the signal peptide of fibroblast growth factor 4 (FGF4). Results Solubility (up to 10 mg/ml) and final yields of E. coli produced iCP-SOCS3 (60 mg/L) were sufficient for clinical development; whereas, CP-SOCS3 suffered from low solubility and yields (0.4 mg/ml and 0.2 mg/L, respectively). Cellular uptake of fluorescent iCP-SOCS3 was enhanced by 25-fold as compared to CP-SOCS3 as assessed by flow cytometry, and delivery and persistence of the protein in a variety of tissues (heart, lung, spleen, kidney, liver and brain) were similarly increased. In addition, aMTD promoted the cell-to-cell transfer of SOCS3 protein, and the transferred iCP-SOCS3 was biochemically active as assessed by inhibition of IFN-γ-induced phosphorylation of STAT1 and STAT3 with concomitant suppression of cancer-associated phenotypes in cultured cells and in mice bearing human tumor xenografts (90% inhibition of proliferation and 84% inhibition of migration in PANC-1 cells, and 85% suppression of pancreatic tumor growth, respectively). Conclusion These results provide further validation of MITT as a platform to develop protein-based biotherapeutics, and suggest the use of iCP-SOCS3 as a novel protein therapy against tumors dependent on JAK/STAT signaling.
Citation Format: Youngsil Choi, Seulmee Shin, Kuysook Lee, Daewoong Jo. Intracellular protein therapy with SOCS3 utilizing aMTD-enabled macromolecule intracellular transduction technology (MITT) [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 LB-006.
Abstract LB-007: Intracellular delivery of SOCS3 suppresses pancreatic cancer progression
Authors: Lee Kuysook , Shin Seulmee , Hwang Yunseo , Choi Youngsil , Jo Daewoong 
Published: August 2018

Abstract
Background JAK/STAT signaling, regulated by SOCS family proteins, plays a role in the progression of the inflammation, growth and metastasis of pancreatic adenocarcinoma. SOCS3 suppresses JAK kinase activity and degrades the activated cytokine receptor complex, and its loss contributes to pancreatic cancer development. We have developed advanced macromolecule transduction domains (aMTDs) to deliver recombinant proteins into to cells and tissues. In principle, aMTD enabled SOCS3 could provide an effective treatment for pancreatic tumors that depends on JAK/STAT signaling for growth or survival. Objective The present study utilized aMTD sequences to develop improved cell-permeable (iCP-) SOCS3 as a protein-based biotherapeutic against pancreatic cancer. Method Improved cell-permeable SOCS3 proteins, with and without a 12-amino acid aMTD sequence, were expressed at high yields in E. coli. The proteins also contained a solubilization domain (SD) to enhance the solubility of the recombinant proteins in physiological buffers. Results The iCP-SOCS3 exhibited high solubility, cell-/tissue-permeability and biological activity in pancreatic cancer cell lines (BxPC3, HPAC and PANC-1) with decreased endogenous SOCS3 levels, hyperactived JAK/STAT signaling and hypermethylated socs3 promoter region. The iCP-SOCS3 suppressed cancer-associated phenotypes (i.e., up to 90% inhibition of cell viability, 52% induction of apoptosis, 14.8% inhibition of cell cycle progression, 54% inhibition of migration and 61.5% inhibition of invasion) in pancreatic cancer cells dependent on elevated JAK/STAT signaling, whereas, these effects were not observed in non-cancer cells (H6c7, pancreatic duct epithelial cell) or with proteins lacking the aMTD sequence. In contrast, gemcitabine, a conventional pancreatic cancer therapeutic, showed undesirable cytotoxic effects including decreased cell viability and induced apoptosis in non-cancer cells. Furthermore, iCP-SOCS3 (30 mg/kg) induced tumor regression (-168% at day 18) in PANC-1 CDX model. In BxPC-3 orthotopic CDX model, iCP-SOCS3 suppressed tumor growth by 98% which was significantly higher compared to the suppressed tumor growth induced by gemcitabine (77%). Correlating to the tumor suppression, iCP-SOCS3 significantly decreased the secretion of TNF-α and the expression of p-STAT1, p-STAT3 and VEGF in tumor tissues. Conclusion These results provide further evidence that SOCS3 can function as a tumor suppressor and intracellular delivery of SOCS3 with iCP-SOCS3 may provide a novel protein therapy against pancreatic cancer.
Citation Format: Kuysook Lee, Seulmee Shin, Yunseo Hwang, Youngsil Choi, Daewoong Jo. Intracellular delivery of SOCS3 suppresses pancreatic cancer 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 LB-007.
Abstract LB-009: Intracellular delivery with SOCS3 suppresses angiogenesis in tumors
Authors: Jung Yukdong , Lee Kuysook , Choi Youngsil , Jo Daewoong 
Published: August 2018

Abstract
Introduction: Suppressor of cytokine signaling 3 (SOCS3) is a negative feedback regulator in the Janus kinase/signal transducer and activator of transduction (JAK/STAT) signaling pathway, in which it suppresses inflammation and cancer. The JAK/STAT pathway is activated in some tumors thus enhancing the expression of genes involved in proliferation and angiogenesis, notably, vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), angiopoietin-1 and phospoinositide 3-kinase (PI3K)-protein kinase B (AKT).
Objective: The aim of this study is to develop an improved cell-permeable (iCP)-SOCS3 and determine its applicability as an anti-angiogenic agent to treat cancer.
Methods: The iCP-SOCS3 was expressed in E. coli as a 37 kDa recombinant protein fused to an advanced macromolecule transduction domain (aMTD) and humanized solubilization domain (SD). Uptake of a fluorescently tagged iCP-SOCS3 by cells and tissues was monitored by flow cytometry and confocal microscopy. Biological and biochemical activities of iCP-SOCS3 were determined by in vitro angiogenesis assays and changes in biomarker expression and anti-tumor activities were analyzed in mice bearing human tumor xenografts.
Results: The iCP-SOCS3 improved on an earlier generation of CP-SOCS3 fused to FGF4-derived membrane translocating motif (MTM) with regard to solubility, yield, delivery into cells and tissues and biological activity. The iCP-SOCS3 suppressed proliferation of human umbilical vein endothelial cells (HUVECs) by 60% and migration by 54% via inhibition of AKT/mTOR signaling pathway. Moreover, iCP-SOCS3 inhibited invasion of EC cells in PANC-1 and U87MG conditioned medium by 38% and 95%, respectively. In tube formation assay, iCP-SOCS3 reduced the number of branch points by 79% and tube length by 60% compared to the vehicle. In chick chorioallantoic membrane (CAM) assay, iCP-SOCS3 significantly inhibited angiogenesis by 83% in dose dependent manner compared to the reference drug, retinoic acid. The anti-angiogenic activity of iCP-SOCS3 was also shown in matrigel plug assay. The new vessel formation induced by U87-MG glioblastoma cells was significantly reduced by the treatment of iCP-SOCS3. In PANC-1, U87-MG and HepG2 cell-derived xenograft models, iCP-SOCS3 suppressed expression of pro-angiogenic factors including VEGF, ERK and angiopoietin 1.
Conclusion: These results validate iCP-SOCS3 targets in promoting angiogenesis and suggest ways to reduce angiogenesis in various types of tumors.
Citation Format: Yukdong Jung, Kuysook Lee, Youngsil Choi, Daewoong Jo. Intracellular delivery with SOCS3 suppresses angiogenesis in tumors [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 LB-009.
Abstract LB-036: Targeting the WNT-pathway in 3D-cultures of pancreatic cancer biopsies
Authors: Herpers Bram , Basten Sander , Yan Kuan , Dijkmans Thomas , Schueler Julia , Giesemann Torsten , Price Leo S. 
Published: August 2018

Abstract
Creating patient-derived xenograft (PDX) models from human cancers allows bio-banking, propagation, characterization and drug testing on a large panel of tumor variants. We developed 3D in vitro culture methods for PDX model-derived cell isolates. Each individual model was subjected to a wide panel of culture medium formulations, amongst which some media contain WNT-pathway stimulating components. We identified an absolute dependency of a pancreatic in vitro PDX tumor to the WNT-signaling potentiating factor R-Spondin, while other pancreas tumors showed little or no dependency. To further study the dependency of pancreatic PDX tumors on the functionality and signaling of the WNT-pathway, we exposed in vitro PDX cultures to increasing doses of anti-FZD7, anti-FZD8 and to the Porcupine inhibitor LGK974, which blocks WNT palmitoylation and therefore secretion. We also applied the same approach to establish cultures from patient-derived pancreatic cancer biopsies. Our findings are that the WNT-pathway is functional and contributing to pancreas cancer outgrowth and morphological differentiation to different extents in different patient-models. Our in vitro culture system in combination with 3D phenotypic analysis allows in-detail study of the WNT-pathway, as well as by supplementation or omission of secreted factors, addition of small molecule inhibitors and by exposure to antibodies. These in vitro models can be used to test novel compounds and antibodies targeting the WNT pathway and underscore the WNT pathway as a potential target in pancreatic cancer.
Citation Format: Bram Herpers, Sander Basten, Kuan Yan, Thomas Dijkmans, Julia Schueler, Torsten Giesemann, Leo S. Price. Targeting the WNT-pathway in 3D-cultures of pancreatic cancer biopsies [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 LB-036.
Abstract LB-051: An Astragalus-based Chinese herbal medicine extraction inhibits cancer stem cell growth and sensitizes of drug-resistant human non-small cell lung cancer cells for targeted therapy
Authors: Huang Tse-Hung , Hsu Hsuan-Min , Huang Chi-Ying F. 
Published: August 2018

Abstract
Chinese herbal medicine has been used extensively as a complementary intervention for many diseases including cancer, based on its relatively lower cytotoxicity and side-effects. The major criticism of its clinical use remains the scientific based evidence. Here, we described a precision-based translational approach to identify and validate an Astragalus-based herbal medicine (YQ1) as an antagonist of non-small cell lung cancer. Using various big data analysis approaches, we found that the L1000 profiles of YQ1was associated with anti-cancer stem cell signatures. It suggested that treatment of YQ1 might reduce cancer stem cell population. Experimentally, YQ1 treatment resulted in the significant reduction in lung cancer stem cell properties, including side population, sphere formation, stemness marker expression, and drug resistance. YQ1 may have anti-cancer stem cell ability via suppressing Wnt/β-catenin signaling to inhibit the self-renewal or/and proliferation of lung cancer stem cells. Notably, the combination of YQ1 and gefitinib significantly suppresses tumorigenesis in vivo, reflected by the down-regulation of EGFR signaling and β-catenin. Collectively, we provided preclinical evidence for using YQ1 as a cancer stem cell inhibitor to potentiate anti-tumor effects when combined with tyrosine-kinase inhibitors to significantly improve therapeutic efficacy in NSCLC patients.
Citation Format: Tse-Hung Huang, Hsuan-Min Hsu, Chi-Ying F. Huang. An Astragalus-based Chinese herbal medicine extraction inhibits cancer stem cell growth and sensitizes of drug-resistant human non-small cell lung cancer cells for targeted 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 LB-051.
Abstract LB-055: Discoidin domain receptor 1 (DDR1) ablation promotes tissue fibrosis and hypoxia to induce aggressive, basal-like breast cancer
Authors: Drain Allison P. , Takai Ken , Lawson Devon A. , Littlepage Laurie E. , Karpuj Marcela , Kessenbrock Kai , Le Annie , Inoue Kenichi , Weaver Valerie M. , Werb Zena 
Published: August 2018

Abstract
Breast cancer studies implicate the discoidin domain receptor 1 (DDR1) in both pro- and anti-tumor roles. DDR1 is often overexpressed in breast tumors and its activation by collagen promotes tumor cell invasion and potentiates metastatic colonization by cell lines. However, DDR1 transfection into low DDR1 expressing tumor cells hinders cell migration. Moreover, low expression of DDR1 correlates with higher grade human tumors and associates with worse prognosis. To better understand the role of DDR1 in breast tumor progression and metastasis, we established a DDR1 knockout and crossed it into the MMTV-PyMT mouse. DDR1 loss compromised luminal cell adhesion and expanded the basal/myoepithelial population in adult mice. PyMT DDR1-/- tumors grew faster and had increased lung metastasis both in size and number of metastatic colonies. The DDR1-/- tumors broadly exhibited a more basal-like phenotype with enhanced fibrosis and were more hypoxic. Furthermore, these tumors contained more CD90+ CD24+ tumor cells and more K8+ K14+ cells surrounding necrotic regions. Taken together these data suggest DDR1 loss confers a growth advantage favoring basal cell expansion via impaired luminal adhesion and enhanced basal differentiation leading to more aggressive, basal-like disease.
Citation Format: Allison P. Drain, Ken Takai, Devon A. Lawson, Laurie E. Littlepage, Marcela Karpuj, Kai Kessenbrock, Annie Le, Kenichi Inoue, Valerie M. Weaver, Zena Werb. Discoidin domain receptor 1 (DDR1) ablation promotes tissue fibrosis and hypoxia to induce aggressive, basal-like breast 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 LB-055.
Abstract LB-084: Dasatinib reduces tumor growth in xenograft models derived from human lung tumors with YES1 overexpression
Authors: Garmendia Irati , Bértolo Cristina , Ferrer Irene , Pajares María J , Ajona Daniel , Paz-Ares Luis , Pio Ruben , Montuenga Luis M , Agorreta Jackeline 
Published: August 2018

Abstract
Introduction Lung cancer accounts for 13.2% of all new cancer cases and is the leading cause of cancer-related death worldwide. For a reduction of mortality rates, new personalized therapies are needed. The great challenge is to identify the alterations responsible for the tumor malignant phenotype which should be targeted by specific therapeutic strategies. The aim of our study is to evaluate the use of YES1 kinase amplification and overexpression as a companion biomarker to predict sensitivity to dasatinib using patient-derived xenograft models. Previous results Our previous experiments showed that downregulation of YES1 expression by specific siRNAs impairs cell proliferation and survival in lung cancer cell lines with amplification and high expression of YES1 (YES1High). Inhibition of YES1 in cell lines with normal copy number and low expression (YES1Low) had no effect on cell proliferation. Moreover, YES1 downregulation induced apoptosis and inhibited invasion only in YES1High cells, and YES1 overexpression promoted proliferation in lung cancer cell lines.
Methods Eight patient-derived xenograft mouse models were used: two YES1High adenocarcinomas (ADC), two YES1Low ADC, two YES1High squamous cell carcinomas (SCC) and two YES1Low SCC. Small fragments of xenografted tumors (2-3 mm in diameter) were subcutaneously implanted into the right flank of 7-8-week-old BALB/c-Rag2−/−-IL2γc−/− immunodeficient mice. Mice were treated with dasatinib (60 mg/kg) or citric acid daily by oral gavage. Tumor volume was measured twice a week. Tumor volumes were calculated by the formula (LxW2)/2.
Results In those tumors harboring YES1 amplification (YES1High), dasatinib significantly reduced tumor growth. This treatment induced tumor response in both ADC and SCC PDX models. On the contrary, in tumors with low expression of YES1, dasatinib did not affect tumor growth in ADC or SCC PDX models.
Conclusions Dasatinib treatment significantly reduces tumor growth in YES1High PDX models but not in YES1Low models, providing support to our hypothesis that YES1 expression may be a useful companion biomarker for dasatinib as NSCLC treatment.
Citation Format: Irati Garmendia, Cristina Bértolo, Irene Ferrer, María J Pajares, Daniel Ajona, Luis Paz-Ares, Ruben Pio, Luis M Montuenga, Jackeline Agorreta. Dasatinib reduces tumor growth in xenograft models derived from human lung tumors with YES1 overexpression [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 LB-084.
Abstract LB-096: A gain-of-function screen identifies drivers of drug resistance in BRAFV600E melanoma cell lines
Authors: Feddersen Charlotte R. , Schillo Jacob L. , Vaughn Hayley R. , Voigt Andrew P. , Zhu Eliot Y. , Wadsworth Lexy S. , Stipp Christopher S. , Dupuy Adam J. 
Published: August 2018

Abstract
Melanoma is the deadliest form of skin cancer, and until recently patients had few treatment options upon diagnosis. Fortunately, this lack of treatment options changed with the discovery of vemurafenib, a highly selective kinase inhibitor that specifically targets the BRAFV600E mutant protein, found in ~50% of all melanoma cases. Initially, the kinase inhibitor provided complete or partial response to over 50% of patients and prolonged progression-free survival. However, the majority of patients relapse once tumors acquire resistance to vemurafenib. Some of the genetic resistance mechanisms are known, including BRAF amplification, expression of truncated BRAF, and RAS mutation. However, a significant portion of vemurafenib resistance cases is unexplained. While drug resistance can be delayed by combining vemurafenib with cobimetinib, a MEK inhibitor, mechanisms of resistance to this drug combination are not well characterized.
We performed a gain-of-function screen to identify drivers of drug resistance to vemurafenib or vemurafenib combined with cobimetinib. Briefly, A375 melanoma cells were engineered to express the hyperactive transposase SB100. These cells were then subsequently transfected with the mutagenic T2Onc3 transposon to produce a population of mutagenized A375 cells. Three weeks of vemurafenib treatment selected for cells with the acquired ability to grow in the presence of drug. Candidate driver mutations were then identified by profiling the sites of transposon insertion in drug-resistant cells to find genes that were recurrently over-expressed by transposon insertions. This led to the discovery of five genes that act as drivers of vemurafenib resistance when overexpressed in the human melanoma cell line A375. Candidate genes were then validated in multiple melanoma cell lines by assessing cell growth and viability in varying concentrations of vemurafenib.
An analogous screen was performed in A375 to identify drivers of resistance to the combination of vemurafenib and cobimetinib. In addition, we are assessing the diversity of drug resistance mechanisms by performing mutagenesis in three additional human melanoma cell lines.
The characterization of the novel drug resistance pathways identified here will improve treatment outcomes for patients with BRAFV600E mutant melanoma.
Citation Format: Charlotte R. Feddersen, Jacob L. Schillo, Hayley R. Vaughn, Andrew P. Voigt, Eliot Y. Zhu, Lexy S. Wadsworth, Christopher S. Stipp, Adam J. Dupuy. A gain-of-function screen identifies drivers of drug resistance in BRAFV600E melanoma cell lines [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 LB-096.