bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract B37: RNAi and CRISPR/Cas9-based in vivo models for drug discovery
Authors: Premsrirut Prem K. , Wang Chia-Lin , Yang Yu-ting , Pelossof Rafii , Leslie Christina , Fellmann Christof , Dow Lukas , Zuber Johannes , Lowe Scott 
Published: May 2018

Abstract
With the advent of RNAi and more recently CRISPR/Cas9 technologies, the speed and precision by which genetically engineered mouse models of cancer can be created is unprecedented. Having previously engineered a miRE scaffold for enhanced shRNA processing and now with the introduction of a powerful new SplashRNA algorithm for accurate shRNA prediction, we have brought RNAi technology to its peak by increasing potency and reducing off-target effects, such that it can be effectively exploited to preclinically mimic drug therapy and even combination therapy not only in vitro but also in live mice. Here, we take advantage of Cas9-expressing mice and demonstrate that in situ delivery of sgRNAs can lead to somatic mutagenesis to promote rapid tumorigenesis in mice. By combining this approach with inducible and reversible RNAi-mediated gene silencing to mimic drug therapy in the same mice, we now have an advanced platform to perform target validation and toxicity assessment of novel candidate targets in vivo. Here, we showcase how synergizing our RNAi and CRISPR/Cas9 genetic toolbox will help facilitate cancer drug discovery research and increase our confidence in predicting drug responses in humans into a new era.
Citation Format: Prem K. Premsrirut, Chia-Lin Wang, Yu-ting Yang, Rafii Pelossof, Christina Leslie, Christof Fellmann, Lukas Dow, Johannes Zuber, Scott Lowe. RNAi and CRISPR/Cas9-based in vivo models for drug discovery [abstract]. In: Proceedings of the AACR Special Conference: Advances in Modeling Cancer in Mice: Technology, Biology, and Beyond; 2017 Sep 24-27; Orlando, Florida. Philadelphia (PA): AACR; Cancer Res 2018;78(10 Suppl):Abstract nr B37.
Abstract IA07: Deconstructing p53 pathways in tumor suppression
Authors: Mello Stephano Spano , Bieging-Rolett Kathryn , Kaiser Alyssa , Valente Elizabeth Joy , Raj Nitin , McClendon Jacob , Flowers Brittany Maria , Morgens David Warren , Bassik Michael Cory , Attardi Laura Donatella 
Published: May 2018

Abstract
The p53 transcription factor is a critical tumor suppressor in humans and mice. Despite this essential function, the molecular pathways through which p53 acts in tumor suppression remain enigmatic. To define the transcriptional programs through which p53 suppresses carcinogenesis, we have taken combined mouse genetic and genomic approaches. We previously generated a panel of p53 knock-in mouse strains expressing mutants in the first (p5325,26), second (p5353,54), or both (p5325,26,53,54) of two amino-terminal transcriptional activation domains (TADs). In terms of transcriptional activity, we have discovered that p5325,26 is severely impaired for transactivation of the majority of canonical p53 target genes (e.g. p21, Puma, Noxa), but retains the ability to activate a set of primarily novel p53 target genes, while p5325,26,53,54 lacks transactivation activity altogether. The p5353,54 mutant, in contrast, is uncompromised for transactivation of p53 target genes. The p5325,26,53,54 mutant is completely defective in tumor suppression, underscoring the importance of transcriptional activation for p53-mediated tumor suppression. Intriguingly, the p5325,26 mutant retains full activity in suppressing a variety of cancers, indicating that efficient transactivation of most canonical p53 target genes is dispensable for tumor suppression. As p5325,26 activates only a subset of p53-inducible genes, yet retains tumor-suppressor activity, it has helped pinpoint a small set of novel, direct p53-inducible tumor suppression-associated target genes (TSAGs) whose functions we are currently interrogating through genetic screens. Additionally, we have made the surprising discovery that the p5353,54 mutant suppresses pancreatic cancer more efficiently than wild-type p53, and that this capacity correlates with the ability of p5353,54 to hyperactivate a subset of p53 target genes. Indeed, our analysis of these genes has uncovered Ptpn14, a direct p53 target gene critical for mediating p53 function in suppressing pancreatic cancer cell growth. Ptpn14 encodes a protein tyrosine phosphatase that negatively regulates the oncoprotein Yap in both human and mouse cells. These studies thus reveal a p53-Ptpn14-Yap axis in pancreatic cancer. Together, these strategies will help to elaborate the transcriptional networks fundamental for p53 function in tumor suppression.
Citation Format: Stephano Spano Mello, Kathryn Bieging-Rolett, Alyssa Kaiser, Elizabeth Joy Valente, Nitin Raj, Jacob McClendon, Brittany Maria Flowers, David Warren Morgens, Michael Cory Bassik, Laura Donatella Attardi. Deconstructing p53 pathways in tumor suppression [abstract]. In: Proceedings of the AACR Special Conference: Advances in Modeling Cancer in Mice: Technology, Biology, and Beyond; 2017 Sep 24-27; Orlando, Florida. Philadelphia (PA): AACR; Cancer Res 2018;78(10 Suppl):Abstract nr IA07.
Abstract IA14: Lessons from modeling neural tumors in mice
Authors: Xie Xuan-Hua , Alcantara Sheila , Laks Dan , Shi Yufeng , Parada Luis F. 
Published: May 2018

Abstract
We have used genetically engineered mouse models to study GBM. In particular, through mutation of glioma relevant genes (P53; Pten; Nf1) we have examined the scope of the cell of origin for these tumors. We find that as cells in the adult neural stem cell lineage progress, their capacity to undergo glioma formation is largely compromised though cellular phenotypes arise as a consequence of mutation. We find that in the subventricular zone, only quiescent stem cells and early progenitors can readily form GBM following mutation. Our search for additional cells of origin has further identified additional source cells for these tumors. GBM can arise from the OPC lineage following the same mutations, and these tumors have a distinct signature that can be identified among the TCGA GBM data sets. Thus the cell of origin is constrained to stem-like cells and tumor characteristics include signatures revealing the lineage of origin. We have further examined the properties of tumor growth and transplantation and find that these tumors grow in hierarchical fashion with a relatively small and quiescent cell population at the apex. These “cancer stem cells” can be identified and isolated. New single-cell sequencing technology promises to reveal many previous secrets from these cells. Understanding how a stem cell transforms into a cancer stem cell may provide novel approaches for therapeutic targeting.
Citation Format: Xuan-Hua Xie, Sheila Alcantara, Dan Laks, Yufeng Shi, Luis F. Parada. Lessons from modeling neural tumors in mice [abstract]. In: Proceedings of the AACR Special Conference: Advances in Modeling Cancer in Mice: Technology, Biology, and Beyond; 2017 Sep 24-27; Orlando, Florida. Philadelphia (PA): AACR; Cancer Res 2018;78(10 Suppl):Abstract nr IA14.
Abstract PR11: Investigating lung cancer cells-of-origin using three-dimensional organoid cultures
Authors: Wong Kwok-Kin , Kim Carla , Brainson Christine Fillmore 
Published: May 2018

Abstract
There are many distinct epithelial cell types in the adult lung that could give rise to non-small cell lung cancer. These cell types include tracheal basal stem cells, tracheal club cells, distal lung bronchioalveolar stem cells (BASCs), and distal lung alveolar type 2 cells (AT2). My laboratory is using mouse models to test directly which lung stem and progenitor cells are most fit to serve as cells-of-origin of lung adenocarcinoma and lung squamous cell carcinoma. Previously, we demonstrated that tracheal club cells and BASCs are the likely cells-of-origin for KRASG12D; Lkb1-null adenosquamous tumors. The same strategy is now being used to test which cells can give rise to adenocarcinomas using the LSL:EGFRT90M/L858R (LSL=lox-stop-lox) mouse strain, or squamous cell carcinoma using the Lkb1flox/flox; Ptenflox/flox mouse strain. Briefly, tracheas and lungs from Cre-naïve mice are enzymatically dissociated, and fluorescence activated cell sorting (FACS) is used to enrich for four distinct populations: NGFR+/Sca1+ tracheal basal cells, NGFR-/Sca1+ tracheal club cells, Sca1+ distal lung BASCs, and Sca1- distal lung AT2 cells. Each population is then incubated with either adeno-GFP control virus or adeno-Cre virus to induce EGFR T790M/L858R expression or delete both alleles of Lkb1 and Pten. Three dimensional air-liquid-interface cultures are then used to expand the populations as organoids. For EGFR organoids, both basal and club cells give rise to larger, faster-growing bronchiolar organoids after EGFR is induced. AT2 cells similarly give rise to much larger alveolar organoids after EGFR induction, and BASCs change from giving rise to both bronchiolar and alveolar organoids to nearly exclusively alveolar. For Lkb1/Pten organoids, all four populations can tolerate deletion of the two genes, but it is only the basal cell and club cell organoids that progress to a squamous morphology in vitro. While both BASC and AT2 organoids continue to grow, they appear phenotypically normal. Transplant assays to assess malignancy and RNA-seq and ChIP-seq studies to explore gene transcription and epigenetic changes are under way. These experiments will shed light on the cells of the adult lung that give rise to genotype-specific tumors, and will aid us to better classify lung cancer heterogeneity from a cell-of-origin perspective.
Citation Format: Kwok-Kin Wong, Carla Kim, Christine Fillmore Brainson. Investigating lung cancer cells-of-origin using three-dimensional organoid cultures [abstract]. In: Proceedings of the AACR Special Conference: Advances in Modeling Cancer in Mice: Technology, Biology, and Beyond; 2017 Sep 24-27; Orlando, Florida. Philadelphia (PA): AACR; Cancer Res 2018;78(10 Suppl):Abstract nr PR11.
Subcellular Localization of Antigen in Keratinocytes Dictates Delivery of CD4+ T-cell Help for the CTL Response upon Therapeutic DNA Vaccination into the Skin
Authors: Bąbała Nikolina , Bovens Astrid , de Vries Evert , Iglesias-Guimarais Victoria , Ahrends Tomasz , Krummel Matthew F. , Borst Jannie , Bins Adriaan D. 
Published: May 2018

Abstract
In a mouse model of therapeutic DNA vaccination, we studied how the subcellular localization of vaccine protein impacts antigen delivery to professional antigen-presenting cells and efficiency of CTL priming. Cytosolic, membrane-bound, nuclear, and secretory versions of ZsGreen fluorescent protein, conjugated to MHC class I and II ovalbumin (OVA) epitopes, were expressed in keratinocytes by DNA vaccination into the skin. ZsGreen-OVA versions reached B cells in the skin-draining lymph node (dLN) that proved irrelevant for CTL priming. ZsGreen-OVA versions were also actively transported to the dLN by dendritic cells (DC). In the dLN, vaccine proteins localized to classical (c)DCs of the migratory XCR1+ and XCR− subtypes, and—to a lesser extent—to LN-resident cDCs. Secretory ZsGreen-OVA induced the best antitumor CTL response, even though its delivery to cDCs in the dLN was significantly less efficient than for other vaccine proteins. Secretory ZsGreen-OVA protein proved superior in CTL priming, because it led to in vivo engagement of antigen-loaded XCR1+, but not XCR1−, cDCs. Secretory ZsGreen-OVA also maximally solicited CD4+ T-cell help. The suboptimal CTL response to the other ZsGreen-OVA versions was improved by engaging costimulatory receptor CD27, which mimics CD4+ T-cell help. Thus, in therapeutic DNA vaccination into the skin, mere inclusion of helper epitopes does not ensure delivery of CD4+ T-cell help for the CTL response. Targeting of the vaccine protein to the secretory route of keratinocytes is required to engage XCR1+ cDC and CD4+ T-cell help and thus to promote CTL priming. Cancer Immunol Res; 6(7); 835–47. ©2018 AACR.
IL22 Promotes Kras-Mutant Lung Cancer by Induction of a Protumor Immune Response and Protection of Stemness Properties
Authors: Khosravi Nasim , Caetano Mauricio S. , Cumpian Amber M. , Unver Nese , De la Garza Ramos Cynthia , Noble Oscar , Daliri Soudabeh , Hernandez Belinda J. , Gutierrez Berenice A. , Evans Scott E. , Hanash Samir , Alekseev Andrei M. , Yang Yi , Chang Seon Hee , Nurieva Roza , Kadara Humam , Chen Jichao , Ostrin Edwin J. , Moghaddam Seyed Javad 
Published: May 2018

Abstract
Somatic KRAS mutations are the most common oncogenic variants in lung cancer and are associated with poor prognosis. Using a Kras-induced lung cancer mouse model, CC-LR, we previously showed a role for inflammation in lung tumorigenesis through activation of the NF-κB pathway, along with induction of interleukin 6 (IL6) and an IL17-producing CD4+ T-helper cell response. IL22 is an effector molecule secreted by CD4+ and γδ T cells that we previously found to be expressed in CC-LR mice. IL22 mostly signals through the STAT3 pathway and is thought to act exclusively on nonhematopoietic cells with basal IL22 receptor (IL22R) expression on epithelial cells. Here, we found that higher expression of IL22R1 in patients with KRAS-mutant lung adenocarcinoma was an independent indicator of poor recurrence-free survival. We then showed that genetic ablation of Il22 in CC-LR mice (CC-LR/IL22KO mice) caused a significant reduction in tumor number and size. This was accompanied by significantly lower tumor cell proliferation, angiogenesis, and STAT3 activation. Il22 ablation was also associated with significant reduction in lung-infiltrating inflammatory cells and expression of protumor inflammatory cytokines. Conversely, this was accompanied with increased antitumor Th1 and cytotoxic CD8+ T-cell responses, while suppressing the protumor immunosuppressive T regulatory cell response. In CC-LR/IL22KO mice, we found significantly reduced expression of core stemness genes and the number of prototypical SPC+CCSP+ stem cells. Thus, we conclude that IL22 promotes Kras-mutant lung tumorigenesis by driving a protumor inflammatory microenvironment with proliferative, angiogenic, and stemness contextual cues in epithelial/tumor cells. Cancer Immunol Res; 6(7); 788–97. ©2018 AACR.
Bidirectional Connections between Depression and Ataxia Severity in Spinocerebellar Ataxia Type 3 Patients
Authors: Lin Min-Ting , Yang Jin-Shan , Chen Ping-Ping , Qian Mei-Zhen , Lin Hui-Xia , Chen Xiao-Ping , Shang Xian-Jin , Wang Dan-Ni , Chen Yu-Chao , Jiang Bin , Chen Yi-Jun , Chen Wan-Jin , Wang Ning , Gan Shi-Rui 
Published: May 2018

<b><i>Background:</i></b> Spinocerebellar ataxia type 3 (SCA3), which is the most common subtype of SCA worldwide, exhibits common neuropsychological symptoms such as depression. However, the contribution of depression to the severity of SCA3 has not yet been thoroughly investigated. <b><i>Methods:</i></b> The present study investigated the prevalence of depression using Beck depression inventory in 104 molecularly conﬁrmed SCA3 patients from China. The putative risk factors for depression and whether the depression could affect the severity of ataxia were established by multivariable linear regression models. <b><i>Results:</i></b> The frequency of depression in the study subjects was 57.69% (60/104), which was higher than that in SCA3 patients from a subset of other populations. The gender (<i>p</i> = 0.03) and severity (<i>p</i> &#x3c; 0.01) of ataxia were those risk factors that could affect depression. Conversely, depression (<i>p</i> &#x3c; 0.01) together with the duration (<i>p </i>&#x3c; 0.01) of SCA3 could also play a positive role in the severity of ataxia. <b><i>Conclusions:</i></b> The extremely common depression results from motor disability caused by ataxia; it also affects the disease severity of SCA3. These findings suggested that depression was a part of neurodegeneration in SCA3 and necessitated intensive focus and interventions while caring for SCA3 patients.
Variability and Confidence Intervals for the Mean of Climate Data with Short- and Long-Range Dependence
Authors: Bowers Matthew C. , Tung Wen-wen 
Published: May 2018

AbstractThis paper presents an adaptive procedure for estimating the variability and determining error bars as confidence intervals for climate mean states by accounting for both short- and long-range dependence. While the prevailing methods for quantifying the variability of climate means account for short-range dependence, they ignore long memory, which is demonstrated to lead to underestimated variability and hence artificially narrow confidence intervals. To capture both short- and long-range correlation structures, climate data are modeled as fractionally integrated autoregressive moving-average processes. The preferred model can be selected adaptively via an information criterion and a diagnostic visualization, and the estimated variability of the climate mean state can be computed directly from the chosen model. The procedure was demonstrated by determining error bars for four 30-yr means of surface temperatures observed at Potsdam, Germany, from 1896 to 2015. These error bars are roughly twice the width as those obtained using prevailing methods, which disregard long memory, leading to a substantive reinterpretation of differences among mean states of this particular dataset. Despite their increased width, the new error bars still suggest that a significant increase occurred in the mean temperature state of Potsdam from the 1896–1925 period to the most recent period, 1986–2015. The new wider error bars, therefore, communicate greater uncertainty in the mean state yet present even stronger evidence of a significant temperature increase. These results corroborate a need for more meticulous consideration of the correlation structures of climate data—especially of their long-memory properties—in assessing the variability and determining confidence intervals for their mean states.
Burnout in Otolaryngology–Head and Neck Surgery: A Single Academic Center Experience
Authors: Geelan‐Hansen Katie , Anne Samantha , Benninger Michael S. 
Published: May 2018

Burnout in modern medicine is becoming more recognized and researched. The objective in this study is to evaluate burnout in a tertiary care academic institution and compare results among faculty, trainees, and advanced practice practitioners (APPs) in a cross‐sectional survey using the Maslach Burnout Inventory. Fifty‐two surveys were distributed; 44 participants completed the survey (85%): 25 staff physicians (57%), 14 resident physicians (32%), and 5 nurse practitioners (11%). Staff physicians had low emotional exhaustion, moderate depersonalization, and low result for reduced personal accomplishments; trainees reported low emotional exhaustion, high depersonalization, and moderate reduced personal accomplishment; and nurse practitioners reported moderate on all 3 dimensions. There is overall low burnout in this tertiary care academic center of otolaryngologist providers and no difference in rates among the different groups (trainees, APPs, staff). Measures addressing specific deficiencies among dimensions of burnout would be helpful to prevent disintegration of physician satisfaction into burnout.
VLA-4 mediated adhesion of melanoma cells on the blood–brain barrier is the critical cue for melanoma cell intercalation and barrier disruption
Authors: García-Martín Ana B , Zwicky Pascale , Gruber Thomas , Matti Christoph , Moalli Federica , Stein Jens V , Francisco David , Enzmann Gaby , Levesque Mitchell P , Hewer Ekkehard , Lyck Ruth 
Published: May 2018

Melanoma is the most aggressive skin cancer in humans. One severe complication is the formation of brain metastasis, which requires extravasation of melanoma cells across the tight blood–brain barrier (BBB). Previously, VLA-4 has been assigned a role for the adhesive interaction of melanoma cells with non-BBB endothelial cells. However, the role of melanoma VLA-4 for breaching the BBB remained unknown. In this study, we used a mouse in vitro BBB model and imaged the shear resistant arrest of melanoma cells on the BBB. Similar to effector T cells, inflammatory conditions of the BBB increased the arrest of melanoma cells followed by a unique post-arrest behavior lacking immediate crawling. However, over time, melanoma cells intercalated into the BBB and compromised its barrier properties. Most importantly, antibody ablation of VLA-4 abrogated melanoma shear resistant arrest on and intercalation into the BBB and protected the BBB from barrier breakdown. A tissue microarray established from human brain metastasis revealed that indeed a majority of 92% of all human melanoma brain metastases stained VLA-4 positive. We propose VLA-4 as a target for the inhibition of brain metastasis formation in the context of personalized medicine identifying metastasizing VLA-4 positive melanoma.
The effects of dihydroartemisinin on inflammatory bowel disease-related bone loss in a rat model
Authors: Ge Xingtao , Chen Zhijian , Xu Zhenjie , Lv Fang , Zhang Kewei , Yang Yu 
Published: May 2018

Bone loss is one of the important extra-intestinal manifestations in patients with inflammatory bowel diseases (IBDs). Compounds derived from natural products have been used to treat IBDs. However, the role of natural products on IBD-induced bone loss is not completely clarified. In the present study, we observed the effects of dihydroartemisinin (DHA), an antimalaria drug, on IBD and IBD-induced bone loss in a rat model. Chronic IBD model was established in Sprague-Dawley rats by giving them 2.5% dextran sodium sulfate in drinking water. DHA was given by intraperitoneal injection. Blood, colon, and bone samples were collected for biomarker assay and histological analysis. There was an obvious increase in tumor necrotic factor (TNF) α and receptor activator of nuclear factor (NF)-kB ligand (RANKL), and decrease in procollagen type 1 N-terminal propeptide (P1NP) level in IBD groups compared with the normal control (p < 0.05). The disease activity score of IBD rats was significantly higher than the control (p < 0.01). Obvious decrease in disease activity score, TNFα, and RANKL level and increase in P1NP were observed in DHA-treated IBD rats. Bone loss, shown as the decrease in bone mineral density, bone volume fraction, and trabecular number and increase in trabecular separation were observed in IBD rats compared with control (p < 0.01). DHA treatment obviously abolished the bone loss, in particular in the high-dose group (p < 0.05). DHA treatment also inhibited the excessive osteoclast formation; RANKL protein expression; and RANK, TRAF6, Fra-1, NFATc1 mRNA expression induced by IBD. Our data indicated that DHA may be a potential therapeutic agent for IBD and IBD-induced bone loss. Impact statement Bone loss is one of the important extra-intestinal manifestations in patients with inflammatory bowel diseases (IBDs). Studies have shown that compounds derived from natural products are useful in the treatment of IBDs. However, few studies have investigated the role of compounds derived from natural products in treatment of osteoporosis in IBDs. The current study aimed to show the effects of dihydroartemisinin (DHA), antimalaria drug, on bone loss in a rat model of IBD. The findings showed that DHA intervention dose dependently protected against bone loss in IBD rats by inhibiting tumor necrotic factor α production and osteoclast formation. These findings highlights that DHA may be beneficial for bone health in those patients with IBD.