tbuckley/cabot-search huggingface dataset:
527. Exploiting Retroviral Recombination for the Delivery of Full-Length Dystrophin cDNA
Authors: John R. Counsell, Zeinab Asgarian, Jinhong Meng, Veronica Ferrer, Steven J. Howe, Francesco Muntoni, Jennifer E. Morgan, Olivier Danos
Journal: Molecular Therapy (2015)

Duchenne muscular dystrophy (DMD) is caused by loss of dystrophin expression in patient muscle cells. Restoration of dystrophin expression could alleviate the symptoms of DMD, although the large size of full-length dystrophin cDNA (11.1kb) and the restricted packaging capacity of viral vectors limit the application of standard gene transfer methods. Retroviral vectors have a pseudo-diploid RNA genome which can co-package two distinct RNA genomes to form a heterozygous virion. Template-switching between co-packaged strands is a common event during HIV-1 reverse transcription, which can lead to recombination and the production of chimeric proviruses. Template-switching can be influenced by a number of cis and trans-acting factors such as structures in the viral packaging sequences and mutations of reverse transcriptase. In this work, the recombinogenic characteristics of HIV-1 based lentiviral vectors have been exploited for the purpose of reconstituting full-length dystrophin cDNA from overlapping 5’ and 3’ portions of the full-length coding sequence. The vector has been initially optimised using a small bicistronic cassette to evaluate the recombination efficiency in standard lentiviral vectors compared with those harbouring mutations in the HIV-1 packaging sequence and reverse transcriptase. Interestingly, the results suggested that a standard lentivirus vector could deliver a full-length, recombined provirus with greater efficiency than vectors harbouring mutations to cis and trans factors. A multiplicity of infection (MOI) of 40 heterozygous vector particles per target cell was sufficient to obtain a reconstituted full-length Neomycin-IRES-GFP provirus in 40% of transduced HeLa cells. When the viral RNA components were further modified to render functional provirus synthesis dependent on recombination (schematic displayed in figure 1), the vector was capable of reconstituting a full-length dystrophin-gfp fusion protein in HEK293t cells. Flow cytometry analysis showed that heterozygous vectors produced 0.1% GFP-positive cells following a MOI50 vector dose. Furthermore, genomic DNA extracted from these cells contained a full-length dystrophin provirus, which was detected by nested PCR and sequenced by clonal analysis. This work reveals that retroviral recombination offers a means to circumvent retroviral packaging limitations and may be of use in dystrophin gene therapy following further optimisation.
S127 Effect of cannabis smoking on the development of bullous lung disease: a structured literature review
Authors: L Ribeiro, P Ind
Journal: Thorax (2016)

<h3>Background</h3> With increasing cannabis use, physicians need to know more about its respiratory effects. However, there are few long term studies of cannabis smoking, mostly due to legality issues and the confounding effects of tobacco. <h3>Aims</h3> We reviewed the effect of chronic cannabis use on bullous lung disease. <h3>Methods</h3> 18 out of 69 English-language publications, prior to April 2016, from MEDLINE, Scopus, and Web of Science databases, which reported bullous lung disease in cannabis users, were examined. Case reports and case series were included. <h3>Results</h3> The only cross-sectional study reported an increase in the rates of macroscopic emphysema in tobacco only (17 of 92) and tobacco + cannabis smokers (15 of 91), but not in cannabis only smokers (1 of 75) compared to non-smokers.1 The remaining case series and case reports described a total of 56 marijuana smokers presenting with bullous lung disease, often with pneumothorax and predominantly upper lobe involvement (Table 1). Concurrent tobacco smoking was present in all but 3 cases. The majority of cases reported heavy cannabis use, though direct comparison was difficult due to variation in usage measurements. All 4 case series that measured lung function reported normal findings. <h3>Conclusions</h3> While the clinical association of cannabis smoking and peripheral lung bullae is well recognised (and consequently often not reported) there is scant documentation in the literature correlating marijuana smoking with bullous lung disease. <h3>Reference</h3> Aldington S, <i>et al</i>. Effect of cannabis on pulmonary structure, function and symptoms. <i>Thorax</i>. 2007;<b>62</b>(12):1058–63.
Prognostic Significance of FLT3 Mutations in Pediatric Non-Promyelocytic Acute Myeloid Leukemia.
Authors: Hyoung Jin Kang, Seung Hee Hong, In Ho Kim, Byung-Kiu Park, Kyou Sup Han, Han Ik Cho, Hee Young Shin, Hyo Seop Ahn
Journal: Blood (2004)
Small Cell Lung Cancer Exhibits Frequent Inactivating Mutations in the Histone Methyltransferase KMT2D/MLL2 : CALGB 151111 (Alliance)
Authors: Arnaud Augert, Qing Zhang, Breanna Bates, Min Cui, Xiaofei Wang, Gary Wildey, Afshin Dowlati, David MacPherson
Journal: Journal of Thoracic Oncology (2016)
181. Optimization of In Vivo Delivery of Baculovirus Gene Therapy Vectors for the Treatment of Alpha-1 Antitrypsin Deficiency
Authors: Laura vanLieshout, Jondavid de Jong, Jacob vanVloten, Sarah K. Wootton
Journal: Molecular Therapy (2016)

Alpha-1 antitrypsin deficiency (AATD) is a single-gene disorder commonly associated with adult onset lung diseases, including emphysema, chronic obstructive pulmonary disease and airway inflammation. Alpha-1 antitrypsin (AAT) is produced primarily in hepatocytes and is subsequently secreted into the bloodstream and travels to the lung where it inhibits neutrophil elastase. Unregulated neutrophil elastase activity in individuals with AATD causes degradation of various components of the extracellular matrix producing damage to alveolar and airway epithelial cells. Furthermore, mutated AAT folds incorrectly causing intracellular aggregation within hepatocytes, which can result in liver damage in addition to pulmonary symptoms. The development of gene therapy strategies has been a popular approach for the treatment of AATD due to monogenic nature of this genetic disease. Recently, baculovirus based vectors have garnered attention for possible application in gene therapy due to multiple factors. Baculoviruses can tolerate large gene insertions (>38kb), are capable of transducing mammalian cells but not replicating or integrating into host chromosomes and finally, humans lack pre-existing immunity to these insect viruses. Although there are many characteristics that make baculovirus vectors attractive for use in gene therapy, there is little in vivo data on bio-distribution and tissue tropism in mammalian models. As AATD is a disease affecting both the liver and the lung, we examined several delivery routes of a baculovirus vector expressing the human placental alkaline phosphatase reporter gene in C57BL/6 mice. To target the lung we investigated intranasal and intratracheal administration and for liver delivery intravenous, intrahepatic and intraportal vein injections were utilized. Lung delivery of wild type baculovirus yielded little transduction, however intrahepatic injection resulted in moderate liver transduction as well as unexpected spread to other tissues. Different formulations of the baculovirus vector were examined for pulmonary delivery including a variety of viscoeleastic gels. Additionally, pharmacological agents such as clodronate liposomes to deplete macrophages and cobra venom factor to ablate complement activity were examined in an effort to bolster transduction.
Abstract 4039: Inhibition of Pak4 blocks growth of breast cancer cells
Authors: Chetan K. Rane, William Senapedis, Erkan Baloglu, Sharon Shacham, Audrey G. Minden
Journal: Cancer Research (2015)

Abstract The p21-activated kinase family of proteins promotes cell survival and plays an important role in cell proliferation, cell cycle regulation and cell shape determination. There are six mammalian PAK proteins which can be subdivided into two groups by sequence homology and mode of activation- Group A Paks consisting of Pak 1, 2 and 3 and Group B Paks consisting of Pak 4, 5 and 6. There is a growing list of evidence that PAK proteins are overexpressed in many cancer types, including colon, lung, and breast cancer. PAK4 is frequently overproduced in breast cancer, including Her2 positive and triple negative breast cancers, while it is expressed at low levels in normal mammary tissue. Our preliminary evidence showed that untransformed mouse mammary epithelial cells have low levels of PAK4 that become elevated when these cells are transformed by Her2 overexpression. Furthermore, PAK4 overexpressed in mammary epithelial cells leads to oncogenic transformation and tumorigenesis in mice. We hypothesize that PAK4 plays an important role in promoting mammary tumorigenesis, and that it can serve as an effective drug target for breast cancer treatment. To test this hypothesis, we have analyzed two orally available PAK4 Allosteric Modulators (PAM; KPT-8752 and KPT-9274), which reduce the steady state PAK4 protein level in cancer cells. We observed that when we treat breast cancer cells with KPT-8752, we severely block the cell growth. KPT-8752, is effective both in Her2 positive and triple negative breast cancer cells. Our results indicate that Pak4 is an important regulator of breast cancer cell growth. Experiments studying the effects of PAMs (KPT8752 and KPT9274) on tumorigenesis in animal models of breast cancer are ongoing. Citation Format: Chetan K. Rane, William Senapedis, Erkan Baloglu, Sharon Shacham, Audrey G. Minden. Inhibition of Pak4 blocks growth of breast cancer cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4039. doi:10.1158/1538-7445.AM2015-4039
GP surgeries will get 1500 more pharmacists after successful pilot
Authors: Zosia Kmietowicz
Journal: BMJ (2016)

GP surgeries in England are due to get a further 1500 pharmacists working on their premises by 2020-21, after a successful pilot scheme.

NHS England said that applications would open next month for £100m (€118m; $123m) of funding to expand the scheme, which was launched in July 2015. The initial response was so overwhelming that, in November 2015, NHS England doubled the project’s funding from £15m to £31m.1

This year 490 clinical pharmacists have been working in about 650 practices …
Abstract LB-039: 1,25 dihydroxyvitamin D3 and cisplatin combination modulate p73 in bladder cancer
Authors: Brittany L. Bunch, Anna Woloszynska-Read, Donald L. Trump, Candace S. Johnson
Journal: Cancer Research (2015)

Abstract Cisplatin-based combination chemotherapy is the standard approach to therapy of advanced bladder cancer. While there is a substantial initial response rate and occasional complete response, long term control is not common. There is a need to develop new therapeutic approaches to bladder cancer. 1,25 dihydroxyvitamin D3 (1,25D3), the active metabolite of vitamin D, enhances the anti-tumor effects of cisplatin in preclinical bladder cancer models. We evaluated the mechanism of 1,25D3 potentiation of cisplatin cytotoxicity through in vitro studies in bladder cancer cell lines. Our studies suggest that enhanced cytotoxicity is mediated through modulation of p73. The ratio of the pro-apoptotic TAp73 isoform to the anti-apoptotic ΔNp73 isoform is important in determining cellular response to cisplatin. Our studies demonstrate that pretreatment with 1,25D3 (0.5 uM) for 24 hrs followed by cisplatin (0.75 ug/ml) for 48 hrs increases the ratio of TA/ΔNp73 mRNA transcripts 2-fold in the T24 bladder cancer cell line, and increases TAp73's transcriptional target, BAX approximately 3-fold compared to cisplatin alone. Protein levels of p73 and BAX, are also increased with 1,25D3 pretreatment followed by cisplatin in T24 cells as determined by western blot. Using a TransAm p53 binding assay after p53 immunodepletion, cisplatin treatment was found to decrease TAp73 functional ability to bind DNA by approximately half, although this has not yet been determined to be statistically significant. 1,25D3 pretreatment followed by cisplatin prevents the decrease in TAp73 DNA binding found after cisplatin alone. These data suggest an increase in TAp73 pro-apoptotic functional abilities resulting from 1,25D3 pretreatment followed by cisplatin in T24 cells. These findings suggest that 1,25D3 may have potential to be used in combination with cisplatin to increase the apoptotic response. Further studies are being performed to determine the requirement of TAp73 in 1,25D3 potentiation of cisplatin cytotoxicity by using T24 cells transfected with TAp73 shRNA in assays previously used to determine synergism, ie. MTT, clonogenic, and apoptosis assays. Supported by NCI grants CA067267 and CA016056. Citation Format: Brittany L. Bunch, Anna Woloszynska-Read, Donald L. Trump, Candace S. Johnson. 1,25 dihydroxyvitamin D3 and cisplatin combination modulate p73 in bladder cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr LB-039. doi:10.1158/1538-7445.AM2015-LB-039
Allogeneic Hematopoietic Stem Cell Transplantation (HCT) in First Remission Improves Outcome Irrespective of FLT3-ITD Allelic Burden Among Patients with Acute Myeloid Leukemia and FLT3-ITD Mutation
Authors: Betul Oran, Richard E. Champlin, Jorge E. Cortes, Marcos de Lima, Xuemei Wang, Hsiang-Chun Chen, Farhad Ravandi, Stefan O. Ciurea, Hagop M. Kantarjian, Gautam Borthakur
Journal: Blood (2014)
Characterisation of the minimal area of homozygous 13q14.3 deletion in B-CLL and isolation of candidate tumour suppressor genes.
Authors: M Corcoran, R Mullenbach, R Ibbotson, O Rasool, A Iyengar, D Grander, R Chapman, N Yankovsky, [No Value] Wu, M Merup, G Juliusson, G Gahrton, [No Value] Brodyansky, C Buys, E Zabarovsky, D Oscier, S Einhorn
Journal: Blood (1996)
Abstract 1433: CDKL2 promotes epithelial-mesenchymal transition and breast cancer progression
Authors: Linna Li, Jeffrey Chang, Guangwei Du, Robert Amato, Wenliang Li
Journal: Cancer Research (2015)

Abstract The epithelial-mesenchymal transition (EMT) confers mesenchymal properties on epithelial cells and has been closely associated with the acquisition of aggressive traits by epithelial cancer cells. To identify novel regulators of EMT, we carried out cDNA screens that covered 500 human kinases. Subsequent characterization of candidate kinases led us to uncover cyclin-dependent kinase-like 2 (CDKL2) as a novel potent promoter for EMT and breast cancer progression. CDKL2-expressing human mammary gland epithelial cells displayed enhanced mesenchymal traits and stem cell-like phenotypes, which was acquired through activating a ZEB1/E-cadherin/beta-catenin positive feedback loop and regulating CD44 mRNA alternative splicing to promote conversion of CD24-high cells to CD44-high cells. Furthermore, CDKL2 enhanced primary tumor formation and metastasis in a breast cancer xenograft model. Notably, CDKL2 is expressed significantly higher in mesenchymal human breast cancer cell lines than in epithelial lines, and its over-expression/amplification in human breast cancers is associated with shorter disease-free survival. Taken together, our study uncovered a major role for CDKL2 in promoting EMT and breast cancer progression. Citation Format: Linna Li, Jeffrey Chang, Guangwei Du, Robert Amato, Wenliang Li. CDKL2 promotes epithelial-mesenchymal transition and breast cancer progression. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1433. doi:10.1158/1538-7445.AM2015-1433
Syria violence is condemned as evacuation begins
Authors: Anne Gulland
Journal: BMJ (2016)

A coalition of health groups has condemned the Syrian government for turning doctors into “targets.”

Speaking on behalf of a group of non-governmental organisations at a press conference in Turkey, Mohamad Katoub, of the Syrian American Medical Society, condemned the Syrian and Russian governments’ attacks on eastern Aleppo and the failure of the international community to intervene.

He said, “We condemn the Syrian government for waging a war against its people, for turning medics into targets, and for denying humanitarian aid to besieged populations.” …
OP54HETEROPHILIC INTERACTION OF CD62E AND CD15 MEDIATES ADHESION OF METASTATIC NON-SMALL CELL LUNG CANCER CELLS TO BRAIN ENDOTHELIUM
Authors: Samah A. Jassam, Zaynah Maherally, James R. Smith, Keyoumars Ashkan, Helen L. Fillmore, Geoffrey J. Pilkington
Journal: Neuro-Oncology (2015)

INTRODUCTION: CD15, a cell adhesion molecule which is overexpressed in various cancers, is thought to be a key player in non-central nervous system (CNS) metastasis. However, the role of CD15 in brain metastasis is largely unexplored. CD15, a trisaccharide adhesion molecule, is over-expressed in non-CNS metastatic cancers. The role of CD15/CD62E interaction in lung to brain metastasis is still unexplored. The aim of this study is to therefore provide a better understanding of CD15/CD62E interaction and its role in adhesion in brain metastasis. METHOD: Expression of CD62E and CD15 were evaluated in brain microvascular endothelial cells (hCMEC/D3), primary (A549 and COR-L105) and metastatic NSCLC cells (NCI-H1299, SEBTA-001 and SEBTA-005) using flow cytometry (FC), immunocytochemistry (ICC) and Western blotting (WB). Adhesion assays under static conditions and physiological flow (shear stress 2.5 dyn/cm2) conditions were conducted. Immunoblocking of CD15 was performed to investigate its role in adhesion. RESULTS: Low levels of CD15 was seen on hCMEC/D3 (19.69%) compared to metastatic NSCLC cells (NCI-H1299: 79%, SEBTA-001: 54% and SEBTA-005: 39%) and primary NSCLC (COR-L105: 31% and A549: 23%) cells (P < 0.001). CD62E expression was high in hCMEC/D3 cells activated with TNF-α. Metastatic NSCLC adhered most strongly to hCMEC/D3 compared to primary NSCLC cells. CD15 immunoblocking blunted this adhesion under static and shear stress conditions (p < 0.001), confirming the correlation between CD15 and cerebral metastasis. CONCLUSION: This study enhances understanding of lung cancer cell-brain endothelial adhesion and suggests that CD15 may play a role in adhesion in concert with TNF-α activation of its binding partner CD62E.
Abstract 2887: Epigenetic silencing of the DNA repair genes, BRCA1 and MLH1, induced by hypoxic stress in a pathway dependent on the histone demethylase, LSD1
Authors: Yuhong Lu, Adrian Chu, Narendra Wajapeyee, Mitchell S. Turker, Peter M. Glazer
Journal: Cancer Research (2015)

Abstract Hypoxia is a key microenviromental stress in solid tumors that is associated with poor prognosis. It has been indicated that hypoxia induces genetic instability through multiple mechanisms. Previously, we have shown that BRCA1 and RAD51, in the homology dependent repair (HDR) pathway, and MLH1 and MSH2, key components of MMR, are down-regulated at the transcriptional level in response to hypoxia. Here we show that hypoxia also drives epigenetic modification of the BRCA1 and MLH1 promoters, with decreased H3K4 methylation as a key repressive modification produced by the lysine specific histone demethylase, LSD1. In cells containing the BRCA1 promoter driving a selectable HPRT gene, long-term silencing of the promoter was observed following exposure to hypoxic stress. The silenced clones showed decreased H3K4 methylation and decreased H3K9 acetylation in the BRCA1 promoters, consistent with the acute effects of hypoxic stress. Similarly, we also found that hypoxia causes durable long-term silencing of the MLH1 promoter in a pathway that requires LSD1. Knockdown of LSD1 or its co-repressor, CoREST, also prevents the re-silencing (and associated cytosine DNA methylation) of the endogenous MLH1 promoter in RKO colon cancer cells following transient reactivation by treatment with the DNA methyltransferase inhibitor 5-aza-2′-deoxycytidine (5-aza-dC). These results suggest that hypoxia is a driving force for long-term silencing of DNA repair genes, including BRCA1 and MLH1, thereby promoting genome instability and tumor progression. Citation Format: Yuhong Lu, Adrian Chu, Narendra Wajapeyee, Mitchell S. Turker, Peter M. Glazer. Epigenetic silencing of the DNA repair genes, BRCA1 and MLH1, induced by hypoxic stress in a pathway dependent on the histone demethylase, LSD1. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2887. doi:10.1158/1538-7445.AM2015-2887
High Risk Adva­nced Stage Hodgkin Lymphoma Is Well Controlled with 2 Cycles of Escalated Beacopp Followed By 4 Cycles of ABVD in Patients Who Rapidly Achieve Metabolic CR on Interim PET/CT Scan
Authors: Meirav Kedmi, Arie Apel, Tima Davidson, Elinor Goshen, Yaron Davidovitz, Miriam Berkowicz, Itai Levi, Eldad J. Dann, Aaron Polliack, Arnon Nagler, Abraham Avigdor
Journal: Blood (2014)
Innate Immunity in Idiopathic Thrombocytopenic Purpura (ITP)
Authors: Akio Saito, Akihiko Yokohama, Hirotaka Nakahashi, Kohtaro Toyama, Takeki Mitsui, Yoko Hashimoto, Hiromi Koiso, Takayuki Saitoh, Hideki Uchiumi, Hiroshi Handa, Morio Sawamura, Masamitsu Karasawa, Hirokazu Murakami, Yoshihisa Nojima, Norifumi Tsukamoto
Journal: Blood (2008)
Meta Analysis of Grade 3 and/or 4 Toxicities in Low-Grade Non-Hodgkin Lymphoma (LG-NHL) Patients Receiving Maintenance Rituximab (MR) Therapy: Impact of Schedule on Toxicity
Authors: Chadi Nabhan, Dana Villines, Tina V. Valdez, Michele Ghielmini, Shu-Fang Hsu Schmitz, Sonali M. Smith
Journal: Blood (2011)
3D Modeling of Novel Transforming JAK Mutations in T-Cell Acute Lymphoblastic Leukemia Reveals Altered Pseudokinase-Kinase Domain Interactions That Result in Constitutive JAK Kinase Activity
Authors: Kirsten Canté-Barrett, Joost CM Uitdehaag, Jessica GCAM Buijs-Gladdines, Wilco K Smits, Rogier C Buijsman, Guido JR Zaman, Rob Pieters, Jules PP Meijerink
Journal: Blood (2015)