High efficiency of alphaviral gene transfer in combination with 5-fluorouracil in a mouse mammary tumor model

Copyright: Copyright 2014 Elsevier B.V., All rights reserved.Background: The combination of virotherapy and chemotherapy may enable efficient tumor regression that would be unachievable using either therapy alone. In this study, we investigated the efficiency of transgene delivery and the cytotoxic effects of alphaviral vector in combination with 5-fluorouracil (5-FU) in a mouse mammary tumor model (4 T1).Methods: Replication-deficient Semliki Forest virus (SFV) vectors carrying genes encoding fluorescent proteins were used to infect 4 T1 cell cultures treated with different doses of 5-FU. The efficiency of infection was monitored via fluorescence microscopy and quantified by fluorometry. The cytotoxicity of the combined treatment with 5-FU and alphaviral vector was measured using an MTT-based cell viability assay. In vivo experiments were performed in a subcutaneous 4 T1 mouse mammary tumor model with different 5-FU doses and an SFV vector encoding firefly luciferase.Results: Infection of 4 T1 cells with SFV prior to 5-FU treatment did not produce a synergistic anti-proliferative effect. An alternative treatment strategy, in which 5-FU was used prior to virus infection, strongly inhibited SFV expression. Nevertheless, in vivo experiments showed a significant enhancement in SFV-driven transgene (luciferase) expression upon intratumoral and intraperitoneal vector administration in 4 T1 tumor-bearing mice pretreated with 5-FU: here, we observed a positive correlation between 5-FU dose and the level of luciferase expression.Conclusions: Although 5-FU inhibited SFV-mediated transgene expression in 4 T1 cells in vitro, application of the drug in a mouse model revealed a significant enhancement of intratumoral transgene synthesis compared with 5-FU untreated mice. These results may have implications for efficient transgene delivery and the development of potent cancer treatment strategies using alphaviral vectors and 5-FU.publishersversionPeer reviewe

Tumour therapy using cytokine-expressing semliki forest virus vectors

THESIS 7937Semliki Forest Virus (SFV) vector is a transient RNA based suicidal expression vector system and has been previously used as a potential anti-cancer agent. Recently, a new enhanced SFV vector has been developed, pSFV10-E. Cells transfected with this vector yield up to ten times more foreign protein than cells transfected with original (non-enhanced) expression vector pSFV10. The two IL-12 gene subunits were cloned from mouse splenocytes and inserted into the pSFV10-E and pSFV10 vectors. Both the pSFV-mIL-12 constructs were characterised for their expression levels qualitatively and quantitatively in BHK-21 and K-BALB cells. The secretion of biologically active mIL-12 was confirmed by inducing splenocytes for IFN-y production using IL-12 expressed by BHK-21 and K-BALB cells. K-BALB, CT26, and 4T1 tumours are the experimental tum our models in our study

Data for: 3403472

The crucial challenge experienced by the present executives is how to effectively manage and drive the multigenerational task force in today’s competitive and dynamic business world. For the very first time in the history, the current workforce consists of all the different generations in various role of the organisation (Zeneke et al 2000) .These cohorts cling to different workplace values (Lyons et al,2007), personalities (Twenge and Campbell,2008),expectations towards work preferences(Twenge 2010).Employees of different generations are rooted with uneven expectations (what they value) in terms of opportunities for growth and advancement; learning and sharing; rewards and recognition; autonomy and formal relationship ;professional involvement and commitment; concern towards customers and society ; affinity towards technology and adaptability to change .Effectively managing these variations among the different generation cohort will result in higher efficiency, employee productivity and innovation (Kupperschmidt 2000,Koyan 2007).Obviously the other side is reflected by, improper coordination and inefficiency in employee productivity due to misunderstandings and miscommunications ,(Fyock 1990) .Previous literature (Turkiewicz 2000,Bradford 1993,Adams 2000) reports that due to the post industrial development along with drastic changes in the information technology Gen X and Gen Y are equipped with technology enabled skills that Baby Boomers lack. In spite of which all the three generations compete for resources and positions (Raines 1997). Academics and researchers have focused publications to educate HR executives to adopt to the requirements of the demands of the present Gen X and Gen Y employees(Alsop,2008;Gorman et al.,2004;Hershalter and Epstein,2010) .Failure to cope with the change have resulted in unprofitable practices leading to employee issues such as increased attrition rate, employee disengagement and so on(Hurst and Good,2009) Hence, the need of the hour for the executives is to effectively manage this diversified task force to cut an edge in the present competitive business world. Previous research in the field of workplace values have focused on advertising personnel, business school professors, CPA’s ,manufacturing, hospitality and insurance sectors .The objective of the present research is to explore the variations in workplace values held by the employees in Public and Private Banks representing different generational cohorts in Indian scenario . Hence our research evidence might encourage employers to titivate their HR systems to be able to effectively attract, engage and retain their best talents. An overview of the various generational cohorts and their workplace values are explained followed by an empirical analysis. The successive parts highlight the result and discussions

The pyrrolo-1,5-benzoxazepine,PBOX-6, inhibits the growth of breast cancer cells in vitro independent of estrogen receptor status and inhibits breast tumor growth in vivo

Members of a novel series of pyrrolo-1,5-benzoxazepine (PBOX) compounds have been shown to induce apoptosis in a number of human leukemia cell lines of different haematological lineage, suggesting their potential as anti-cancer agents. In this study, we sought to determine if PBOX-6, a well characterised member of the PBOX series of compounds, is also an effective inhibitor of breast cancer growth. Two estrogen receptor (ER)-positive (MCF-7 and T-47-D) and two ER-negative (MDA-MB-231 and SK-BR-3) cell lines were examined. The 3,4,5-dimethylthiazol-2-yl-2,5-diphenyl-tetrazolium bromide (MTT) assay was used to determine reduction in cell viability. PBOX-6 reduced the cell viability of all four cell lines tested, regardless of ER status, with IC(50) values ranging from 1.0 to 2.3 microM. PBOX-6 was most effective in the SK-BR-3 cells, which express high endogenous levels of the HER-2 oncogene. Overexpression of the HER-2 oncogene has been associated with aggressive disease and resistance to chemotherapy. The mechanism of PBOX-6-induced cell death was due to apoptosis, as indicated by the increased proportion of cells in the pre-G1 peak and poly(ADP-ribose) polymerase (PARP) cleavage. Moreover, intratumoural administration of PBOX-6 (7.5 mg/kg) significantly inhibited tumour growth in vivo in a mouse mammary carcinoma model (p=0.04, n=5, Student's t-test). Thus, PBOX-6 could be a promising anti-cancer agent for both hormone-dependent and -independent breast cancers.</p

Antagonistic inflammatory phenotypes dictate tumor fate and response to immune checkpoint blockade

© 2020 The Author(s). Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)Inflammation can support or restrain cancer progression and the response to therapy. Here, we searched for primary regulators of cancer-inhibitory inflammation through deep profiling of inflammatory tumor microenvironments (TMEs) linked to immune-dependent control in mice. We found that early intratumoral accumulation of interferon gamma (IFN-γ)-producing natural killer (NK) cells induced a profound remodeling of the TME and unleashed cytotoxic T cell (CTL)-mediated tumor eradication. Mechanistically, tumor-derived prostaglandin E2 (PGE2) acted selectively on EP2 and EP4 receptors on NK cells, hampered the TME switch, and enabled immune evasion. Analysis of patient datasets across human cancers revealed distinct inflammatory TME phenotypes resembling those associated with cancer immune control versus escape in mice. This allowed us to generate a gene-expression signature that integrated opposing inflammatory factors and predicted patient survival and response to immune checkpoint blockade. Our findings identify features of the tumor inflammatory milieu associated with immune control of cancer and establish a strategy to predict immunotherapy outcomes.This work was supported by a Cancer Research UK Institute Award (A19258) to S.Z. We thank colleagues at CRUK Manchester Institute core facilities, in particular, Biological Resource Unit, Transgenic Breeding, Molecular Biology, Histology, and Flow Cytometry. E.B. was supported by an EMBO long-term fellowship ( ALTF-69-2016 ) and an EMBO advanced fellowship ( aALTF-638-2018 ). C.P.B. was funded by the National Institute for Health Research Manchester Biomedical Research Centre . G.J. was supported by a scholarship from The Society of Swedish Engineers in Great Britain . K.W.-B. was supported by a doctoral studentship from the Medical Research Council . C.R.e.S. was supported by The Francis Crick Institute , which receives core funding from Cancer Research UK ( FC001136 ), the UK Medical Research Council ( FC001136 ), and the Wellcome Trust ( FC001136 ), by an ERC Advanced Investigator grant ( AdG 268670 ), by a Wellcome Investigator Award ( WT106973MA ), and by a prize from the Louis-Jeantet Foundation . N.G. was supported by an Imperial Confidence in Concept Scheme ( RSRO_P71752 ). D.M.D. was supported by a Wellcome Trust Investigator Award ( 110091/Z/15/Z ).info:eu-repo/semantics/publishedVersio