Increased resistance to proteasome inhibitors in multiple myeloma mediated by cIAP2 overexpression - implications for a combinatorial treatment
ABSTRACT: In this study we wanted to investigate the role of cIAP2 in drug resistance, the plausible mechanisms underlying this resistance and possible strategies to overcome this by combinatorial treatment in human MM cell lines containing a TRAF3 mutation Gene expression profiling of the human MM cell line LP-1 under 6 different experimental perturbations: untreated or treated with bortezomib (10nM), 2 different timepoints (6 and 24 hours) and cIAP2 overexpression or endogenous cIAP2 level.
Project description:Multiple Myeloma (MM) is cancer in the antibody-producing plasma cells. It comprises 1 percent of all hematological malignancies. MM is incurable and fatal. The proteasome inhibitor bortezomib has improved treatment significantly, but inherent and acquired resistance remains a problem. Glutathione (GSH) is an important red-ox buffer in eukaryotic cells. In this experiment we investigate how GSH affects bortezomib-induced gene expression changes
Project description:Bortezomib is a proteasome inhibitor used in severel different hematological malignancies. Resistance to this drug is still poorly understood. In order get more insight in the resistance mechanism, we developed several bortezomib resistant subclones of the CCRF-CEM T-ALL cell line. On these subclones comparative Genome hybridization (arrayCGH) for DNA copy number analysis gene expression and micro-RNA expression arrays were performed. We performed gene expression microarray analysis on four different bortezomib resistant subclones of the CCRF-CEM cell line. The resistant subclones were compaired to treated and untreated the parental CCRF-CEM wildtype cell line.
Project description:Bortezomib is a proteasome inhibitor used in severel different hematological malignancies. Resistance to this drug is still poorly understood. In order get more insight in the resistance mechanism, we developed several bortezomib resistant subclones of the THP-1 monocytic/macrophage cell line. On these subclones expression arrays were performed. We performed expression array three different bortezomib resistant subclones of the THP-1 cell line. The resistant subclones were spotted against the parental THP-1 wildtype cell line.
Project description:Copy number and Gene expression profiling of HT-29 wild-type and bortezomib resistant cell lines Identification of mechanisms of bortezomib resistance Gene expression differences between HT-29 cell line variants
Project description:Copy number and Gene expression profiling of HT-29 wild-type and bortezomib resistant cell lines Identification of mechanisms of bortezomib resistance Copy number differences between HT-29 cell line variants and a HapMap control population of 9 female samples were identified using the Agilent 1M Human CGH microarray
Project description:Proteasome inhibitors are important chemotherapeutics in the treatment of multiple myeloma, but they are currently used empirically as no markers of sensitivity have been validated. We have identified expression of tight junction protein (TJP) 1 as being associated with sensitivity of plasma cells in vitro and in vivo to proteasome inhibitors. TJP1 suppressed expression of genes in the major histocompatibility class II region, including two catalytically active immunoproteasome subunits, thereby decreasing proteasome activity, a critical determinant of proteasome inhibitor sensitivity. This occurred through suppression by TJP1 of signaling through the epidermal growth factor receptor/Janus kinase 1/signal transducer and activator of transcription 3 pathway. In the clinic, high TJP1 expression in myeloma patients was associated with a significantly higher likelihood of responding to bortezomib, and with a longer time-to-progression after treatment. Taken together, these data support the use of TJP1 as a biomarker of sensitivity and resistance to proteasome inhibitors. To further elucidate mechanisms of bortezomib resistance, we developed human-derived multiple myeloma cell lines with a 4-fold or greater resistance to bortezomib. Then total RNA for bortezomib resistant (BR) and wild type (WT) was extracted and used for comparison by gene expression profiling.
Project description:To better understand the immunosuppressor mechanism of ptaquiloside in splenic NK cells and the reversion of this effect by selenium, we have employed whole genome microarray expression profile to identify genes associated with immunosuppression. Among 89 genes induced by ptaquiloside treatment in splenic NK cells only two genes (Mt1 and Mt2) were identified as related with its immunosuppressor effect. Moreover this augmented expression of Mt1 and Mt2 was totally abrogated by selenium co-treatment. These results were confirmed by flow cytometry in splenic cells harvested from other six mice and treated in vitro for 1 hour with ptaquiloside and/or selenium. Twenty mice were separated randomly into four groups as Control (water), Pt (ptaquiloside 5.3 mg/kg), PtSe (ptaquiloside 5.3 mg/kg and selenium 1.3 mg/kg) and Se (selenium 1.3 mg/kg) and were treated daily by gavage for 14 days. After treatment, untouched NK cells were isolated using the NK cell isolation kit, LS columns, and QuadroMACS cell separator system (Miltenyi Biotec, Inc.) to perform RNA isolation and whole-genome gene expression profile. Thereby, five independent experiments were performed per group using different donors for each experiment. To confirm the increase of metallothionein protein induced by ptaquiloside, splenic cells were harvested from other six mice and treated in vitro for 1 hour with ptaquiloside [4.4 µg/mL] and/or selenium [0.1 mM] and analyzed by flow cytometry.
Project description:We show that exposure of artificial human skin tissue to intense, picosecond-duration THz pulses affects expression levels of numerous genes associated with non-melanoma skin cancers, psoriasis and atopic dermatitis. Genes affected by intense THz pulses include nearly half of the epidermal differentiation complex (EDC) members. EDC genes, which are mapped to the chromosomal human region 1q21, encode for proteins that partake in epidermal differentiation and are often overexpressed in conditions such as psoriasis and skin cancer. In nearly all the genes differentially expressed by exposure to intense THz pulses, the induced changes in transcription levels are opposite to disease-related changes. Total RNA from exposed artificial human skin tissues to picosecond-duration broadband (0.2–2.5 THz) THz pulses with 1 kHz repetition rate, 1/e2 spot-size diameter of 1.5 mm and pulse energies of either 1.0 μJ or 0.1 μJ. For comparison, we have exposed skin tissues to UVA pulses (400 nm, 0.1 ps, 0.024 μJ).
Project description:A LysM Receptor-like Kinase Mediates Chitin Perception and Fungal Resistance in Arabidopsis; Jinrong Wan,1 Xuecheng Zhang,1 David Neece,2 Katrina M. Ramonell,3 Steve Clough,2,4 Sung-yong Kim,1 Minviluz Stacey,1 and Gary Stacey1*; 1Division of Plant Sciences, National Center for Soybean Biotechnology, C.S. Bond Life Sciences Center, University of Missouri-Columbia, Columbia, MO 65211, USA; 2Department of Crop Sciences, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA; 3Department of Biological Sciences, University of Alabama, Tuscaloosa, AL 35487, USA; 4US Department of Agriculture, Soybean/Maize Germplasm, Pathology and Genetics Research, Urbana, IL 61801, USA; *To whom correspondence should be addressed. E-mail: firstname.lastname@example.org; Abstract: Chitin, a polymer of N-acetyl-D-glucosamine, is found in fungal cell walls, but not in plants. Plant cells are capable of perceiving chitin fragments (chitooligosaccharides) to trigger various defense responses. We identified a LysM receptor-like protein (AtLysM RLK1) that is required for the perception of chitooligosaccharides in Arabidopsis. Mutation of this gene blocked the induction of almost all chitooligosaccharide-responsive genes (CRGs) and led to more susceptibility to fungal pathogens, but not to a bacterial pathogen. In addition, exogenously applied chitooligosaccharides enhanced resistance against both fungal and bacterial pathogens in the wild-type plants, but not in the mutant. Together, our data strongly suggest AtLysM RLK1 is the chitin receptor or a key part of the receptor complex and chitin is a PAMP (pathogen-associated molecular pattern) in fungi recognized by the receptor leading to the induction of plant innate immunity against fungal pathogens. Since LysM RLKs were also recently shown to be critical for the perception of the rhizobial lipo-chitin Nod signals, our data suggest that LysM RLKs not just recognize friendly symbiotic rhizobia (via their lipo-chitin Nod signals), but also hostile fungal pathogens (via their cell wall chitin). These data suggest a possible evolutionary relationship between the perception mechanisms of Nod signals and chitin by plants. Experiment Overall Design: wild type Col-0 and chitin receptor mutants treated with or without chitooctaose