Design and Optimization of Selective Protein Kinase C θ (PKCθ) Inhibitors for the Treatment of Autoimmune Diseases

Jiménez, Juan-Miguel; Boyall, Dean; Brenchley, Guy; Collier, Philip N.; Davis, Christopher J.; Fraysse, Damien; Keily, Shazia B.; Henderson, Jaclyn L.; Miller, Andrew T.; Pierard, Françoise Y. T. M.; Settimo, Luca; Twin, Heather; Bolton, Claire; Curnock, Adam P.; Chiu, Peter; Tanner, Adam; Young, Stephen

doi:10.1021/jm301465a

Cited by 41 publications

(37 citation statements)

References 45 publications

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“…7b), which lack transcriptional memory (unpublished observation). We found that the PKC inhibitor C2737 abrogated the enhanced protein levels of NFAT in the nucleus on RS (Fig. 7c).…”

Section: Resultsmentioning

confidence: 69%

Priming of transcriptional memory responses via the chromatin accessibility landscape in T cells

Hardy

Sutton

et al. 2017

Sci Rep

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Memory T cells exhibit transcriptional memory and “remember” their previous pathogenic encounter to increase transcription on re-infection. However, how this transcriptional priming response is regulated is unknown. Here we performed global FAIRE-seq profiling of chromatin accessibility in a human T cell transcriptional memory model. Primary activation induced persistent accessibility changes, and secondary activation induced secondary-specific opening of previously less accessible regions associated with enhanced expression of memory-responsive genes. Increased accessibility occurred largely in distal regulatory regions and was associated with increased histone acetylation and relative H3.3 deposition. The enhanced re-stimulation response was linked to the strength of initial PKC-induced signalling, and PKC-sensitive increases in accessibility upon initial stimulation showed higher accessibility on re-stimulation. While accessibility maintenance was associated with ETS-1, accessibility at re-stimulation-specific regions was linked to NFAT, especially in combination with ETS-1, EGR, GATA, NFκB, and NR4A. Furthermore, NFATC1 was directly regulated by ETS-1 at an enhancer region. In contrast to the factors that increased accessibility, signalling from bHLH and ZEB family members enhanced decreased accessibility upon re-stimulation. Interplay between distal regulatory elements, accessibility, and the combined action of sequence-specific transcription factors allows transcriptional memory-responsive genes to “remember” their initial environmental encounter.

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“…7b), which lack transcriptional memory (unpublished observation). We found that the PKC inhibitor C2737 abrogated the enhanced protein levels of NFAT in the nucleus on RS (Fig. 7c).…”

Section: Resultsmentioning

confidence: 69%

Priming of transcriptional memory responses via the chromatin accessibility landscape in T cells

Hardy

Sutton

et al. 2017

Sci Rep

View full text Add to dashboard Cite

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“…S5A and S5B in the supplemental material). Given that previous studies in the context of the immune system have established that PKC-signals to and activates NF-B family members (31, 72), we next addressed the impact of PKC-on NF-B activity in our system by utilizing a novel, highly selective ATP-competitive PKC-specific inhibitor, compound 27 (C27) (75). As anticipated, C27 abrogated PKCactivity both in our in vitro PKC-activity assay (Fig.…”

Section: Pkc-activity Is Required For Chromatin Accessibility Of Key mentioning

confidence: 82%

Chromatinized Protein Kinase C-θ Directly Regulates Inducible Genes in Epithelial to Mesenchymal Transition and Breast Cancer Stem Cells

Zafar

Hardy

et al. 2014

Molecular and Cellular Biology

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g Epithelial to mesenchymal transition (EMT) is activated during cancer invasion and metastasis, enriches for cancer stem cells (CSCs), and contributes to therapeutic resistance and disease recurrence. Signal transduction kinases play a pivotal role as chromatin-anchored proteins in eukaryotes. Here we report for the first time that protein kinase C-theta (PKC-) promotes EMT by acting as a critical chromatin-anchored switch for inducible genes via transforming growth factor ␤ (TGF-␤) and the key inflammatory regulatory protein NF-B. Chromatinized PKC-exists as an active transcription complex and is required to establish a permissive chromatin state at signature EMT genes. Genome-wide analysis identifies a unique cohort of inducible PKC--sensitive genes that are directly tethered to PKC-in the mesenchymal state. Collectively, we show that cross talk between signaling kinases and chromatin is critical for eliciting inducible transcriptional programs that drive mesenchymal differentiation and CSC formation, providing novel mechanisms to target using epigenetic therapy in breast cancer. Epithelial to mesenchymal transition (EMT) is a key step in cancer progression and the process of metastasis that creates a reservoir for cancer stem cells (CSCs) and is associated with highly aggressive traits (1-4). As well as driving metastasis, these CSCs, or "precursor" metastatic cells, play a pivotal role in therapeutic resistance and relapse in breast cancer patients (5, 6). Breast CSCs are a distinct subpopulation of mesenchymal cells that possess several important features, namely, expression of key surface markers (CD44 high and CD24 low ) (7), a distinct transcriptome (3), the ability to form spherical colonies in suspension cultures (termed mammospheres) (8), and enhanced resistance to chemotherapy (9, 10) and ionizing radiation (11)(12)(13)(14).Eukaryotes utilize the chromatin landscape as its epigenetic template within the nucleus of living cells in order to promote inducible gene transcription in response to environmental signals. Highly compacted chromatin structures are enriched in nucleosomes and are transcriptionally silent, and a net loss of nucleosomes from gene-specific regulatory regions increases chromatin accessibility and initiates context-specific transcriptional programs. In addition to ATP-dependent chromatin remodeling and exchange of histone variants with canonical histones, histone modifications are thought to alter gene expression, by changing chromatin structure and/or by providing a platform that promotes binding of transcriptional regulators (15-23).Novel classes of chromatin-associated enzymes that play critical roles in modulating chromatin structure within the human genome have recently been discovered. In particular, signaling kinases can act as chromatin regulators of inducible gene transcription in both higher and lower eukaryotes by two distinct mechanisms: relaying signals from the cytoplasm to the nucleus and direct association with chromatin-bound transcription complexes at activated tar...

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“…Taken together, these evidences validate PKCθ as a particularly attractive target to selectively manipulate T eff cell functions that are relevant to pathogenesis of different diseases, including, as our results suggested, muscular dystrophy. Noteworthy, a large effort is now devoted to the synthesis of pharmacological PKCθ inhibitors, to be used for anti-inflammatory therapies, and several PKCθ inhibitors have been identified and reported in recent literature (Boschelli et al, 2010, Chand et al, 2012, Curnock et al, 2014, Cywin et al, 2007, George et al, 2015, Hage-Sleiman et al, 2015, Jimenez et al, 2013, Katoh et al, 2016). Some of them bind selectively PKCθ while others are non-selective in nature.…”

Section: Introductionmentioning

confidence: 99%

Pharmacological Inhibition of PKCθ Counteracts Muscle Disease in a Mouse Model of Duchenne Muscular Dystrophy

et al. 2017

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Inflammation plays a considerable role in the progression of Duchenne Muscular Dystrophy (DMD), a severe muscle disease caused by a mutation in the dystrophin gene. We previously showed that genetic ablation of Protein Kinase C θ (PKCθ) in mdx, the mouse model of DMD, improves muscle healing and regeneration, preventing massive inflammation. To establish whether pharmacological targeting of PKCθ in DMD can be proposed as a therapeutic option, in this study we treated young mdx mice with the PKCθ inhibitor Compound 20 (C20). We show that C20 treatment led to a significant reduction in muscle damage associated with reduced immune cells infiltration, reduced inflammatory pathways activation, and maintained muscle regeneration. Importantly, C20 treatment is efficient in recovering muscle performance in mdx mice, by preserving muscle integrity. Together, these results provide proof of principle that pharmacological inhibition of PKCθ in DMD can be considered an attractive strategy to modulate immune response and prevent the progression of the disease.Research in contextDuchenne muscular dystrophy (DMD) is a severe muscle disease affecting 1:3500 male births. DMD is caused by a mutation in dystrophin gene, coding for a protein required for skeletal and cardiac muscle integrity. Lack of a functional dystrophin is primarily responsible for the muscle eccentric contraction-induced muscle damage, observed in dystrophic muscle. However, inflammation plays a considerable role in the progression of DMD. Glucocorticoids, which have anti-inflammatory properties, are being used to treat DMD with some success; however, long term treatment with these drugs induces muscle atrophy and wasting, outweighing their benefit. The identification of specific targets for anti-inflammatory therapies is one of the ongoing therapeutic options. Although blunting inflammation would not be a “cure” for the disease, the emerging clue is that multiple strategies, addressing different aspects of the pathology, which may eventually converge, may be successful. In this context, we previously showed that genetic ablation of Protein Kinase C θ (PKCθ), an enzyme known to be involved in immune response, in mdx, the mouse model of DMD, improves muscle healing and regeneration, preventing massive inflammation. To establish whether pharmacological targeting of PKCθ in DMD can be proposed as a therapeutic option, in this study we treated young mdx mice with the PKCθ inhibitor Compound 20 (C20). We show that C20 treatment led to a significant reduction in muscle damage associated with reduced immune cells infiltration, reduced inflammatory pathways activation, and maintained muscle regeneration. Importantly, C20 treatment is efficient in recovering muscle performance in mdx mice, by preserving muscle integrity. Together, these results provide proof of principle that pharmacological inhibition of PKCθ in DMD can be considered an attractive strategy to modulate immune response and prevent the progression of the disease.

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Design and Optimization of Selective Protein Kinase C θ (PKCθ) Inhibitors for the Treatment of Autoimmune Diseases

Cited by 41 publications

References 45 publications

Priming of transcriptional memory responses via the chromatin accessibility landscape in T cells

Priming of transcriptional memory responses via the chromatin accessibility landscape in T cells

Chromatinized Protein Kinase C-θ Directly Regulates Inducible Genes in Epithelial to Mesenchymal Transition and Breast Cancer Stem Cells

Pharmacological Inhibition of PKCθ Counteracts Muscle Disease in a Mouse Model of Duchenne Muscular Dystrophy

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