Chemical target and pathway toxicity mechanisms defined in primary human cell systems

Berg, Ellen L.; Yang, Jian; Melrose, Jennifer; Nguyen, Dat; Privat, Sylvie; Rosler, Elen S.; Kunkel, Eric J.; Ekins, Sean

doi:10.1016/j.vascn.2009.10.001

Cited by 76 publications

(93 citation statements)

References 49 publications

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“…Having demonstrated the ability of RN486 to potently block both BCR-and FcR-mediated biological responses in the tailored cellular assays for immunoreceptor signaling, we tested the compound in a panel of more complex primary human cell systems, namely the Bio-MAP Systems, that would permit us to not only further define the mechanisms of action of the compound, but also study its potential for causing off-target effects (Berg et al, 2010). The BioMAP Systems panel consists of 12 primary human monoculture or cocultures stimulated with various proinflammatory or immunomodulatory stimuli, including PBMC ϩ endothelial cell cocultures stimulated with LPS (LPS System), or superantigen (cocktail of ligands for T cell receptor) (SAg System), and PBMC ϩ B cell coculture stimulated with anti-IgM and mild TCR ligands (BT System).…”

Section: Resultsmentioning

confidence: 99%

“…To determine the profile of RN486 in more complex primary human cell systems, we had the compound tested at 0.01, 0.1, 1, and 10 M in the BioMAP Systems platform (BioSeek LLC, South San Francisco, CA) across 12 BioMAP Systems containing early passage primary human cells cultured alone or as cocultures and stimulated with various proinflammatory or immunomodulatory stimuli. These systems have been described previously (Berg et al, 2010) Resources, 1996), the Association for the Assessment and Accreditation of Laboratory Animal Care International, and American Veterinary Medical Association Guidelines on Euthanasia (2007). In vivo procedures conducted at Ricerca Biosciences, LLC (Taipei, Taiwan) were approved by its Institutional Animal Care and Use Committee and performed according to the same guidelines as those followed at Hoffmann-La Roche.…”

Section: Enzymatic Activity and Binding Assaysmentioning

confidence: 99%

“…Although still limited in their complexity compared with disease tissues, the BioMAP Systems are designed to model disease environments in vitro by exposing primary human cells, mostly in coculture, to combinations of disease-related stimuli. More importantly, these systems have been extensively studied with a broad range of therapeutics and shown to have the capacity for identifying mechanisms of drug actions and potential toxicities (Berg et al, 2006(Berg et al, , 2010. In these assays, RN486 selectively inhibited the function of B cells in a BCR and TCR ligand-stimulated B cell/PBMC coculture without affecting the function of other cell types including T cells in the same culture and macrophages stimulated with LPS.…”

Section: Btk Inhibitors For Rheumatoid Arthritis 99mentioning

confidence: 99%

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RN486, a Selective Bruton's Tyrosine Kinase Inhibitor, Abrogates Immune Hypersensitivity Responses and Arthritis in Rodents

Kim²,

Postelnek³

et al. 2012

J Pharmacol Exp Ther

127

119

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Genetic mutation and pharmacological inhibition of Bruton's tyrosine kinase (Btk) both have been shown to prevent the development of collagen-induced arthritis (CIA) in mice, providing a rationale for the development of Btk inhibitors for treating rheumatoid arthritis (RA). In the present study, we characterized a novel Btk inhibitor, 6-cyclopro-, in vitro and in rodent models of immune hypersensitivity and arthritis. We demonstrated that RN486 not only potently and selectively inhibited the Btk enzyme, but also displayed functional activities in human cell-based assays in multiple cell types, blocking Fc receptor cross-linking-induced degranulation in mast cells (IC 50 ϭ 2.9 nM), Fc␥ receptor engagement-mediated tumor necrosis factor ␣ production in monocytes (IC 50 ϭ 7.0 nM), and B cell antigen receptor-induced expression of an activation marker, CD69, in B cells in whole blood (IC 50 ϭ 21.0 nM). RN486 displayed similar functional activities in rodent models, effectively preventing type I and type III hypersensitivity responses. More importantly, RN486 produced robust anti-inflammatory and boneprotective effects in mouse CIA and rat adjuvant-induced arthritis (AIA) models. In the AIA model, RN486 inhibited both joint and systemic inflammation either alone or in combination with methotrexate, reducing both paw swelling and inflammatory markers in the blood. Together, our findings not only demonstrate that Btk plays an essential and conserved role in regulating immunoreceptor-mediated immune responses in both humans and rodents, but also provide evidence and mechanistic insights to support the development of selective Btk inhibitors as small-molecule disease-modifying drugs for RA and potentially other autoimmune diseases.

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Section: Resultsmentioning

confidence: 99%

Section: Enzymatic Activity and Binding Assaysmentioning

confidence: 99%

Section: Btk Inhibitors For Rheumatoid Arthritis 99mentioning

confidence: 99%

See 1 more Smart Citation

RN486, a Selective Bruton's Tyrosine Kinase Inhibitor, Abrogates Immune Hypersensitivity Responses and Arthritis in Rodents

Kim²,

Postelnek³

et al. 2012

J Pharmacol Exp Ther

127

119

View full text Add to dashboard Cite

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“…BioMAP assays were performed as previously described (26,27). Data for CBP30 were compared with pan-BET inhibitors (JQ1, I-BET, PFI-1, and I-BET151) (10)(11)(12)(13) …”

Section: Methodsmentioning

confidence: 99%

“…The biological activity of CBP30 was explored using a panel of 12 stimulated primary human cell types (BioMAP) (26,27). This analysis revealed broad antiinflammatory activity but no pronounced cytotoxicity at concentrations up to 10 μM.…”

Section: Cbp30 Preferentiallymentioning

confidence: 99%

CBP30, a selective CBP/p300 bromodomain inhibitor, suppresses human Th17 responses

Hammitzsch

Tallant

Fedorov

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

212

200

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Th17 responses are critical to a variety of human autoimmune diseases, and therapeutic targeting with monoclonal antibodies against IL-17 and IL-23 has shown considerable promise. Here, we report data to support selective bromodomain blockade of the transcriptional coactivators CBP (CREB binding protein) and p300 as an alternative approach to inhibit human Th17 responses. We show that CBP30 has marked molecular specificity for the bromodomains of CBP and p300, compared with 43 other bromodomains. In unbiased cellular testing on a diverse panel of cultured primary human cells, CBP30 reduced immune cell production of IL-17A and other proinflammatory cytokines. CBP30 also inhibited IL-17A secretion by Th17 cells from healthy donors and patients with ankylosing spondylitis and psoriatic arthritis. Transcriptional profiling of human T cells after CBP30 treatment showed a much more restricted effect on gene expression than that observed with the pan-BET (bromo and extraterminal domain protein family) bromodomain inhibitor JQ1. This selective targeting of the CBP/p300 bromodomain by CBP30 will potentially lead to fewer side effects than with the broadly acting epigenetic inhibitors currently in clinical trials.CBP/p300 | bromodomain | epigenetic inhibitors | Th17 | ankylosing spondylitis

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ToxCast: Predicting Toxicity Potential Through High‐Throughput Bioactivity Profiling

Houck

Richard

Judson

et al. 2013

High‐Throughput Screening Methods in Toxicity Testing

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Chemical target and pathway toxicity mechanisms defined in primary human cell systems

Cited by 76 publications

References 49 publications

RN486, a Selective Bruton's Tyrosine Kinase Inhibitor, Abrogates Immune Hypersensitivity Responses and Arthritis in Rodents

RN486, a Selective Bruton's Tyrosine Kinase Inhibitor, Abrogates Immune Hypersensitivity Responses and Arthritis in Rodents

CBP30, a selective CBP/p300 bromodomain inhibitor, suppresses human Th17 responses

ToxCast: Predicting Toxicity Potential Through High‐Throughput Bioactivity Profiling

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