Extracellular Ion Channel Inhibitor Antibodies

Naylor, Jacqueline; Beech, David J

doi:10.2174/1877381800901010036

Cited by 16 publications

(18 citation statements)

References 37 publications

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“…Furthermore, we show that these fully human anti-hOrai1 mAbs reduce I CRAC currents in a recombinant stable expression system, decrease Ca 21 influx and cytokine secretion in Jurkat cells, and attenuate NFAT transcriptional activity. Our results validate our monoclonal antibody strategy for targeting diverse classes of ion channels selectively (Naylor and Beech, 2009), and further support the concept of antagonizing CRAC as a potential therapeutic approach for autoimmune disease.…”

Section: Introductionsupporting

confidence: 77%

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Generation and Characterization of Fully Human Monoclonal Antibodies Against Human Orai1 for Autoimmune Disease

Lin

Elliott

Colombero

et al. 2013

J Pharmacol Exp Ther

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Calcium entry into T cells following antigen stimulation is crucial for nuclear factor of activated T cells (NFAT)-mediated T cell activation. The movement of calcium is mediated by calcium release-activated calcium (CRAC) channels. There are two key components of this channel: Orai1 is the pore-forming subunit located in the plasma membrane, and stromal interaction molecule 1 (STIM1) functions as a Ca 21 sensor in the endoplasmic reticulum. A subset of human patients carry mutations in either STIM1 or Orai1 that affect protein function or expression, resulting in defective store-operated Ca 21 influx and CRAC channel function, and impaired T cell activation. These patients suffer from a hereditary form of severe combined immune deficiency syndrome, highlighting the importance of the CRAC channel for T lymphocyte function in humans. Since autoreactive T cells play an important role in the development of autoimmune diseases such as rheumatoid arthritis, multiple sclerosis, and organ transplantation, Orai1 becomes an attractive therapeutic target for ameliorating autoimmune disease. We developed a novel approach to inhibiting CRAC function by generating high-affinity fully human monoclonal antibodies to human Orai1. These antibodies inhibited I CRAC current, store-operated Ca 21 influx, NFAT transcription, and cytokine release. These fully human antibodies to human Orai1 may represent a novel therapeutic approach for the treatment of autoimmunity.

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Section: Introductionsupporting

confidence: 77%

“…This suggests that binding by the antibody may play a role in the conformational change of TM4, thus restoring a closed state to the channel. How this would be specifically accomplished will require (Naylor and Beech, 2009). The reported antibodies were generated to 20-mer peptides of the turret region of the ion channels in question.…”

Section: Discussionmentioning

confidence: 99%

Generation and Characterization of Fully Human Monoclonal Antibodies Against Human Orai1 for Autoimmune Disease

Lin

Elliott

Colombero

et al. 2013

J Pharmacol Exp Ther

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“…In all, these studies suggest that TRPA1 is a potential target for novel therapeutics (McMahon and Wood, 2006;Viana and FerrerMontiel, 2009;Lapointe and Altier, 2011;Strassmaier and Bakthavatchalam, 2011;Meseguer et al, 2014). Based on the published feasibility (Xu et al, 2005;Klionsky et al, 2006;Gómez-Varela et al, 2007;Naylor and Beech, 2009;Sun and Li, 2013), we attempted to generate monoclonal antagonist antibodies to TRPA1. Here, we report the generation and pharmacological characterization of monoclonal antibodies (mAbs) that antagonize cold, exogenous, and endogenous ligand activation of human TRPA1.…”

Section: Introductionmentioning

confidence: 99%

Mouse Monoclonal Antibodies to Transient Receptor Potential Ankyrin 1 Act as Antagonists of Multiple Modes of Channel Activation

Lee

Wang

Padaki

et al. 2014

J Pharmacol Exp Ther

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The transient receptor potential ankyrin 1 (TRPA1) channel has been implicated in different pathophysiologies that include asthma, cough, itch, and inflammatory pain. Agonists of TRPA1 such as mustard oil and its key component allyl isothiocyanate (AITC) cause pain and neurogenic inflammation in humans and pain behaviors in rodents. Hence, TRPA1 antagonists are being pursued as potential therapeutics. With the goal of generating monoclonal antibodies (mAbs) to human TRPA1 that could act as selective antagonists, we immunized mice with a variety of antigens expressing TRPA1 channels. After generation of hybridomas, the hybridoma conditioned media were screened to identify the mAbs that bind TRPA1 channels by a flow cytometry assay utilizing U2OS or Chinese hamster ovary (CHO) cells stably expressing TRPA1. The purified IgGs from the hybridomas that showed selective binding to TRPA1 were evaluated for antagonism in agonist-induced 45 Ca 21 uptake assays using CHO-TRPA1 cells. Several of the mAbs showed concentration-dependent inhibition of AITC and cold (4°C) activation of TRPA1. The most potent mAb, 2B10, had IC 50 values of approximately 260 and 90 nM in the two assays, respectively. These antagonist mAbs also blocked osmotically activated TRPA1 as well as activation by an endogenous agonist (4-oxo-2-nonenal). In summary, we generated mouse mAbs against TRPA1 that act as antagonists of multiple modes of TRPA1 activation.

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“…136 One strategy for generating functional ion channel antibodies has been to target the third extracellular (E3) loop of the channel (Fig. 2b) (successes and practicalities reviewed in Naylor and Beech 137 ). Xu et al 138 applied E3 targeting to the transient receptor potential (TRP) Ca 2+ channel member TRPC5 and the voltage-gated sodium channel Na V 1.5 to generate target-specific polyclonal inhibitors, while Yang et al 139 used a 14-amino acid E3 peptide from the external end of the human Kv1.3 pore region to generate a polyclonal antibody by immunization.…”

Section: Ion Channel Targetsmentioning

confidence: 99%

New Horizons in Therapeutic Antibody Discovery: Opportunities and Challenges versus Small-Molecule Therapeutics

Smith¹

2015

SLAS Discovery

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Antibody drugs have become an increasingly significant component of the therapeutic landscape. Their success has been driven by some of their unique properties, in particular their very high specificity and selectivity, in contrast to the offtarget liabilities of small molecules (SMs). Antibodies can bring additional functionality to the table with their ability to interact with the immune system, and this can be further manipulated with advances in antibody engineering. This review summarizes what antibody therapeutics have achieved to date and what opportunities and challenges lie ahead. The target landscape for large molecules (LMs) versus SMs and some of the challenges for antibody drug development are discussed. Effective penetration of membrane barriers and intracellular targeting is one challenge, particularly across the highly resistant blood-brain barrier. The expanding pipeline of antibody-drug conjugates offers the potential to combine SM and LM modalities in a variety of creative ways, and antibodies also offer exciting potential to build bi-and multispecific molecules. The ability to pursue more challenging targets can also be further exploited but highlights the need for earlier screening in functional cell-based assays. I discuss how this might be addressed given the practical constraints imposed by high-throughput screening sample type and process differences in antibody primary screening.

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Extracellular Ion Channel Inhibitor Antibodies

Cited by 16 publications

References 37 publications

Generation and Characterization of Fully Human Monoclonal Antibodies Against Human Orai1 for Autoimmune Disease

Generation and Characterization of Fully Human Monoclonal Antibodies Against Human Orai1 for Autoimmune Disease

Mouse Monoclonal Antibodies to Transient Receptor Potential Ankyrin 1 Act as Antagonists of Multiple Modes of Channel Activation

New Horizons in Therapeutic Antibody Discovery: Opportunities and Challenges versus Small-Molecule Therapeutics

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