Characterization of a membrane binding loop leads to engineering botulinum neurotoxin B with improved therapeutic efficacy

Yin, Linxiang; Masuyer, Geoffrey; Zhang, Sicai; Zhang, Jie; Miyashita, Shin-Ichiro; Burgin, David; Lovelock, Laura; Coker, Shu-Fen; Fu, Tian‐Min; Stenmark, Pål; Dong, Min

doi:10.1371/journal.pbio.3000618

Cited by 19 publications

(16 citation statements)

References 86 publications

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“…Murine synaptotagmin I had a 10-fold lower affinity, but still a higher affinity than human synaptotagmin I. Consequently, within 48 h, no detectable amounts of BoNT/B were taken up by the SIMA reporter cells (Figure 4) that expressed both synaptotagmin II and, to a larger extent, synaptotagmin I (Figure 3). By contrast, a mutant BoNT/B, in which two amino acids had been replaced to increase the binding to human synaptotagmin I and II named BoNT/B-MY [22,25] inhibited the stimulus-dependent reporter release with a similar IC50 as BoNT/A (Figure 2) and was readily taken up into the SIMA reporter cells (Figure 4). Thus, in contrast to in vitro or in vivo assays that determine the inhibition of muscle contraction of wild-type mice [26] which react to BoNT/B and BoNT/B-MY with similar sensitivity, the SIMA reporter cell assay distinguished between the low potency of wild-type BoNT/B and the higher potency of BoNT/B-MY, and hence, more closely reflected the human-relevant potencies.…”

Section: Discussionmentioning

confidence: 99%

Discrimination of the Activity of Low-Affinity Wild-Type and High-Affinity Mutant Recombinant BoNT/B by a SIMA Cell-Based Reporter Release Assay

Neuschäfer-Rube

Pathe-Neuschäfer-Rube

Püschel

2022

Toxins

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Botulinum neurotoxin (BoNT) is used for the treatment of a number of ailments. The activity of the toxin that is isolated from bacterial cultures is frequently tested in the mouse lethality assay. Apart from the ethical concerns inherent to this assay, species-specific differences in the affinity for different BoNT serotypes give rise to activity results that differ from the activity in humans. Thus, BoNT/B is more active in mice than in humans. The current study shows that the stimulus-dependent release of a luciferase from a differentiated human neuroblastoma–based reporter cell line (SIMA-hPOMC1-26-Gluc) was inhibited by clostridial and recombinant BoNT/A to the same extent, whereas both clostridial and recombinant BoNT/B inhibited the release to a lesser extent and only at much higher concentrations, reflecting the low activity of BoNT/B in humans. By contrast, the genetically modified BoNT/B-MY, which has increased affinity for human synaptotagmin, and the BoNT/B protein receptor inhibited luciferase release effectively and with an EC50 comparable to recombinant BoNT/A. This was due to an enhanced uptake into the reporter cells of BoNT/B-MY in comparison to the recombinant wild-type toxin. Thus, the SIMA-hPOMC1-26-Gluc cell assay is a versatile tool to determine the activity of different BoNT serotypes providing human-relevant dose-response data.

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Section: Discussionmentioning

confidence: 99%

Discrimination of the Activity of Low-Affinity Wild-Type and High-Affinity Mutant Recombinant BoNT/B by a SIMA Cell-Based Reporter Release Assay

Neuschäfer-Rube

Pathe-Neuschäfer-Rube

Püschel

2022

Toxins

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“…Similarly, results from an in vitro binding study with synaptosomes indicate that RTP004 increased maximal percentage binding of BoNTA heavy chain to rat brain nerve terminals [ 27 ]. Enhanced binding to nerve terminals and extracellular matrix elements may facilitate the localization of BoNTA and reduce diffusion from the injection site [ 28 ], a finding that was observed in a study of DAXI and onabotulinumtoxinA in mice [ 18 ]. In that study, a DAXI dose of 2.5 times that of the onabotulinumtoxinA dose was required to see comparable diffusion.…”

Section: Background On Daximentioning

confidence: 99%

Overview of DaxibotulinumtoxinA for Injection: A Novel Formulation of Botulinum Toxin Type A

Solish

Carruthers

Kaufman³

et al. 2021

Drugs

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Botulinum toxin type A (BoNTA) products are widely used for therapeutic and aesthetic indications, but there is a need for longer-lasting treatments that maintain symptom relief between injections and reduce the frequency of re-treatment. DaxibotulinumtoxinA for Injection (DAXI) is a novel BoNTA product containing highly purified 150-kDa core neurotoxin and is the first to be formulated with a proprietary stabilizing excipient peptide (RTP004) instead of human serum albumin. The positively charged RTP004 has been shown to enhance binding of the neurotoxin to neuronal surfaces, which may enhance the likelihood of neurotoxin internalization. DAXI produces robust, extended efficacy across both aesthetic and therapeutic indications. In an extensive glabellar lines clinical program, DAXI showed a high degree of efficacy, a consistent median time to loss of none or mild glabellar line severity of 24 weeks, and median time until return to baseline of up to 28 weeks. In adults with cervical dystonia, DAXI at 125 U and 250 U significantly improved Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS) total scores, with a median duration of efficacy of 24 and 20 weeks, respectively, which compares favorably with the 12–14 weeks’ duration reported for approved BoNTA products. Overall, DAXI was well tolerated, and the consistent extended duration of effect suggests that DAXI has the potential to improve the management of both aesthetic and therapeutic conditions.

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“…For instance, Yin et al generated a "gain-of-function" mutation by replacing two non-aromatic residues at an extended loop in the C-terminal receptor-binding domain of BoNT/B (85). This engineering BoNT/B showed enhanced binding to neuronal membrane, enhanced efficacy in paralyzing muscles, and lowered systemic diffusion (85). In addition, BoNTs were designed as novel analgesic by utilizing the ability of BoNT to cleave SNARE complexes (83).…”

Section: Future Perspectivementioning

confidence: 99%

Botulinum Neurotoxin Therapy for Depression: Therapeutic Mechanisms and Future Perspective

Liu

Luo

2021

Front. Psychiatry

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Depression is one of the most common mental disorders, which causes global burden. Antidepressants and psychotherapies are the mainstay of treatment for depression, which have limited efficacy. Thus, alternative approaches for preventing and treating depression are urgently required. Recent clinical trials and preclinical researches have clarified that peripheral facial injection of botulinum neurotoxin type A (BoNT/A) is a rapid, effective and relative safe therapy for improving some symptoms of depression. Despite its safety and efficacy, the underlying therapeutic mechanisms of BoNT/A for depression remains largely unclear. In the present review, we updated and summarized the clinical and preclinical evidence supporting BoNT/A therapy for the treatment of depression. We further discussed the potential mechanisms underlying therapeutic effects of BoNT/A on depression. Notably, we recently identified that the anti-depressant effects of BoNT/A associated with up-regulation of 5-HT levels and brain-derived neurotrophic factor (BDNF) expression in the hippocampus in a preclinical mouse model. In summary, these studies suggest that BoNT/A therapy is a potential effective and safe intervention for the management of depression. However, fundamental questions remain regarding the future prospects of BoNT/A therapy, including safety, efficacy, dose-response relationships, identification of potential predictors of response, and the precise mechanisms underlying BoNT/A therapy.

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Characterization of a membrane binding loop leads to engineering botulinum neurotoxin B with improved therapeutic efficacy

Cited by 19 publications

References 86 publications

Discrimination of the Activity of Low-Affinity Wild-Type and High-Affinity Mutant Recombinant BoNT/B by a SIMA Cell-Based Reporter Release Assay

Discrimination of the Activity of Low-Affinity Wild-Type and High-Affinity Mutant Recombinant BoNT/B by a SIMA Cell-Based Reporter Release Assay

Overview of DaxibotulinumtoxinA for Injection: A Novel Formulation of Botulinum Toxin Type A

Botulinum Neurotoxin Therapy for Depression: Therapeutic Mechanisms and Future Perspective

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