Design of modified botulinum neurotoxin A1 variants with a shorter persistence of paralysis and duration of action

Scheps, Daniel; Paz, Manuela López de la; Jurk, Marcel; Hofmann, Fred; Frevert, Jürgen

doi:10.1016/j.toxicon.2017.09.006

Cited by 12 publications

(13 citation statements)

References 37 publications

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“…Since both N-terminal and C-terminal regions of LC have been implicated in LC membrane localization ( 10 ), the LPH region may contribute to localization, but it is not anticipated to be sufficient for intracellular LC targeting to the membrane. Other factors that have been suggested to affect BoNT LC stability inside the neuronal cell include ubiquitination and deubiquitination, tyrosine phosphorylation, a dileucine motif, and a threonine (T420) in the C-terminal portion of the LC, which is conserved between subtypes A1 and A3 ( 6 , 7 , 34 – 36 ). Future experiments will be needed to determine whether there is a common underlying molecular mechanism, such as intracellular localization, or multiple mechanisms that can determine BoNT LC persistence in neuronal cells.…”

Section: Discussionmentioning

confidence: 99%

The Light Chain Defines the Duration of Action of Botulinum Toxin Serotype A Subtypes

Pellett

Bradshaw

Tepp

et al. 2018

mBio

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Botulinum neurotoxin (BoNT) is the causative agent of botulism and a widely used pharmaceutical to treat a variety of neurological diseases. BoNTs are 150-kDa protein toxins organized into heavy chain (HC) and light chain (LC) domains linked by a disulfide bond. The HC selectively binds to neurons and aids cell entry of the enzymatically active LC. There are seven immunological BoNT serotypes (A to G); each serotype includes genetic variants, termed subtypes. Only two subtypes, BoNT/A1 and BoNT/B1, are currently used as therapeutics. BoNT serotype A (BoNT/A) subtypes A2 to A8 show distinct potency, duration of action, and pathology relative to BoNT/A1. Specifically, BoNT/A3 possesses shorter duration of action and elicits distinct symptoms in mice at high toxin doses. In this report, we analyzed the roles of LC and HC of BoNT/A3 for duration of action, neuronal cell entry, and mouse pathology by using clostridium-derived recombinant hybrid BoNTs consisting of reciprocal LC and HC (BoNTA1/A3 and BoNTA3/A1). Hybrid toxins were processed in their expression host to a dichain BoNT consisting of LC and HC linked via a disulfide bond. The LC and HC defined BoNT potency in mice and BoNT toxicity for cultured neuronal cells, while the LC defined the duration of BoNT action in cell and mouse models. Protein alignment identified a previously unrecognized region within the LC subtype A3 (LC/A3) relative to the other LC serotype A (LC/A) subtypes (low primary acid homology [LPH]) that correlated to intracellular LC localization. This study shows the utility of recombinant hybrid BoNTs with new therapeutic potential, while remaining sensitive to antitoxins and therapies to native BoNT.

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Section: Discussionmentioning

confidence: 99%

The Light Chain Defines the Duration of Action of Botulinum Toxin Serotype A Subtypes

Pellett

Bradshaw

Tepp

et al. 2018

mBio

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“…Surprisingly, in this study, when the extensor digitorum brevis was injected with a combination of both BoNT/A and E, recovery was similar to that observed with BoNT/E alone; this is not consistent with the findings in a number of animal experiments, nor with the understanding that BoNT/A light chain survives in the neuronal cytosol and results in ongoing paralysis, even after the BoNT/E has worn off [ 19 , 20 , 21 ]. Short acting BoNTs, such as E and F, have potential clinical applications in therapeutic areas where a significantly shorter duration of response (3–6 weeks) when compared to that of BoNT/A (3–4 months) is required, for example, in orthopedics and rehabilitation medicine [ 22 ]. Recently, a biotechnology company, Bonti, announced that it is developing a BoNT/E product, EB-001, in aesthetic and therapeutic indications.…”

Section: Alternative Serotypes Broadening the Therapeutic Landscamentioning

confidence: 99%

The Expanding Therapeutic Utility of Botulinum Neurotoxins

Fonfría

Maignel

Lezmi

et al. 2018

Toxins

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Botulinum neurotoxin (BoNT) is a major therapeutic agent that is licensed in neurological indications, such as dystonia and spasticity. The BoNT family, which is produced in nature by clostridial bacteria, comprises several pharmacologically distinct proteins with distinct properties. In this review, we present an overview of the current therapeutic landscape and explore the diversity of BoNT proteins as future therapeutics. In recent years, novel indications have emerged in the fields of pain, migraine, overactive bladder, osteoarthritis, and wound healing. The study of biological effects distal to the injection site could provide future opportunities for disease-tailored BoNT therapies. However, there are some challenges in the pharmaceutical development of BoNTs, such as liquid and slow-release BoNT formulations; and, transdermal, transurothelial, and transepithelial delivery. Innovative approaches in the areas of formulation and delivery, together with highly sensitive analytical tools, will be key for the success of next generation BoNT clinical products.

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“…Second, an engineering approach has been taken to modify the pharmacological properties of native toxins by specific mutations. For example, a mutated BoNT/A1 has been created with faster onset and a shorter duration of action than BoNT/A1 wild type [ 60 ], opening the way to design BoNT variants with novel and useful properties.…”

Section: Intracerebral Bonts: Future Directionsmentioning

confidence: 99%

Exploiting Botulinum Neurotoxins for the Study of Brain Physiology and Pathology

Caleo

Restani

2018

Toxins

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Botulinum neurotoxins are metalloproteases that specifically cleave N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins in synaptic terminals, resulting in a potent inhibition of vesicle fusion and transmitter release. The family comprises different serotypes (BoNT/A to BoNT/G). The natural target of these toxins is represented by the neuromuscular junction, where BoNTs block acetylcholine release. In this review, we describe the actions of botulinum toxins after direct delivery to the central nervous system (CNS), where BoNTs block exocytosis of several transmitters, with near-complete silencing of neural networks. The use of clostridial neurotoxins in the CNS has allowed us to investigate specifically the role of synaptic activity in different physiological and pathological processes. The silencing properties of BoNTs can be exploited for therapeutic purposes, for example to counteract pathological hyperactivity and seizures in epileptogenic brain foci, or to investigate the role of activity in degenerative diseases like prion disease. Altogether, clostridial neurotoxins and their derivatives hold promise as powerful tools for both the basic understanding of brain function and the dissection and treatment of activity-dependent pathogenic pathways.

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Design of modified botulinum neurotoxin A1 variants with a shorter persistence of paralysis and duration of action

Cited by 12 publications

References 37 publications

The Light Chain Defines the Duration of Action of Botulinum Toxin Serotype A Subtypes

The Light Chain Defines the Duration of Action of Botulinum Toxin Serotype A Subtypes

The Expanding Therapeutic Utility of Botulinum Neurotoxins

Exploiting Botulinum Neurotoxins for the Study of Brain Physiology and Pathology

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