Botulinum neurotoxin type C protease induces apoptosis in differentiated human neuroblastoma cells

Rust, Aleksander; Leese, Charlotte; Binz, Thomas; Davletov, Bazbek

doi:10.18632/oncotarget.8903

Cited by 25 publications

(16 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We were surprised to find that BoNT/C mutants exert different neurodegenerative effects in cultured neurons. This effect was previously attributed to cleavage of SNAP-25 and syntaxin-1A/1B, and to the complete elimination of one of them [ 40 , 41 ]. However, we found that in vitro neurodegeneration triggered by BoNT/C-wt and BoNT/C α-51 occurs even if a small portion of syntaxin-1A/1B is resistant to cleavage.…”

Section: Discussionmentioning

confidence: 93%

“…BoNT/C is long known to cause neurodegeneration in neuronal cultures [ 39 – 41 ], and this effect can be monitored by staining neurofilament-200 (NF200), a major component of axon cytoskeleton. After 12 hours of incubation, BoNT/C-wt caused a substantial loss of NF200 and the formation of varicosities, also filled with cleaved SNAP-25, all along the neurites ( Fig 2 ).…”

Section: Resultsmentioning

confidence: 99%

“…In conclusion, this work highlights that minimal changes can functionally impact BoNT biological activity, and suggests that inspection of structure-activity relationships may be used to generate tailor-made toxins with ad hoc pharmacological properties to improve the present applications and expand the clinical landscape of BoNT pharmacotherapy [ 41 , 46 , 64 , 65 ].…”

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Botulinum neurotoxin C mutants reveal different effects of syntaxin or SNAP-25 proteolysis on neuromuscular transmission

et al. 2017

Self Cite

View full text Add to dashboard Cite

Botulinum neurotoxin serotype C (BoNT/C) is a neuroparalytic toxin associated with outbreaks of animal botulism, particularly in birds, and is the only BoNT known to cleave two different SNARE proteins, SNAP-25 and syntaxin. BoNT/C was shown to be a good substitute for BoNT/A1 in human dystonia therapy because of its long lasting effects and absence of neuromuscular damage. Two triple mutants of BoNT/C, namely BoNT/C S51T/R52N/N53P (BoNT/C α-51) and BoNT/C L200W/M221W/I226W (BoNT/C α-3W), were recently reported to selectively cleave syntaxin and have been used here to evaluate the individual contribution of SNAP-25 and syntaxin cleavage to the effect of BoNT/C in vivo. Although BoNT/C α-51 and BoNT/C α-3W toxins cleave syntaxin with similar efficiency, we unexpectedly found also cleavage of SNAP-25, although to a lesser extent than wild type BoNT/C. Interestingly, the BoNT/C mutants exhibit reduced lethality compared to wild type toxin, a result that correlated with their residual activity against SNAP-25. In spite of this, a local injection of BoNT/C α-51 persistently impairs neuromuscular junction activity. This is due to an initial phase in which SNAP-25 cleavage causes a complete blockade of neurotransmission, and to a second phase of incomplete impairment ascribable to syntaxin cleavage. Together, these results indicate that neuroparalysis of BoNT/C at the neuromuscular junction is due to SNAP-25 cleavage, while the proteolysis of syntaxin provides a substantial, but incomplete, neuromuscular impairment. In light of this evidence, we discuss a possible clinical use of BoNT/C α-51 as a botulinum neurotoxin endowed with a wide safety margin and a long lasting effect.

show abstract

Section: Discussionmentioning

confidence: 93%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Botulinum neurotoxin C mutants reveal different effects of syntaxin or SNAP-25 proteolysis on neuromuscular transmission

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

“…Figure 1B shows that no cleavage was detected when the cells were incubated with concentrations as high as 30 nM BoNT/B. However, when BoNT/B entry was facilitated by Lipofectamine 3000 ( Rust et al, 2016 ), we observed dose-dependent disappearance of VAMP2, indicating that BoNT/B can cleave VAMP2 but is unable to enter N2A cells. Following BoNT/B-induced cleavage there were no VAMP2 breakdown products ( Figure 1C ) most likely due to their immediate degradation by intracellular clearance mechanisms ( Foran et al, 2003b ).…”

Section: Resultsmentioning

confidence: 91%

A Cell Line for Detection of Botulinum Neurotoxin Type B

et al. 2017

Self Cite

View full text Add to dashboard Cite

Botulinum neurotoxins (BoNTs) type A and type B are commonly used as biopharmaceutics for neurological diseases, uniquely allowing months-long paralysis of target muscles. Their exquisite neuronal specificity is conferred by a multistep process of binding, internalization, cytosolic escape and cleavage of the neuron-specific proteins, SNAP-25 and vesicle-associated membrane proteins (VAMPs), ultimately to inhibit secretion of neurotransmitters. Currently the mouse lethality bioassay is the only available method for quality control testing of VAMP-cleaving botulinum products. Refined assays for botulinum product testing are urgently needed. Specifically, in vitro replacement assays which can account for all steps of BoNT intoxication are in high demand. Here, we describe a novel SiMa cell-based approach where re-engineering of the VAMP molecule allows detection of all BoNT/B intoxication steps using a luminescent enzymatic reaction with sensitivity comparable to mouse LD50 bioassay. The presented one-step enzyme-linked immunosorbent assay meets 3Rs (replacement, reduction, and refinement of the use of animals) objectives, is user-friendly and will accelerate development of new botulinum drugs. The sensitive enzymatic reporter cell line could also be adapted for the detection of toxin activity during the manufacture of botulinum and tetanus vaccines.

show abstract

“…In three studies, investigators injected BoNT into a malignant tumor and demonstrated cellular apoptosis and reduction of tumor size [57][58][59] (Table 3). In another six studies, adding BoNT-A to cancer cell cultures reduced cell growth, induced apoptosis, and inhibited mitosis in various cancer cell lines: Prostate, breast, colon, and pancreatic tumors [60][61][62][63][64][65]. In one study, transfection of insulin secreting cells by BoNT-A reduced insulin secretion, suggesting a potential for treatment of insulinomas [66].…”

Section: The Effects Of Botulinum Neurotoxins Injections On Malignantmentioning

confidence: 99%

Botulinum Neurotoxins and Cancer—A Review of the Literature

Mittal

Jabbari

2020

Toxins

View full text Add to dashboard Cite

Botulinum neurotoxins (BoNT) possess an analgesic effect through several mechanisms including an inhibition of acetylcholine release from the neuromuscular junction as well as an inhibition of specific pain transmitters and mediators. Animal studies have shown that a peripheral injection of BoNTs impairs the release of major pain transmitters such as substance P, calcitonin gene related peptide (CGRP) and glutamate from peripheral nerve endings as well as peripheral and central neurons (dorsal root ganglia and spinal cord). These effects lead to pain relief via the reduction of peripheral and central sensitization both of which reflect important mechanisms of pain chronicity. This review provides updated information about the effect of botulinum toxin injection on local pain caused by cancer, painful muscle spasms from a remote cancer, and pain at the site of cancer surgery and radiation. The data from the literature suggests that the local injection of BoNTs improves muscle spasms caused by cancerous mass lesions and alleviates the post-operative neuropathic pain at the site of surgery and radiation. It also helps repair the parotid damage (fistula, sialocele) caused by facial surgery and radiation and improves post-parotidectomy gustatory hyperhidrosis. The limited literature that suggests adding botulinum toxins to cell culture slows/halts the growth of certain cancer cells is also reviewed and discussed.

show abstract

Botulinum neurotoxin type C protease induces apoptosis in differentiated human neuroblastoma cells

Cited by 25 publications

References 28 publications

Botulinum neurotoxin C mutants reveal different effects of syntaxin or SNAP-25 proteolysis on neuromuscular transmission

Botulinum neurotoxin C mutants reveal different effects of syntaxin or SNAP-25 proteolysis on neuromuscular transmission

A Cell Line for Detection of Botulinum Neurotoxin Type B

Botulinum Neurotoxins and Cancer—A Review of the Literature

Contact Info

Product

Resources

About