The aminoglycoside G418 hinders de novo prion infection in cultured cells

Arshad, Hamza; Patel, Zeel; Mehrabian, Mohadeseh; Bourkas, Matthew E.C.; Al-Azzawi, Zaid A.M.; Schmitt‐Ulms, Gerold; Watts, Joel C.

doi:10.1016/j.jbc.2021.101073

Cited by 9 publications

(15 citation statements)

References 88 publications

(105 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To observe a 50% difference in sample means with a power of 0.80 and an α‐value of 0.05, a power calculation reveals that a minimum sample size of 3 is required, assuming an average standard deviation of 20%. Thus, a minimum of 3–4 independent replicates per condition were utilized, which is consistent with our previous studies performed using similar experimental paradigms (Arshad et al, 2021; Bourkas et al, 2019).…”

Section: Methodssupporting

confidence: 74%

“…Untagged recombinant BVPrP (residues 23–231) containing either the M109 or I109 polymorphism, as well as their counterparts containing the G127V substitution, were expressed and purified in E. coli Rosetta2(DE3) cells as previously described for recombinant hamster PrP(23‐231) (Bourkas et al, 2019). Recombinant mouse PrP(23‐230) was produced similarly (Arshad et al, 2021). Purified recombinant PrP was stored at −80℃ in 10 mM sodium phosphate buffer, pH 5.8.…”

Section: Methodsmentioning

confidence: 99%

“…The plate was then placed in a BMG CLARIOstar plate reader and was incubated at 37℃ for 83.25 h with cycles of shaking (1 min) and rest (4 min). ThT readings (excitation 444 ± 5 nm; emission: 485 ± 5 nm) were taken every 5 minutes, and the lag phases were quantified as previously described (Arshad et al, 2021). Samples that failed to aggregate were assigned a lag phase of 83.25 h.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

A single protective polymorphism in the prion protein blocks cross‐species prion replication in cultured cells

Arshad

Patel

Amano

et al. 2022

Journal of Neurochemistry

Self Cite

View full text Add to dashboard Cite

Prions are infectious protein aggregates that cause a variety of fatal neurodegenerative conditions in humans and animals, collectively referred to as prion diseases (Colby & Prusiner, 2011;Scheckel & Aguzzi, 2018;Watts et al., 2006). The naturally occurring prion diseases include scrapie in sheep, chronic wasting disease in cervids, as well as Creutzfeldt-Jakob disease, Gerstmann-Sträussler-Scheinker disease, fatal familial insomnia, and kuru in humans. The infectious proteins underlying these diseases are composed of an endogenously expressed protein called the cellular prion protein (PrP C ) (Oesch

show abstract

Section: Methodssupporting

confidence: 74%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

A single protective polymorphism in the prion protein blocks cross‐species prion replication in cultured cells

Arshad

Patel

Amano

et al. 2022

Journal of Neurochemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…Because of its potential therapeutic implications, the most intriguing finding in this study was the observation that PrP C levels in a human co-culture model of neurons and astrocytes can be reduced by more than 50% when cells are exposed to non-toxic levels of CGs, a class of compounds that is well-characterized in its clinical use following decades of research and use in patients with heart disease. Note that the mechanism through which CGs cause this diminution of steady-state PrP C levels appears unrelated to the way another aminoglycoside, G418 (geneticin), impairs the de novo infection of cells with prions in the absence of an influence on PrP C levels or its localization [68].…”

Section: Plos Onementioning

confidence: 99%

Cardiac glycoside-mediated turnover of Na, K-ATPases as a rational approach to reducing cell surface levels of the cellular prion protein

et al. 2022

Self Cite

View full text Add to dashboard Cite

It is widely anticipated that a reduction of brain levels of the cellular prion protein (PrPC) can prolong survival in a group of neurodegenerative diseases known as prion diseases. To date, efforts to decrease steady-state PrPC levels by targeting this protein directly with small molecule drug-like compounds have largely been unsuccessful. Recently, we reported Na,K-ATPases to reside in immediate proximity to PrPC in the brain, unlocking an opportunity for an indirect PrPC targeting approach that capitalizes on the availability of potent cardiac glycosides (CGs). Here, we report that exposure of human co-cultures of neurons and astrocytes to non-toxic nanomolar levels of CGs causes profound reductions in PrPC levels. The mechanism of action underpinning this outcome relies primarily on a subset of CGs engaging the ATP1A1 isoform, one of three α subunits of Na,K-ATPases expressed in brain cells. Upon CG docking to ATP1A1, the ligand receptor complex, and PrPC along with it, is internalized by the cell. Subsequently, PrPC is channeled to the lysosomal compartment where it is digested in a manner that can be rescued by silencing the cysteine protease cathepsin B. These data signify that the repurposing of CGs may be beneficial for the treatment of prion disorders.

show abstract

“…While, another derivative of AmB, MS-8209, failed to show a direct effect on PrP Sc [ 208 ]. Recently, aminoglycoside G418 (Geneticin) has been reported to interfere with the de novo processes involved in earlier stages of prion infection, without affecting the level or localization of PrP [ 209 ].…”

Section: Antibioticsmentioning

confidence: 99%

Prion therapeutics: Lessons from the past

Shim

Sharma

2022

Prion

View full text Add to dashboard Cite

Prion diseases are a group of incurable zoonotic neurodegenerative diseases (NDDs) in humans and other animals caused by the prion proteins. The abnormal folding and aggregation of the soluble cellular prion proteins (PrP C ) into scrapie isoform (PrP Sc ) in the Central nervous system (CNS) resulted in brain damage and other neurological symptoms. Different therapeutic approaches, including stalling PrP C to PrP Sc conversion, increasing PrP Sc removal, and PrP C stabilization, for which a spectrum of compounds, ranging from organic compounds to antibodies, have been explored. Additionally, a non-PrP targeted drug strategy using serpin inhibitors has been discussed. Despite numerous scaffolds being screened for anti-prion activity in vitro , only a few were effective in vivo and unfortunately, almost none of them proved effective in the clinical studies, most likely due to toxicity and lack of permeability. Recently, encouraging results from a prion-protein monoclonal antibody, PRN100, were presented in the first human trial on CJD patients, which gives a hope for better future for the discovery of other new molecules to treat prion diseases. In this comprehensive review, we have re-visited the history and discussed various classes of anti-prion agents, their structure, mode of action, and toxicity. Understanding pathogenesis would be vital for developing future treatments for prion diseases. Based on the outcomes of existing therapies, new anti-prion agents could be identified/synthesized/designed with reduced toxicity and increased bioavailability, which could probably be effective in treating prion diseases.

show abstract

The aminoglycoside G418 hinders de novo prion infection in cultured cells

Cited by 9 publications

References 88 publications

A single protective polymorphism in the prion protein blocks cross‐species prion replication in cultured cells

A single protective polymorphism in the prion protein blocks cross‐species prion replication in cultured cells

Cardiac glycoside-mediated turnover of Na, K-ATPases as a rational approach to reducing cell surface levels of the cellular prion protein

Prion therapeutics: Lessons from the past

Contact Info

Product

Resources

About