Genetically engineered cellular models of prion propagation

Arshad, Hamza; Watts, Joel C.

doi:10.1007/s00441-022-03630-z

Cited by 6 publications

(5 citation statements)

References 145 publications

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“…In conclusion, overexpression of PrP C was not enough to confer differentiated ReN cultures susceptible to infection with sCJD MM1, VV2, RML or 263K prions. While we could not determine the explanation for the resistance of ReN cells to prion infection, our findings reflect the current lack of any immortalized human cell line that are permissive to infection with naturally occurring human prion isolates [ 10 ]. It remains a significant challenge to develop novel in vitro models of prion infection that are capable of replicating a range of prion strains including those in humans, are susceptible to prion-induced toxicity, and are amenable to genetic manipulations.…”

Section: Discussionmentioning

confidence: 81%

“…This is particularly true for human prion isolates that have been notoriously difficult to culture. Indeed, despite numerous efforts over the years, an immortalized neuron-like cellular model has never successfully replicated naturally occurring human prion isolates [ 10 ]. Certain cell culture systems that propagate prions are often used to examine PrP Sc trafficking, but are mostly limited to isolates of mouse-adapted scrapie and usually do not reproduce prion toxicity [ 11 , 12 ].…”

Section: Introductionmentioning

confidence: 99%

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Differentiated cultures of an immortalized human neural progenitor cell line do not replicate prions despite PrP ^C overexpression

Slota

Wang

Lusansky

et al. 2023

Prion

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Prions are misfolded proteins that accumulate within the brain in association with a rare group of fatal and infectious neurological disorders in humans and animals. A current challenge to research is a lack of in vitro model systems that are compatible with a wide range of prion strains, reproduce prion toxicity, and are amenable to genetic manipulations. In an attempt to address this need, here we produced stable cell lines that overexpress different versions of PrP C through lentiviral transduction of immortalized human neural progenitor cells (ReN VM). Differentiated cultures made from the neural progenitor cell lines overexpressed PrP C within 3D spheroid-like structures of TUBB3 + neurons and we observed evidence that PrP C modulates formation of these structures, consistent with PrP C ’s role in neurogenesis. However, through repeated measurements of amyloid seeding activity in 6-week time course experiments, we failed to observe any evidence of prion replication within the differentiated ReN cultures following challenge with four prion isolates (human sCJD subtypes MM1 and VV2, and rodent adapted scrapie strains RML and 263K). We attributed amyloid seeding activity detected within the cultures to residual inoculum and concluded that PrP C overexpression was insufficient to confer permissiveness of ReN cultures to prion infection. While our ReN cell prion infection model was unsuccessful, additional efforts to develop cellular models of human prion disease are highly warranted.

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Section: Discussionmentioning

confidence: 81%

Section: Introductionmentioning

confidence: 99%

Differentiated cultures of an immortalized human neural progenitor cell line do not replicate prions despite PrP ^C overexpression

Slota

Wang

Lusansky

et al. 2023

Prion

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“…Cultured cells have been used for many years in the field for studying the cell biology of prion propagation and testing potential anti-prion compounds. Arshad and Watts will discuss how using CRISPR/Cas9 technologies recently allowed to expand the range of prions and prion strains to study (Arshad and Watts 2022). One drawback such models usually have is that they do not provide a neurodegenerative phenotype.…”

Section: Old and New Model Systemsmentioning

confidence: 99%

New developments in prion disease research

Gilch

Schätzl

2023

Cell Tissue Res

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Prion diseases are fatal and infectious neurodegenerative disorders and prototypic conformational diseases. They are caused by the structural conversion of the cellular prion protein (PrP C ) into the pathological PrP Sc isoform. Examples are scrapie in sheep, bovine spongiform encephalopathy (BSE) in cattle, chronic wasting disease (CWD) in cervids, and Creutzfeldt-Jacob disease (CJD) in humans. The concept that a protein can constitute an infectious agent is now widely accepted. Even more, prion-like features of proteins playing central roles in the pathogenesis of other neurodegenerative diseases were described in recent years. Prions are transmissible within and sometimes between species. This was highlighted by the BSE epidemic that devastated entire economies and resulted in infections of humans and several other species. Whereas BSE was widely contained by an international package of complementary measures, another animal prion disease is currently taking center stage: CWD. Prion research traditionally uses a highly multi-disciplinary approach, from structural biology to field research. This special issue highlights recent developments and discusses progress and remaining challenges. Leading researchers describe new concepts, experimental models, and technologies that drive prion research. Reviews grouped in chapters describe recent advances in the understanding of the infectious unit and the molecular determinants of prion strains and new findings in the cell biology of prions. Experts discuss the modern diagnosis of prion diseases and its influence on the diagnosis of other neurodegenerative disorders. Finally, therapeutic targets and strategies for therapy and prophylaxis are described, although a final cure for these devastating diseases is not yet at the horizon.

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“…Cultured cell models, on the other hand, represent a more tractable system for studying prion replication as they provide the benefit of using intact cells that express membrane anchored PrP C with its correct post-translational modifications (38). CRISPR/Cas9 gene editing has expanded the range of prion strains that can be studied in cultured cells (39). For instance, CAD5 cells, which can be infected with several different strains of mouse prions (40, 41), can be ablated for endogenous MoPrP (CAD5-PrP -/- cells) and then used as a blank canvass to express PrPs from different species (42, 43).…”

Section: Introductionmentioning

confidence: 99%

The Molecular Determinants of a Universal Prion Acceptor

Arshad,

Patel,

Al-Azzawi

et al. 2024

Preprint

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In prion diseases, the species barrier limits the transmission of prions from one species to another. However, cross-species prion transmission is remarkably efficient in bank voles, and this phenomenon can be recapitulated in mice by expression of the bank vole prion protein (BVPrP). The molecular determinants of BVPrP's ability to function as a universal or near-universal acceptor for prions remain incompletely defined. Building on our finding that cultured cells expressing BVPrP can replicate both mouse and hamster prion strains, we conducted a systematic analysis to identify key residues in BVPrP that permit cross-species prion replication. Consistent with previous findings, we demonstrate that residues N155 and N170 of BVPrP, which are absent in mouse PrP but present in hamster PrP, are critical for cross-species prion replication. Additionally, BVPrP residues V112, I139, and M205, which are absent in hamster PrP but present in mouse PrP, are also required to enable replication of both mouse and hamster prions. Unexpectedly, we found that residues E227 and S230 near the C-terminus of BVPrP severely restrict the accumulation of prions following cross-species prion challenge, suggesting that they may have evolved to counteract the inherent propensity of BVPrP to misfold. PrP variants with an enhanced ability to replicate both mouse and hamster prions displayed accelerated spontaneous aggregation kinetics in vitro. These findings suggest that BVPrP's unusual properties are governed by a key set of amino acids and that the enhanced misfolding propensity of BVPrP may enable cross-species prion replication.

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Genetically engineered cellular models of prion propagation

Cited by 6 publications

References 145 publications

Differentiated cultures of an immortalized human neural progenitor cell line do not replicate prions despite PrP ^C overexpression

Differentiated cultures of an immortalized human neural progenitor cell line do not replicate prions despite PrP ^C overexpression

New developments in prion disease research

The Molecular Determinants of a Universal Prion Acceptor

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Genetically engineered cellular models of prion propagation

Cited by 6 publications

References 145 publications

Differentiated cultures of an immortalized human neural progenitor cell line do not replicate prions despite PrP C overexpression

Differentiated cultures of an immortalized human neural progenitor cell line do not replicate prions despite PrP C overexpression

New developments in prion disease research

The Molecular Determinants of a Universal Prion Acceptor

Contact Info

Product

Resources

About

Differentiated cultures of an immortalized human neural progenitor cell line do not replicate prions despite PrP ^C overexpression

Differentiated cultures of an immortalized human neural progenitor cell line do not replicate prions despite PrP ^C overexpression