Chronic wasting disease prion infection of differentiated neurospheres

Iwamaru, Yoshifumi; Mathiason, Candace K.; Telling, Glenn C.; Hoover, Edward A.

doi:10.1080/19336896.2017.1336273

Cited by 8 publications

(5 citation statements)

References 16 publications

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“…Later analysis showed that these cells are most likely brain tissue-derived fibroblasts. More recently, differentiated neurospheres from elk PrP-overexpressing mice were shown to be susceptible to infection with CWD prions 49 . Although not involving single cell cloning, this is a fairly complex strategy that requires growing primary cultures from transgenic mice and hence limits its applicability.…”

Section: Discussionmentioning

confidence: 99%

Gene-edited murine cell lines for propagation of chronic wasting disease prions

Walia

Lee

et al. 2019

Sci Rep

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Prions cause fatal infectious neurodegenerative diseases in humans and animals. Cell culture models are essential for studying the molecular biology of prion propagation. Defining such culture models is mostly a random process, includes extensive subcloning, and for many prion diseases few or no models exist. One example is chronic wasting disease (CWD), a highly contagious prion disease of cervids. To extend the range of cell models propagating CWD prions, we gene-edited mouse cell lines known to efficiently propagate murine prions. Endogenous prion protein (PrP) was ablated in CAD5 and MEF cells, using CRISPR-Cas9 editing. PrP knock-out cells were reconstituted with mouse, bank vole and cervid PrP genes by lentiviral transduction. Reconstituted cells expressing mouse PrP provided proof-of-concept for re-established prion infection. Bank voles are considered universal receptors for prions from a variety of species. Bank vole PrP reconstituted cells propagated mouse prions and cervid prions, even without subcloning for highly susceptible cells. Cells reconstituted with cervid PrP and infected with CWD prions tested positive in prion conversion assay, whereas non-reconstituted cells were negative. This novel cell culture platform which is easily adjustable and allows testing of polymorphic alleles will provide important new insights into the biology of CWD prions.

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Section: Discussionmentioning

confidence: 99%

Gene-edited murine cell lines for propagation of chronic wasting disease prions

Walia

Lee

et al. 2019

Sci Rep

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“…The development of ex vivo models allowed relatively fast detection of prions, including low-titer prion infectivity, with partial recapitulation of prion pathogenesis and ability to test anti-prion compounds [ 106 , 107 ]. Differentiated neurospheres from Tg(ElkPrP)5037 +/– mice overexpressing elk PrP-amplified CWD prions successfully within three weeks post-infection [ 108 ]. The prion organotypic slice culture assay (POSCA) was developed by the Aguzzi group in 2008 [ 109 ].…”

Section: Availability Of Models For Studying Cwd Prionsmentioning

confidence: 99%

Gene-Edited Cell Models to Study Chronic Wasting Disease

Thapa

Winkens

Tahir

et al. 2022

Viruses

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Prion diseases are fatal infectious neurodegenerative disorders affecting both humans and animals. They are caused by the misfolded isoform of the cellular prion protein (PrPC), PrPSc, and currently no options exist to prevent or cure prion diseases. Chronic wasting disease (CWD) in deer, elk and other cervids is considered the most contagious prion disease, with extensive shedding of infectivity into the environment. Cell culture models provide a versatile platform for convenient quantification of prions, for studying the molecular and cellular biology of prions, and for performing high-throughput screening of potential therapeutic compounds. Unfortunately, only a very limited number of cell lines are available that facilitate robust and persistent propagation of CWD prions. Gene-editing using programmable nucleases (e.g., CRISPR-Cas9 (CC9)) has proven to be a valuable tool for high precision site-specific gene modification, including gene deletion, insertion, and replacement. CC9-based gene editing was used recently for replacing the PrP gene in mouse and cell culture models, as efficient prion propagation usually requires matching sequence homology between infecting prions and prion protein in the recipient host. As expected, such gene-editing proved to be useful for developing CWD models. Several transgenic mouse models were available that propagate CWD prions effectively, however, mostly fail to reproduce CWD pathogenesis as found in the cervid host, including CWD prion shedding. This is different for the few currently available knock-in mouse models that seem to do so. In this review, we discuss the available in vitro and in vivo models of CWD, and the impact of gene-editing strategies.

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“…As expected from previous studies with stem cells, neurospheres were found to express higher levels of PrP once differentiated [ 97 ]. Recently, neurospheres from tg5037 mice expressing elk PrP were shown to be capable of propagating PrP Sc from both elk and deer CWD inoculum [ 99 ]. Neurospheres are therefore infectable with a wide array of prions of different strains and species.…”

Section: Neurospheresmentioning

confidence: 99%

“…As with primary cultures, neurospheres infected with Chandler, ME7, or 22L prions exhibit cytopathic changes, including increased membrane permeability, death of astrocytes, and detachment of from the coverslips that they were grown on. The use of a pan-caspase inhibitor was effective in mitigating these changes [ 99 ].…”

Section: Neurospheresmentioning

confidence: 99%

“…The use of specific types of media and growth factors can also have a large effect on PrP Sc levels. Specifically, Iwamaru et al, found that neurospheres cultured in bFGF had much more PrP Sc deposition as compared with those grown in FBS [ 99 ]. Kocisko et al, found that Rov9 cells cultured in optimum produced four times the PrP Sc of those cultured in MEM [ 20 ].…”

Section: Enhancing Prion Propagation In Model Systemsmentioning

confidence: 99%

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From Cell Culture to Organoids-Model Systems for Investigating Prion Strain Characteristics

Pineau

Sim

2021

Biomolecules

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Prion diseases are the hallmark protein folding neurodegenerative disease. Their transmissible nature has allowed for the development of many different cellular models of disease where prion propagation and sometimes pathology can be induced. This review examines the range of simple cell cultures to more complex neurospheres, organoid, and organotypic slice cultures that have been used to study prion disease pathogenesis and to test therapeutics. We highlight the advantages and disadvantages of each system, giving special consideration to the importance of strains when choosing a model and when interpreting results, as not all systems propagate all strains, and in some cases, the technique used, or treatment applied, can alter the very strain properties being studied.

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Chronic wasting disease prion infection of differentiated neurospheres

Cited by 8 publications

References 16 publications

Gene-edited murine cell lines for propagation of chronic wasting disease prions

Gene-edited murine cell lines for propagation of chronic wasting disease prions

Gene-Edited Cell Models to Study Chronic Wasting Disease

From Cell Culture to Organoids-Model Systems for Investigating Prion Strain Characteristics

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