Distinct neuronal alterations distinguish two subtypes of sporadic Creutzfeldt-Jakob disease with shared dysfunctional pathways
Katie Williams, Bradley R. Groveman, Simote T. Foliaki, Brent Race, Arielle Hay, Ryan O. Walters, Tina Thomas, Gianluigi Zanusso, James A. Carroll, Cathryn L. Haigh
Katie Williams, Bradley R. Groveman, Simote T. Foliaki, Brent Race, Arielle Hay, Ryan O. Walters, Tina Thomas, Gianluigi Zanusso, James A. Carroll, Cathryn L. Haigh
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Research Article Cell biology Infectious disease Neuroscience

Distinct neuronal alterations distinguish two subtypes of sporadic Creutzfeldt-Jakob disease with shared dysfunctional pathways

Abstract

Prion diseases are a family of transmissible, neurodegenerative conditions caused by misfolded proteins called prions. Human cerebral organoids can be infected with prions from sporadic Creutzfeldt-Jakob Disease (sCJD) brain tissue. Initial experiments indicated that the cerebral organoids may be able to differentiate biological properties of different sCJD subtypes. If so, it would be possible to investigate the pathogenic similarities and differences. Herein, we investigated multiple infections of cerebral organoids with 2 sCJD subtypes, comparing hallmark features of disease as well as neuronal function and health. Our results show that, while all infections produced seeding-capable prion protein (PrP), which increased from 90–180 days after infection, a sCJD subtype preference for protease-resistant PrP deposition was observed. Both subtypes caused substantial electrophysiological dysfunction in the infected organoids, which appeared uncoupled from PrP deposition. Neuronal dysfunction was associated with changes in neurotransmitter receptors that differed between the subtypes but produced the same outcome of a shift from inhibitory toward excitatory neurotransmission. Further changes indicated shared deficits in mitochondrial dynamics, and subtype influenced alterations in intracellular signaling pathways, cytoskeletal structure, and the extracellular matrix. We conclude that cerebral organoids demonstrate both common mitochondrial deficits and sCJD subtype–specific changes in neurotransmission and organoid architecture.

Authors

Katie Williams, Bradley R. Groveman, Simote T. Foliaki, Brent Race, Arielle Hay, Ryan O. Walters, Tina Thomas, Gianluigi Zanusso, James A. Carroll, Cathryn L. Haigh

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Figure 1

Prion infection of the cerebral organoids.

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Prion infection of the cerebral organoids. (A) Schematic showing the exp...
(A) Schematic showing the experimental approach of multiple infections and organoid batches. Created with Biorender. (B and C) RT-QuIC analysis of the organoids at 90 (blue dots) and 175–180 (orange dots) dpi, showing the percentage of positive wells per reaction (B) and the reciprocal of the time to threshold (C). Dotted lines indicate the threshold above which the reaction is considered positive. (D) Western blotting following proteinase K digest for PrP of organoids harvested at 90 and 180 dpi. Each lane is from a single organoid. (E) Densitometric quantification of C. (F) MV1 and MV2 infection densitometry in D combined by infection. (G) Example PrP IHC images of the NBH and infected organoids. Scale bars: 500 μm for whole organoids;100 μm for 20× magnified fields. The boxed region is shown magnified to the right. On all plots, individual points indicate single organoids, **P < 0.01, ****P < 0.0001, Mann Whitney U test.