Proteasome mutations associated with CANDLE syndrome cause altered neuronal development by dysregulating polyamine synthesis
Clayton W. Winkler, Benjamin Schwarz, Katie Williams, Sara Alehashemi, Simote T. Foliaki, Joseph Snow, Lisa Joseph, Audrey Thurm, Christopher L. Friend, Gwendolyn Cooper, Eric Bohrnsen, Farzana Bhuyan, Nathan T. Brandes, Ruin Moaddel, Manfred Boehm, Guibin Chen, Cole D. Kimzey, Bibiana Bielekova, Joanna Kocot, Peter Kosa, Cathryn L. Haigh, Raphaela Goldbach-Mansky, Karin E. Peterson
Clayton W. Winkler, Benjamin Schwarz, Katie Williams, Sara Alehashemi, Simote T. Foliaki, Joseph Snow, Lisa Joseph, Audrey Thurm, Christopher L. Friend, Gwendolyn Cooper, Eric Bohrnsen, Farzana Bhuyan, Nathan T. Brandes, Ruin Moaddel, Manfred Boehm, Guibin Chen, Cole D. Kimzey, Bibiana Bielekova, Joanna Kocot, Peter Kosa, Cathryn L. Haigh, Raphaela Goldbach-Mansky, Karin E. Peterson
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Research Article Genetics Metabolism Neuroscience

Proteasome mutations associated with CANDLE syndrome cause altered neuronal development by dysregulating polyamine synthesis

Abstract

Loss-of-function mutations in PSMB8/β5i and other components of the 20S proteasome result in multiorgan diseases, such as Chronic atypical neutrophilic dermatosis with lipodystrophy and elevated temperature (CANDLE) syndrome. Neurocognitive dysfunction associated with CANDLE suggests that proteasomal mutations may impact neuronal function and development early in life. We generated cerebral organoids (COs) from induced pluripotent stem cells (iPSCs) made from patients with CANDLE. The COs from CANDLE iPSCs exhibited impaired neuronal development when compared with COs from healthy control iPSCs. Impaired neuronal maturation in CANDLE COs was correlated with increased polyamines, which were also elevated in CSF from patients with CANDLE. The proteasome-regulated ornithine decarboxylase (ODC), the rate limiting enzyme in polyamine biosynthesis, was elevated in CANDLE neurons. Inhibition of ODC reversed polyamine overproduction and repaired neuronal maturation in CANDLE COs, suggesting a potential therapeutic avenue for intervention. These findings demonstrate that dysfunction of the proteasome affects neuronal development through overproduction of polyamines via dysregulation of ODC and offer insight into potential therapeutic strategies for CNS-related proteasomal dysfunction.

Authors

Clayton W. Winkler, Benjamin Schwarz, Katie Williams, Sara Alehashemi, Simote T. Foliaki, Joseph Snow, Lisa Joseph, Audrey Thurm, Christopher L. Friend, Gwendolyn Cooper, Eric Bohrnsen, Farzana Bhuyan, Nathan T. Brandes, Ruin Moaddel, Manfred Boehm, Guibin Chen, Cole D. Kimzey, Bibiana Bielekova, Joanna Kocot, Peter Kosa, Cathryn L. Haigh, Raphaela Goldbach-Mansky, Karin E. Peterson

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Figure 7

Impact of DFMO on metabolism and neuronal development in CANDLE COs.

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Impact of DFMO on metabolism and neuronal development in CANDLE COs. (A)...
(A) Comparison of 100 μM DFMO-treated CANDLE COs versus vehicle-treated CANDLE CO metabolites via paired t test by on the average metabolite levels (CO technical n = 12, biological n = 3 CAN). Metabolites annotated as being associated with amine metabolism (purple dots), Acetyl-CoA metabolism (light blue dots), and nucleobase metabolism (magenta dots) are indicated. (B) Change in putrescine signal in COs relative to the average of the vehicle control (VC) by donor. An unpaired t test was used to compare groups. *P < 0.05, **P < 0.01). (C) Metabolites that changed in response to DFMO treatment in CANDLE COs via a paired t test with P < 0.1 organized by pathway. Node color reflects the fold change due to DFMO and the size reflects the significance of that change by patient as indicated in the associated legend. (D and E) Quantification of IHC labeling of SOX2 and TUJ1 analyzed on sections from n = 10–12 vehicle-treated and n = 12 DFMO-treated CANDLE COs. Data are plotted as percent of each signal within each tissue section using symbols described in Supplemental Table 1 to indicate COs generated from different iPSCs. Data in both D and E were analyzed by a 2-way ANOVA with Šidák’s multiple comparisons test. *P < 0.05, ****P < 0.0001. Representative IHC labeling of the mature neuron marker TUJ1 in F VC-treated and (G) DFMO-treated CAN2 COs.