A Stem Cell-Based Screening Platform Identifies Compounds that Desensitize Motor Neurons to Endoplasmic Reticulum Stress

Thams, Sebastian; Lowry, Emily; Larraufie, Marie Hélène; Spiller, Krista; Li, Hai; Williams, Damian J.; Hoang, Phuong Thi; Jiang, Elise; Williams, Luis A.; Sandoe, Jackson; Eggan, Kevin; Lieberam, Ivo; Kanning, Kevin C.; Stockwell, Brent R.; Henderson, Christopher E.; Wichterle, Hynek

doi:10.1016/j.ymthe.2018.10.010

Cited by 46 publications

(46 citation statements)

References 100 publications

(147 reference statements)

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“…CPA blocks sarco/endoplasmic reticulum Ca 2+ -ATPase (SERCA), the Ca 2+ -pump in endoplasmic reticulum (ER) membrane that helps to maintain Ca 2+ balance and ER homeostasis, and thus has a broader effect. Notably, CPA was reported to accelerate the degeneration of SpMNs expressing hSOD1 G93A mutant, suggesting that CPA treatment may reveal ALS-relevant differential vulnerability between iCrMNs and iSpMNs (Thams et al, 2019).…”

Section: Resultsmentioning

confidence: 99%

Stem cell-derived cranial and spinal motor neurons reveal proteostatic differences between ALS resistant and sensitive motor neurons

Fujiki

Iannitelli

et al. 2019

eLife

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In amyotrophic lateral sclerosis (ALS) spinal motor neurons (SpMN) progressively degenerate while a subset of cranial motor neurons (CrMN) are spared until late stages of the disease. Using a rapid and efficient protocol to differentiate mouse embryonic stem cells (ESC) to SpMNs and CrMNs, we now report that ESC-derived CrMNs accumulate less human (h)SOD1 and insoluble p62 than SpMNs over time. ESC-derived CrMNs have higher proteasome activity to degrade misfolded proteins and are intrinsically more resistant to chemically-induced proteostatic stress than SpMNs. Chemical and genetic activation of the proteasome rescues SpMN sensitivity to proteostatic stress. In agreement, the hSOD1 G93A mouse model reveals that ALS-resistant CrMNs accumulate less insoluble hSOD1 and p62-containing inclusions than SpMNs. Primary-derived ALS-resistant CrMNs are also more resistant than SpMNs to proteostatic stress. Thus, an ESC-based platform has identified a superior capacity to maintain a healthy proteome as a possible mechanism to resist ALS-induced neurodegeneration.

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Section: Resultsmentioning

confidence: 99%

Stem cell-derived cranial and spinal motor neurons reveal proteostatic differences between ALS resistant and sensitive motor neurons

Fujiki

Iannitelli

et al. 2019

eLife

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“…However, neither genetic inhibition of the UPR via ablation of PERK, nor genetic UPR enhancement via ablation of GADD34, had a beneficial effect in mutant SOD1 mice (Dzhashiashvili et al, 2019 ). More recent studies in SOD1-iPSC and mouse models demonstrated that MNs are more sensitive to ER stress and identified a number of modifiers, including TUDCA, a bile acid derivative which is currently undergoing clinical trials in ALS (Thams et al, 2019 ; Paganoni et al, 2020 ).…”

Section: Discussionmentioning

confidence: 99%

The Role of Mitochondrial Dysfunction and ER Stress in TDP-43 and C9ORF72 ALS

Dafinca

Barbagallo

Talbot

2021

Front. Cell. Neurosci.

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Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease of the motor system with complex determinants, including genetic and non-genetic factors. Despite this heterogeneity, a key pathological signature is the mislocalization and aggregation of specific proteins in the cytoplasm, suggesting that convergent pathogenic mechanisms focusing on disturbances in proteostasis are important in ALS. In addition, many cellular processes have been identified as potentially contributing to disease initiation and progression, such as defects in axonal transport, autophagy, nucleocytoplasmic transport, ER stress, calcium metabolism, the unfolded protein response and mitochondrial function. Here we review the evidence from in vitro and in vivo models of C9ORF72 and TDP-43-related ALS supporting a central role in pathogenesis for endoplasmic reticulum stress, which activates an unfolded protein response (UPR), and mitochondrial dysfunction. Disruption in the finely tuned signaling between the ER and mitochondria through calcium ions may be a crucial trigger of mitochondrial deficits and initiate an apoptotic signaling cascade, thus acting as a point of convergence for multiple upstream disturbances of cellular homeostasis and constituting a potentially important therapeutic target.

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“…Loss of microtubule mass or altered microtubule dynamics in axons and dendrites are major contributors to neurodegenerative diseases such as ALS, Parkinson's disease, Alzheimer's disease, and several tauopathies (59). A recent screen of a ~100 small molecules identified compounds protective of ER stress in a chemicallyinduced ER stress model of ALS (SOD1 G93A ) neurodegeneration (60). Future studies will determine whether compounds that affect microtubule dynamics are able to prevent disease-relevant phenotypes in cellular models of neurodegenerative diseases.…”

Section: Discussionmentioning

confidence: 99%

Precision genetic cellular models identify therapies protective against endoplasmic reticulum stress

Lebedeva

Wagner

Sahdeo

et al. 2020

Preprint

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Congenital disorders of glycosylation (CDG) and deglycosylation (CDDG) are a collection of rare pediatric disorders with symptoms that range from mild to life threatening. They typically affect multiple organ systems and usually present with neurological abnormalities including hypotonia, cognitive impairment, and intractable seizures. Several genes have been implicated in the thirty-six types of CDG, but currently NGLY1 is the only known CDDG gene. A common biological mechanism among CDG types and in CDDG is endoplasmic reticulum (ER) stress. Here, we develop two isogenic human cellular models of CDG (PMM2, the most prevalent type of CDG, and DPAGT1) and of the only CDDG (NGYL1) in an effort to identify drugs that can alleviate ER stress. Systematic phenotyping identified elevated ER stress and autophagy levels among other cellular and morphological phenotypes in each of the cellular models. We screened a complex drug library for compounds able to correct aberrant morphological phenotypes in each of the models using an agnostic phenotypic cell painting assay based on >300 cellular features. The image-based screen identified multiple candidate compounds able to correct aberrant morphology, and we show a subset of these are able to correct cellular and molecular defects in each of the models. These results provide new directions for the treatment of rare diseases of glycosylation and deglycosylation and a framework for new drug screening paradigms for more common neurodegenerative diseases characterized by ER stress.Here we utilize a morphological profiling and screening paradigm to identify agents that protect against the cellular stresses resulting from CDG and CDDG causal mutations. We focus specifically on mutations in PMM2 and DPAGT1, and in NGLY1, which causes the only reported CDDG (6). We used genetic engineering to generate clinically relevant CDG and CDDG genotypes in a karyotypically normal human cell line (hTERT RPE-1) in order to create cellular models amenable to mutation-specific phenotype identification. The hTERT RPE-1 line was selected to allow for morphology-based high content imaging screens to identify phenotypes that are consequences of ER stress. These CDG and CDDG cell lines were used in high-content small molecule screens to identify compounds that revert the imaging phenotypes caused by these mutations. Specifically, we selected 1049 annotated compounds for screening representing a broad chemical space and multiple target classes. In order to validate the performance of the screen, we selected 16 compounds that were ranked amongst the best at phenotype reversion in the screen (protective compounds) and 10 compounds that did not affect aberrant phenotypes (non-active negative control compounds). We then evaluated these compounds in assays designed to test how well they revert mutational phenotypes in our three cellular models. Results Establishment of precise human cellular models of CDG and CDDGGenome editing was used to generate hTERT RPE-1 cell lines that mimic genotypes associated with...

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A Stem Cell-Based Screening Platform Identifies Compounds that Desensitize Motor Neurons to Endoplasmic Reticulum Stress

Cited by 46 publications

References 100 publications

Stem cell-derived cranial and spinal motor neurons reveal proteostatic differences between ALS resistant and sensitive motor neurons

Stem cell-derived cranial and spinal motor neurons reveal proteostatic differences between ALS resistant and sensitive motor neurons

The Role of Mitochondrial Dysfunction and ER Stress in TDP-43 and C9ORF72 ALS

Precision genetic cellular models identify therapies protective against endoplasmic reticulum stress

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