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

An, Disi; Fujiki, Ryosuke; Iannitelli, Dylan E.; Smerdon, John W.; Maity, Shuvadeep; Rose, Matthew F.; Gelber, Alon; Wanaselja, Elizabeth K; Yagudayeva, Ilona; Lee, Joun Y.; Vogel, Christine; Wichterle, Hynek; Engle, Elizabeth C.; Mazzoni, Esteban O.

doi:10.7554/elife.44423

Cited by 30 publications

(37 citation statements)

References 77 publications

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“…Previously, several groups have suggested therapeutic target molecules 10 , 44 , 45 , but our transcriptome analysis results did not confirm those findings ( Supplementary Data Fig. 4D ).…”

Section: Discussioncontrasting

confidence: 93%

Multi-omic analysis of selectively vulnerable motor neuron subtypes implicates altered lipid metabolism in ALS

Lee

Park

et al. 2021

Nat Neurosci

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Amyotrophic lateral sclerosis (ALS) is a devastating disorder in which motor neurons degenerate, the causes of which remain unclear. In particular, the basis for selective vulnerability of spinal motor neurons (sMNs) and resistance of ocular motor neurons (oMNs) to degeneration in ALS has yet to be elucidated. Here, we applied comparative multi-omics analysis of human induced pluripotent stem cell (hiPSC)-derived sMNs and oMNs to identify shared metabolic perturbations in inherited and sporadic ALS sMNs, revealing dysregulation in lipid metabolism and its related genes. Targeted metabolomics studies confirmed such findings in sMNs of 17 ALS ( SOD1 , C9ORF72 , TDP43 and sporadic) hiPSC lines, identifying elevated levels of arachidonic acid (AA). Pharmacological reduction of AA levels was sufficient to reverse ALS-related phenotypes in both human sMNs and in vivo in Drosophila and SOD1 G93A mouse models. Collectively, these findings pinpoint a catalytic step of lipid metabolism as a potential therapeutic target for ALS.

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“…Previously, several groups have suggested therapeutic target molecules 10 , 44 , 45 , but our transcriptome analysis results did not confirm those findings ( Supplementary Data Fig. 4D ).…”

Section: Discussioncontrasting

confidence: 93%

Multi-omic analysis of selectively vulnerable motor neuron subtypes implicates altered lipid metabolism in ALS

Lee

Park

et al. 2021

Nat Neurosci

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“…Moreover, it has been reported that oculomotor and trochlear MNs highly express PHOX2A ( Allodi et al., 2019 ), which is structurally similar to PHOX2B, and reported to form heterodimers with PHOX2B in vitro ( Di Lascio et al., 2016 ). Oculomotor and trochlear motor neurons show a better survival rate than spinal cord MNs in ALS ( An et al., 2019 ), thus high PHOX2A expression in these cells might serve to protect them from cell death associated with ALS. In this study, expression of PHOX2A was also downregulated in TARDBP mutant axons (RNA-seq; fold change = 0.1085, p = 0.0104, Table S5 ); interestingly, the expression of PHOX2A protein in mutant MNs was indistinguishable from that detected in the WT (data not shown).…”

Section: Discussionmentioning

confidence: 99%

Reduced PHOX2B stability causes axonal growth impairment in motor neurons with TARDBP mutations

et al. 2021

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Amyotrophic lateral sclerosis (ALS) is an adult-onset incurable motor neuron (MN) disease. The reasons for selective MN vulnerability in ALS are unknown. Axonal pathology is among the earliest signs of ALS. We searched for novel modulatory genes in human MN axon shortening affected by TARDBP mutations. In transcriptome analysis of RNA present in the axon compartment of human-derived induced pluripotent stem cell (iPSC)-derived MNs, PHOX2B (paired-like homeobox protein 2B) showed lower expression in TARDBP mutant axons, which was consistent with axon qPCR and in situ hybridization. PHOX2B mRNA stability was reduced in TARDBP mutant MNs. Furthermore, PHOX2B knockdown reduced neurite length in human MNs. Finally, phox2b knockdown in zebrafish induced short spinal axons and impaired escape response. PHOX2B is known to be highly express in other types of neurons maintained after ALS progression. Collectively, TARDBP mutations induced loss of axonal resilience, which is an important ALS-related phenotype mediated by PHOX2B downregulation.

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“…Since iPSCs can be differentiated to motor neurons with high efficiency, resulting in a relatively homogeneous population of cells, experiments such as ribosome footprinting will be much more straightforward in cell-based models, where ribosomes can be rapidly isolated from the bulk culture. It is even possible to generate specific sub-populations of motor neurons, such as cranial motor neurons, or limb-innervating lateral motor column motor neurons ( Amoroso et al, 2013 ; An et al, 2019 ). This is additional degree of differentiation is potentially useful given that CMT2D primarily affects spinal alpha motor neurons.…”

Section: Human Cell-based Models Of Trna Synthetase-associated Neuropathymentioning

confidence: 99%

An Integrated Approach to Studying Rare Neuromuscular Diseases Using Animal and Human Cell-Based Models

Hines

Lutz

Murray

et al. 2022

Front. Cell Dev. Biol.

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As sequencing technology improves, the identification of new disease-associated genes and new alleles of known genes is rapidly increasing our understanding of the genetic underpinnings of rare diseases, including neuromuscular diseases. However, precisely because these disorders are rare and often heterogeneous, they are difficult to study in patient populations. In parallel, our ability to engineer the genomes of model organisms, such as mice or rats, has gotten increasingly efficient through techniques such as CRISPR/Cas9 genome editing, allowing the creation of precision human disease models. Such in vivo model systems provide an efficient means for exploring disease mechanisms and identifying therapeutic strategies. Furthermore, animal models provide a platform for preclinical studies to test the efficacy of those strategies. Determining whether the same mechanisms are involved in the human disease and confirming relevant parameters for treatment ideally involves a human experimental system. One system currently being used is induced pluripotent stem cells (iPSCs), which can then be differentiated into the relevant cell type(s) for in vitro confirmation of disease mechanisms and variables such as target engagement. Here we provide a demonstration of these approaches using the example of tRNA-synthetase-associated inherited peripheral neuropathies, rare forms of Charcot-Marie-Tooth disease (CMT). Mouse models have led to a better understanding of both the genetic and cellular mechanisms underlying the disease. To determine if the mechanisms are similar in human cells, we will use genetically engineered iPSC-based models. This will allow comparisons of different CMT-associated GARS alleles in the same genetic background, reducing the variability found between patient samples and simplifying the availability of cell-based models for a rare disease. The necessity of integrating mouse and human models, strategies for accomplishing this integration, and the challenges of doing it at scale are discussed using recently published work detailing the cellular mechanisms underlying GARS-associated CMT as a framework.

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

Cited by 30 publications

References 77 publications

Multi-omic analysis of selectively vulnerable motor neuron subtypes implicates altered lipid metabolism in ALS

Multi-omic analysis of selectively vulnerable motor neuron subtypes implicates altered lipid metabolism in ALS

Reduced PHOX2B stability causes axonal growth impairment in motor neurons with TARDBP mutations

An Integrated Approach to Studying Rare Neuromuscular Diseases Using Animal and Human Cell-Based Models

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