Axon guidance deficits in a human sensory neuron model of Fabry disease

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The study used induced pluripotent stem cells from an isogenic GLA knockout line (Fabry disease; p.S364del) and a healthy control to differentiate into sensory neuron-like cells, then compared them using transcriptional and proteomic analyses and microfluidic neurite assays. Fabry disease sensory neurons showed dysregulated disease-related pathways, including axon guidance alterations at both the RNA and protein level, and microfluidic assays demonstrated shorter neurite length. AGAL enzyme supplementation reduced Gb3 accumulation and lowered protein levels of ephrin 5A (EFNA5) and glycoprotein M6A (GPM6A), but did not restore the transcriptomic state. The paper relates to endometriosis/adenomyosis research insofar as it models neuropathic mechanisms relevant to chronic pelvic pain phenotypes, but it does not explicitly discuss endometriosis or adenomyosis; it was included in the corpus via a keyword match in the upstream search index.

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Abstract

Fabry disease (FD) is a rare genetic galactosidase alpha ( GLA ) gene associated lysosomal disorder caused by alpha-galactosidase A (AGAL) deficiency, leading to sphingolipid (globotriaosylceramide, Gb3) accumulation in multiple tissues. Burning pain due to small fiber neuropathy is an early symptom with great impact on health- related quality of life. The pathophysiological role of Gb3 accumulations in sensory neurons of the dorsal root ganglia is incompletely understood. We have differentiated induced pluripotent stem cells of an isogenic GLA knockout line (p.S364del, hemizygous) and its healthy control into sensory neurons to model FD in vitro . We have compared both lines on transcriptional and proteomic level and investigated the effects of AGAL enzyme supplementation. FD sensory neurons showed dysregulation of disease-related pathways, including axon guidance at both RNA and protein level and microfluidic assays revealed shorter neurite length. While AGAL did not restore the transcriptomic state, it reduced Gb3 accumulation and lowered protein ephrin 5A and glycoprotein M6A level. These findings highlight axon guidance alterations in an isogenic human FD sensory model, with potential implications for early central and peripheral innervation in small fiber neuropathy.
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Abstract Fabry disease (FD) is a rare genetic galactosidase alpha (GLA) gene associated lysosomal disorder caused by alpha-galactosidase A (AGAL) deficiency, leading to sphingolipid (globotriaosylceramide, Gb3) accumulation in multiple tissues. Burning pain due to small fiber neuropathy is an early symptom with great impact on health- related quality of life. The pathophysiological role of Gb3 accumulations in sensory neurons of the dorsal root ganglia is incompletely understood. We have differentiated induced pluripotent stem cells of an isogenic GLA knockout line (p.S364del, hemizygous) and its healthy control into sensory neurons to model FD in vitro. We have compared both lines on transcriptional and proteomic level and investigated the effects of AGAL enzyme supplementation. FD sensory neurons showed dysregulation of disease-related pathways, including axon guidance at both RNA and protein level and microfluidic assays revealed shorter neurite length. While AGAL did not restore the transcriptomic state, it reduced Gb3 accumulation and lowered protein ephrin 5A and glycoprotein M6A level. These findings highlight axon guidance alterations in an isogenic human FD sensory model, with potential implications for early central and peripheral innervation in small fiber neuropathy. Competing Interest Statement The authors have declared no competing interest. Footnotes Abbreviations AGAL, alpha-galactosidase A ; BCA, bicinchoninic acid assay; bNGF, beta-nerve growth factor; CTRL, healthy control line; DE, differentially expressed; DRG, dorsal root ganglia; EFNA5, ephrin 5A; EGA, European genome-phenome archive; FCS, fetal bovine serum; FD, Fabry disease; Gb3, globotriaosylceramide; GLA, galactosidase alpha; GPM6A, glycoprotein M6-A; GSEA, gene set enrichment analysis; iF, feeder-like cells; iPSC, induced pluripotent stem cell; iSN, iPSC derived sensory-like neuron; IsoFD, isogenic FD cell line; ORA, over-representation analysis; P2RX3, purinergic receptor P2X 3; padj, adjusted P-value; PFA, paraformaldehyde; PRPH, peripherin; ROI, region of interest; SCN9A, sodium voltage-gated channel alpha subunit 9; SFN, small fiber neuropathy; TPM, transcript per million; TRPA1, transient receptor potential cation channel subfamily A member 1; TRPV1, transient receptor potential cation channel subfamily V member 1.

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