Glutaric Aciduria type I (GA1) is a rare neurometabolic disorder caused by mutations in the GDCH gene encoding for glutaryl-CoA dehydrogenase (GCDH) in the catabolic pathway of lysine, hydroxylysine and tryptophan. GCDH deficiency leads to increased concentrations of glutaric acid (GA) and 3-hydroxyglutaric acid (3-OHGA) in body fluids and tissues. These metabolites are the main triggers of brain damage. Mechanistic studies supporting neurotoxicity in mouse models have been conducted. However, the different vulnerability to some stressors between mouse and human brain cells reveals the need to have a reliable human neuronal model to study GA1 pathogenesis. In the present work we generated a GCDH knockout (KO) in the human neuroblastoma cell line SH-SY5Y by CRISPR/Cas9 technology. SH-SY5Y-GCDH KO cells accumulate GA, 3-OHGA, and glutarylcarnitine when exposed to lysine overload. GA or lysine treatment triggered neuronal damage in GCDH deficient cells. SH-SY5Y-GCDH KO cells also displayed features of GA1 pathogenesis such as increased oxidative stress vulnerability. Restoration of the GCDH activity by gene replacement rescued neuronal alterations. Thus, our findings provide a human neuronal cellular model of GA1 to study this disease and show the potential of gene therapy to rescue GCDH deficiency.
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We thank Paula del Río for providing us with the plasmid pCCL.PGK.FANCA.WPRE. We are indebted to the Flow Cytometry Platform of IDIBAPS for technical help.
AM-B is recipient of a PIF-Salut predoctoral contract from Generalitat de Catalunya, Spain. MG-M is recipient of a Sara Borrell contract CD21/00019 from Instituto de Salud Carlos III (ISCIII). This work was supported by grants to CF from the CIVP19A5949-Fundación Ramon Areces, Merck Sharp Dohme España S.A, ACCI-CIBERER (ER21P2AC737) and PID2020-119692RB-C22 Spanish Ministerio de Ciencia e Innovación, with partial support from the Generalitat de Catalunya SGR2021/01169 and SGR2021/01423. It also acknowledges the support of CHARLIE Consortium, a project supported by ISCIII under the frame of E-Rare-3, the ERA-Net for Research on Rare Diseases, EJPRD grant nr: 825575. CIBERER is an initiative of the ISCIII. We also acknowledge the support of CERCA Program/Generalitat de Catalunya. This work was developed at the Centro Esther Koplowitz, Barcelona, Spain.
The authors declare no competing interests.
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Mateu-Bosch, A., Segur-Bailach, E., García-Villoria, J. et al. Modeling Glutaric Aciduria Type I in human neuroblastoma cells recapitulates neuronal damage that can be rescued by gene replacement. Gene Ther (2023). https://doi.org/10.1038/s41434-023-00428-8