Translatome analysis of tuberous sclerosis complex 1 patient-derived neural progenitor cells reveals rapamycin-dependent and independent alterations / Pauline MARTIN ; Francis ROBERT ; Krzysztof J. SZKOP ; Nicholas E. REDMOND ; Srirupa BHATTACHARYYA ; Jennifer WANG ; Shan CHEN ; Roberta L. BEAUCHAMP ; Irene NOBELI ; Jerry PELLETIER ; Ola LARSSON ; Vijaya RAMESH in Molecular Autism, 14 (2023)  

Public ISBD [article]  inMolecular Autism > 14 (2023) . - 39 p. Titre : Translatome analysis of tuberous sclerosis complex 1 patient-derived neural progenitor cells reveals rapamycin-dependent and independent alterations Type de document : Texte imprimé et/ou numérique Auteurs : Pauline MARTIN, Auteur ; Francis ROBERT, Auteur ; Krzysztof J. SZKOP, Auteur ; Nicholas E. REDMOND, Auteur ; Srirupa BHATTACHARYYA, Auteur ; Jennifer WANG, Auteur ; Shan CHEN, Auteur ; Roberta L. BEAUCHAMP, Auteur ; Irene NOBELI, Auteur ; Jerry PELLETIER, Auteur ; Ola LARSSON, Auteur ; Vijaya RAMESH, Auteur Article en page(s) : 39 p. Langues : Anglais (eng) Mots-clés : Humans  *Tuberous Sclerosis/genetics/metabolism  Tumor Suppressor Proteins/genetics  Sirolimus/pharmacology  *Autism Spectrum Disorder  Mechanistic Target of Rapamycin Complex 1  Stem Cells/metabolism  Autism spectrum disorder  Early neurodevelopment  Neural progenitor cells  Polysome profiling  Rmc-6272  Tsc1  Translatome  Tuberous sclerosis complex  mTORC1 Index. décimale : PER Périodiques Résumé : BACKGROUND: Tuberous sclerosis complex (TSC) is an inherited neurocutaneous disorder caused by mutations in the TSC1 or TSC2 genes, with patients often exhibiting neurodevelopmental (ND) manifestations termed TSC-associated neuropsychiatric disorders (TAND) including autism spectrum disorder (ASD) and intellectual disability. Hamartin (TSC1) and tuberin (TSC2) proteins form a complex inhibiting mechanistic target of rapamycin complex 1 (mTORC1) signaling. Loss of TSC1 or TSC2 activates mTORC1 that, among several targets, controls protein synthesis by inhibiting translational repressor eIF4E-binding proteins. Using TSC1 patient-derived neural progenitor cells (NPCs), we recently reported early ND phenotypic changes, including increased cell proliferation and altered neurite outgrowth in TSC1-null NPCs, which were unaffected by the mTORC1 inhibitor rapamycin. METHODS: Here, we used polysome profiling, which quantifies changes in mRNA abundance and translational efficiencies at a transcriptome-wide level, to compare CRISPR-edited TSC1-null with CRISPR-corrected TSC1-WT NPCs generated from one TSC donor (one clone/genotype). To assess the relevance of identified gene expression alterations, we performed polysome profiling in postmortem brains from ASD donors and age-matched controls. We further compared effects on translation of a subset of transcripts and rescue of early ND phenotypes in NPCs following inhibition of mTORC1 using the allosteric inhibitor rapamycin versus a third-generation bi-steric, mTORC1-selective inhibitor RMC-6272. RESULTS: Polysome profiling of NPCs revealed numerous TSC1-associated alterations in mRNA translation that were largely recapitulated in human ASD brains. Moreover, although rapamycin treatment partially reversed the TSC1-associated alterations in mRNA translation, most genes related to neural activity/synaptic regulation or ASD were rapamycin-insensitive. In contrast, treatment with RMC-6272 inhibited rapamycin-insensitive translation and reversed TSC1-associated early ND phenotypes including proliferation and neurite outgrowth that were unaffected by rapamycin. CONCLUSIONS: Our work reveals ample mRNA translation alterations in TSC1 patient-derived NPCs that recapitulate mRNA translation in ASD brain samples. Further, suppression of TSC1-associated but rapamycin-insensitive translation and ND phenotypes by RMC-6272 unveils potential implications for more efficient targeting of mTORC1 as a superior treatment strategy for TAND. En ligne : https://dx.doi.org/10.1186/s13229-023-00572-3 Permalink : https://www.cra-rhone-alpes.org/cid/opac_css/index.php?lvl=notice_display&id=518 [article] Translatome analysis of tuberous sclerosis complex 1 patient-derived neural progenitor cells reveals rapamycin-dependent and independent alterations [Texte imprimé et/ou numérique] / Pauline MARTIN, Auteur ; Francis ROBERT, Auteur ; Krzysztof J. SZKOP, Auteur ; Nicholas E. REDMOND, Auteur ; Srirupa BHATTACHARYYA, Auteur ; Jennifer WANG, Auteur ; Shan CHEN, Auteur ; Roberta L. BEAUCHAMP, Auteur ; Irene NOBELI, Auteur ; Jerry PELLETIER, Auteur ; Ola LARSSON, Auteur ; Vijaya RAMESH, Auteur . - 39 p. Langues : Anglais (eng) in Molecular Autism > 14 (2023) . - 39 p. Mots-clés : Humans  *Tuberous Sclerosis/genetics/metabolism  Tumor Suppressor Proteins/genetics  Sirolimus/pharmacology  *Autism Spectrum Disorder  Mechanistic Target of Rapamycin Complex 1  Stem Cells/metabolism  Autism spectrum disorder  Early neurodevelopment  Neural progenitor cells  Polysome profiling  Rmc-6272  Tsc1  Translatome  Tuberous sclerosis complex  mTORC1 Index. décimale : PER Périodiques Résumé : BACKGROUND: Tuberous sclerosis complex (TSC) is an inherited neurocutaneous disorder caused by mutations in the TSC1 or TSC2 genes, with patients often exhibiting neurodevelopmental (ND) manifestations termed TSC-associated neuropsychiatric disorders (TAND) including autism spectrum disorder (ASD) and intellectual disability. Hamartin (TSC1) and tuberin (TSC2) proteins form a complex inhibiting mechanistic target of rapamycin complex 1 (mTORC1) signaling. Loss of TSC1 or TSC2 activates mTORC1 that, among several targets, controls protein synthesis by inhibiting translational repressor eIF4E-binding proteins. Using TSC1 patient-derived neural progenitor cells (NPCs), we recently reported early ND phenotypic changes, including increased cell proliferation and altered neurite outgrowth in TSC1-null NPCs, which were unaffected by the mTORC1 inhibitor rapamycin. METHODS: Here, we used polysome profiling, which quantifies changes in mRNA abundance and translational efficiencies at a transcriptome-wide level, to compare CRISPR-edited TSC1-null with CRISPR-corrected TSC1-WT NPCs generated from one TSC donor (one clone/genotype). To assess the relevance of identified gene expression alterations, we performed polysome profiling in postmortem brains from ASD donors and age-matched controls. We further compared effects on translation of a subset of transcripts and rescue of early ND phenotypes in NPCs following inhibition of mTORC1 using the allosteric inhibitor rapamycin versus a third-generation bi-steric, mTORC1-selective inhibitor RMC-6272. RESULTS: Polysome profiling of NPCs revealed numerous TSC1-associated alterations in mRNA translation that were largely recapitulated in human ASD brains. Moreover, although rapamycin treatment partially reversed the TSC1-associated alterations in mRNA translation, most genes related to neural activity/synaptic regulation or ASD were rapamycin-insensitive. In contrast, treatment with RMC-6272 inhibited rapamycin-insensitive translation and reversed TSC1-associated early ND phenotypes including proliferation and neurite outgrowth that were unaffected by rapamycin. CONCLUSIONS: Our work reveals ample mRNA translation alterations in TSC1 patient-derived NPCs that recapitulate mRNA translation in ASD brain samples. Further, suppression of TSC1-associated but rapamycin-insensitive translation and ND phenotypes by RMC-6272 unveils potential implications for more efficient targeting of mTORC1 as a superior treatment strategy for TAND. En ligne : https://dx.doi.org/10.1186/s13229-023-00572-3 Permalink : https://www.cra-rhone-alpes.org/cid/opac_css/index.php?lvl=notice_display&id=518

---

1 (1 - 1 / 1)