Loss of the Chr16p11.2 ASD candidate gene QPRT leads to aberrant neuronal differentiation in the SH-SY5Y neuronal cell model
Abstract Background Altered neuronal development is discussed as the underlying pathogenic mechanism of autism spectrum disorders (ASD). Copy number variations of 16p11.2 have recurrently been identified in individuals with ASD. Of the 29 genes within this region, quinolinate phosphoribosyltransfera...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2018-11-01
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| Series: | Molecular Autism |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13229-018-0239-z |
| _version_ | 1818433261637992448 |
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| author | Denise Haslinger Regina Waltes Afsheen Yousaf Silvia Lindlar Ines Schneider Chai K. Lim Meng-Miao Tsai Boyan K. Garvalov Amparo Acker-Palmer Nicolas Krezdorn Björn Rotter Till Acker Gilles J. Guillemin Simone Fulda Christine M. Freitag Andreas G. Chiocchetti |
| author_facet | Denise Haslinger Regina Waltes Afsheen Yousaf Silvia Lindlar Ines Schneider Chai K. Lim Meng-Miao Tsai Boyan K. Garvalov Amparo Acker-Palmer Nicolas Krezdorn Björn Rotter Till Acker Gilles J. Guillemin Simone Fulda Christine M. Freitag Andreas G. Chiocchetti |
| author_sort | Denise Haslinger |
| collection | DOAJ |
| description | Abstract Background Altered neuronal development is discussed as the underlying pathogenic mechanism of autism spectrum disorders (ASD). Copy number variations of 16p11.2 have recurrently been identified in individuals with ASD. Of the 29 genes within this region, quinolinate phosphoribosyltransferase (QPRT) showed the strongest regulation during neuronal differentiation of SH-SY5Y neuroblastoma cells. We hypothesized a causal relation between this tryptophan metabolism-related enzyme and neuronal differentiation. We thus analyzed the effect of QPRT on the differentiation of SH-SY5Y and specifically focused on neuronal morphology, metabolites of the tryptophan pathway, and the neurodevelopmental transcriptome. Methods The gene dosage-dependent change of QPRT expression following Chr16p11.2 deletion was investigated in a lymphoblastoid cell line (LCL) of a deletion carrier and compared to his non-carrier parents. Expression of QPRT was tested for correlation with neuromorphology in SH-SY5Y cells. QPRT function was inhibited in SH-SY5Y neuroblastoma cells using (i) siRNA knockdown (KD), (ii) chemical mimicking of loss of QPRT, and (iii) complete CRISPR/Cas9-mediated knock out (KO). QPRT-KD cells underwent morphological analysis. Chemically inhibited and QPRT-KO cells were characterized using viability assays. Additionally, QPRT-KO cells underwent metabolite and whole transcriptome analyses. Genes differentially expressed upon KO of QPRT were tested for enrichment in biological processes and co-regulated gene-networks of the human brain. Results QPRT expression was reduced in the LCL of the deletion carrier and significantly correlated with the neuritic complexity of SH-SY5Y. The reduction of QPRT altered neuronal morphology of differentiated SH-SY5Y cells. Chemical inhibition as well as complete KO of the gene were lethal upon induction of neuronal differentiation, but not proliferation. The QPRT-associated tryptophan pathway was not affected by KO. At the transcriptome level, genes linked to neurodevelopmental processes and synaptic structures were affected. Differentially regulated genes were enriched for ASD candidates, and co-regulated gene networks were implicated in the development of the dorsolateral prefrontal cortex, the hippocampus, and the amygdala. Conclusions In this study, QPRT was causally related to in vitro neuronal differentiation of SH-SY5Y cells and affected the regulation of genes and gene networks previously implicated in ASD. Thus, our data suggest that QPRT may play an important role in the pathogenesis of ASD in Chr16p11.2 deletion carriers. |
| first_indexed | 2024-12-14T16:18:17Z |
| format | Article |
| id | doaj.art-ce0cb06f639d43dd818685fcf52ff225 |
| institution | Directory Open Access Journal |
| issn | 2040-2392 |
| language | English |
| last_indexed | 2024-12-14T16:18:17Z |
| publishDate | 2018-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Molecular Autism |
| spelling | doaj.art-ce0cb06f639d43dd818685fcf52ff2252022-12-21T22:54:52ZengBMCMolecular Autism2040-23922018-11-019111710.1186/s13229-018-0239-zLoss of the Chr16p11.2 ASD candidate gene QPRT leads to aberrant neuronal differentiation in the SH-SY5Y neuronal cell modelDenise Haslinger0Regina Waltes1Afsheen Yousaf2Silvia Lindlar3Ines Schneider4Chai K. Lim5Meng-Miao Tsai6Boyan K. Garvalov7Amparo Acker-Palmer8Nicolas Krezdorn9Björn Rotter10Till Acker11Gilles J. Guillemin12Simone Fulda13Christine M. Freitag14Andreas G. Chiocchetti15Department of Child and Adolescent Psychiatry, Psychosomatics and Psychotherapy, University Hospital Frankfurt, JW Goethe University FrankfurtDepartment of Child and Adolescent Psychiatry, Psychosomatics and Psychotherapy, University Hospital Frankfurt, JW Goethe University FrankfurtDepartment of Child and Adolescent Psychiatry, Psychosomatics and Psychotherapy, University Hospital Frankfurt, JW Goethe University FrankfurtDepartment of Child and Adolescent Psychiatry, Psychosomatics and Psychotherapy, University Hospital Frankfurt, JW Goethe University FrankfurtInstitute of Experimental Cancer Research in PediatricsFaculty of Medicine and Health Sciences, Macquarie UniversityNeuropathology, University of GiessenNeuropathology, University of GiessenInstitute of Cell Biology and Neuroscience and Buchmann Institute for Molecular Life Sciences (BMLS), JW Goethe University of FrankfurtGenXPro GmbHGenXPro GmbHNeuropathology, University of GiessenFaculty of Medicine and Health Sciences, Macquarie UniversityInstitute of Experimental Cancer Research in PediatricsDepartment of Child and Adolescent Psychiatry, Psychosomatics and Psychotherapy, University Hospital Frankfurt, JW Goethe University FrankfurtDepartment of Child and Adolescent Psychiatry, Psychosomatics and Psychotherapy, University Hospital Frankfurt, JW Goethe University FrankfurtAbstract Background Altered neuronal development is discussed as the underlying pathogenic mechanism of autism spectrum disorders (ASD). Copy number variations of 16p11.2 have recurrently been identified in individuals with ASD. Of the 29 genes within this region, quinolinate phosphoribosyltransferase (QPRT) showed the strongest regulation during neuronal differentiation of SH-SY5Y neuroblastoma cells. We hypothesized a causal relation between this tryptophan metabolism-related enzyme and neuronal differentiation. We thus analyzed the effect of QPRT on the differentiation of SH-SY5Y and specifically focused on neuronal morphology, metabolites of the tryptophan pathway, and the neurodevelopmental transcriptome. Methods The gene dosage-dependent change of QPRT expression following Chr16p11.2 deletion was investigated in a lymphoblastoid cell line (LCL) of a deletion carrier and compared to his non-carrier parents. Expression of QPRT was tested for correlation with neuromorphology in SH-SY5Y cells. QPRT function was inhibited in SH-SY5Y neuroblastoma cells using (i) siRNA knockdown (KD), (ii) chemical mimicking of loss of QPRT, and (iii) complete CRISPR/Cas9-mediated knock out (KO). QPRT-KD cells underwent morphological analysis. Chemically inhibited and QPRT-KO cells were characterized using viability assays. Additionally, QPRT-KO cells underwent metabolite and whole transcriptome analyses. Genes differentially expressed upon KO of QPRT were tested for enrichment in biological processes and co-regulated gene-networks of the human brain. Results QPRT expression was reduced in the LCL of the deletion carrier and significantly correlated with the neuritic complexity of SH-SY5Y. The reduction of QPRT altered neuronal morphology of differentiated SH-SY5Y cells. Chemical inhibition as well as complete KO of the gene were lethal upon induction of neuronal differentiation, but not proliferation. The QPRT-associated tryptophan pathway was not affected by KO. At the transcriptome level, genes linked to neurodevelopmental processes and synaptic structures were affected. Differentially regulated genes were enriched for ASD candidates, and co-regulated gene networks were implicated in the development of the dorsolateral prefrontal cortex, the hippocampus, and the amygdala. Conclusions In this study, QPRT was causally related to in vitro neuronal differentiation of SH-SY5Y cells and affected the regulation of genes and gene networks previously implicated in ASD. Thus, our data suggest that QPRT may play an important role in the pathogenesis of ASD in Chr16p11.2 deletion carriers.http://link.springer.com/article/10.1186/s13229-018-0239-zAutism16p11.2Quinolinate phosphoribosyltransferaseQuinolinic acidKynurenineCRISPR/Cas9 |
| spellingShingle | Denise Haslinger Regina Waltes Afsheen Yousaf Silvia Lindlar Ines Schneider Chai K. Lim Meng-Miao Tsai Boyan K. Garvalov Amparo Acker-Palmer Nicolas Krezdorn Björn Rotter Till Acker Gilles J. Guillemin Simone Fulda Christine M. Freitag Andreas G. Chiocchetti Loss of the Chr16p11.2 ASD candidate gene QPRT leads to aberrant neuronal differentiation in the SH-SY5Y neuronal cell model Molecular Autism Autism 16p11.2 Quinolinate phosphoribosyltransferase Quinolinic acid Kynurenine CRISPR/Cas9 |
| title | Loss of the Chr16p11.2 ASD candidate gene QPRT leads to aberrant neuronal differentiation in the SH-SY5Y neuronal cell model |
| title_full | Loss of the Chr16p11.2 ASD candidate gene QPRT leads to aberrant neuronal differentiation in the SH-SY5Y neuronal cell model |
| title_fullStr | Loss of the Chr16p11.2 ASD candidate gene QPRT leads to aberrant neuronal differentiation in the SH-SY5Y neuronal cell model |
| title_full_unstemmed | Loss of the Chr16p11.2 ASD candidate gene QPRT leads to aberrant neuronal differentiation in the SH-SY5Y neuronal cell model |
| title_short | Loss of the Chr16p11.2 ASD candidate gene QPRT leads to aberrant neuronal differentiation in the SH-SY5Y neuronal cell model |
| title_sort | loss of the chr16p11 2 asd candidate gene qprt leads to aberrant neuronal differentiation in the sh sy5y neuronal cell model |
| topic | Autism 16p11.2 Quinolinate phosphoribosyltransferase Quinolinic acid Kynurenine CRISPR/Cas9 |
| url | http://link.springer.com/article/10.1186/s13229-018-0239-z |
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