Purine nucleoside phosphorylase deficiency induces p53-mediated intrinsic apoptosis in human induced pluripotent stem cell-derived neurons
Abstract Purine nucleoside phosphorylase (PNP) is an important enzyme in the purine degradation and salvage pathway. PNP deficiency results in marked T lineage lymphopenia and severe immunodeficiency. Additionally, PNP-deficient patients and mice suffer from diverse non-infectious neurological abnor...
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Nature Portfolio
2022-05-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-10935-0 |
| _version_ | 1818161645491322880 |
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| author | Michael Tsui Jeremy Biro Jonathan Chan Weixian Min Kerry Dobbs Luigi D. Notarangelo Eyal Grunebaum |
| author_facet | Michael Tsui Jeremy Biro Jonathan Chan Weixian Min Kerry Dobbs Luigi D. Notarangelo Eyal Grunebaum |
| author_sort | Michael Tsui |
| collection | DOAJ |
| description | Abstract Purine nucleoside phosphorylase (PNP) is an important enzyme in the purine degradation and salvage pathway. PNP deficiency results in marked T lineage lymphopenia and severe immunodeficiency. Additionally, PNP-deficient patients and mice suffer from diverse non-infectious neurological abnormalities of unknown etiology. To further investigate the cause for these neurologic abnormalities, induced pluripotent stem cells (iPSC) from two PNP-deficient patients were differentiated into neurons. The iPSC-derived PNP-deficient neurons had significantly reduced soma and nuclei volumes. The PNP-deficient neurons demonstrated increased spontaneous and staurosporine-induced apoptosis, measured by cleaved caspase-3 expression, together with decreased mitochondrial membrane potential and increased cleaved caspase-9 expression, indicative of enhanced intrinsic apoptosis. Greater expression of tumor protein p53 was also observed in these neurons, and inhibition of p53 using pifithrin-α prevented the apoptosis. Importantly, treatment of the iPSC-derived PNP-deficient neurons with exogenous PNP enzyme alleviated the apoptosis. Inhibition of ribonucleotide reductase (RNR) in iPSC derived from PNP-proficient neurons with hydroxyurea or with nicotinamide and trichostatin A increased the intrinsic neuronal apoptosis, implicating RNR dysfunction as the potential mechanism for the damage caused by PNP deficiency. The findings presented here establish a potential mechanism for the neurological defects observed in PNP-deficient patients and reinforce the critical role that PNP has for neuronal viability. |
| first_indexed | 2024-12-11T16:21:04Z |
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| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-11T16:21:04Z |
| publishDate | 2022-05-01 |
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| spelling | doaj.art-ea144a7cc5a540f8acdc8dcc639a1e1f2022-12-22T00:58:51ZengNature PortfolioScientific Reports2045-23222022-05-0112111410.1038/s41598-022-10935-0Purine nucleoside phosphorylase deficiency induces p53-mediated intrinsic apoptosis in human induced pluripotent stem cell-derived neuronsMichael Tsui0Jeremy Biro1Jonathan Chan2Weixian Min3Kerry Dobbs4Luigi D. Notarangelo5Eyal Grunebaum6Developmental and Stem Cell Biology Program, Hospital for Sick ChildrenDevelopmental and Stem Cell Biology Program, Hospital for Sick ChildrenDevelopmental and Stem Cell Biology Program, Hospital for Sick ChildrenDevelopmental and Stem Cell Biology Program, Hospital for Sick ChildrenLaboratory of Clinical Immunology and Microbiology, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of HealthLaboratory of Clinical Immunology and Microbiology, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of HealthDevelopmental and Stem Cell Biology Program, Hospital for Sick ChildrenAbstract Purine nucleoside phosphorylase (PNP) is an important enzyme in the purine degradation and salvage pathway. PNP deficiency results in marked T lineage lymphopenia and severe immunodeficiency. Additionally, PNP-deficient patients and mice suffer from diverse non-infectious neurological abnormalities of unknown etiology. To further investigate the cause for these neurologic abnormalities, induced pluripotent stem cells (iPSC) from two PNP-deficient patients were differentiated into neurons. The iPSC-derived PNP-deficient neurons had significantly reduced soma and nuclei volumes. The PNP-deficient neurons demonstrated increased spontaneous and staurosporine-induced apoptosis, measured by cleaved caspase-3 expression, together with decreased mitochondrial membrane potential and increased cleaved caspase-9 expression, indicative of enhanced intrinsic apoptosis. Greater expression of tumor protein p53 was also observed in these neurons, and inhibition of p53 using pifithrin-α prevented the apoptosis. Importantly, treatment of the iPSC-derived PNP-deficient neurons with exogenous PNP enzyme alleviated the apoptosis. Inhibition of ribonucleotide reductase (RNR) in iPSC derived from PNP-proficient neurons with hydroxyurea or with nicotinamide and trichostatin A increased the intrinsic neuronal apoptosis, implicating RNR dysfunction as the potential mechanism for the damage caused by PNP deficiency. The findings presented here establish a potential mechanism for the neurological defects observed in PNP-deficient patients and reinforce the critical role that PNP has for neuronal viability.https://doi.org/10.1038/s41598-022-10935-0 |
| spellingShingle | Michael Tsui Jeremy Biro Jonathan Chan Weixian Min Kerry Dobbs Luigi D. Notarangelo Eyal Grunebaum Purine nucleoside phosphorylase deficiency induces p53-mediated intrinsic apoptosis in human induced pluripotent stem cell-derived neurons Scientific Reports |
| title | Purine nucleoside phosphorylase deficiency induces p53-mediated intrinsic apoptosis in human induced pluripotent stem cell-derived neurons |
| title_full | Purine nucleoside phosphorylase deficiency induces p53-mediated intrinsic apoptosis in human induced pluripotent stem cell-derived neurons |
| title_fullStr | Purine nucleoside phosphorylase deficiency induces p53-mediated intrinsic apoptosis in human induced pluripotent stem cell-derived neurons |
| title_full_unstemmed | Purine nucleoside phosphorylase deficiency induces p53-mediated intrinsic apoptosis in human induced pluripotent stem cell-derived neurons |
| title_short | Purine nucleoside phosphorylase deficiency induces p53-mediated intrinsic apoptosis in human induced pluripotent stem cell-derived neurons |
| title_sort | purine nucleoside phosphorylase deficiency induces p53 mediated intrinsic apoptosis in human induced pluripotent stem cell derived neurons |
| url | https://doi.org/10.1038/s41598-022-10935-0 |
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