Identification of Targets from LRRK2 Rescue Phenotypes
Parkinson’s disease (PD) is an age-dependent neurodegenerative condition. Leucine-rich repeat kinase 2 (LRRK2) mutations are the most frequent cause of sporadic and autosomal dominant PD. The exact role of LRRK2 protective variants (R1398H, N551K) together with a pathogenic mutant (G2019S) in aging...
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MDPI AG
2021-01-01
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| author | Joanne Toh Ling Ling Chua Patrick Ho Edwin Sandanaraj Carol Tang Hongyan Wang Eng King Tan |
| author_facet | Joanne Toh Ling Ling Chua Patrick Ho Edwin Sandanaraj Carol Tang Hongyan Wang Eng King Tan |
| author_sort | Joanne Toh |
| collection | DOAJ |
| description | Parkinson’s disease (PD) is an age-dependent neurodegenerative condition. Leucine-rich repeat kinase 2 (LRRK2) mutations are the most frequent cause of sporadic and autosomal dominant PD. The exact role of LRRK2 protective variants (R1398H, N551K) together with a pathogenic mutant (G2019S) in aging and neurodegeneration is unknown. We generated the following myc-tagged UAS-LRRK2 transgenic Drosophila: LRRK2 (WT), N551K, R1398H, G2019S single allele, and double-mutants (N551K/G2019S or R1398H/G2019S). The protective variants alone were able to suppress the phenotypic effects caused by the pathogenic LRRK2 mutation. Next, we conducted RNA-sequencing using mRNA isolated from dopaminergic neurons of these different groups of transgenic <i>Drosophila.</i> Using pathway enrichment analysis, we identified the top 10 modules (<i>p</i> < 0.05), with “LRRK2 in neurons in Parkinson’s disease” among the candidates. Further dissection of this pathway identified the most significantly modulated gene nodes such as eEF1A2, ACTB, eEF1A, and actin cytoskeleton reorganization. The induction of the pathway was successfully restored by the R1398H protective variant and R1398H-G2019S or N551K-G2019S rescue experiments. The oxidoreductase family of genes was also active in the pathogenic mutant and restored in protective and rescue variants. In summary, we provide in vivo evidence supporting the neuroprotective effects of LRRK2 variants. RNA sequencing of dopaminergic neurons identified upregulation of specific gene pathways in the <i>Drosophila</i> carrying the pathogenic variant, and this was restored in the rescue phenotypes. Using protective gene variants, our study identifies potential new targets and provides proof of principle of a new therapeutic approach that will further our understanding of aging and neurodegeneration in PD. |
| first_indexed | 2024-03-10T13:29:03Z |
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| institution | Directory Open Access Journal |
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| language | English |
| last_indexed | 2024-03-10T13:29:03Z |
| publishDate | 2021-01-01 |
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| series | Cells |
| spelling | doaj.art-6f6d4e20230f45c4b9a82332ef5571e12023-11-21T08:28:34ZengMDPI AGCells2073-44092021-01-011017610.3390/cells10010076Identification of Targets from LRRK2 Rescue PhenotypesJoanne Toh0Ling Ling Chua1Patrick Ho2Edwin Sandanaraj3Carol Tang4Hongyan Wang5Eng King Tan6Department of Research, National Neuroscience Institute, SGH Campus, Singapore 169856, SingaporeDepartment of Research, National Neuroscience Institute, SGH Campus, Singapore 169856, SingaporeDepartment of Research, National Neuroscience Institute, SGH Campus, Singapore 169856, SingaporeDepartment of Research, National Neuroscience Institute, SGH Campus, Singapore 169856, SingaporeDepartment of Research, National Neuroscience Institute, Singapore 308433, SingaporeNeuroscience and Behavioral Disorders Program, Duke NUS Medical School, Singapore 169857, SingaporeDepartment of Neurology, National Neuroscience Institute Singapore, Singapore 308433, SingaporeParkinson’s disease (PD) is an age-dependent neurodegenerative condition. Leucine-rich repeat kinase 2 (LRRK2) mutations are the most frequent cause of sporadic and autosomal dominant PD. The exact role of LRRK2 protective variants (R1398H, N551K) together with a pathogenic mutant (G2019S) in aging and neurodegeneration is unknown. We generated the following myc-tagged UAS-LRRK2 transgenic Drosophila: LRRK2 (WT), N551K, R1398H, G2019S single allele, and double-mutants (N551K/G2019S or R1398H/G2019S). The protective variants alone were able to suppress the phenotypic effects caused by the pathogenic LRRK2 mutation. Next, we conducted RNA-sequencing using mRNA isolated from dopaminergic neurons of these different groups of transgenic <i>Drosophila.</i> Using pathway enrichment analysis, we identified the top 10 modules (<i>p</i> < 0.05), with “LRRK2 in neurons in Parkinson’s disease” among the candidates. Further dissection of this pathway identified the most significantly modulated gene nodes such as eEF1A2, ACTB, eEF1A, and actin cytoskeleton reorganization. The induction of the pathway was successfully restored by the R1398H protective variant and R1398H-G2019S or N551K-G2019S rescue experiments. The oxidoreductase family of genes was also active in the pathogenic mutant and restored in protective and rescue variants. In summary, we provide in vivo evidence supporting the neuroprotective effects of LRRK2 variants. RNA sequencing of dopaminergic neurons identified upregulation of specific gene pathways in the <i>Drosophila</i> carrying the pathogenic variant, and this was restored in the rescue phenotypes. Using protective gene variants, our study identifies potential new targets and provides proof of principle of a new therapeutic approach that will further our understanding of aging and neurodegeneration in PD.https://www.mdpi.com/2073-4409/10/1/76neurodegenerationParkinson’s diseaseRNA sequencing<i>Drosophila melanogaster</i>LRRK2 |
| spellingShingle | Joanne Toh Ling Ling Chua Patrick Ho Edwin Sandanaraj Carol Tang Hongyan Wang Eng King Tan Identification of Targets from LRRK2 Rescue Phenotypes Cells neurodegeneration Parkinson’s disease RNA sequencing <i>Drosophila melanogaster</i> LRRK2 |
| title | Identification of Targets from LRRK2 Rescue Phenotypes |
| title_full | Identification of Targets from LRRK2 Rescue Phenotypes |
| title_fullStr | Identification of Targets from LRRK2 Rescue Phenotypes |
| title_full_unstemmed | Identification of Targets from LRRK2 Rescue Phenotypes |
| title_short | Identification of Targets from LRRK2 Rescue Phenotypes |
| title_sort | identification of targets from lrrk2 rescue phenotypes |
| topic | neurodegeneration Parkinson’s disease RNA sequencing <i>Drosophila melanogaster</i> LRRK2 |
| url | https://www.mdpi.com/2073-4409/10/1/76 |
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