Novel Poly-Arginine Peptide R18D Reduces α-Synuclein Aggregation and Uptake of α-Synuclein Seeds in Cortical Neurons
Background/Objectives: The role of α-synuclein (α-syn) pathology in Parkinson’s disease (PD) is well established; however, effective therapies remain elusive. Two mechanisms central to PD neurodegeneration are the intracellular aggregation of misfolded α-syn and the uptake of α-syn aggregates into n...
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2025-01-01
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| author | Emma C. Robinson Anastazja M. Gorecki Samuel R. Pesce Vaishali Bagda Ryan S. Anderton Bruno P. Meloni |
| author_facet | Emma C. Robinson Anastazja M. Gorecki Samuel R. Pesce Vaishali Bagda Ryan S. Anderton Bruno P. Meloni |
| author_sort | Emma C. Robinson |
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| description | Background/Objectives: The role of α-synuclein (α-syn) pathology in Parkinson’s disease (PD) is well established; however, effective therapies remain elusive. Two mechanisms central to PD neurodegeneration are the intracellular aggregation of misfolded α-syn and the uptake of α-syn aggregates into neurons. Cationic arginine-rich peptides (CARPs) are an emerging class of molecule with multiple neuroprotective mechanisms of action, including protein stabilisation. This study characterised both intracellular α-syn aggregation and α-syn uptake in cortical neurons in vitro. Thereafter, this study examined the therapeutic potential of the neuroprotective CARP, R18D (18-mer of D-arginine), to prevent the aforementioned PD pathogenic processes through a cell-free thioflavin-T (ThT) assay and in cortical neurons. Methods: To induce intracellular α-syn aggregation, rat primary cortical neurons were exposed to α-syn seed (0.14 μM) for 2 h to allow uptake of the protein, followed by R18D treatment (0.0625, 0.125, 0.25, 0.5 μM), and a subsequent measurement of α-syn aggregates 48 h later using a homogenous time-resolved fluorescence (HTRF) assay. To assess neuronal uptake, α-syn seeds were covalently labelled with an Alexa-Fluor 488 fluorescent tag, pre-incubated with R18D (0.125, 0.25, 0.5 μM), and then exposed to cortical neurons for 24 h and assessed via confocal microscopy. Results: It was demonstrated that R18D significantly reduced both intracellular α-syn aggregation and α-syn seed uptake in neurons by 37.8% and 77.7%, respectively. Also, R18D reduced the aggregation of α-syn monomers in the cell-free assay. Conclusions: These findings highlight the therapeutic potential of R18D to inhibit key α-syn pathological processes and PD progression. |
| format | Article |
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| institution | Kabale University |
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| language | English |
| publishDate | 2025-01-01 |
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| series | Biomedicines |
| spelling | doaj-art-4737a9ff08d841379f13bd95f1cd5c3f2025-01-24T13:24:05ZengMDPI AGBiomedicines2227-90592025-01-0113112210.3390/biomedicines13010122Novel Poly-Arginine Peptide R18D Reduces α-Synuclein Aggregation and Uptake of α-Synuclein Seeds in Cortical NeuronsEmma C. Robinson0Anastazja M. Gorecki1Samuel R. Pesce2Vaishali Bagda3Ryan S. Anderton4Bruno P. Meloni5Perron Institute for Neurological and Translational Science, Nedlands 6009, AustraliaSchool of Health Sciences, University of Notre Dame, Fremantle 6106, AustraliaCentre for Neuromuscular and Neurological Disorders, University of Western Australia, Nedlands 6009, AustraliaPerron Institute for Neurological and Translational Science, Nedlands 6009, AustraliaSchool of Health Sciences, University of Notre Dame, Fremantle 6106, AustraliaPerron Institute for Neurological and Translational Science, Nedlands 6009, AustraliaBackground/Objectives: The role of α-synuclein (α-syn) pathology in Parkinson’s disease (PD) is well established; however, effective therapies remain elusive. Two mechanisms central to PD neurodegeneration are the intracellular aggregation of misfolded α-syn and the uptake of α-syn aggregates into neurons. Cationic arginine-rich peptides (CARPs) are an emerging class of molecule with multiple neuroprotective mechanisms of action, including protein stabilisation. This study characterised both intracellular α-syn aggregation and α-syn uptake in cortical neurons in vitro. Thereafter, this study examined the therapeutic potential of the neuroprotective CARP, R18D (18-mer of D-arginine), to prevent the aforementioned PD pathogenic processes through a cell-free thioflavin-T (ThT) assay and in cortical neurons. Methods: To induce intracellular α-syn aggregation, rat primary cortical neurons were exposed to α-syn seed (0.14 μM) for 2 h to allow uptake of the protein, followed by R18D treatment (0.0625, 0.125, 0.25, 0.5 μM), and a subsequent measurement of α-syn aggregates 48 h later using a homogenous time-resolved fluorescence (HTRF) assay. To assess neuronal uptake, α-syn seeds were covalently labelled with an Alexa-Fluor 488 fluorescent tag, pre-incubated with R18D (0.125, 0.25, 0.5 μM), and then exposed to cortical neurons for 24 h and assessed via confocal microscopy. Results: It was demonstrated that R18D significantly reduced both intracellular α-syn aggregation and α-syn seed uptake in neurons by 37.8% and 77.7%, respectively. Also, R18D reduced the aggregation of α-syn monomers in the cell-free assay. Conclusions: These findings highlight the therapeutic potential of R18D to inhibit key α-syn pathological processes and PD progression.https://www.mdpi.com/2227-9059/13/1/122Parkinson’s diseaseα-synucleincationic arginine-rich peptidesR18Dprimary cortical neuronsα-synuclein seeds |
| spellingShingle | Emma C. Robinson Anastazja M. Gorecki Samuel R. Pesce Vaishali Bagda Ryan S. Anderton Bruno P. Meloni Novel Poly-Arginine Peptide R18D Reduces α-Synuclein Aggregation and Uptake of α-Synuclein Seeds in Cortical Neurons Biomedicines Parkinson’s disease α-synuclein cationic arginine-rich peptides R18D primary cortical neurons α-synuclein seeds |
| title | Novel Poly-Arginine Peptide R18D Reduces α-Synuclein Aggregation and Uptake of α-Synuclein Seeds in Cortical Neurons |
| title_full | Novel Poly-Arginine Peptide R18D Reduces α-Synuclein Aggregation and Uptake of α-Synuclein Seeds in Cortical Neurons |
| title_fullStr | Novel Poly-Arginine Peptide R18D Reduces α-Synuclein Aggregation and Uptake of α-Synuclein Seeds in Cortical Neurons |
| title_full_unstemmed | Novel Poly-Arginine Peptide R18D Reduces α-Synuclein Aggregation and Uptake of α-Synuclein Seeds in Cortical Neurons |
| title_short | Novel Poly-Arginine Peptide R18D Reduces α-Synuclein Aggregation and Uptake of α-Synuclein Seeds in Cortical Neurons |
| title_sort | novel poly arginine peptide r18d reduces α synuclein aggregation and uptake of α synuclein seeds in cortical neurons |
| topic | Parkinson’s disease α-synuclein cationic arginine-rich peptides R18D primary cortical neurons α-synuclein seeds |
| url | https://www.mdpi.com/2227-9059/13/1/122 |
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