Abstract
Purpose: Fingolimod (FTY720) is widely used in the management of multiple sclerosis and is known to upregulate Akt and Erk neuroprotective survival signalling pathways in the neurons and astrocytes. The drug protects neurons against ischemic stroke and has also been shown to protect the retina against damage caused by experimental glaucoma. In this study, we aimed to investigate the molecular changes caused by fingolimod in the retina and different regions of the brain under normal conditions.
Methods: CBA/CaHArc mice were treated with the FTY720 drug (7.5 mg/kg i.p weekly) or vehicle control for 2 months. Retina and brain tissues (frontal cortex, hippocampus, cerebellum) were harvested and examined from drug treated and control mice (n=10). A multiplexed proteomics using chemical isobaric tandem mass tags (TMTs) using LC/MS was carried out. Detailed functional and protein-protein interaction analyses were performed using Ingenuity pathway analysis, STRING and Panther computational tools.
Results: FTY720 administration lead to differential proteomics changes in various regions of the central nervous system (CNS) particularly in the frontal cortex and cerebellum. Mass spectrometry revealed 29, 10, 1272 and 667 upregulated and 16, 22, 1361, 1265 downregulated proteins in the retina, hippocampus, frontal cortex and cerebellum respectively out of a total of >6000 proteins identified in each case. Computational analysis demonstrated several of the proteins associated with neuroprotective signalling were upregulated (e.g; Sphingosine 1 phosphate receptor 1 (S1PR1) p< 0.0001; phosphatidylinositol 3-Kinase < 0.02; insulin receptor < 0.001; serpin a3k < 0.00001) while markers associated with pro-inflammatory and apoptotic pathways (Caspase 1< 0.007; caspase 3 < 0.01; Tumour necrosis factor (TNF) alpha < 0.0001; tissue plasminogen activator (tPA) < 0.02; interleukin 1R < 0.001; interleukin enhancer binding factor 2 (ILF2) < 0.03) were downregulated.
Conclusions: This study for the first time provides a comprehensive profile of proteomics changes in different CNS regions under normal conditions upon chronic fingolimod treatment. Protein quantification and computational analysis highlight that fingolimod not only promotes suppression of pro-inflammatory pathways, but also up-regulates neuroprotective pathways.
Disclosure
Vivek Gupta: Nothing to disclose.
Mehdi Mirzaei: Nothing to disclose.
Alexander Klistorner: Nothing to disclose.
Stuart Graham: Nothing to disclose.
Source of funding: Novartis Pharmaceuticals Australia.
Methods: CBA/CaHArc mice were treated with the FTY720 drug (7.5 mg/kg i.p weekly) or vehicle control for 2 months. Retina and brain tissues (frontal cortex, hippocampus, cerebellum) were harvested and examined from drug treated and control mice (n=10). A multiplexed proteomics using chemical isobaric tandem mass tags (TMTs) using LC/MS was carried out. Detailed functional and protein-protein interaction analyses were performed using Ingenuity pathway analysis, STRING and Panther computational tools.
Results: FTY720 administration lead to differential proteomics changes in various regions of the central nervous system (CNS) particularly in the frontal cortex and cerebellum. Mass spectrometry revealed 29, 10, 1272 and 667 upregulated and 16, 22, 1361, 1265 downregulated proteins in the retina, hippocampus, frontal cortex and cerebellum respectively out of a total of >6000 proteins identified in each case. Computational analysis demonstrated several of the proteins associated with neuroprotective signalling were upregulated (e.g; Sphingosine 1 phosphate receptor 1 (S1PR1) p< 0.0001; phosphatidylinositol 3-Kinase < 0.02; insulin receptor < 0.001; serpin a3k < 0.00001) while markers associated with pro-inflammatory and apoptotic pathways (Caspase 1< 0.007; caspase 3 < 0.01; Tumour necrosis factor (TNF) alpha < 0.0001; tissue plasminogen activator (tPA) < 0.02; interleukin 1R < 0.001; interleukin enhancer binding factor 2 (ILF2) < 0.03) were downregulated.
Conclusions: This study for the first time provides a comprehensive profile of proteomics changes in different CNS regions under normal conditions upon chronic fingolimod treatment. Protein quantification and computational analysis highlight that fingolimod not only promotes suppression of pro-inflammatory pathways, but also up-regulates neuroprotective pathways.
Disclosure
Vivek Gupta: Nothing to disclose.
Mehdi Mirzaei: Nothing to disclose.
Alexander Klistorner: Nothing to disclose.
Stuart Graham: Nothing to disclose.
Source of funding: Novartis Pharmaceuticals Australia.
| Original language | English |
|---|---|
| Article number | EP1679 |
| Pages (from-to) | 882-883 |
| Number of pages | 2 |
| Journal | Multiple Sclerosis Journal |
| Volume | 23 |
| Issue number | 3 suppl |
| DOIs | |
| Publication status | Published - Oct 2017 |
| Event | MSPARIS2017: 7th Joint ECTRIMS - ACTRIMS Meeting - Paris, France Duration: 25 Oct 2017 → 28 Oct 2018 Conference number: 7th |