Glaucoma is one of the leading causes of vision loss in the world and it is characterized by the dysfunction of retinal ganglion cells (RGCs). An early pathomechanism of glaucoma is the degeneration of the axons of RGCs and finding new treatments that could prevent axon degeneration is of great interest. Increasing the concentration of coenzyme nicotinamide adenine dinucleotide (NAD) has been shown to be axon protective in a number of neurodegenerative systems. Increasing NAD could potentially be achieved by increasing the catalytic properties of the terminal enzyme involved in the cytosolic production of neuronal NAD, nicotinamide mononucleotide adenylyltransferase 2 (NMNAT2). NMNAT2 is thus an ideal therapeutic target. Polyphenol A (PA), which is a polyphenol that will not be disclosed, has been demonstrated to be NAD-boosting through positive modulation of NMNAT2. The aim was to develop a cell-based assay for screening PA and 12 novel analogues of PA for their NAD-boosting effects in brain cortex, retinal and liver cells isolated from C57BL/6J mice. The protocol involved using a bioluminescence assay for which optimization of variables such as cell concentration, substrate (nicotinamide) concentration, PA concentration and incubation time was performed. The method development resulted in a one-day protocol for testing PA and its analogues in cortical cells. PA and several of its analogues exhibited NAD-boosting effects. This protocol along with the results from the screening can be further used for the development of novel drugs that could prevent glaucoma and other axon and neurodegenerations.
| Identifer | oai:union.ndltd.org:UPSALLA1/oai:DiVA.org:uu-419896 |
| Date | January 2020 |
| Creators | Jöe, Melissa |
| Publisher | Uppsala universitet, Institutionen för farmaceutisk biovetenskap |
| Source Sets | DiVA Archive at Upsalla University |
| Language | English |
| Detected Language | English |
| Type | Student thesis, info:eu-repo/semantics/bachelorThesis, text |
| Format | application/pdf |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | UPTEC K, 1650-8297 ; 20023 |
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