Messenger RNA based therapeutics as a novel curative approach for disorders of neurodegeneration the example of Niemann Pick type C

Specific Aim 1: NPC1 skin fibroblasts from our NPC1 patient repository will be corrected using Moderna's RNA based therapy. Successful correction will be evaluated by Western blot, analysis of cholesterol trafficking (Filipin stain) and measurement of complex lipids including oxysterols in supernatant of fibroblasts and cell extracts. All analyses will be done in my laboratory through already established methods. Specific Aim 2: To identify best route of administration (i.p., s.c., i.v. in combination with or without i.t.) of tagged messenger RNA based therapeutics in 20 wildtype mice that are bred on the same background as the NPC1 knock-out mice. Tissue distribution of tagged messenger RNA will be studied in relation to the route of administration with particular emphasis on the brain. Specific Aim 3a: To use messenger RNA based therapeutics in a double blind, placebo controlled, cross over design in a cohort of newborn 12 npc1 knockout and 12 wildtype mice using the route of administration determined in Aim 1. Primary endpoints include plasma oxysterol levels, filipin stain (cholesterol accumulation) in liver, skin and purkinje cells and europsychologial testing. Secondary endpoints include transcriptome, methylome, metabolome and biomarker levels that are currently in development. Results will inform whether early intervention may be able to prevent disease manifestations, which may be important for future newborn screening for NPC1 in humans.Specific Aim 3b: To use messenger RNA based therapeutics in a double blind, placebo controlled, cross over design in a cohort of adult 12 npc1 knockout and 12 wildtype mice. Primary endpoints include plasma oxysterol levels, filipin stain (cholesterol accumulation) in liver, skin and purkinje cells and neuropsychologial testing. Secondary endpoints include transcriptome, methylome, metabolome and biomarker levels that are currently in development. Results will inform about the effect of late intervention on disease progression.