Alpha-synuclein (?-Syn) is one of the most abundant proteins in the CNS that is known to be a major player in the neurodegeneration observed in Parkinson?s disease. We show that stroke (transient focal ischemia) upregulates ?-Syn protein expression and nuclear translocation in neurons of adult rodents and humans. We further show that knockdown or knockout of ?-Syn significantly decreases the infarction and promotes better neurological recovery in rodents subjected to focal ischemia. Based on these exciting new leads, in this proposal we wish to test the therapeutic potential of targeting ?-Syn in post-stroke brain by following the criteria set by the Stroke Treatment Academic Industry Roundtable (STAIR) consortium.
Aim 1 is to evaluate the window of therapeutic opportunity, effect of sex, age, route of administration and toxicity of ?-Syn siRNA therapy following focal ischemia in rodents. We further observed that a microRNA called miR-7a potently targets ?-Syn. Importantly miR-7a showed an inverse relation to ?-Syn (miR-7a levels were down-regulated while ?-Syn levels were upregulated after stroke). Hence, we will test the efficacy of replenishing miR-7a in the post-stroke brain to repress ?-Syn and thus decrease brain damage. Testing alternate approaches to target a protein gives better opportunities for future clinical translation. Hence, miR-7a mimic therapy will serve as an alternate approach to ?-Syn siRNA therapy.
Aim 2 is to evaluate the window of therapeutic opportunity, effect of sex, age, route of administration, toxicity and long-term effects of miR-7a mimic after focal ischemia. The mechanisms that contribute to ?-Syn-mediated secondary brain damage after stroke are not well understood. We demonstrate that ?-Syn protein formed in excess in brain during the acute phase after stroke oligomerizes and forms aggregates with time. We further show that ?-Syn promotes brain damage by multiple pathologic mechanisms including mitochondrial fission. In chronic neurodegeneration, ?-Syn is known to act as an essential scaffolding molecule for the activation of GSK-3? and the subsequent Tau hyperphosphorylation that leads to activation of Drp1 which promotes mitochondrial fission. In preliminary studies we observed increased phosphorylation of GSK-3?, Tau and Drp1.
In Aim 3, we will test if ?-Syn promotes post-ischemic mitochondrial fission and brain damage by involving GSK-3? and Tau. The long-term goal of these studies is to evaluate if targeting ?-Syn is a viable option for stroke therapy in both males and females and at different ages.

Public Health Relevance

?-Synuclein is a protein that plays a major role in promoting neurodegeneration in Parkinson?s disease. We observed that ?-Synuclein is also induced in brain after stroke. Hence, we will test the therapeutic efficacy of preventing ?-Synuclein in young/adult and middle-aged male and female rodents to identify the window of therapeutic opportunity, toxicity, best route. We will also study a molecular mechanism that is thought to mediate ?-Synuclein toxicity after stroke. The long-term goal is to develop a therapy to prevent post-stroke brain damage and neurological dysfunction.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS101960-05
Application #
10109156
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Bosetti, Francesca
Project Start
2017-04-01
Project End
2022-02-28
Budget Start
2021-03-01
Budget End
2022-02-28
Support Year
5
Fiscal Year
2021
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Neurosurgery
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Verma, Rajkumar; Ritzel, Rodney M; Harris, Nia M et al. (2018) Inhibition of miR-141-3p Ameliorates the Negative Effects of Poststroke Social Isolation in Aged Mice. Stroke 49:1701-1707
Kim, Joonki; Kang, Sung-Wook; Mallilankaraman, Karthik et al. (2018) Transcriptome analysis reveals intermittent fasting-induced genetic changes in ischemic stroke. Hum Mol Genet 27:1497-1513
Mehta, Suresh L; Vemuganti, Raghu (2018) Ischemic Stroke Alters the Expression of the Transcribed Ultraconserved Regions of the Genome in Rat Brain. Stroke 49:1024-1028