Downregulation of striatal Ca1.3 inhibits the escalation of levodopa-induced dyskinesia in male and female parkinsonian rats of advanced age

Authors

Margaret E. Caulfield, Department of Translational Neuroscience, College of Human Medicine, Michigan State University, 400 Monroe Ave NW, Grand Rapids, MI 49503, USA.
Molly J. Vander Werp, Department of Translational Neuroscience, College of Human Medicine, Michigan State University, 400 Monroe Ave NW, Grand Rapids, MI 49503, USA.
Jennifer A. Stancati, Department of Translational Neuroscience, College of Human Medicine, Michigan State University, 400 Monroe Ave NW, Grand Rapids, MI 49503, USA.
Timothy J. Collier, Department of Translational Neuroscience, College of Human Medicine, Michigan State University, 400 Monroe Ave NW, Grand Rapids, MI 49503, USA.
Caryl E. Sortwell, Department of Translational Neuroscience, College of Human Medicine, Michigan State University, 400 Monroe Ave NW, Grand Rapids, MI 49503, USA.
Ivette M. Sandoval, Parkinson's Disease Research Unit, Department of Translational Neuroscience, Barrow Neurological Institute, Phoenix, AZ 85013, USA.
Fredric P. Manfredsson, Parkinson's Disease Research Unit, Department of Translational Neuroscience, Barrow Neurological Institute, Phoenix, AZ 85013, USA.Follow
Kathy Steece-Collier, Department of Translational Neuroscience, College of Human Medicine, Michigan State University, 400 Monroe Ave NW, Grand Rapids, MI 49503, USA. Electronic address: collie68@msu.edu.

Document Type

Article

Abstract

In the past 25 years, the prevalence of Parkinson's disease (PD) has nearly doubled. Age remains the primary risk factor for PD and as the global aging population increases this trend is predicted to continue. Even when treated with levodopa, the gold standard dopamine (DA) replacement therapy, individuals with PD frequently develop therapeutic side effects. Levodopa-induced dyskinesia (LID), a common side effect of long-term levodopa use, represents a significant unmet clinical need in the treatment of PD. Previously, in young adult (3-month-old) male parkinsonian rats, we demonstrated that the silencing of Ca1.3 (Cacan1d) L-type voltage-gated calcium channels via striatal delivery of rAAV-Ca1.3-shRNA provides uniform protection against the induction of LID, and significant reduction of established severe LID. With the goal of more closely replicating a clinical demographic, the current study examined the effects of Ca1.3-targeted gene therapy on LID escalation in male and female parkinsonian rats of advanced age (18-month-old at study completion). We tested the hypothesis that silencing aberrant Ca1.3 channel activity in the parkinsonian striatum would prevent moderate to severe dyskinesia with levodopa dose escalation. To test this hypothesis, 15-month-old male and female F344 rats were rendered unilaterally parkinsonian and primed with low-dose (3-4 mg/kg) levodopa. Following the establishment of stable, mild dyskinesias, rats received an intrastriatal injection of either the Cacna1d-specific rAAV-Ca1.3-shRNA vector (CAV-shRNA), or the scramble control rAAV-SCR-shRNA vector (SCR-shRNA). Daily (M-Fr) low-dose levodopa was maintained for 4 weeks during the vector transduction and gene silencing window followed by escalation to 6 mg/kg, then to 12 mg/kg levodopa. SCR-shRNA-shRNA rats showed stable LID expression with low-dose levodopa and the predicted escalation of LID severity with increased levodopa doses. Conversely, complex behavioral responses were observed in aged rats receiving CAV-shRNA, with approximately half of the male and female subjects-therapeutic 'Responders'-demonstrating protection against LID escalation, while the remaining half-therapeutic 'Non-Responders'-showed LID escalation similar to SCR-shRNA rats. Post-mortem histological analyses revealed individual variability in the detection of Cacna1d regulation in the DA-depleted striatum of aged rats. However, taken together, male and female therapeutic 'Responder' rats receiving CAV-shRNA had significantly less striatal Cacna1d in their vector-injected striatum relative to contralateral striatum than those with SCR-shRNA. The current data suggest that mRNA-level silencing of striatal Ca1.3 channels maintains potency in a clinically relevant in vivo scenario by preventing dose-dependent dyskinesia escalation in rats of advanced age. As compared to the uniform response previously reported in young male rats, there was notable variability between individual aged rats, particularly females, in the current study. Future investigations are needed to derive the sex-specific and age-related mechanisms which underlie variable responses to gene therapy and to elucidate factors which determine the therapeutic efficacy of treatment for PD.

Keywords

Adeno-associated virus (AAV), Aging, Ca(V)1.3 channels, Gene therapy, Levodopa-induced dyskinesia, Parkinson's disease, Striatum, Viral vectors

Medical Subject Headings

Rats; Male; Female; Animals; Levodopa (adverse effects); Down-Regulation; Rats, Sprague-Dawley; Rats, Inbred F344; Dyskinesia, Drug-Induced (metabolism); Dopamine; Parkinson Disease (drug therapy); RNA, Small Interfering; Antiparkinson Agents (pharmacology); Oxidopamine

Publication Date

6-1-2023

Publication Title

Neurobiology of disease

E-ISSN

1095-953X

Volume

181

First Page

106111

PubMed ID

37001610

Digital Object Identifier (DOI)

10.1016/j.nbd.2023.106111

Recommended Citation

Caulfield, Margaret E.; Vander Werp, Molly J.; Stancati, Jennifer A.; Collier, Timothy J.; Sortwell, Caryl E.; Sandoval, Ivette M.; Manfredsson, Fredric P.; and Steece-Collier, Kathy, "Downregulation of striatal Ca1.3 inhibits the escalation of levodopa-induced dyskinesia in male and female parkinsonian rats of advanced age" (2023). Translational Neuroscience. 2250.
https://scholar.barrowneuro.org/neurobiology/2250