Akriti Srivastava, PhD

Pronouns

She/Her/Hers

Rank

Research Fellow

Institution

MGH

BWH-MGH Title

Postdoc fellow

Department

Neurology

Authors

Akriti Srivastava*, Ning Xia, Rachit Bakshi, Michael A. Schwarzschild, Grace F. Crotty

Vitamin B12 inhibits LRRK2 kinase activity in neuronal cells overexpressing the G2019S variant pathogenic for Parkinson’s disease

WMSS 2022

Giving an opportunity to women in Science to discuss their ideas and achievements is a great push and motivation for women like me who are beginning their career in the field. Interacting with women who have carved a niche for themselves in science and also who are progressing towards their goal would inspire me even more to conduct meaningful research. My research goal is to explore the therapeutic benefits of dietary compounds or lifestyle changes in Parkinson’s disease that can delay disease progression in the patients and could be promising candidates for future clinical trials.

Background

The G2019S missense mutation in Leucine-rich repeat kinase (LRRK2) gene is associated with kinase hyperactivity and increased risk of PD. Recently vitamin B12 was reported to allosterically inhibit LRRK2 kinase activity. We aim to investigate whether vitamin B12 dose-dependently inhibits LRRK2 kinase activity in neuronal cells overexpressing wild-type (WT) or G2019S LRRK2.

Methods

Human neuroblastoma SHSY-5Y cells were transfected with WT and G2019S LRRK2 using BacMam viral gene delivery system and exposed to vitamin B12 at different concentrations. After 24h their protein was isolated. Levels of p-LRRK2 (S935), LRRK2, Rab10, and p-Rab10 (T73) were analyzed in both the groups. The effects of vitamin B12 on cytotoxicity induced by PD-associated toxins MPP+ and rotenone were also assessed using MTS assay.

Results

p-LRRK2 (S935)/LRRK2 and p-Rab10 (T73)/Rab10 ratios were significantly lower in G2019S LRRK2-overexpressing cells treated with B12, compared to untreated controls. In WT LRRK2-overexpressing cells however, no significant effect of B12 was observed. Vitamin B12 also significantly reduced MPP+ and rotenone toxicity in SHSY-5Y cells.

Conclusions

Vitamin B12 significantly reduced markers of LRRK2 kinase activity in G2019S LRRK2-overexpressing SHSY-5Y cells. Additionally, B12 improved cell viability of oxidative toxin-exposed cells, supporting its neuroprotective potential. These preliminary findings encourage further evaluation of vitamin B12 in disease-modifying therapy for LRRK2 PD.