Journal of Medicinal Chemistry. 2020 Dec 21
Daniel J. Apicco, Evgeny Shlevkov, Catherine L. Nezich, David T. Tran, Edward Guilmette, Justin W. Nicholatos, Collin M. Bantle, Yi Chen, Kelly E. Glajch, Neeta A. Abraham, Lan T. Dang, G. Campbell Kaynor, Ellen A. Tsai, Khanh-Dung H. Nguyen, Joost Groot, YuTing Liu, Andreas Weihofen, Jessica A. Hurt, Heiko Runz, and Warren D. Hirsta
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AAV vector packaging Plasmids were then shipped to Packgene Biotech (Worcester, MA) for cloning into AAV entry vectors under the CAG promoter followed by packaging into AAV using the PHP.B serotype. Request Quote

Research Field: CNS

AAV Serotype: PHP.B

Dose: Adeno-associated viruses (AAVs) were injected into mice on postnatal day 0 (P0). Pups were anesthetized on ice and were then injected intracerebroventricularly (i.c.v.) with 4 μl virus (1E11 GCs) mixed with fast green dye (<1% total).


Inositol-1,4,5-triphosphate (IP3) kinase B (ITPKB) is a ubiquitously expressed lipid kinase that inactivates IP3, a secondary messenger that stimulates calcium release from the endoplasmic reticulum (ER). Genome-wide association studies have identified common variants in the ITPKB gene locus associated with reduced risk of sporadic Parkinson’s disease (PD). Here, we investigate whether ITPKB activity or expression level impacts PD phenotypes in cellular and animal models. In primary neurons, knockdown or pharmacological inhibition of ITPKB increased levels of phosphorylated, insoluble α-synuclein pathology following treatment with α-synuclein preformed fibrils (PFFs). Conversely, ITPKB overexpression reduced PFF-induced α-synuclein aggregation. We also demonstrate that ITPKB inhibition or knockdown increases intracellular calcium levels in neurons, leading to an accumulation of calcium in mitochondria that increases respiration and inhibits the initiation of autophagy, suggesting that ITPKB regulates α-synuclein pathology by inhibiting ER-to-mitochondria calcium transport. Furthermore, the effects of ITPKB on mitochondrial calcium and respiration were prevented by pretreatment with pharmacological inhibitors of the mitochondrial calcium uniporter complex, which was also sufficient to reduce α-synuclein pathology in PFF-treated neurons. Taken together, these results identify ITPKB as a negative regulator of α-synuclein aggregation and highlight modulation of ER-to-mitochondria calcium flux as a therapeutic strategy for the treatment of sporadic PD.

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