Citation
- Authors: Srivastava V. et al.
- Year: 2023
- Journal: Autophagy 19 474-504
- Applications: in vitro / DNA, siRNA / INTERFERin, jetOPTIMUS
- Cell type: SH-SY5Y
Description: Human neuroblastoma cells
Known as:
Method
SHSY-5Y cells were plated in a 6-well tissue culture plate at a density of 1,000,000 cells per well in complete growth media (DMEM high glucose media, 2% L-glutamine, 10% FBS and 1% tetracycline) and incubated at 37°C for 24 h to 70% confluence. Then, the cells were transiently transfected with the cox8-mCherry-EGFP reporter [88] (Addgene, 78,520; deposited by David Chan) according to the jetOPTIMUS (Polyplus, 117–07) instructions.
siRNA transfection was performed using INTERFERin (Polyplus, 101,000,028) following the manufacturers’ protocols. In brief, SH-SY5Y cells were incubated for 24 h with the siRNA transfection complexes (total concentration of 25 nM, i.e., 8.33 nM of each diRNA duplex). PINK1 depletion was confirmed by qPCR. Six independent experiments, each in triplicates were performed.
Abstract
Impaired mitophagy is a primary pathogenic event underlying diverse aging-associated diseases such as Alzheimer and Parkinson diseases and sarcopenia. Therefore, augmentation of mitophagy, the process by which defective mitochondria are removed, then replaced by new ones, is an emerging strategy for preventing the evolvement of multiple morbidities in the elderly population. Based on the scaffold of spermidine (Spd), a known mitophagy-promoting agent, we designed and tested a family of structurally related compounds. A prototypic member, 1,8-diaminooctane (VL-004), exceeds Spd in its ability to induce mitophagy and protect against oxidative stress. VL-004 activity is mediated by canonical aging genes and promotes lifespan and healthspan in C. elegans. Moreover, it enhances mitophagy and protects against oxidative injury in rodent and human cells. Initial structural characterization suggests simple rules for the design of compounds with improved bioactivity, opening the way for a new generation of agents with a potential to promote healthy aging.