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Merck

Inhibition of class I PI3K enhances chaperone-mediated autophagy.

The Journal of cell biology (2020-10-14)
S Joseph Endicott, Zachary J Ziemba, Logan J Beckmann, Dennis N Boynton, Richard A Miller
要旨

Chaperone-mediated autophagy (CMA) is the most selective form of lysosomal proteolysis, where individual peptides, recognized by a consensus motif, are translocated directly across the lysosomal membrane. CMA regulates the abundance of many disease-related proteins, with causative roles in neoplasia, neurodegeneration, hepatosteatosis, and other pathologies relevant to human health and aging. At the lysosomal membrane, CMA is inhibited by Akt-dependent phosphorylation of the CMA regulator GFAP. The INS-PI3K-PDPK1 pathway regulates Akt, but its role in CMA is unclear. Here, we report that inhibition of class I PI3K or PDPK1 activates CMA. In contrast, selective inhibition of class III PI3Ks does not activate CMA. Isolated liver lysosomes from mice treated with either of two orally bioavailable class I PI3K inhibitors, pictilisib or buparlisib, display elevated CMA activity, and decreased phosphorylation of lysosomal GFAP, with no change in macroautophagy. The findings of this study represent an important first step in repurposing class I PI3K inhibitors to modulate CMA in vivo.

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ジメチルスルホキシド, Hybri-Max, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%
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OptiPrep 密度勾配媒体, used for cell and subcellular organelle isolation
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デキサメタゾン, powder, BioReagent, suitable for cell culture, ≥97%
Roche
cOmplete、ミニ、プロテアーゼ阻害剤カクテル, Tablets provided in a glass vial
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アデノシン5′-三リン酸 二ナトリウム塩 水和物, 99%
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Hoechst 33258 溶液, 1 mg/mL in H2O, ≥98.0% (HPLC)
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バフィロマイシン A1 レディーメイド溶液, 0.16 mM in DMSO, from Streptomyces griseus
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Leupeptin, Hemisulfate, Synthetic, Synthetic version of the microbial-derived inhibitor Leupeptin against trypsin-like and cysteine protease activities.