Drugs that mirror the cellular effects of starvation mimics are considered promising therapeutics for common metabolic disorders, such as obesity, liver steatosis, and for ageing. Starvation, or caloric restriction, is known to activate the transcription factor EB (TFEB), a master regulator of lipid metabolism and lysosomal biogenesis and function. Here, we report a nanotechnology-enabled high-throughput screen to identify small-molecule agonists of TFEB and discover three novel compounds that promote autophagolysosomal activity. The three lead compounds include the clinically approved drug, digoxin; the marine-derived natural product, ikarugamycin; and the synthetic compound, alexidine dihydrochloride, which is known to act on a mitochondrial target. Mode of action studies reveal that these compounds activate TFEB via three distinct Ca2+-dependent mechanisms. Formulation of these compounds in liver-tropic biodegradable, biocompatible nanoparticles confers hepatoprotection against diet-induced steatosis in murine models and extends lifespan of Caenorhabditis elegans. These results support the therapeutic potential of small-molecule TFEB activators for the treatment of metabolic and age-related disorders.
Fluorescent molecules are highly susceptible to their local environment. Thus, a fluorescent molecule near a plasmonic nanoparticle can experience changes in local electric field and local density of states that reshape its intrinsic emission spectrum. By avoiding ensemble averaging while simultaneously measuring the super-resolved position of the fluorophore and its emission spectrum, single-molecule hyperspectral imaging is uniquely suited to differentiate changes in the spectrum from heterogeneous ensemble effects. Thus, we uncover for the first time single-molecule fluorescence emission spectrum reshaping upon near-field coupling to individual gold nanoparticles using hyperspectral super-resolution fluorescence imaging, and we resolve this spectral reshaping as a function of the nanoparticle/dye spectral overlap and separation distance. We find that dyes bluer than the plasmon resonance maximum are red-shifted and redder dyes are blue-shifted. The primary vibronic peak transition probabilities shift to favor secondary vibronic peaks, leading to effective emission maxima shifts in excess of 50 nm, and we understand these light-matter interactions by combining super-resolution hyperspectral imaging and full-field electromagnetic simulations.
