Esterified Trehalose Analogues Protect Mammalian Cells from Heat Shock

Abstract

Trehalose is a disaccharide produced by many organisms to better enable them to survive environmental stresses, including heat, cold, desiccation, and reactive oxygen species. Mammalian cells do not naturally biosynthesize trehalose; however, when introduced into mammalian cells, trehalose provides protection from damage associated with freezing and drying. One of the major difficulties in using trehalose as a cellular protectant for mammalian cells is the delivery of this disaccharide into the intracellular environment; mammalian cell membranes are impermeable to the hydrophilic sugar trehalose. A panel of cell-permeable trehalose analogues, in which the hydrophilic hydroxyl groups of trehalose are masked as esters, have been synthesized and the ability of these analogues to load trehalose into mammalian cells has been evaluated. Two of these analogues deliver millimolar concentrations of free trehalose into a variety of mammalian cells. Critically, Jurkat cells incubated with these analogues show improved survival after heat shock, relative to untreated Jurkat cells. The method reported herein thus paves the way for the use of esterified analogues of trehalose as a facile means to deliver high concentrations of trehalose into mammalian cells for use as a cellular protectant.

Quantification of the Disaccharide Trehalose from Biological Samples: A Comparison of Analytical Methods

“…Neither the enzymatic trehalose assay nor the HPLC-RID-based assay can differentiate between these different disaccharides. Esterified trehalose analogues have also recently been used to deliver trehalose into mammalian cells; for trehalose quantification, one study used the enzymatic trehalose assay and another study used the LC–MS/MS-based assay. , The concentrations of esterified trehalose analogues from these cells could also be directly quantified using the LC–MS/MS-based assay; it is unlikely that the enzymatic trehalose assay or the HPLC-RID-based assay could detect these analogues.…”

“…The disaccharide trehalose, or α- d -glucose­(1→1)­α- d -glucose (Figure a), acts as a remarkable cellular protectant for many different organisms, including bacteria, fungi, plants, insects, and eukaryotic microorganisms. − When subjected to extreme conditions, such as cold, heat, desiccation, or reactive oxygen species, these organisms biosynthesize high concentrations of both intra- and extracellular trehalose, which allows them to better survive these environmental stresses. − For example, the desiccation-tolerant plant, Selaginella lepidophylla, accumulates intracellular trehalose at levels up to 12% of its dry weight during periods of drought . High concentrations of trehalose are also found in Saccharomyces cerevisiae (a strain of yeast) that are subjected to heating; yeast mutants that are defective in the genes that encode for trehalose biosynthesis are unable to produce trehalose upon heat shock and are much less resistant to heating than wild-type yeast. , Mammals do not naturally produce trehalose; however, delivery of trehalose into mammalian cells improves survival rates after freezing, drying, or heat shock. − Furthermore, the administration of exogenous trehalose has been shown to provide neuroprotective effects in animal models of Huntington’s disease, Parkinson’s disease, and amytrophic lateral sclerosis. − The ability of trehalose to protect cells from damage, along with its lack of cellular toxicity, has generated interest in using this disaccharide as a general cellular protectant. ,− ,− …”

Dry storage of mammalian spermatozoa and cells: state-of-the-art and possible future directions

“…Given that the amount of trehalose loaded into spermatozoa by diffusion or phase transition is limited, it is worthy exploring alternative strategies. One promising strategy may be the use of trehalose analogues, in which the hydrophilic hydroxyl groups of trehalose are turned into esters (Bragg et al 2017). Another promising permeable version of trehalose is trehalose hexaacetate, a molecule synthesised several years ago that is lipophilic and can readily cross the membrane (Abazari et al 2015).…”

Water Effect on Peroxyoxalate Kinetics and Mechanism for Oxalic Esters with Distinct Reactivities

“…Besides, the peroxyoxalate system can be applied in different fields of pharmacology and diagnosis. Since malignant cells produce increased amounts of hydrogen peroxide, formulations based on oxalates can be used to show the foci of tumor growth and other inflammatory processes (10–13). A wide variety of organic compounds can be determined through the peroxyoxalate system, including cholesterol, glucose, lactose, bilirubin, oxalate, formaldehyde, formic acid, L‐amino acid, polyamines and acetylcholine (14–17).…”