Lactotrehalose, an analog of trehalose, increases energy metabolism without promoting clostridioides difficile infection in mice

Authors: ZHANG, YIMING - University Of Washington Medical School; SHAIKH, NURMOHAMMAD - Washington University School Of Medicine; FEREY, JEREMIE - Washington University School Of Medicine; WANKHADE, UMESH - University Arkansas For Medical Sciences (UAMS); CHINTAPALLI, SREE - University Arkansas For Medical Sciences (UAMS); HIGGINS, CASSANDRA - Washington University School Of Medicine; CROWLEY, JAN - Washington University School Of Medicine; HEITMEIER, MONIQUE - Washington University School Of Medicine; STOTHARD, ALICYN - Central Michigan University; MIHI, BELGACEM - Washington University School Of Medicine; GOOD, MISTY - Washington University School Of Medicine; HIGASHIYAMA, TAKANOBU - Research & Development, Inc; SWARTS, BENJAMIN - Central Michigan University; HRUZ, PAUL - Washington University School Of Medicine; SHANKAR, KARTIK - University Arkansas For Medical Sciences (UAMS); TARR, PHILLIP - Washington University School Of Medicine; DEBOSCH, BRIAN - Washington University School Of Medicine

Submitted to: Gastroenterology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/26/2019
Publication Date: 12/12/2019
Citation: Zhang, Y., Shaikh, N., Ferey, J.L., Wankhade, U.D., Chintapalli, S.V., Higgins, C.B., Crowley, J.R., Heitmeier, M.R., Stothard, A.I., Mihi, B., Good, M.R., Higashiyama, T., Swarts, B.M., Hruz, P.W., Shankar, K., Tarr, P.I., DeBosch, B.J. 2019. Lactotrehalose, an analog of trehalose, increases energy metabolism without promoting clostridioides difficile infection in mice. Gastroenterology. 158(5):1402-1416.e2. https://doi.org/10.1053/j.gastro.2019.11.295.
DOI: https://doi.org/10.1053/j.gastro.2019.11.295

Interpretive Summary: Obesity-associated disorders such as non alcoholic fatty liver disease (NAFLD) are on the rise in last few decades. There is a need to understand the molecular underpinnings in order to prevent NAFLD, mitigate obesity and to improve metabolic health. The potential influence of dietary factors as a tool to prevent disease is worth considering, an in particular fibers and other components that can modify the normal population of bacteria in the gut ("gut microbiome"). In the current study, one such factor, lactotrehalose, was tested in a mouse model of fatty liver. A related sugar, trehalose, is a disaccharide that may promote expansion of harmful bacteria such as Clostridioides difficile ribotypes that metabolize trehalose via an enzyme called trehalose-6-phosphate hydrolase (treA). Using the trehalase-resistant analogue of trehalose (lactotrehalose) in the diet of mice was found to have beneficial metabolic effects without expanding harmful bacteria such as C difficile. Lactotrehalose and trehalose each blocked glucose transport, and induced a gene expression pattern associated with fasting in primary liver cells (hepatocytes). Mice given lactotrehalose while on obesity- and NAFLD-promoting high-fructose diet did not develop metabolic anomalies, and had increased whole-body energy expenditure, compared with mice given trehalose or water. Lactotrehalose reduced fecal markers of microbial branched chain amino acid biosynthesis and increased expression of microbial genes that regulate insulin signaling. In conclusion, lactotrehalose improves metabolic parameters, compared with trehalose, without increasing the abundance or virulence of C difficile. Trehalase-resistant trehalose analogues can be useful tools to learn about mechanisms to thwart NAFLD development, and these mechanisms may be shared, in part, with select foods that are part of a healthy diet pattern.

Technical Abstract: Trehalose is a disaccharide that might be used in treatment of cardiometabolic diseases. However, trehalose consumption promotes expansion of Clostridioides difficile ribotypes that metabolize trehalose via trehalose-6-phosphate hydrolase (treA). Furthermore, brush border and renal trehalases can reduce the efficacy of trehalose by cleaving it into monosaccharides. We investigated whether a trehalase-resistant analogue of trehalose (lactotrehalose) has the same metabolic effects of trehalose without expanding C difficile. We performed studies with HEK293 and Caco2 cells, primary hepatocytes from mice, and human intestinal organoids. Glucose transporters were overexpressed in HEK293 cells and glucose transport was quantified. Primary hepatocytes were cultured with or without trehalose or lactotrehalose and gene expression patterns were analyzed. C57B6/J mice were given oral antibiotics and trehalose or lactotrehalose in drinking water, or only water (control), followed by gavage with the virulent C difficile ribotype 027 (CD027); fecal samples were analyzed for ToxA or ToxB by ELISA. Other mice were given trehalose or lactotrehalose in drinking water for 2 days before placement on a chow or 60% fructose diet for 10 days. Liver tissues were collected and analyzed by histologic, serum biochemical, and RNAseq, autophagic flux, and thermogenesis analyses. We quantified portal trehalose and lactotrehalose bioavailability by gas chromatography mass spectrometry. Fecal microbiomes were analyzed by 16S rRNA sequencing and principal component analyses. Lactotrehalose and trehalose each blocked glucose transport in HEK293 cells, and induced a gene expression pattern associated with fasting in primary hepatocytes. Compared with mice on the chow diet, mice on the high-fructose diet had increased circulating cholesterol, higher ratios of liver weight:body weight, hepatic lipid accumulation (steatosis), and liver gene expression patterns of carbohydrate-responsive de novo lipogenesis. Mice given lactotrehalose while on the high-fructose diet did not develop any of these features and had increased whole-body caloric expenditure compared with mice given trehalose or water and fed a high-fructose diet. Livers from mice given lactotrehalose had increased transcription of genes that regulate mitochondrial energy metabolism compared with liver from mice given trehalose or controls. Lactotrehalose was bioavailable in venous and portal circulation and fecal samples. Lactotrehalose reduced fecal markers of microbial branched chain amino acid biosynthesis and increased expression of microbial genes that regulate insulin signaling. In mice given antibiotics followed by CD027, neither lactotrehalose nor trehalose increased levels of the bacteria or its toxin in stool—in fact, trehalose reduced the abundance of CD027 in stool. Lactotrehalose and trehalose reduced markers of inflammation in rectal tissue following CD027 infectio Lactotrehalose is a trehalase-resistant analogue that increases metabolic parameters, compared with trehalose, without increasing the abundance or virulence of C difficile strain CD027. Trehalase-resistant trehalose analogues might be developed as next-generation fasting-mimetics for treatment of diabetes and nonalcoholic fatty liver disease.