P08 Mapping intermediary metabolic enzymes within human hair follicles
Chloe Farren, Martin Jürgens, Lisa Wanke, Georgeu Garrick, Bessam Farjo, Nilofer Farjo, Jörn Michael Völker, Erik Schulze zur Wiesche, Adrian Biddle, Michael Philpott, Rosalind HannenAbstract
Introduction and aims
Healthy hair follicles (HFs) engage in aerobic glycolysis, with preferential metabolism of glucose to lactate, despite oxygen presence; with 10% of total glucose oxidized to CO2. However, location and expression of key intermediary metabolic proteins within the different compartments of HF remains to be determined. The aim of this study is to investigate the expression profile of key enzymes involved within aerobic glycolysis and mitochondrial respiration within the HF.
Methods
Immunofluorescence staining was used to explore the expression profile of key metabolic proteins within different compartments of human scalp skin. Glycolytic proteins investigated included: hexokinase II (HKII), glucose-6-phosphate-dehydrogenase, phosphofructokinase platelet (PFKP) and liver/muscle (PFKL/M). Aerobic glycolytic proteins investigated were: lactate dehydrogenase-α (LDHA), and monocarboxylate transporter 1 (MCT1). We also investigated pyruvate dehydrogenase-E1-α (PDHA1), which links glycolysis to the tricarboxylic acid (TCA) cycle, and TCA cycle proteins: acetyl-CoA transferase (ACOA), and isocitrate dehydrogenase; n = 3.
Results
All metabolic proteins investigated were expressed throughout the different HF compartments. The DP had low expression of metabolic proteins apart from PFKP and IDH1/2, which had moderate expression levels; suggesting low levels of metabolic activity. The germinative epithelium and matrix highly expressed HKII, PFKL/M and MCT1, with moderate-to-high expression of LDHA indicating active aerobic glycolysis in both compartments. The cortex had high expression of glycolytic proteins, PDHA1 and ACOA, but not LDHA, which suggests a preference for mitochondrial respiration. The inner root sheath had high expression of all proteins excluding PFKP, suggesting high metabolic active. The outer root sheath had a high expression of mitochondrial and transporter proteins.
Conclusions
These data suggest that metabolism is not uniform across the HF; that patterns and rates of metabolism may differ between compartments depending on function. We are now carrying out targeted and untargeted 13C metabolomic studies on ex vivo HF, and the Seahorse-XF-Pro Analyzer on hair-derived cells to investigate these findings further.