title: Sex-comparative study of mouse cerebellum physiology under adult-onset hypothyroidism: The significance of GC-MS metabolomic data normalization in meta-analysis.
abstract: A systematic data quality validation and normalization strategy is an important component of the omic profile meta-analysis, ensuring comparability of the profiles and exclusion of experimental biases from the derived biological conclusions. In this study, we present the normalization methodology applied on the sets of cerebellum gas chromatography-mass spectrometry metabolic profiles of 124days old male and female animals in an adult-onset-hypothyroidism (AOH) mouse model before combining them into a sex-comparative analysis. The employed AOH model concerns the monitoring of the brain physiology of Balb/cJ mice after eight-week administration of 1%w/v KClO4in the drinking water, initiated on the 60th day of their life. While originating from the same animal study, the tissues of the two sexes were processed and their profiles acquired and analyzed at different time periods. Hence, the previously published profile set of male mice was first re-annotated based on the presently available resources. Then, after being validated as acquired under the same analytical conditions, both profiles sets were corrected for derivatization biases and filtered for low-confidence measurements based on the same criteria. The final normalized 73-metabolite profiles contribute to the currently few available omic datasets of the AOH effect on brain molecular physiology, especially with respect to sex differentiation. Multivariate statistical analysis indicated one (unknown) and three (succinate, benzoate, myristate) metabolites with significantly higher and lower, respectively, cerebellum concentration in the hypothyroid compared to the euthyroid female mice. The respective numbers for the males were two and 24. Comparison of the euthyroid cerebellum metabolic profiles between the two sexes indicated 36 metabolites, including glucose, myo- and scyllo-inositol, with significantly lower concentration in the females versus the males. This implies that the female mouse cerebellum has been conditioned to smaller changes in its metabolic activity with respect to the pathways involving these metabolites compared to the male animals. In conclusion, our study indicated a much subtler AOH effect on the cerebellum metabolic activity of the female compared to the male mice. The leaner metabolic profile of the female mouse cerebellum was suggested as a potential factor contributing to this phenomenon.
authors.affiliation: Metabolic Engineering and Systems Biology Laboratory, Institute of Chemical Engineering Sciences, Foundation for Research and Technology-Hellas (FORTH/ICE-HT), Patras, Greece; School of Medicine, University of Patras, Greece.
authors.forename: Christoniki
authors.initials: C
authors.lastname: Maga-Nteve
authors.affiliation: Metabolic Engineering and Systems Biology Laboratory, Institute of Chemical Engineering Sciences, Foundation for Research and Technology-Hellas (FORTH/ICE-HT), Patras, Greece; Human & Animal Physiology Laboratory, Department of Biology, University of Patras, Greece.
authors.forename: Catherine G
authors.initials: CG
authors.lastname: Vasilopoulou
authors.affiliation: Metabolic Engineering and Systems Biology Laboratory, Institute of Chemical Engineering Sciences, Foundation for Research and Technology-Hellas (FORTH/ICE-HT), Patras, Greece.
authors.forename: Caterina
authors.initials: C
authors.lastname: Constantinou
authors.affiliation: Human & Animal Physiology Laboratory, Department of Biology, University of Patras, Greece.
authors.forename: Marigoula
authors.initials: M
authors.lastname: Margarity
authors.affiliation: Metabolic Engineering and Systems Biology Laboratory, Institute of Chemical Engineering Sciences, Foundation for Research and Technology-Hellas (FORTH/ICE-HT), Patras, Greece; Departments of Chemical & Biomolecular Engineering and Bioengineering, University of Maryland, College Park, MD 20742, USA. Electronic address: mklapa@iceht.forth.gr.
authors.forename: Maria I
authors.initials: MI
authors.lastname: Klapa
mesh_terms: D000818:Animals; D001835:Body Weight; D002531:Cerebellum / Q000737:chemistry / Q000378:metabolism* / Q000503:physiopathology; D016000:Cluster Analysis; D004195:Disease Models, Animal; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D007037:Hypothyroidism / Q000378:metabolism* / Q000503:physiopathology; D008297:Male; D055442:Metabolome; D055432:Metabolomics / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D025341:Principal Component Analysis; D012737:Sex Factors; D049490:Systems Biology
keywords: Adult-onset hypothyroidism (AOH); Balb/cJ mouse cerebellum; GC–MS metabolomic data validation and normalization; Mammalian brain metabolism; Metabolic network analysis; Systems biology