Metabolomic Approaches to Study Chemical Exposure-Related Metabolism Alterations in Mammalian Cell Cultures

Abstract

Biological organisms are constantly exposed to an immense repertoire of molecules that cover environmental or food-derived molecules and drugs, triggering a continuous flow of stimuli-dependent adaptations. The diversity of these chemicals as well as their concentrations contribute to the multiplicity of induced effects, including activation, stimulation, or inhibition of physiological processes and toxicity. Metabolism, as the foremost phenotype and manifestation of life, has proven to be immensely sensitive and highly adaptive to chemical stimuli. Therefore, studying the effect of endo- or xenobiotics over cellular metabolism delivers valuable knowledge to apprehend potential cellular activity of individual molecules and evaluate their acute or chronic benefits and toxicity. The development of modern metabolomics technologies such as mass spectrometry or nuclear magnetic resonance spectroscopy now offers unprecedented solutions for the rapid and efficient determination of metabolic profiles of cells and more complex biological systems. Combined with the availability of well-established cell culture techniques, these analytical methods appear perfectly suited to determine the biological activity and estimate the positive and negative effects of chemicals in a variety of cell types and models, even at hardly detectable concentrations. Metabolic phenotypes can be estimated from studying intracellular metabolites at homeostasis in vivo, while in vitro cell cultures provide additional access to metabolites exchanged with growth media. This article discusses analytical solutions available for metabolic phenotyping of cell culture metabolism as well as the general metabolomics workflow suitable for testing the biological activity of molecular compounds. We emphasize how metabolic profiling of cell supernatants and intracellular extracts can deliver valuable and complementary insights for evaluating the effects of xenobiotics on cellular metabolism. We note that the concepts and methods discussed primarily for xenobiotics exposure are widely applicable to drug testing in general, including endobiotics that cover active metabolites, nutrients, peptides and proteins, cytokines, hormones, vitamins, etc.

Simulate the Dynamic Interactions Across Nutritional Supplements, the Polybiome, and the Biochemical Approach Dependent on Patient-Derived Gastrointestinal Knowledge

“…Concerns about the pharmacological effects of endogenous phytonutrients and genetically altered microbes nevertheless remain in large numbers. Research methods that are now being used to examine the effects of food ingredients on inflammatory disorders, bioaccumulation interactions between food components and the human body, the examination of the intestinal poly-biome, and the potential antibacterial capabilities of phytochemicals, such as phytonutrients vs. malignancy, all rely heavily on supplying natural biological molecules to a variety of mammalian species using two-dimensional carcinoma models [45][46][47][48][49][50][51][52][53][54][55][56]. The pharmacological effects of different classes of drugs cannot be assigned to their original components but must be attributed to their metabolites [57][58][59].…”

Phytochemicals as Regulators of Tumor Glycolysis and Hypoxia Signaling Pathways: Evidence from In Vitro Studies

“…One of the applications of cell culture metabolomics is deciphering the antitumoral molecular mechanisms of phytochemicals by exposing cancer cells to various doses of dietary compounds. The experimental design should consider several key aspects which must be taken into account: the objectives of the study (targeted/untargeted strategy, pathway reconstruction, enrichment analysis, biomarker identification), the availability of appropriate analytical equipment and standard operating procedures, the availability of various metabolite standards, in vitro models (primary cells, cell lines, spent culture media), the cell culture growth conditions, the tested phytochemicals (extracts or standard compounds), the physical and chemical properties of tested phytochemicals—solubility, stability, doses and exposure time, the selection of appropriate data analysis tools [ 28 ].…”

Mass Spectrometry-Based Cellular Metabolomics: Current Approaches, Applications, and Future Directions

“…Routine testing in food laboratories currently depends on traditional and time-consuming culture methods. Given the short shelf life of meat products, a rapid metabolomics-based analytical method could assist in the development of routine diagnostic testing for foodborne pathogens. Xenobiotics: The growing numbers of xenobiotics resulting from technological advances drives the need for testing their impact on living organisms in order to comprehend their impact on physiology. , Hence, the comprehension of the interactions of xenobiotics with organisms as well as mechanisms of action becomes crucial in toxicology as well as pharmacotherapy. , Metals and organic compounds are common pollutants. Exposure to even low amounts of heavy metals can disrupt normal central nervous system development .…”

“…Xenobiotics: The growing numbers of xenobiotics resulting from technological advances drives the need for testing their impact on living organisms in order to comprehend their impact on physiology. , Hence, the comprehension of the interactions of xenobiotics with organisms as well as mechanisms of action becomes crucial in toxicology as well as pharmacotherapy. , Metals and organic compounds are common pollutants. Exposure to even low amounts of heavy metals can disrupt normal central nervous system development .…”