Analysis of in vitro toxicity of five cell-penetrating peptides by metabolic profiling.
Cell-penetrating peptides (CPPs) are promising candidates for safe and efficient delivery vectors for a wide range of cargoes. However, any compound that is internalized into cells may affect the cell homeostasis. The plethora of possible biological responses makes large scale "omics" studies appealing approaches for hunting any unsuspected side-effects and evaluate the toxicity of drug candidates. Here we have compared the alterations in cytosolic metabolome of CHO cells caused by five representatives of the most common CPPs: transportan (TP), penetratin (pAntp), HIV Tat derived peptide (pTat), nonaarginine (R(9)) and model amphipathic peptide (MAP). Analysis was done by liquid chromatography-mass spectrometry techniques, principal component analysis and heatmap displays. Results showed that the intracellular metabolome was the most affected by TP followed by pTat and MAP. Only minor changes could be associated with pAntp or R(9) treatment. The cells could recover from a treatment with 5 microM TP, but no recovery was observed at higher concentration. Both metabolomic and control experiments showed that TP affected cellular redox potential, depleted energy and the pools of purines and pyrimidines. In conclusion, we have performed a metabolomic analysis comparing the safety of cell-penetrating peptides and demonstrate the toxicity of one of them.
Kilk
Kalle
K
Department of Biochemistry, Medical Faculty, University of Tartu, The Centre of Excellence for Translational Medicine, 50411 Tartu, Estonia. kalle.kilk@ut.ee
Mahlapuu
Riina
R
Soomets
Ursel
U
Langel
Ulo
U
D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002352:Carrier Proteins / Q000737:chemistry / Q000494:pharmacology / Q000633:toxicity*; D002463:Cell Membrane Permeability; D006224:Cricetinae; D003412:Cricetulus; D004337:Drug Carriers / Q000737:chemistry / Q000494:pharmacology / Q000633:toxicity*; D005978:Glutathione / Q000378:metabolism; D055432:Metabolomics; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology / Q000633:toxicity*; D010539:Permeability / Q000187:drug effects
D002352:Carrier Proteins; D004337:Drug Carriers; D010455:Peptides; D005978:Glutathione