Discovery and targeted proteomics on cutaneous biopsies infected by Borrelia to investigate Lyme disease.
Fiche publication
Date publication
février 2015
Auteurs
Membres identifiés du Cancéropôle Est :
Pr LIPSKER Dan
Tous les auteurs :
Schnell G, Boeuf A, Westermann B, Jaulhac B, Lipsker D, Carapito C, Boulanger N, Ehret-Sabatier L
Lien Pubmed
Résumé
Lyme disease is the most important vector-borne disease in the Northern hemisphere and represents a major public health challenge with insufficient means of and reliable diagnosis. Skin is rarely investigated in proteomics but constitutes in the case of Lyme disease the key interface where the pathogens can enter, persist and multiply. Therefore, we investigated proteomics on skin samples to detect Borrelia proteins directly in cutaneous biopsies in a robust and specific way. We first set up a discovery Ge-LC-MS/MS approach on a murine model infected by B. burgdorferi sensu stricto that allowed the identification of 25 Borrelia proteins among more than 1300 mouse proteins. Then we developed a targeted Ge-LC-SRM assay to detect 9/33 Borrelia proteins/peptides in mice skin tissue samples using heavy labeled synthetic peptides. We successfully transferred this assay from the mouse model to human skin biopsies - naturally infected by Borrelia - and we were able to detect two Borrelia proteins: OspC and flagellin. Considering the extreme variability of OspC, we developed an extended SRM assay to target a large set of variants. This assay afforded the detection of nine peptides belonging to either OspC or flagellin in human skin biopsies. We further shortened the sample preparation and showed that Borrelia is detectable in mouse and human skin biopsies by directly using a liquid digestion followed by LC-SRM analysis without any prefractionation. This study thus shows that a targeted SRM approach is a promising tool for the early direct diagnosis of Lyme disease with high sensitivity (
Référence
Mol Cell Proteomics. 2015 Feb 24. pii: mcp.M114.046540.