Abstract 4586: MRM-based targeted analysis of potential cancer markers in blood
Notice bibliographique
Résumé
Abstract Introduction. Our previous discovery phase work using iTRAQ analysis on tissue homogenates from endometrial carcinoma, glioblastoma, head and neck squamous cell carcinoma have resulted in a number of potential markers for each of the types of cancer. A number of these potential markers were subsequently verified using conventional molecular biology approaches on the same as well as additional cohorts of samples. Recent studies using a new variant of the iTRAQ reagent, mTRAQ, in combination with multiple reaction monitoring (MRM) approaches on a hybrid triple quadrupole linear ion trap instrument also showed that the differential expression levels of a number of these potential markers could be observed in archived formalin fixed paraffin embedded tissue. We are therefore extending this approach to the detection and verification of differential expression of some of these potential markers in plasma. Methods. Serum or plasma samples from individuals diagnosed with EmCa or GBM or who had no known forms of cancer were studied. Each sample was individually depleted of the twenty most abundant proteins using an immunocapture column (Proteoprep 20, Sigma Aldrich). Samples were then enzymatically digested and labeled with one of the three versions of mTRAQ reagents. Data obtained during the discovery phase was used to direct the choice of MRM transitions. Transitions were generated for proteotypic peptides from each of the proteins of interest, each of which was targeted using a minimum of 2 peptides and 3 transitions per labeled peptide. Samples were mixed after labeling and analyzed using 2D LC-MRM analysis. An intermediate immunocapture step prior to digestion was incorporated to enrich for lower abundance proteins. Results: One of the potential markers of interest, clusterin, was successfully detected in sera from the three categories of patients, EmCa, GBM and non-malignant, however, no differential expression was observed. PIGR, which was another of the potential markers for EmCa, was observed exclusively in the serum from an EmCa patient. This confirmed its presence in sera and suggested that it might be possible to screen for its expression level therein. A third potential marker, Chaperonin 10, not initially observed using the direct approach described above, was detected after performing an immunocapture step prior to digestion and LC-MRM analysis. Current efforts are focused on titrating the antibody required to ensure any differential expression in plasma is accurately reflected in the results of the differential analyses. Conclusions: Our results have proved that the tissue-to-serum approach is feasible and that the differential expression of some potential markers detected in tissue are reflected in serum or plasma levels. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4586.
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Comment cette classification a été obtenuedéplier
Prédiction distillée sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Apprise à partir de 10 348 étiquettes directes de Codex et de 10 348 étiquettes directes de Gemma. Le mode candidate est l'union des têtes enseignantes seuillées; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont ni des étiquettes humaines ni des étiquettes directes de modèles de pointe.
Scores Codex et Gemma par catégorie
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,001 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,001 | 0,002 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,001 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,011 | 0,000 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule tête enseignante, pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».