MALDI imaging-based classification of hepatocellular carcinoma and non-malignant lesions in fibrotic liver tissue

Abstract

Histopathologic differentiation of nodular lesions in cirrhotic liver is difficult even for experienced hepatopathologists especially regarding diagnosis of hepatocellular carcinoma (HCC) in biopsies. For this reason, new tissue markers are needed to reinforce histopathologic decision-making. With advances in molecular techniques, proteomic analysis may help to confirm the diagnosis of HCC. Matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI IMS) is a powerful technology which allows to determine and to localize proteins directly in tissue sections. Using MALDI IMS proteomic patterns of cryosections with lesions of HCC (n = 15) and non-malignant fibrotic liver tissue (n = 11) were investigated to establish a classification model of HCC, which was validated in an independent set of tissue to distinguish HCC (n = 10) from regenerative nodules (n = 8). By correlating generated mass spectrometric images with the histology of the tissue sections we found that the expression of 4 proteins as indicated by m/z 6274, m/z 6647, m/z 6222 and m/z 6853 was significantly higher in HCC tissue than in non-tumorous liver tissue. The generated classification model based on the most significant 3 differentially expressed proteins allowed a reliable prediction of benign and malignant lesions in fibrotic liver tissue with a sensitivity and specificity of 90 % in the validation set. The identified MALDI IMS proteomic signature can be diagnostically helpful to allow simplifying the diagnostic process and minimize the risks of delays in establishing the objective final diagnosis and initiating treatment of patients with HCC.

Zusammenfassung

Die histopathologische Differenzierung nodulärer Läsionen in der zirrhotischen Leber ist für Pathologen schwierig, insbesondere in Biopsiematerial. Es werden daher neue Gewebemarker benötigt, um die histopathologische Entscheidung zu stützen. Die bildgebende Massenspektrometrie (Matrix-assisted laser desorption/ionization imaging mass spectrometry – MALDI-IMS) ist eine leistungsfähige Technologie, die es ermöglicht, Proteine direkt im Gewebe zu detektieren und zu lokalisieren, und es ggf. erlaubt, die Diagnose eines hepatozellulären Karzinoms (HCC) zu bestätigen.

In dieser Arbeit wurden proteomische Muster in Kryostatschnitten von Läsionen des HCC (n = 15) und von nicht-malignem fibrotischem Lebergewebe (n = 11) mittels MALDI-IMS untersucht. Durch die Korrelation der erzeugten massenspektrometrischen Bilder mit der Histologie des entsprechenden Gewebeschnitts konnten 4 Proteine mit den m/z-Werten 6274, 6647, 6222 und 6853 identifiziert werden, die eine signifikant höhere Expression im HCC-Gewebe im Vergleich zur Fibrose aufweisen. Basierend auf 3 dieser Proteine mit der höchsten Signifikanz wurde ein Klassifikationsmodell erstellt und an einer unabhängigen Gruppe von Gewebeproben des HCC (n = 10) und benigner Regeneratknoten (n = 8) validiert. Mit diesem Klassifikationsmodell konnte eine verlässliche Vorhersage zwischen benignen und malignen Läsionen im fibrotischen Lebergewebe mit einer Sensitivität und Spezifität von 90 % getroffen werden. Diese identifizierte MALDI-IMS-Proteomsignatur kann zukünftig hilfreich sein, um eine endgültige und objektive Erstellung der Diagnose zu vereinfachen und so das Risiko von Verzögerungen bei der Diagnosebestimmung sowie der Therapieeinleitung von HCC-Patienten zu minimieren.

Schlüsselwörter

hepatozelluläres Karzinom - bildgebende Massenspektrometrie - MALDI IMS - Leberkrebs - Proteomik

Key words

hepatocellular carcinoma - imaging mass spectrometry - MALDI IMS - liver cancer - proteomics

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