Abstract P5-05-04: MALDI mass spectrometry imaging profile of low molecular metabolites in breast carcinoma tissues embedded in frozen tissue microarray
Authors: Torata N , Kubo M , Miura D , Ohuchida K , Miyazaki T , Fujimura Y , Hayakawa E , Kai M , Oda Y , Mizumoto K , Hashizume M , Nakamura M 
Published: May 2016

[Background]

Metabolomics is now widely utilized for searching disease markers or identification of new drug targets. In common method, tissue samples originated in human resected specimens are stored by formalin-fixed, paraffin-embedded (FFPE) blocks. However, these samples are inadequate to measure low molecular metabolites or lipids. Furthermore, extraction process that is required for conventional mass-spectrometry causes the loss of information on the spatial localization of the metabolites. In this study, we directly analyzed breast carcinoma tissues embedded in frozen tissue microarray (fTMA) using MALDI mass spectrometry imaging (MALDI-MSI). With our original method, we could obtain profiles of low molecular metabolite and mapping images of several tissues at one time.

[Method]

Six fTMA blocks were constructed by 119 breast tissues (carcinoma 84, normal 35) from 99 patients and sectioned at 10 um thickness. MADLI-MSI were performed by AXIMA Confidence (Shimadzu, Japan) with 9-aminoacridine as a matrix (m/z range:50∼1000, Negative Ion mode). Carcinoma and normal area in individual tissues were confirmed by H&E staining of slide grasses after MADLI-MSI analysis. Acquired MSI data were processed with the freely available software BioMap (Novartis, Switzerland).

[Result]

We could detected 1,915 peaks derive from endogenous metabolite by direct tissue MALDI-MSI analysis of breast carcinoma fTMA. Among them, 185 peaks that could be commonly detected were subjected to further analysis. Among these peaks, we could identify 18 metabolites related to energy metabolism such as ATP. By comparison of metabolite profiles obtained from carcinoma with normal tissues, we found that the energy charge (EC; which is related to ATP, ADP and AMP concentrations) and the sum of adenosine phosphate compound intensities (AXP) were significantly higher than that of normal tissue (EC; T : N = 0.56 : 0.35, AXP; 17453 : 2066, p&lt;0.0001), but there were no significant difference with lymph node metastasis, tumor histological type and tumor size. In comparison with tumor subtype, higher EC was observed in ER(+)/ Her2(-) tumor than others but AXP showed no significant among all subtypes including Ki-67 labeling index.

[Conclusion]

A combination of fTMA and MALDI-MSI is promising approach for biomarker discovery because it can achieve high throughput metabolic mapping without obvious artifact or other problem. In this study, even though high EC value were indicated in carcinoma tissue than normal but newly biomarker candidate was indeterminate at this moment. Identification of the candidates of novel carcinoma tissue biomarker is now underway.

Citation Format: Torata N, Kubo M, Miura D, Ohuchida K, Miyazaki T, Fujimura Y, Hayakawa E, Kai M, Oda Y, Mizumoto K, Hashizume M, Nakamura M. MALDI mass spectrometry imaging profile of low molecular metabolites in breast carcinoma tissues embedded in frozen tissue microarray. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P5-05-04.