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Ambient ionization mass spectrometric analysis of human surgical specimens to distinguish renal cell carcinoma from healthy renal tissue

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Abstract

Touch spray-mass spectrometry (TS-MS) is an ambient ionization technique (ionization of unprocessed samples in the open air) that may find intraoperative applications in quickly identifying the disease state of cancerous tissues and in defining surgical margins. In this study, TS-MS was performed on fresh kidney tissue (∼1–5 cm3), within 1 h of resection, from 21 human subjects afflicted by renal cell carcinoma (RCC). The preliminary diagnostic value of TS-MS data taken from freshly resected tissue was evaluated. Principal component analysis (PCA) of the negative ion mode (m/z 700–1000) data provided the separation between RCC (16 samples) and healthy renal tissue (13 samples). Linear discriminant analysis (LDA) on the PCA-compressed data estimated sensitivity (true positive rate) and specificity (true negative rate) of 98 and 95 %, respectively, based on histopathological evaluation. The results indicate that TS-MS might provide rapid diagnostic information in spite of the complexity of unprocessed kidney tissue and the presence of interferences such as urine and blood. Desorption electrospray ionization-MS imaging (DESI-MSI) in the negative ionization mode was performed on the tissue specimens after TS-MS analysis as a reference method. The DESI imaging experiments provided phospholipid profiles (m/z 700–1000) that also separated RCC and healthy tissue in the PCA space, with PCA-LDA sensitivity and specificity of 100 and 89 %, respectively. The TS and DESI loading plots indicated that different ions contributed most to the separation of RCC from healthy renal tissue (m/z 794 [PC 34:1 + Cl]− and 844 [PC 38:4 + Cl]− for TS vs. m/z 788 [PS 36:1 − H]− and 810 [PS 38:4 − H]− for DESI), while m/z 885 ([PI 38:4 − H]−) was important in both TS and DESI. The prospect, remaining hurdles, and future work required for translating TS-MS into a method of intraoperative tissue diagnosis are discussed.

Touch spray-mass spectrometry used for lipid profiling of fresh human renal cell carcinoma. Left) Photograph of the touch spray probe pointed at the MS inlet. Right) Average mass spectra of healthy renal tissue (blue) and RCC (red)

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Acknowledgments

The research reported in this publication was supported by the National Institute of Biomedical Imaging and Bioengineering of the National Institutes of Health under award number R21EB015722. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. K. A. Kerian gratefully recognizes the funding support from the Purdue University Center for Cancer Research SIRG Graduate Research Assistantship Award that funded him for this study.

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Authors and Affiliations

  1. Department of Chemistry, Center for Analytical Instrumentation Development and Purdue University Center for Cancer Research, Purdue University, West Lafayette, IN, 47907, USA

    Clint M. Alfaro, Alan K. Jarmusch, Valentina Pirro, Kevin S. Kerian & R. Graham Cooks

  2. Department of Urology, Indiana University School of Medicine, Indiana University Melvin and Bren Simon Cancer Center, Indianapolis, IN, 46202, USA

    Timothy A. Masterson

  3. Department of Pathology and Laboratory Medicine, Indiana University School of Medicine, Indiana University Melvin and Bren Simon Cancer Center, Indianapolis, IN, 46202, USA

    Liang Cheng

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  1. Clint M. Alfaro
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Correspondence to R. Graham Cooks.

Ethics declarations

The research involving human subjects was conducted in compliance with the ethical guidelines of the approved Institutional Review Board protocols at the Indiana University School of Medicine (study # 1205008669R004) and Purdue University (study # 1203011967). The renal cell carcinoma samples were obtained from 21 human subjects after they provided written informed consent to participate in the research study.

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The authors declare that they have no conflict of interest.

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Alfaro, C.M., Jarmusch, A.K., Pirro, V. et al. Ambient ionization mass spectrometric analysis of human surgical specimens to distinguish renal cell carcinoma from healthy renal tissue. Anal Bioanal Chem 408, 5407–5414 (2016). https://doi.org/10.1007/s00216-016-9627-4

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  • DOI: https://doi.org/10.1007/s00216-016-9627-4

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