254
24.2 Results and Discussion
Moving forwards from these first results, we then applied MALDI-MSI proteomics
approach on real FNAB. At a first instance, challenging technical aspects of this
approach, such as the interference of hemoglobin and the morphological and proteomic stability of samples, were overcome [2]. Then, in accordance with the primary aim of the study [3], we are now working on the implementation of a
MALDI-MSI statistical tool that could be potentially used to assist pathologists in
diagnosis of indeterminate cases. Real thyroid FNAs from hyperplastic and papillary thyroid cancer (PTC) nodules, were collected from patients (San Gerardo
Hospital, Monza, Italy) and transferred into CytoLyt solution, centrifuged and resuspended in PreservCyt solution. Cytospin spots have been positioned onto conductive slides and intact proteins MALDI-MSI analysis was performed within the
m/z 3000–20,000 range. Regions of interest (ROIs) containing pathological areas
(thyrocytes clusters) were comprehensively annotated by pathologist. This pilot
study underlines the feasibility of the MALDI-MSI approach to spatially localize
proteins in cancer cells area and its ability to discriminate, based on the proteomic
profile, hyperplastic and PTC real FNA [4].
The present study introduces an original methodological approach to build a
proteomic diagnostic tool in thyroid cytopathology by taking advantage of MALDIMSI technology for the characterization of indeterminate for malignancy thyroid
nodules.
Acknowledgments This work was funded thanks to the “Associazione Italiana per la Ricerca sul
Cancro” - AIRC MFAG Grant 2016 Id.18445.
References
1. Mosele N, Smith A, Galli M, Pagni F, Magni F (2017) MALDI-MSI analysis of cytological smears: the study of thyroid cancer. Methods Mol Biol 1618:37–47. https://doi.
org/10.1007/978-1-4939-7051-3_5
2. Piga I, Capitoli G, Denti V, Tettamanti S, Smith A, Chinello C, Stella M, Leni D, Garancini M,
Galimberti S, Magni F, Pagni F (2019) The management of haemoglobin interference for the
MALDI-MSI proteomics analysis of thyroid fine needle aspiration biopsies. J Anal Bioanal
Chem 411(20):5007–5012. https://doi.org/10.1007/s00216-019-01908-w
3. Piga I, Capitoli G, Tettamanti S, Denti V, Smith A, Chinello C, Stella M, Leni D, Garancini
M, Galimberti S, Magni F, Pagni F (2019) Feasibility study for the MALDI-MSI analysis of
thyroid fine needle aspiration biopsies: evaluating the morphological and proteomic stability
over time. Proteomics Clin Appl 13(1):e1700170. https://doi.org/10.1002/prca.201700170
4. Capitoli G, Piga I, Galimberti S, Leni D, Pincelli AI, Garancini M, Clerici F, Mahajneh A,
Brambilla V, Smith A, Magni F, Pagni F (2019) MALDI-MSI as a complementary diagnostic tool in cytopathology: a pilot study for the characterization of thyroid nodules. Cancers
11(9):pii: E1377. https://doi.org/10.3390/cancers11091377
I. Piga et al.
24.2 Results and Discussion
Moving forwards from these first results, we then applied MALDI-MSI proteomics
approach on real FNAB. At a first instance, challenging technical aspects of this
approach, such as the interference of hemoglobin and the morphological and proteomic stability of samples, were overcome [2]. Then, in accordance with the primary aim of the study [3], we are now working on the implementation of a
MALDI-MSI statistical tool that could be potentially used to assist pathologists in
diagnosis of indeterminate cases. Real thyroid FNAs from hyperplastic and papillary thyroid cancer (PTC) nodules, were collected from patients (San Gerardo
Hospital, Monza, Italy) and transferred into CytoLyt solution, centrifuged and resuspended in PreservCyt solution. Cytospin spots have been positioned onto conductive slides and intact proteins MALDI-MSI analysis was performed within the
m/z 3000–20,000 range. Regions of interest (ROIs) containing pathological areas
(thyrocytes clusters) were comprehensively annotated by pathologist. This pilot
study underlines the feasibility of the MALDI-MSI approach to spatially localize
proteins in cancer cells area and its ability to discriminate, based on the proteomic
profile, hyperplastic and PTC real FNA [4].
The present study introduces an original methodological approach to build a
proteomic diagnostic tool in thyroid cytopathology by taking advantage of MALDIMSI technology for the characterization of indeterminate for malignancy thyroid
nodules.
Acknowledgments This work was funded thanks to the “Associazione Italiana per la Ricerca sul
Cancro” - AIRC MFAG Grant 2016 Id.18445.
References
1. Mosele N, Smith A, Galli M, Pagni F, Magni F (2017) MALDI-MSI analysis of cytological smears: the study of thyroid cancer. Methods Mol Biol 1618:37–47. https://doi.
org/10.1007/978-1-4939-7051-3_5
2. Piga I, Capitoli G, Denti V, Tettamanti S, Smith A, Chinello C, Stella M, Leni D, Garancini M,
Galimberti S, Magni F, Pagni F (2019) The management of haemoglobin interference for the
MALDI-MSI proteomics analysis of thyroid fine needle aspiration biopsies. J Anal Bioanal
Chem 411(20):5007–5012. https://doi.org/10.1007/s00216-019-01908-w
3. Piga I, Capitoli G, Tettamanti S, Denti V, Smith A, Chinello C, Stella M, Leni D, Garancini
M, Galimberti S, Magni F, Pagni F (2019) Feasibility study for the MALDI-MSI analysis of
thyroid fine needle aspiration biopsies: evaluating the morphological and proteomic stability
over time. Proteomics Clin Appl 13(1):e1700170. https://doi.org/10.1002/prca.201700170
4. Capitoli G, Piga I, Galimberti S, Leni D, Pincelli AI, Garancini M, Clerici F, Mahajneh A,
Brambilla V, Smith A, Magni F, Pagni F (2019) MALDI-MSI as a complementary diagnostic tool in cytopathology: a pilot study for the characterization of thyroid nodules. Cancers
11(9):pii: E1377. https://doi.org/10.3390/cancers11091377
I. Piga et al.