Imaging mass spectrometry analysis of renal amyloidosis biopsies reveals protein co-localization with amyloid deposits

Amyloidosis is a heterogeneous group of protein misfolding diseases characterized by deposition of amyloid proteins. The kidney is frequently affected, especially by immunoglobulin light chain (AL) and serum amyloid A (SAA) amyloidosis as the most common subgroups. Current diagnosis relies on histopathological examination, Congo red staining, or electron microscopy. Subtyping is done by immunohistochemistry; however, commercially available antibodies lack specificity. The purpose of this study was to identify and map amyloid proteins in formalin-fixed paraffin-embedded tissue sections using matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI IMS) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis in an integrated workflow. Renal amyloidosis and non-amyloidosis biopsies were processed for histological and MS analysis. Mass spectra corresponding to the congophilic areas were directly linked to the histological and MS images for correlation studies. Peptides for SAA and AL were detected by MALDI IMS associated to Congo red-positive areas. Sequence determination of amyloid peptides by LC-MS/MS analysis provided protein distribution and identification. Serum amyloid P component, apolipoprotein E, and vitronectin proteins were identified in both AA and AL amyloidosis, showing a strong correlation with Congo red-positive regions. Our findings highlight the utility of MALDI IMS as a new method to type amyloidosis in histopathological routine material and characterize amyloid-associated proteins that may provide insights into the pathogenetic process of amyloid formation. Graphical abstract Image correlation between hematoxylin and eosin, histochemistry (Congo red), and MALDI IMS of tissue sections from a patient affected with AA-amyloidosis. Hematoxylin and eosin staining ( HE ) shows glomerular structures ( black arrows ). Amyloid deposits appear in red in the Congo red-stained section in bright light ( CR ), and show an apple-green birefringence under polarized light ( GB , yellow arrows ). MALDI images for m/z = 1456.72, m/z = 1811.89 and m/z = 1156.6 correlate with Congo-red positive areas