Amiloidosis Detection by Mass Spectrometry: Development of a Diagnostic Technique.

MARIA SOLEDAD SAEZ; ELSA MERCEDES NUCIFORA; PABLO POMATA; VICTORIA LUX; SILVA MORENO; PIA VALACCO; MARIA LOURDES POSADAS MARTINEZ

Congreso; Shefiled congress of pathology; 2024

Amiloidosis Detection by Mass Spectrometry: Development of a Diagnostic Technique.Maria Soledad Saez2, Ricardo Neme Tauil1, Melina Chulvi, Mercedes Juarez Araoz2, Maria Pia Valacco1, German Fernández1, Elsa Nucífora2, Patricia Sorroche2, Maria Adela Aguirre2, Pablo Pomata3, Victoria Lux-Lantos3, Silvia Moreno1, Maria Lourdes PosadasMartinez21CEQUIBIEM, IQUIBICEN/CONICET, Biological Chemistry Department, FCEN, UBA.2Hospital Italiano de Buenos Aires.3IBYME, CONICET.e-mail: smoreno@qb.fcen.uba.ar, Background: Amyloidosis, a rare disease impacting 1 in 10,000 individuals globally, arises from misfolded proteins forming insoluble fibrillar deposits in various body regions.Purpose: Identifying these proteins is crucial for appropriate treatment . Amyloid fibers are visible as apple green birefringent color with polarized light in tissue slides, stained with Congo Red. The gold standard for the identification of the amyloid protein, is LMD/MS, which consists of a laser microdissection of the birefringent material and subsequent analysis by tandem mass spectrometry1. Alongside our Hospital, the Institute of Biology and Experimental Medicine (IBYME/Conicet) and CEQUIBIEM, we aim to develop this diagnostic technique to offer it as a service in Argentina and Latin America.Method: The study included pathological samples from various organs affected by amyloidosis: Heart (9), Oral cavity (2), Liver (1), Larynx (1), and Rectum (1), in addition to 3 control Heart samples. These samples were slides stained with Congo Red, and paraffin embedded blocks. Laser microdissection was performed. At CEQUIBIEM, a standardized protocol for protein extraction was established for both microdissected samples (9) and fixed biopsy sections (14). All samples underwent analysis using nano HPLC coupled with Orbitrap (QExactive) mass spectrometry.Results: In 6 of the 9 microdissected samples analyzed, amyloidogenic proteins were detected. Amyloidogenic proteins were detected in all 14 biopsy samples analyzed directly by LC/MS. In all cases, 5 proteins that are described as “signature” that accompany the amyloid fiber (Apolipoprotein A-IV. Apolipoprotein A-I; Apolipoprotein E, Serumamyloid and Vitronectin) were identified. Also, amyloidogenic proteins were detected in abundance in non-microdissected biopsies, suggesting a granular accumulation compared to LMD/MS samples.Conclusions: Further analysis of diverse tissue samples and corresponding controls is necessary, yet initial findings indicate promising prospects for the technique's development and application. 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