Irene Poppi (Presenter)
Clinical Mass Spectrometry, L.U.M., Maggiore Hospital, Bologna, Italy
Bio: Irene Poppi received the degree in Chemistry from the Department of Chemistry and Pharmaceutical Sciences , University of Ferrara (Italy) in 2011. In april 2011, she joined the Department of Chemistry and Pharmaceutical Sciences, Ferrara University, (Italy) as a research fellow in the project: “Functional materials in nanoparticles structures: SAM photoisomerization of azobenzenes by UV spectroscopy / Vis and NMR”. In june 2011, she joined the Department of Life Science and Biotechnology - University of Ferrara, (Italy) as a research fellow in the project:”Characterization of bioactive molecules to counteract human pathogenic bacteria and phytopathogenic bacteria, by liquid chromatography coupled with mass spectrometry". In 2015, she joined the Department Cardio-thoracic-vascular, Faculty of medicine, University of Bologna (Italy), assigned at Central laboratory (Mass Spectrometry Division) S.Orsola Hospital and Maggiore Hospital Bologna (Italy), as a research fellow in the project: “Proteomics of amyloidosis - typing of amyloid proteins by liquid chromatography coupled with mass spectrometry". Her current research interests include Clinical Mass Spectrometry, method development, method validation, Proteomics and Drug Monitoring
Authorship: Irene Poppi (1), Ornella Leone (2), Rita Mancini (1), Candida Cristina Quarta (3), Barbara Corti (2), Claudio Rapezzi (4), Matteo Conti (1)
1.Clinical Mass Spectrometry, L.U.M. Maggiore Hosp. 2.Department of Pathology, S.Orsola-Malpighi Hosp. 3. Cardiology, University College, London 4.Institute of Cardiology, S.Orsola-Malpighi Hosp.
| Short Abstract Amyloidosis is a pathological condition characterized by abnormal deposition of serum proteins in organs and tissues. Specific knowledge of the type of protein deposits in hystologically positive biopsies can be of paramount importance in differential diagnosis and personalized medical treatments. Since AL (light chain amyloid) and ATTR (transthyretin-related amyloid) proteins are most frequently involved in heart amyloidosis, we developed an high sensitivity method for specifically identifying these proteins in heart biopsies with a targeted proteomic approach. The assay has been successfully applied to protein typing in heart histological samples. |
Long Abstract
Amyloidosis refers to the extracellular deposition of fibrils that are composed of low molecular weight subunits (5 to 25 kD) of a variety of serum proteins. Amyloid deposits can occur in a variety of organs (heart, kidney, liver, and autonomic nervous system) [1].The heart is one of the most frequent sites of amyloid deposition. AL (light chain amyloid) and ATTR (transthyretin-related amyloid) proteins are most commonly involved [2]. At histology amyloid fibrils have a characteristic appearance and generate a green birefringence under polarized light when stained with Congo red [3], however knowledge of the type of protein deposits in Congo red positive biopsies is of paramount importance in differential diagnosis andspecific medical treatment.
Few methods for amyloidosis typing have been published so far, employing LMD (Laser Micro Dissection) to select fibrils in histological sections to be analyzed by High Resolution Mass Spectrometry approaches [4].In this work, we developed a novel methodfor sensitivetyping of amyloid proteins by a target LC-MS/MS approach,without prior LMD.
The assay was developed to characterize amyloid deposits in paraffin embedded endomyocardial biopsy fragments weighting about 0.1 mg and/or tissue sections of about 10um thickness. In brief, heart tissue was dewaxed, rehydrated and trypsin digested. Tryptic peptides were separated by reverse phase chromatography (on a Biobasic, Thermo Fisher Scientific - West Palm Beach, FL, USA) with gradient elution. Proteotipic peptides were selected by SRMATLAS (http://www.srmatlas.org/) and SKYLINE (https://skyline.gs.washington.edu/) for AL and ATTR. Their identification was performed onto a Q-Trap 5500 mass spectrometer (Sciex - Concord, Ontario,CAN) operated in positive electrospray ionization (ESI) and MRM detection mode. The assay has been successfully applied to a significant number of histological samples obtained from known AL, ATTR or mixed type amylosis cases. Due to its sensitivity, our data suggest that the assay could be suitable for biomedical research and even for clinical diagnostics applications.
References & Acknowledgements:
[1] William J McKenna, MDet al, “Clinical manifestations and diagnosis of amyloid cardiomyopathy” _ http://www.uptodate.com/
[2] Claudio Rapezzi, MD et al, “Systemic Cardiac Amyloidoses” _ Circulation September 29, 2009
[3] Laura M. Dember“Amyloidosis-Associated Kidney Disease” _ Am SocNephrol17: 3458–3471, 2006
[4] Sun et al _ “The Successful Diagnosis and Typing of Systemic Amyloidosis Using A Microwave- Assisted Filter-Aided Fast Sample Preparation Method and LC/MS/MS Analysis” _ PLOS ONE May 18, 2015
| Description | Y/N | Source |
| Grants | no | |
| Salary | no | |
| Board Member | no | |
| Stock | no | |
| Expenses | no |
IP Royalty: no
| Planning to mention or discuss specific products or technology of the company(ies) listed above: | no |