Periodic Reporting for period 1 - IR4FTD (Discovery of new Frontal Temporal Dementia multimodal spectral markers in biofluids.)
Periodo di rendicontazione: 2023-10-01 al 2025-09-30
Frontotemporal Dementia (FTD), the second most prevalent dementia in individuals under 60, is a rare and challenging disease to diagnose. Unlike other conditions, FTD lacks a single definitive diagnostic test, often requiring a combination of costly or invasive examinations. Its symptoms can also overlap with those of Alzheimer's Disease, further complicating diagnosis. The search for reliable biomarkers is a significant challenge in FTD research. This project proposes a holistic approach, using vibrational spectroscopy to acquire the complete chemical fingerprint in a label-free manner, rather than focusing on individual biomarkers. The central aim is to identify distinct spectroscopic fingerprints for FTD and Alzheimer’s disease (AD) in saliva and plasma samples using multimodal spectroscopy and machine learning. Samples from FTD, AD, and healthy individuals over 45 years of age will be analyzed using Raman, mid-infrared, and near-infrared spectroscopy. The successful outcome of this research could lead to the development of a novel screening tool capable of detecting chemical fingerprints indicative of FTD, AD, and potentially other diseases, significantly improving early detection and diagnosis.
Activities Performed
Optimization of Sample Preparation for Spectroscopy
*Deposited saliva, plasma (and platelets) on various substrates (aluminum foil, Kevley slides, cover glass, stainless steel, (silicon wafers)).
*Compared spectral data from different infrared techniques: ATR, transflection, and Near-Infrared Spectroscopy (NIR).
*Added freeze-drying as a preparation method alongside air drying to enhance long-term sample preservation.
*Selected Silicon wafers as a new substrate compatible with both spectroscopy and X-ray fluorescence.
*Included platelets as an additional biofluid due to their relevance in neurodegenerative biomarkers (TDP-43 for FTD and ALS).
*Investigated effects of cryopreservation on platelets over time (1h, 3h, 6h thawing), using spectroscopy and biological assays.
Preliminary Patient Sample Analysis
*Analyzed 10 plasma samples (6 FTD patients, 4 controls) using multiple techniques (ATR, NIR, TXRF).
*Conducted PCA analysis to differentiate between FTD and control samples — clear separation observed.
Exploratory Work
*Aimed to expand research to more patients but was not completed due to lack of access to additional patients.
Scientific & Technical
*Established a robust protocol for sample preparation involving different substrates and drying techniques.
*Identified low-cost and safe substrates (e.g. silicon wafers) for multimodal analysis, enhancing accessibility.
*Demonstrated feasibility of vibrational spectroscopy to discriminate biofluids in neurodegenerative diseases.
*Cryopreservation study on platelets led to new insights into how thawing time affects molecular content.
Publications & Recognition
2 peer-reviewed publications were produced.
3 peer-reviewed are in preparation.
1 invited chapter in preparation.
1 oral presentation at an International Conference.
Optimization of Sample Preparation for Spectroscopy
*Deposited saliva, plasma (and platelets) on various substrates (aluminum foil, Kevley slides, cover glass, stainless steel, (silicon wafers)).
*Compared spectral data from different infrared techniques: ATR, transflection, and Near-Infrared Spectroscopy (NIR).
*Added freeze-drying as a preparation method alongside air drying to enhance long-term sample preservation.
*Selected Silicon wafers as a new substrate compatible with both spectroscopy and X-ray fluorescence.
*Included platelets as an additional biofluid due to their relevance in neurodegenerative biomarkers (TDP-43 for FTD and ALS).
*Investigated effects of cryopreservation on platelets over time (1h, 3h, 6h thawing), using spectroscopy and biological assays.
Preliminary Patient Sample Analysis
*Analyzed 10 plasma samples (6 FTD patients, 4 controls) using multiple techniques (ATR, NIR, TXRF).
*Conducted PCA analysis to differentiate between FTD and control samples — clear separation observed.
Exploratory Work
*Aimed to expand research to more patients but was not completed due to lack of access to additional patients.
Scientific & Technical
*Established a robust protocol for sample preparation involving different substrates and drying techniques.
*Identified low-cost and safe substrates (e.g. silicon wafers) for multimodal analysis, enhancing accessibility.
*Demonstrated feasibility of vibrational spectroscopy to discriminate biofluids in neurodegenerative diseases.
*Cryopreservation study on platelets led to new insights into how thawing time affects molecular content.
Publications & Recognition
2 peer-reviewed publications were produced.
3 peer-reviewed are in preparation.
1 invited chapter in preparation.
1 oral presentation at an International Conference.
1. Innovative Sample Preparation Protocols for Infrared Spectroscopy
*First comparative study of low-cost substrates (aluminum foil, stainless steel, silicon wafers, Kevley slides, cover glass) for infrared spectroscopy of biofluids (plasma, saliva, platelets).
*Traditional IR studies often rely on expensive and fragile substrates like CaF2; your work breaks new ground by showing these affordable alternatives perform comparably, with broader applicability and lower cost.
*The protocol also reduces instrument time by using pre-dried (air or freeze-dried) samples rather than drop deposition directly on the ATR crystal — an efficiency gain not previously demonstrated systematically.
*This contribution is immediately actionable and broadly beneficial for researchers using vibrational spectroscopy in biomedicine.
2. Integration of Multimodal Analysis on Single Substrate
*Demonstrated that Silicon wafers are compatible with both infrared spectroscopy and X-ray fluorescence (TXRF) — enabling multimodal analysis on the same sample.
*This advancement allows researchers to combine molecular (vibrational) and elemental data, giving richer insights without sample loss — a novel and valuable methodology in biofluid analysis.
3. New Insights into Platelet Cryopreservation and Neurodegeneration Biomarkers
*Revealed that cryopreservation affects platelet biochemistry, particularly over time after thawing, altering macromolecular composition in ways that could affect diagnostic utility.
*This study goes beyond current literature, which has paid limited attention to how storage methods impact the molecular profile of platelets — especially in the context of biomarkers like TDP-43 relevant to ALS and FTD.
*Demonstrated the time-dependent degradation or alteration of biomolecules, a factor crucial for biobanking protocols and diagnostic reliability.
4. Spectroscopic Identification of Lipid Changes in Brain Lesions
*Identified a significant decrease in lipid content in lesion regions of the brain using vibrational spectroscopy — findings consistent with mass spectrometry results and indicative of lipid peroxidation.
*These results provide new mechanistic evidence supporting the hypothesis that demyelination in neurodegenerative disease may be reversed by restoring lipid content/myelination — an idea not fully explored in IR spectroscopy before.
*This offers a potential spectroscopic biomarker for neurodegeneration progression and treatment monitoring.
5. Diagnostic Differentiation Using IR and Elemental Spectroscopy
*Successfully demonstrated clear PCA-based discrimination between plasma samples from FTD patients and controls using multiple spectroscopic techniques (ATR, NIR, TXRF) in a trial set that needs to be tested at higher numbers.
*Although based on a small sample set, this work lays the groundwork for non-invasive, label-free diagnostics, and advances the field toward objective spectral biomarkers.
*First comparative study of low-cost substrates (aluminum foil, stainless steel, silicon wafers, Kevley slides, cover glass) for infrared spectroscopy of biofluids (plasma, saliva, platelets).
*Traditional IR studies often rely on expensive and fragile substrates like CaF2; your work breaks new ground by showing these affordable alternatives perform comparably, with broader applicability and lower cost.
*The protocol also reduces instrument time by using pre-dried (air or freeze-dried) samples rather than drop deposition directly on the ATR crystal — an efficiency gain not previously demonstrated systematically.
*This contribution is immediately actionable and broadly beneficial for researchers using vibrational spectroscopy in biomedicine.
2. Integration of Multimodal Analysis on Single Substrate
*Demonstrated that Silicon wafers are compatible with both infrared spectroscopy and X-ray fluorescence (TXRF) — enabling multimodal analysis on the same sample.
*This advancement allows researchers to combine molecular (vibrational) and elemental data, giving richer insights without sample loss — a novel and valuable methodology in biofluid analysis.
3. New Insights into Platelet Cryopreservation and Neurodegeneration Biomarkers
*Revealed that cryopreservation affects platelet biochemistry, particularly over time after thawing, altering macromolecular composition in ways that could affect diagnostic utility.
*This study goes beyond current literature, which has paid limited attention to how storage methods impact the molecular profile of platelets — especially in the context of biomarkers like TDP-43 relevant to ALS and FTD.
*Demonstrated the time-dependent degradation or alteration of biomolecules, a factor crucial for biobanking protocols and diagnostic reliability.
4. Spectroscopic Identification of Lipid Changes in Brain Lesions
*Identified a significant decrease in lipid content in lesion regions of the brain using vibrational spectroscopy — findings consistent with mass spectrometry results and indicative of lipid peroxidation.
*These results provide new mechanistic evidence supporting the hypothesis that demyelination in neurodegenerative disease may be reversed by restoring lipid content/myelination — an idea not fully explored in IR spectroscopy before.
*This offers a potential spectroscopic biomarker for neurodegeneration progression and treatment monitoring.
5. Diagnostic Differentiation Using IR and Elemental Spectroscopy
*Successfully demonstrated clear PCA-based discrimination between plasma samples from FTD patients and controls using multiple spectroscopic techniques (ATR, NIR, TXRF) in a trial set that needs to be tested at higher numbers.
*Although based on a small sample set, this work lays the groundwork for non-invasive, label-free diagnostics, and advances the field toward objective spectral biomarkers.