Detection of Circulating Serum microRNA/Protein Complexes in ASD Using Functionalized Chips for an Atomic Force
Anna L Kaysheva1, Arina I Isaeva1, Tatyana O Pleshakova1
1Laboratory of Nanobiotechnology, Institute of Biomedical Chemistry, Pogodinskaya St. 10/8, 119121 Moscow, Russia.
Researchers developed a novel method to detect autism spectrum disorder (ASD) biomarkers in blood. This technique uses atomic force microscopy (AFM) to identify specific microRNAs linked to ASD in children.
Area of Science:
- Biomedical research
- Molecular biology
- Neuroscience
Background:
- Circulating microRNAs in blood show significant diagnostic potential for various diseases.
- Mature microRNAs from glial cells and neurons can cross the blood-brain barrier and are detectable in serum of individuals with autism spectrum disorders (ASD).
- These microRNAs are found within macrovesicles and lipoprotein particles in serum.
Purpose of the Study:
- To propose and validate an approach for concentrating and detecting microRNAs associated with ASD development in children.
- To utilize atomic force microscopy (AFM) chips with specific oligonucleotide probes for targeted microRNA capture.
- To differentiate between serum samples from ASD patients and healthy volunteers based on microRNA detection.
Main Methods:
- Developed AFM chips functionalized with oligonucleotide molecular probes specific to target microRNAs.
- Incubated functionalized chips with serum samples from children with ASD and healthy controls.
- Analyzed chip surfaces using atomic force microscopy (AFM) to measure object heights.
- Performed MALDI-TOF-MS analysis to identify protein components associated with captured microRNAs.
Main Results:
- AFM revealed distinct object heights on the chip surfaces: 10 nm for ASD patient samples and 6 nm for healthy volunteers.
- MALDI-TOF-MS identified cell proteins involved in nucleic acid binding, proteasome and nucleosome organization, and potassium channel function.
- Specific microRNAs associated with ASD development were successfully concentrated and detected.
Conclusions:
- The proposed method effectively concentrates and detects microRNAs in protein complexes from serum using AFM chips.
- Distinct physical characteristics (height) of captured complexes can differentiate between ASD patients and healthy individuals.
- Identified proteins provide insights into the molecular mechanisms potentially underlying ASD.
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