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Research
A New Strategy to Uncover Fragile X Proteomic Biomarkers Using the Nascent Proteome of Peripheral Blood Mononuclear Cells (PBMCs)
Olivier Dionne
Sherbrooke University
Corresponding Author
ORCiD: https://orcid.org/0000-0002-0531-9761
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: Fragile X syndrome (FXS) is the most prevalent inherited cause of intellectual disabilities and autism spectrum disorders. FXS result from the loss of expression of the FMRP protein, an RNA binding protein that regulate the expression of key synaptic effectors. FXS is also characterized by a wide array of behavioral, cognitive and metabolic impairments. The severity and penetrance of those comorbidities are extremely variable, meaning that a considerable phenotypic heterogeneity is found among fragile X individuals. Unfortunately, clinicians currently have no tools at their disposal to assay patient’s prognosis upon diagnosis. Since the absence of FMRP was repeatedly associated with an aberrant translational metabolism, we decided to study the nascent proteome in order to screen for potential proteomic biomarkers of FXS.
Method: We used a BONCAT (Bioothogonal Non-canonical Amino Acids Tagging) method coupled to label-free mass spectrometry to purify and quantify nascent proteins of peripheral blood mononuclear cells from 7 fragile X male patients that do not express FMRP and 7 age-matched controls. Candidate biomarkers were confirmed by Western blot.
Results: The proteomic analysis identified several proteins which were either up or downregulated in absence of FMRP in FXS individuals as compared to controls. Eleven of those proteins were considered as potential biomarkers, from which 5 were further validated by Western blot. The gene ontology enrichment analysis highlighted molecular pathway that may contribute to FXS physiopathology.
Conclusions: Our results showed that the nascent proteome is well suited for the discovery of FXS biomarkers. In fact, taking advantage of a key alteration in FXS physiopathology led us to successfully identified 11 potential biomarkers.
Keywords
Fragile X syndrome, biomarkers, proteomics, nascent proteome, BONCAT, label-free mass spectrometry, PBMCs, translational medicine
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This is a list of supplementary files associated with this preprint. Click to download.
- TableS1DionneetCorbin2021BiomarkerResearch.xlsx
Table S1. List of all proteins detected in the total proteome
- TableS2DionneetCorbin2021BiomarkerResearch.xlsx
Table S2. List of all dysregulated proteins identified in the total proteome of fragile X PBMCs.
- TableS3DionneetCorbin2021BiomarkerResearch.xlsx
Table S3. List of all proteins detected in the nascent proteome
- TableS4DionneetCorbin2021BiomarkerResearch.xlsx
Table S4. Statistics associated with the LENS network
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This is a preprint. It has not completed peer review.
Research
A New Strategy to Uncover Fragile X Proteomic Biomarkers Using the Nascent Proteome of Peripheral Blood Mononuclear Cells (PBMCs)
Olivier Dionne
Sherbrooke University
Corresponding Author
ORCiD: https://orcid.org/0000-0002-0531-9761
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 15 Jan, 2021
No community comments so far
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: Fragile X syndrome (FXS) is the most prevalent inherited cause of intellectual disabilities and autism spectrum disorders. FXS result from the loss of expression of the FMRP protein, an RNA binding protein that regulate the expression of key synaptic effectors. FXS is also characterized by a wide array of behavioral, cognitive and metabolic impairments. The severity and penetrance of those comorbidities are extremely variable, meaning that a considerable phenotypic heterogeneity is found among fragile X individuals. Unfortunately, clinicians currently have no tools at their disposal to assay patient’s prognosis upon diagnosis. Since the absence of FMRP was repeatedly associated with an aberrant translational metabolism, we decided to study the nascent proteome in order to screen for potential proteomic biomarkers of FXS.
Method: We used a BONCAT (Bioothogonal Non-canonical Amino Acids Tagging) method coupled to label-free mass spectrometry to purify and quantify nascent proteins of peripheral blood mononuclear cells from 7 fragile X male patients that do not express FMRP and 7 age-matched controls. Candidate biomarkers were confirmed by Western blot.
Results: The proteomic analysis identified several proteins which were either up or downregulated in absence of FMRP in FXS individuals as compared to controls. Eleven of those proteins were considered as potential biomarkers, from which 5 were further validated by Western blot. The gene ontology enrichment analysis highlighted molecular pathway that may contribute to FXS physiopathology.
Conclusions: Our results showed that the nascent proteome is well suited for the discovery of FXS biomarkers. In fact, taking advantage of a key alteration in FXS physiopathology led us to successfully identified 11 potential biomarkers.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6