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Spatiotemporal Proteomic Profiling of Human Cerebral Development
Ugljesa Djuric1, Deivid C Rodrigues2, Ihor Batruch3
1From the ‡Laboratory Medicine and Pathology Program, University Health Network, Toronto, Ontario, M5G 2C4, Canada.
Molecular & Cellular Proteomics : MCP
|July 9, 2017
Summary
Mass spectrometry analysis of human brain tissue and stem cells reveals protein signatures of neurodevelopment. This method efficiently identifies novel biomarkers in archival samples for disease and development research.
Area of Science:
- Neuroscience
- Proteomics
- Stem Cell Biology
Background:
- Human post-mortem central nervous system (CNS) tissue and induced pluripotent stem cell (iPSC)-derived neurons offer complementary models for studying neurodevelopmental protein signatures.
- Advances in formalin-fixed, paraffin-embedded (FFPE) protein isolation enable proteomic analysis of archival tissue, crucial for understanding development and disease.
- Defining protein dynamics during neurodevelopment is essential for identifying biomarkers and understanding neurological disorders.
Purpose of the Study:
- To utilize shotgun label-free quantification (LFQ) mass spectrometry (MS) to profile protein signatures in human iPSC-derived neurons and neural progenitor cells (NPCs).
- To apply these protein signatures to define spatiotemporal protein dynamics in developing human FFPE cerebral tissues.
- To evaluate the potential of high-resolution MS for biomarker discovery in archival FFPE human tissues.
Main Methods:
- Shotgun label-free quantification (LFQ) mass spectrometry (MS) was employed.
- Magnetically enriched human cortical neurons and NPCs derived from iPSCs were profiled.
- High-resolution Q Exactive mass spectrometers were used for proteomic analysis.
Main Results:
- Simultaneous quantification of over 2700 proteins was achieved in single LFQ experiments using high-resolution MS.
- Novel biomarkers and signatures for neural progenitor cell (NPC) maintenance and differentiation were identified.
- An abbreviated proteomic strategy efficiently recovered cytoplasmic, membrane-specific, and synaptic proteins common to both in vivo and in vitro neuronal differentiation.
Conclusions:
- High-resolution MS enables comprehensive proteomic profiling of FFPE human tissues, facilitating biomarker discovery.
- The developed proteomic strategy effectively characterizes protein signatures in both iPSC-derived cells and archival FFPE brain tissue.
- This approach highlights the potential for discovering novel insights into neurodevelopment and disease through proteomic analysis of well-annotated archival samples.
