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Proteomic cell surface phenotyping of differentiating acute myeloid leukemia cells
Andreas Hofmann1, Bertran Gerrits, Alexander Schmidt
1Institute of Molecular Systems Biology, Swiss Federal Institute of Technology (ETH) Zurich, Zurich, Switzerland.
Insights
Mass spectrometry phenotyping reveals over 500 membrane proteins in acute myeloid leukemia models. This advanced technique enhances hematologic malignancy subclassification and identifies new drug targets.
Area of Science:
- Proteomics
- Hematology
- Cancer Biology
Background:
- Immunophenotyping is crucial for diagnosing and monitoring hematologic malignancies.
- Current antibody-based methods have limitations in scope and depth, hindering early diagnosis and subclassification.
- A deeper understanding of cell surface markers is needed for improved clinical outcomes.
Purpose of the Study:
- To develop and apply mass spectrometry-based phenotyping for comprehensive analysis of the leukemia surface proteome.
- To overcome limitations of antibody-based methods in identifying cell surface proteins.
- To identify novel markers for improved subclassification and therapeutic targeting of hematologic malignancies.
Main Methods:
- Utilized Cell Surface Capturing (CSC) technology with mass spectrometry.
- Analyzed two all-trans retinoic acid treated acute myeloid leukemia model systems (HL60 and NB4 cells).
- Performed bioinformatic and hierarchical cluster analysis on extensive proteomic data.
Main Results:
- Achieved unprecedented depth, identifying over 500 membrane proteins, including 137 cell surface CD proteins.
- Successfully revealed known differentiation-induced changes in cell surface proteins.
- Identified novel cell surface proteins with limited prior information.
Conclusions:
- Mass spectrometry-based phenotyping offers an extensive and quantitative view of the leukemia surface proteome.
- This approach significantly improves the subclassification of hematologic malignancies.
- The findings facilitate the identification of new therapeutic targets for leukemia treatment.
Abstract:
Immunophenotyping by flow cytometry or immunohistochemistry is a clinical standard procedure for diagnosis, classification, and monitoring of hematologic malignancies. Antibody-based cell surface phenotyping is commonly limited to cell surface proteins for which specific antibodies are available and the number of parallel measurements is limited. The resulting limited knowledge about cell surface protein markers hampers early clinical diagnosis and subclassification of hematologic malignancies. Here, we describe the mass spectrometry based phenotyping of 2 all-trans retinoic acid treated acute myeloid leukemia model systems at an unprecedented level to a depth of more than 500 membrane proteins, including 137 bona fide cell surface exposed CD proteins. This extensive view of the leukemia surface proteome was achieved by developing and applying new implementations of the Cell Surface Capturing (CSC) technology. Bioinformatic and hierarchical cluster analysis showed that the applied strategy reliably revealed known differentiation-induced abundance changes of cell surface proteins in HL60 and NB4 cells and it also identified cell surface proteins with very little prior information. The extensive and quantitative analysis of the cell surface protein landscape from a systems biology perspective will be most useful in the clinic for the improved subclassification of hematologic malignancies and the identification of new drug targets.
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