A Quantitative Single-Cell Proteomics Approach to Characterize an Acute Myeloid Leukemia Hierarchy
A Quantitative Single-Cell Proteomics Approach to Characterize an Acute Myeloid Leukemia Hierarchy
Abstract In recent years, cellular life science research has experienced a significant shift, moving away from conducting bulk cell interrogation towards single-cell analysis. It is only through single cell analysis that a complete understanding of cellular heterogeneity, and the interplay between various cell types that are fundamental to specific biological phenotypes, can be achieved. Single-cell assays at the protein level have been predominantly limited to targeted, antibody-based methods. However, here we present an experimental and computational pipeline, which establishes a comprehensive single-cell mass spectrometry-based proteomics workflow. By exploiting a leukemia culture system, containing functionally-defined leukemic stem cells, progenitors and terminally differentiated blasts, we demonstrate that our workflow is able to explore the cellular heterogeneity within this aberrant developmental hierarchy. We show our approach is capable to quantifying hundreds of proteins across hundreds of single cells using limited instrument time. Furthermore, we developed a computational pipeline (SCeptre), that effectively clusters the data and permits the extraction of cell-specific proteins and functional pathways. This proof-of-concept work lays the foundation for future global single-cell proteomics studies.
Savickas Simonas、Lechman Eric、Schoof Erwin M.、dem Keller Ulrich auf、Haile James Seymour、Porse Bo T.、Dick John E.、Rapin Nicolas、Gentil Coline
Department of Biotechnology and Biomedicine, Technical University of DenmarkPrincess Margaret Cancer Centre, University Health Network||Department of Molecular Genetics, University of TorontoThe Finsen Laboratory, Rigshospitalet, Faculty of Health Sciences, University of Copenhagen||Biotech Research and Innovation Centre (BRIC), University of Copenhagen||Department of Biotechnology and Biomedicine, Technical University of Denmark||Danish Stem Cell Centre (DanStem), University of Copenhagen||Princess Margaret Cancer Centre, University Health Network||Department of Molecular Genetics, University of TorontoDepartment of Biotechnology and Biomedicine, Technical University of DenmarkThe Finsen Laboratory, Rigshospitalet, Faculty of Health Sciences, University of Copenhagen||Biotech Research and Innovation Centre (BRIC), University of Copenhagen||Danish Stem Cell Centre (DanStem), University of CopenhagenThe Finsen Laboratory, Rigshospitalet, Faculty of Health Sciences, University of Copenhagen||Biotech Research and Innovation Centre (BRIC), University of Copenhagen||Danish Stem Cell Centre (DanStem), University of CopenhagenPrincess Margaret Cancer Centre, University Health Network||Department of Molecular Genetics, University of TorontoThe Finsen Laboratory, Rigshospitalet, Faculty of Health Sciences, University of Copenhagen||Biotech Research and Innovation Centre (BRIC), University of Copenhagen||Danish Stem Cell Centre (DanStem), University of CopenhagenThe Finsen Laboratory, Rigshospitalet, Faculty of Health Sciences, University of Copenhagen||Biotech Research and Innovation Centre (BRIC), University of Copenhagen
医学研究方法基础医学细胞生物学
acute myeloid leukemiacomputational biologyproteomicssingle-cell approachestandem mass tags (TMT)
Savickas Simonas,Lechman Eric,Schoof Erwin M.,dem Keller Ulrich auf,Haile James Seymour,Porse Bo T.,Dick John E.,Rapin Nicolas,Gentil Coline.A Quantitative Single-Cell Proteomics Approach to Characterize an Acute Myeloid Leukemia Hierarchy[EB/OL].(2025-03-28)[2025-05-05].https://www.biorxiv.org/content/10.1101/745679.点此复制
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