A simple, rapid, and practical method for single-cell proteomics based on mass-adaptive coating of synthetic peptides

Li, Siqi; Su, Kaichan; Zhuang, Zhenkun; Qin, Qing; Gao, Lei; Deng, Yamei; Liu, Xuyang; Hou, Guixue; Wang, Longtao; Hao, Piliang; Yang, Huanming; Liu, Siqi; Zhu, Hongming; Ren, Yan

doi:10.1016/j.scib.2021.12.022

Cited by 7 publications

(6 citation statements)

References 15 publications

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“…Single-cell proteomics samples were prepared based on mass-adaptive coating of synthetic peptides as previously reported [ 20 ]. See ESM Methods for further details.…”

Section: Methodsmentioning

confidence: 99%

“…The peptide samples were subjected to LC-MS/MS using a Fusio Lumo Tribrid mass spectrometer (Thermo Scientific, USA) as previously reported [ 20 ]. See ESM Methods for further details.…”

Section: Methodsmentioning

confidence: 99%

“…Thus, there is a great demand for the single-cell proteomics, which can sense and quantify more than 1000 proteins in a single mammalian cell [ 19 ]. The single-cell protein profile of circulating human CD34 + cells obtained from an efficient and straightforward mass-adaptive coating-assisted single-cell proteomics (Mad-CASP) revolutionises our knowledge of different CD34 + cell subsets [ 20 ]. Yet, single-cell proteomics studies of islet cell subpopulations are still lacking.…”

Section: Introductionmentioning

confidence: 99%

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Single-cell RNA sequencing combined with single-cell proteomics identifies the metabolic adaptation of islet cell subpopulations to high-fat diet in mice

Jiang

et al. 2022

Diabetologia

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Aims/hypothesis Islets have complex heterogeneity and subpopulations. Cell surface markers representing alpha, beta and delta cell subpopulations are urgently needed for investigations to explore the compositional changes of each subpopulation in obesity progress and diabetes onset, and the adaptation mechanism of islet metabolism induced by a high-fat diet (HFD). Methods Single-cell RNA sequencing (scRNA-seq) was applied to identify alpha, beta and delta cell subpopulation markers in an HFD-induced mouse model of glucose intolerance. Flow cytometry and immunostaining were used to sort and assess the proportion of each subpopulation. Single-cell proteomics was performed on sorted cells, and the functional status of each alpha, beta and delta cell subpopulation in glucose intolerance was deeply elucidated based on protein expression. Results A total of 33,999 cells were analysed by scRNA-seq and clustered into eight populations, including alpha, beta and delta cells. For alpha cells, scRNA-seq revealed that the Ace2 low subpopulation had downregulated expression of genes related to alpha cell function and upregulated expression of genes associated with beta cell characteristics in comparison with the Ace2 high subpopulation. The impaired function and increased fragility of ACE2 low alpha cells exposure to HFD was further suggested by single-cell proteomics. As for beta cells, the CD81 high subpopulation may indicate an immature signature of beta cells compared with the CD81 low subpopulation, which had robust function. We also found differential expression of Slc2a2 in delta cells and a potentially stronger cellular function and metabolism in GLUT2 low delta cells than GLUT2 high delta cells. Moreover, an increased proportion of ACE2 low alpha cells and CD81 low beta cells, with a constant proportion of GLUT2 low delta cells, were observed in HFD-induced glucose intolerance. Conclusions/interpretation We identified ACE2, CD81 and GLUT2 as surface markers to distinguish, respectively, alpha, beta and delta cell subpopulations with heterogeneous maturation and function. The changes in the proportion and functional status of islet endocrine subpopulations reflect the metabolic adaptation of islets to high-fat stress, which weakened the function of alpha cells and enhanced the function of beta and delta cells to bring about glycaemic homeostasis. Our findings provide a fundamental resource for exploring the mechanisms maintaining each islet endocrine subpopulation’s fate and function in health and disease. Data availability The scRNA-seq analysis datasets from the cu...

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“…Single-cell proteomics samples were prepared based on mass-adaptive coating of synthetic peptides as previously reported [ 20 ]. See ESM Methods for further details.…”

Section: Methodsmentioning

confidence: 99%

“…The peptide samples were subjected to LC-MS/MS using a Fusio Lumo Tribrid mass spectrometer (Thermo Scientific, USA) as previously reported [ 20 ]. See ESM Methods for further details.…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Single-cell RNA sequencing combined with single-cell proteomics identifies the metabolic adaptation of islet cell subpopulations to high-fat diet in mice

Jiang

et al. 2022

Diabetologia

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“…Because the protein content of one cell is small, special experimental conditions must be applied to detect larger number of proteins. Detergents should be avoided and, the pipetting and transfer from one tube to another must be minimized while including a mixture of peptides to reduce plasticware absorption [ 18 , 19 ]. Cells can be sorted by fluorescence-activated cell sorting (FACS) using specific antibodies and the various cell populations can be tagged with specific TMT molecules as a chemical barcode to increase the number of proteins per analysis, thereby increasing sensitivity and throughput.…”

Section: Cell-based Interception and Precision Medicine: A New Medica...mentioning

confidence: 99%

The 37TrillionCells initiative for improving global healthcare via cell-based interception and precision medicine: focus on neurodegenerative diseases

Coulombe,

Durcan,

Bernard

et al. 2024

Mol Brain

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One of the main burdens in the treatment of diseases is imputable to the delay between the appearance of molecular dysfunctions in the first affected disease cells and their presence in sufficient number for detection in specific tissues or organs. This delay obviously plays in favor of disease progression to an extent that makes efficient treatments difficult, as they arrive too late. The development of a novel medical strategy, termed cell-based interception and precision medicine, seeks to identify dysfunctional cells early, when tissue damages are not apparent and symptoms not yet present, and develop therapies to treat diseases early. Central to this strategy is the use of single-cell technologies that allow detection of molecular changes in cells at the time of phenotypical bifurcation from health to disease. In this article we describe a general procedure to support such an approach applied to neurodegenerative disorders. This procedure combines four components directed towards highly complementary objectives: 1) a high-performance single-cell proteomics (SCP) method (Detect), 2) the development of disease experimental cell models and predictive computational models of cell trajectories (Understand), 3) the discovery of specific targets and personalized therapies (Cure), and 4) the creation of a community of collaborating laboratories to accelerate the development of this novel medical paradigm (Collaborate). A global initiative named 37TrillionCells (37TC) was launched to advance the development of cell-based interception and precision medicine.

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“…14 Poly(amino acid)s have structures similar to those of proteins and show good compatibility with cells, tissues and other biological components. 15–17 The biocompatibility, biodegradability and self-assembly behavior of poly(amino acid)s make them have a very wide application prospect in controlled drug release systems. 18 In addition, poly(amino acid)s exhibit different characteristics from ordinary macromolecules due to the existence of secondary structures (α-helix and β-fold).…”

Section: Introductionmentioning

confidence: 99%

Self-assembly behavior and sustained drug release properties of amphiphilic poly(amino acid)s

Wang

Han

et al. 2022

New J. Chem.

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A simple, rapid, and practical method for single-cell proteomics based on mass-adaptive coating of synthetic peptides

Cited by 7 publications

References 15 publications

Single-cell RNA sequencing combined with single-cell proteomics identifies the metabolic adaptation of islet cell subpopulations to high-fat diet in mice

Single-cell RNA sequencing combined with single-cell proteomics identifies the metabolic adaptation of islet cell subpopulations to high-fat diet in mice

The 37TrillionCells initiative for improving global healthcare via cell-based interception and precision medicine: focus on neurodegenerative diseases

Self-assembly behavior and sustained drug release properties of amphiphilic poly(amino acid)s

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