Single cell proteomic analysis defines discrete neutrophil functional states in human glioblastoma

Pranvera Sadiku; Alejandro J. Brenes; Rupert L. Mayer; Leila Reyes; Patricia Coelho; Gabi van Stralen; Ailiang Zhang; Manuel A. Sanchez-Garcia; Emily R. Watts; Imran Liaquat; Andrew J.M. Howden; Ikeoluwa Adekoya; Anuka Boldbaatar; Allan MacRaild; Sarah Risbridger; Gillian M. Morrison; Heather MacPherson; Caroline M. Bruce; Shonna Johnston; Robert Grecian; Fiona A. Murphy; Steven M. Pollard; Paul M. Brennan; Karl Mechtler; Sarah R. Walmsley

doi:10.1038/s41467-025-67367-3

Single cell proteomic analysis defines discrete neutrophil functional states in human glioblastoma

Pranvera Sadiku
, Alejandro J. Brenes (Lead / Corresponding author)
, Rupert L. Mayer
, Leila Reyes
, Patricia Coelho
, Gabi van Stralen
, Ailiang Zhang
, Manuel A. Sanchez-Garcia
, Emily R. Watts
, Imran Liaquat
, Andrew J.M. Howden
, Ikeoluwa Adekoya
, Anuka Boldbaatar
, Allan MacRaild
, Sarah Risbridger
, Gillian M. Morrison
, Heather MacPherson
, Caroline M. Bruce
, Shonna Johnston
, Robert Grecian

Fiona A. Murphy, Steven M. Pollard, Paul M. Brennan, Karl Mechtler, Sarah R. Walmsley (Lead / Corresponding author)

Cell Signalling and Immunology

Research output: Contribution to journal › Article › peer-review

Abstract

Neutrophils are vital innate immune cells shown to infiltrate glioblastomas, however we currently lack the molecular understanding of their functional states within the tumour niche. Given that neutrophils are known to display a prominent discordance between mRNA and protein abundance, we developed ultra-sensitive mini-bulk and single cell proteomic (SCP) workflows to study the heterogeneity of peripheral blood and tumour associated neutrophils (TAN) from patients with glioblastoma. Mini-bulk analysis enabled a deeper protein coverage of circulating immature, mature and TAN populations, defining signatures of maturity and demonstrating that TANs resemble mature circulating neutrophils. Analysis of the SCP data results in the detection of >1100 proteins from a single TAN providing a detailed characterization of neutrophil subsets in glioblastoma. Our approach shows evidence of pathogenic and anti-tumorigenic clusters and discovers cell states invisible to scRNAseq, opening new opportunities to selectively target pro-tumoural neutrophil states.

Original language	English
Article number	621
Journal	Nature Communications
Volume	17
Issue number	1
Early online date	15 Dec 2025
DOIs	https://doi.org/10.1038/s41467-025-67367-3
Publication status	E-pub ahead of print - 15 Dec 2025

ASJC Scopus subject areas

General Chemistry
General Biochemistry,Genetics and Molecular Biology
General
General Physics and Astronomy

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Cite this

Sadiku, P., Brenes, A. J., Mayer, R. L., Reyes, L., Coelho, P., van Stralen, G., Zhang, A., Sanchez-Garcia, M. A., Watts, E. R., Liaquat, I., Howden, A. J. M., Adekoya, I., Boldbaatar, A., MacRaild, A., Risbridger, S., Morrison, G. M., MacPherson, H., Bruce, C. M., Johnston, S., ... Walmsley, S. R. (2026). Single cell proteomic analysis defines discrete neutrophil functional states in human glioblastoma. Nature Communications, 17(1), Article 621. Advance online publication. https://doi.org/10.1038/s41467-025-67367-3

@article{579f167d80a144e8b8d2a179071fbb9e,

title = "Single cell proteomic analysis defines discrete neutrophil functional states in human glioblastoma",

abstract = "Neutrophils are vital innate immune cells shown to infiltrate glioblastomas, however we currently lack the molecular understanding of their functional states within the tumour niche. Given that neutrophils are known to display a prominent discordance between mRNA and protein abundance, we developed ultra-sensitive mini-bulk and single cell proteomic (SCP) workflows to study the heterogeneity of peripheral blood and tumour associated neutrophils (TAN) from patients with glioblastoma. Mini-bulk analysis enabled a deeper protein coverage of circulating immature, mature and TAN populations, defining signatures of maturity and demonstrating that TANs resemble mature circulating neutrophils. Analysis of the SCP data results in the detection of >1100 proteins from a single TAN providing a detailed characterization of neutrophil subsets in glioblastoma. Our approach shows evidence of pathogenic and anti-tumorigenic clusters and discovers cell states invisible to scRNAseq, opening new opportunities to selectively target pro-tumoural neutrophil states.",

author = "Pranvera Sadiku and Brenes, \{Alejandro J.\} and Mayer, \{Rupert L.\} and Leila Reyes and Patricia Coelho and \{van Stralen\}, Gabi and Ailiang Zhang and Sanchez-Garcia, \{Manuel A.\} and Watts, \{Emily R.\} and Imran Liaquat and Howden, \{Andrew J.M.\} and Ikeoluwa Adekoya and Anuka Boldbaatar and Allan MacRaild and Sarah Risbridger and Morrison, \{Gillian M.\} and Heather MacPherson and Bruce, \{Caroline M.\} and Shonna Johnston and Robert Grecian and Murphy, \{Fiona A.\} and Pollard, \{Steven M.\} and Brennan, \{Paul M.\} and Karl Mechtler and Walmsley, \{Sarah R.\}",

note = "Publisher Copyright: {\textcopyright} The Author(s) 2025.",

year = "2025",

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doi = "10.1038/s41467-025-67367-3",

language = "English",

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Sadiku, P, Brenes, AJ, Mayer, RL, Reyes, L, Coelho, P, van Stralen, G, Zhang, A, Sanchez-Garcia, MA, Watts, ER, Liaquat, I, Howden, AJM, Adekoya, I, Boldbaatar, A, MacRaild, A, Risbridger, S, Morrison, GM, MacPherson, H, Bruce, CM, Johnston, S, Grecian, R, Murphy, FA, Pollard, SM, Brennan, PM, Mechtler, K & Walmsley, SR 2026, 'Single cell proteomic analysis defines discrete neutrophil functional states in human glioblastoma', Nature Communications, vol. 17, no. 1, 621. https://doi.org/10.1038/s41467-025-67367-3

TY - JOUR

T1 - Single cell proteomic analysis defines discrete neutrophil functional states in human glioblastoma

AU - Sadiku, Pranvera

AU - Brenes, Alejandro J.

AU - Mayer, Rupert L.

AU - Reyes, Leila

AU - Coelho, Patricia

AU - van Stralen, Gabi

AU - Zhang, Ailiang

AU - Sanchez-Garcia, Manuel A.

AU - Watts, Emily R.

AU - Liaquat, Imran

AU - Howden, Andrew J.M.

AU - Adekoya, Ikeoluwa

AU - Boldbaatar, Anuka

AU - MacRaild, Allan

AU - Risbridger, Sarah

AU - Morrison, Gillian M.

AU - MacPherson, Heather

AU - Bruce, Caroline M.

AU - Johnston, Shonna

AU - Grecian, Robert

AU - Murphy, Fiona A.

AU - Pollard, Steven M.

AU - Brennan, Paul M.

AU - Mechtler, Karl

AU - Walmsley, Sarah R.

PY - 2025/12/15

Y1 - 2025/12/15

N2 - Neutrophils are vital innate immune cells shown to infiltrate glioblastomas, however we currently lack the molecular understanding of their functional states within the tumour niche. Given that neutrophils are known to display a prominent discordance between mRNA and protein abundance, we developed ultra-sensitive mini-bulk and single cell proteomic (SCP) workflows to study the heterogeneity of peripheral blood and tumour associated neutrophils (TAN) from patients with glioblastoma. Mini-bulk analysis enabled a deeper protein coverage of circulating immature, mature and TAN populations, defining signatures of maturity and demonstrating that TANs resemble mature circulating neutrophils. Analysis of the SCP data results in the detection of >1100 proteins from a single TAN providing a detailed characterization of neutrophil subsets in glioblastoma. Our approach shows evidence of pathogenic and anti-tumorigenic clusters and discovers cell states invisible to scRNAseq, opening new opportunities to selectively target pro-tumoural neutrophil states.

AB - Neutrophils are vital innate immune cells shown to infiltrate glioblastomas, however we currently lack the molecular understanding of their functional states within the tumour niche. Given that neutrophils are known to display a prominent discordance between mRNA and protein abundance, we developed ultra-sensitive mini-bulk and single cell proteomic (SCP) workflows to study the heterogeneity of peripheral blood and tumour associated neutrophils (TAN) from patients with glioblastoma. Mini-bulk analysis enabled a deeper protein coverage of circulating immature, mature and TAN populations, defining signatures of maturity and demonstrating that TANs resemble mature circulating neutrophils. Analysis of the SCP data results in the detection of >1100 proteins from a single TAN providing a detailed characterization of neutrophil subsets in glioblastoma. Our approach shows evidence of pathogenic and anti-tumorigenic clusters and discovers cell states invisible to scRNAseq, opening new opportunities to selectively target pro-tumoural neutrophil states.

UR - https://www.scopus.com/pages/publications/105027800442

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DO - 10.1038/s41467-025-67367-3

M3 - Article

C2 - 41397978

AN - SCOPUS:105027800442

SN - 2041-1723

VL - 17

JO - Nature Communications

JF - Nature Communications

IS - 1

M1 - 621

ER -

Single cell proteomic analysis defines discrete neutrophil functional states in human glioblastoma

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