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<p>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 &gt;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.</p>

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

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年12月15日

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  • 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

摘要

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.