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A Multicancer Malignant Pleural Effusion Diagnostic Test Using Hexokinase 2 and Single-Cell Sequencing.

Jie Chen1, Yibin Yang2, Zhuo Wang3

  • 1Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai, China.

Clinical Chemistry
|February 10, 2022
PubMed
Summary

A new hexokinase 2-based sequencing (HK2-seq) method offers a highly sensitive and specific diagnostic tool for malignant pleural effusion (MPE). This advanced technique improves upon traditional cytology for multicancer diagnosis.

Keywords:
HK2-seqhexokinase 2malignant pleural effusionrare tumor cellssingle-cell sequencing

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Area of Science:

  • Oncology
  • Molecular Diagnostics
  • Biomarker Discovery

Background:

  • Malignant pleural effusion (MPE) indicates advanced cancer with a poor prognosis.
  • Current diagnostic methods like pleural effusion cytology have limited sensitivity and high variability.
  • There is a need for improved diagnostic tools for MPE across various cancer types.

Purpose of the Study:

  • To introduce and evaluate a novel hexokinase 2-based sequencing (HK2-seq) method for diagnosing multicancer MPE.
  • To assess the diagnostic performance of HK2-seq compared to conventional cytology.
  • To explore the utility of HK2-seq in identifying tumor characteristics through single-cell analysis.

Main Methods:

  • Developed HK2-seq utilizing hexokinase 2 (HK2) as a marker for increased glycolysis in disseminated tumor cells within pleural effusions.
  • Employed single-cell sequencing to confirm malignancy of HK2-identified high glycolytic tumor cells (hgTCs) by analyzing genome-wide copy number alterations (CNAs).
  • Conducted a prospective cohort study of 111 patients with pleural effusion to evaluate HK2-seq performance.

Main Results:

  • HK2-seq demonstrated high diagnostic performance for MPE across 12 cancer types: 91% sensitivity, 84% specificity, 90% PPV, and 86% NPV.
  • Compared to HK2-seq, conventional pleural effusion cytology showed a significantly lower diagnostic sensitivity of 45%.
  • Single-cell sequencing confirmed tumor origin of hgTCs and identified prognostic or targetable CNAs, including those unique to liquid biopsies.

Conclusions:

  • HK2-seq provides a definitive MPE diagnosis with high accuracy across multiple cancer types.
  • The method shows potential for multicancer detection of circulating tumor cells in various body fluids.
  • HK2-seq enables liquid biopsy-based genomic characterization for enhanced diagnostic information.