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Multigenerational cell tracking of DNA replication and heritable DNA damage.

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This study tracks cell division over generations to reveal how cancer-causing changes create differences between sister cells, impacting genome stability and cell diversity.

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Area of Science:

  • Cellular biology
  • Genomics
  • Cancer research

Background:

  • Cell heterogeneity is fundamental to life, influencing development, tumor evolution, and drug responses.
  • Understanding the origins and propagation of cell-to-cell variation is crucial but challenging.
  • Retrospective analyses of cancer genomics struggle to resolve the emergence and inheritance of cellular heterogeneity.

Purpose of the Study:

  • To elucidate how oncogenic perturbations induce sister cell asymmetry and phenotypic heterogeneity.
  • To develop a framework for dissecting phenotypic plasticity at the single-cell level.
  • To investigate cellular processes relevant to early cancer development.

Main Methods:

  • Multigenerational single-cell tracking using endogenously labeled proteins.
  • Dual CRISPR-based genome editing for simultaneous tracking of DNA replication and heritable DNA lesions.
  • Time-resolved lineage analysis combined with iterative staining for cell cycle and DNA damage markers, and single-cell transcriptomics.

Main Results:

  • Detailed tracking of cell lineage trees up to four generations in asynchronously growing cells.
  • Revealed replication and repair dynamics, damage inheritance, and the emergence of sister cell heterogeneity across multiple generations.
  • Delineated how oncogenic events trigger distinct routes to polyploidization, affecting genome integrity.

Conclusions:

  • The study provides a novel framework for dissecting phenotypic plasticity and cell heterogeneity.
  • Identified mechanisms by which oncogenic perturbations drive cell asymmetry and diverse cellular outcomes.
  • Offers insights into early cellular events during cancer development.