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Transfecting and Nucleofecting Human Induced Pluripotent Stem Cells
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Neutrophils Derived from Genetically Modified Human Induced Pluripotent Stem Cells Circulate and Phagocytose Bacteria

Lisa R Trump1, Ramesh C Nayak1, Abhishek K Singh1,2

  • 1Division of Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.

Stem Cells Translational Medicine
|February 23, 2019
PubMed

Insights

Human induced pluripotent stem cells (iPSC) can generate neutrophils for treating infections in neutropenic patients. Activating AKT signaling in these cells significantly enhances their phagocytic activity and NET formation, improving therapeutic potential.

Area of Science:

  • Stem cell biology
  • Immunology
  • Hematology

Background:

  • Neutropenic patients face high morbidity and mortality from bacterial and fungal infections.
  • Neutrophil transfusions are a potential treatment but limited by short half-life and donor availability.
  • Human induced pluripotent stem cells (iPSC) offer a scalable source for neutrophil production.

Purpose of the Study:

  • To investigate the in vitro and in vivo efficacy of neutrophils derived from human iPSCs.
  • To identify signaling pathways limiting iPSC-derived neutrophil function.
  • To assess the therapeutic potential of enhancing iPSC-derived neutrophil function.

Main Methods:

  • Generation and phenotypic analysis of neutrophils from human iPSCs.
  • Assessment of reactive oxygen species production, phagocytosis, and neutrophil extracellular trap (NET) formation.
  • Analysis of signaling pathways (AKT, ERK, STAT3) in iPSC-derived neutrophils.
  • In vivo studies using a mouse model of bacterial peritonitis.

Main Results:

  • iPSC-derived neutrophils exhibited normal or enhanced reactive oxygen species production but reduced phagocytosis and NET formation compared to peripheral blood neutrophils.
  • Impaired AKT activation was identified in iPSC-derived neutrophil precursors.
  • Restoring AKT activation in iPSC-derived neutrophils significantly improved phagocytic activity and NET formation in vitro.
  • In vivo, iPSC-derived neutrophils migrated effectively, and AKT activation enhanced their phagocytic capacity.

Conclusions:

  • iPSC-derived neutrophils are a promising source for transfusion therapy in neutropenic patients.
  • AKT signaling is crucial for optimal iPSC-derived neutrophil function, particularly phagocytosis and NET formation.
  • Enhancing AKT activation in iPSC-derived neutrophils can significantly improve their therapeutic efficacy against infections.
Keywords:
AKTCirculationHuman induced pluripotent stem cellsNeutrophilsPhagocytosis

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