Microglia-secreted TNF-α affects differentiation efficiency and viability of pluripotent stem cell-derived human dopaminergic precursors

PLoS One. 2023 Sep 26;18(9):e0263021. doi: 10.1371/journal.pone.0263021. eCollection 2023.

Abstract

Disease is a neurodegenerative disorder characterised by the progressive loss of dopaminergic cells of the substantia nigra pars compacta. Even though successful transplantation of dopamine-producing cells into the striatum exhibits favourable effects in animal models and clinical trials; transplanted cell survival is low. Since every transplant elicits an inflammatory response which can affect cell survival and differentiation, we aimed to study in vivo and in vitro the impact of the pro-inflammatory environment on human dopaminergic precursors. We first observed that transplanted human dopaminergic precursors into the striatum of immunosuppressed rats elicited an early and sustained activation of astroglial and microglial cells after 15 days' post-transplant. This long-lasting response was associated with Tumour necrosis factor alpha expression in microglial cells. In vitro, conditioned media from activated BV2 microglial cells increased cell death, decreased Tyrosine hydroxylase-positive cells and induced morphological alterations on human neural stem cells-derived dopaminergic precursors at two differentiation stages: 19 days and 28 days. Those effects were ameliorated by inhibition of Tumour necrosis factor alpha, a cytokine which was previously detected in vivo and in conditioned media from activated BV-2 cells. Our results suggest that a pro-inflammatory environment is sustained after transplantation under immunosuppression, providing a window of opportunity to modify this response to increase transplant survival and differentiation. In addition, our data show that the microglia-derived pro-inflammatory microenvironment has a negative impact on survival and differentiation of dopaminergic precursors. Finally, Tumour necrosis factor alpha plays a key role in these effects, suggesting that this cytokine could be an interesting target to increase the efficacy of human dopaminergic precursors transplantation in Parkinson's Disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation
  • Culture Media, Conditioned / pharmacology
  • Cytokines
  • Dopamine
  • Humans
  • Microglia*
  • Rats
  • Tumor Necrosis Factor-alpha* / pharmacology

Substances

  • Tumor Necrosis Factor-alpha
  • Culture Media, Conditioned
  • Dopamine
  • Cytokines

Grants and funding

This research project was funded by the Agencia Nacional de promoción de la investigación, el desarrollo tecnológico y la innovación (http://www.agencia.mincyt.gob.ar/) through grant PICT-2018-00517 to FJP. Founds were also provided by René Barón Foundation to FJP and the International Society of Neurochemistry -Committee for Aid and Education in Neurochemistry (ISN-CAEN) to SDW. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.