Cvetkovic, CarolinePatel, RajanShetty, AryaHogan, Matthew K.Anderson, MorganBasu, NupurAghlara-Fotovat, SamiraRamesh, SrivathsanSardar, DebosmitaVeiseh, OmidWard, Michael E.Deneen, BenjaminHorner, Philip J.Krencik, Robert2022-09-012022-09-012022Cvetkovic, Caroline, Patel, Rajan, Shetty, Arya, et al.. "Assessing Gq-GPCR–induced human astrocyte reactivity using bioengineered neural organoids." <i>Journal of Cell Biology,</i> 221, no. 4 (2022) Rockefeller University Press: https://doi.org/10.1083/jcb.202107135.https://hdl.handle.net/1911/113167Astrocyte reactivity can directly modulate nervous system function and immune responses during disease and injury. However, the consequence of human astrocyte reactivity in response to specific contexts and within neural networks is obscure. Here, we devised a straightforward bioengineered neural organoid culture approach entailing transcription factor–driven direct differentiation of neurons and astrocytes from human pluripotent stem cells combined with genetically encoded tools for dual cell-selective activation. This strategy revealed that Gq-GPCR activation via chemogenetics in astrocytes promotes a rise in intracellular calcium followed by induction of immediate early genes and thrombospondin 1. However, astrocytes also undergo NF-κB nuclear translocation and secretion of inflammatory proteins, correlating with a decreased evoked firing rate of cocultured optogenetic neurons in suboptimal conditions, without overt neurotoxicity. Altogether, this study clarifies the intrinsic reactivity of human astrocytes in response to targeting GPCRs and delivers a bioengineered approach for organoid-based disease modeling and preclinical drug testing.engThis article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).Journal articlejcb_202107135https://doi.org/10.1083/jcb.202107135