TY - JOUR
T1 - Assessing Gq-GPCR–induced human astrocyte reactivity using bioengineered neural organoids
AU - Cvetkovic, Caroline
AU - Patel, Rajan
AU - Shetty, Arya
AU - Hogan, Matthew K.
AU - Anderson, Morgan
AU - Basu, Nupur
AU - Aghlara-Fotovat, Samira
AU - Ramesh, Srivathsan
AU - Sardar, Debosmita
AU - Veiseh, Omid
AU - Ward, Michael E.
AU - Deneen, Benjamin
AU - Horner, Philip J.
AU - Krencik, Robert
N1 - Funding Information:
Research reported in this publication was supported by the National Institute on Aging of the National Institutes of Health (NIH) under Award Number R21AG064567. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Research was also supported by Mission Connect (a program of TIRR Foundation; 019-114), The Michael J. Fox Foundation for Parkinson’s Research (17871), and Cancer Prevention and Research Institute of Texas Award (RP200655). We thank the RNA Core and Center for Bioenergetics at Houston Methodist Research Institute for technical support, Dr. Gillian Hamilton and Dr. Yi-Lan Weng at Houston Methodist Research Institute for helpful discussions, Bushra Biba at Novogene Corporation for assistance with biostatistics, and Dr. Sung Yun Jung and Antrix Jain at the Mass Spectrometry Proteomics Core at Baylor College of Medicine (BCM) for their assistance. The BCM Mass Spectrometry Proteomics Core is supported by the Dan L. Duncan Comprehensive Cancer Center NIH award (P30 CA125123), Cancer Prevention and Research Institute of Texas Core Facility Award (RP170005), and NIH High End Instrument award (S10 OD026804).
Funding Information:
Research reported in this publication was supported by the National Institute on Aging of the National Institutes of Health (NIH) under Award Number R21AG064567. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Research was also supported by Mission Connect (a program of TIRR Foundation; 019-114), The Michael J. Fox Foundation for Parkinson’s Research (17871), and Cancer Prevention and Research Institute of Texas Award (RP200655). We thank the RNA Core and Center for Bioenergetics at Houston Methodist Research Institute for technical support, Dr. Gillian Hamilton and Dr. Yi-Lan Weng at Houston Methodist Research Institute for helpful discussions, Bushra Biba at Novogene Corporation for assistance with biostatistics, and Dr. Sung Yun Jung and Antrix Jain at the Mass Spectrometry Proteomics Core at Baylor College of Medicine (BCM) for their assistance. The BCM Mass Spectrometry Proteomics Core is supported by the Dan L. Duncan Comprehensive Cancer Center NIH award (P30 CA125123), Cancer Prevention and Research Institute of Texas Core Facility Award (RP170005), and NIH High End Instrument award (S10 OD026804). The authors declare no competing interests.
Publisher Copyright:
© 2022 Cvetkovic et al.
PY - 2022/2/9
Y1 - 2022/2/9
N2 - Astrocyte 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.
AB - Astrocyte 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.
KW - Cell signaling
KW - Neuroscience
KW - Stem cells
KW - Bioengineering
KW - Cell Line
KW - Synaptophysin/metabolism
KW - Reproducibility of Results
KW - Spheroids, Cellular/drug effects
KW - Astrocytes/metabolism
KW - Pluripotent Stem Cells/metabolism
KW - Humans
KW - Neural Stem Cells/drug effects
KW - Adenosine Triphosphate/pharmacology
KW - Organoids/metabolism
KW - Calcium/metabolism
KW - GTP-Binding Protein alpha Subunits, Gq-G11/metabolism
KW - Glial Fibrillary Acidic Protein/metabolism
KW - Neurons/metabolism
KW - Receptors, G-Protein-Coupled/metabolism
KW - Inflammation/pathology
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U2 - 10.1083/jcb.202107135
DO - 10.1083/jcb.202107135
M3 - Article
C2 - 35139144
AN - SCOPUS:85125005054
SN - 0021-9525
VL - 221
JO - Journal of Cell Biology
JF - Journal of Cell Biology
IS - 4
M1 - e202107135
ER -