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Endocrine Abstracts (2022) 84 OP13-67 | DOI: 10.1530/endoabs.84.OP-13-67

ETA2022 Oral Presentations Oral Session 13: Basic 3 (5 abstracts)

A single cell atlas of the T3-responsive transcriptome during early cortical neurogenesis in human cerebral organoids

Robert Opitz 1 , Valeria Fernandez-Vallone 2 , Matthias Megges 3 , Audrey Amber Julie Bresser 4 , Benedikt Obermayer-Wasserscheid 5 , Harald Stachelscheid 6 & Peter Kühnen 3


1Charité Universitätsmedizin Berlin, 3) Institute of Experimental Pediatric Endocrinology, Charité – Universitätsmedizin, Institut für Experimentelle Pädiatrische Endokrinologie, Berlin, Germany; 2Berlin Institute of Health, Core Unit Stem Cells, Berlin, Germany; 3Charité Universitätsmedizin Berlin, Institut für Experimentelle Pädiatrische Endokrinologie, Berlin, Germany; 4Charité Universitätsmedizin Berlin, Institute of Experimental Pediatric Endocrinology, Charité Berlin, Germany, Institut für Experimentelle Pädiatrische Endokrinologie, Berlin, Germany; 5Berlin Institute of Health, Bih Core Unit Bioinformatics, Charité – Universitätsmedizin Berlin, Core Unit Bioinformatics, Berlin, Germany; 6Berlin Institute of Health, Bih Stem Cell Core, Berlin Institute of Heath at Charité – Universitätsmedizin Berlin, Core Unit Stem Cells, Berlin, Germany


Perturbation of thyroid hormone (TH) function during prenatal brain development can cause a spectrum of neurological disorders as evident in pediatric patients with untreated congenital hypothyroidism or patients with mutations in THRA and SLC16A2 genes. However, scarcity and inaccessibility of human brain tissue from early developmental stages has been a major roadblock for experimental approaches to decipher local TH action in early development. Advanced in vitro models including human cerebral organoids (hCOs) hold great promises of accelerating our understanding of early human brain development and the hormonal signals that govern the complex processes underlying cortical neurogenesis. In this study, we differentiated hCOs from human induced pluripotent stem cells (hiPSC) in media containing either 1.5 nM T3 (BASAL group) or 20 nM T3 (CHRONIC group) for 9 weeks. Starting at day 13 of differentiation, hCOs were harvested at various developmental stages and analyzed by single cell RNA-seq for T3 treatment effects on the transcriptome. In addition, a subset of hCOs from the BASAL group was acutely treated with 50 nM T3 for 48 h (PULSE) at various stages and similarly analyzed by single cell RNA-seq. To avoid hiPSC line-inherent bias, all treatments were replicated for three different hiPSC lines resulting in 27 multiplexed single cell libraries. Global analysis of all cells collected during this study identified 31 neural cell clusters. Following cluster annotation, we could assign more than 95% of all cells to a major lineage trajectory from neural stem cell state towards excitatory neurons. Our single cell atlas comprised major cortical cell types including various radial glia cell (RGC) populations, three intermediate progenitor (IP) subtypes, as well as nine excitatory neuron subtypes. We defined developmental and T3-responsive expression profiles for genes related to TH action with cell type-specific resolution. For a subset of genes, smFISH was used to correlate spatial mRNA expression patterns with the laminar cell type distribution. We identified partially overlapping gene signatures in response to acute and chronic T3 treatment. Cell type- and developmental stage-specific gene set enrichments highlighted T3 effects on metabolic pathways in RGC, cell cycle regulation in IP and synapse function in neurons. T3-induced gene signatures in hCOs showed limited overlap with published mouse data sets. Chronic T3 treatment did not cause differentiation of aberrant cell types but altered the relative proportion of neuronal cell types in late stage hCOs. This comprehensive atlas of the T3-responsive transcriptome will serve as a unique resource to propel the use of hCOs in delineating the regulatory logic of local TH action in human brain development.

Volume 84

44th Annual Meeting of the European Thyroid Association (ETA) 2022

Brussels, Belgium
10 Sep 2022 - 13 Sep 2022

European Thyroid Association 

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