Profiling Cell Type-Specific Gene Regulatory Regions in Human Cortical Organoids

Schütze, Theresa M.; Bölicke, Nora; Sameith, Katrin; Albert, Mareike

doi:10.1007/978-1-0716-2720-4_2

Cited by 4 publications

(7 citation statements)

References 42 publications

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“…Since EPIREGULIN promoted changes in the cellular composition of the mNcx (Figure 2), which would be expected to bias any gene expression analysis, we performed immuno-fluorescent activated cell sorting (FACS) to isolate neural cell populations. Specifically, RG, IPs and neurons (Figure S4A, B) were isolated based on immuno-staining for the nuclear markers Sox2 and Tbr2 (Florio et al, 2015;Schütze et al, 2023), and neurons based on expression of GFP in the Tubb3::GFP mouse reporter line (Attardo et al, 2008). Isolated cell populations were validated by expression of the marker genes Sox2 and Prom1 (Prominin 1/CD133) for aRG, Eomes (Tbr2) for IPs, and Dcx (Doublecortin) and Tubb3 (Tuj1) for neurons (Figure S4C,D).…”

Section: Addition Of Epiregulin Does Not Induce Major Gene Expression...mentioning

confidence: 99%

“…The sex of embryos was not determined, as it is not likely to be of relevance for the results obtained in the present study. Mouse NSCs were isolated from E11.5/12.5 embryos as previously described (Schmitz et al, 2011;Schütze et al, 2023).…”

Section: Micementioning

confidence: 99%

“…Human cortical organoids were generated using the previously generated human induced pluripotent stem cell (iPSC) line CRTDi004-A (https://hpscreg.eu/cell-line/CRTDi004-A) (Völkner et al, 2022), derived from a healthy donor. The human iPSC line was maintained on Matrigel-coated (Corning, 354277) culture dishes in mTeSR1 and passaged using TrypLE Express enzyme (Schütze et al, 2023). Gorilla organoids were created using the previously generated gorilla iPSC line GC1 (Wunderlich et al, 2014).…”

Section: Human and Primate Induced Pluripotent Stem Cell Linesmentioning

confidence: 99%

“…Mouse NSCs were plated on 6-well plates coated with poly-D-lysine and laminin at a density of 105 cells/mm² and grown in NSC medium which contains a 1:1 mixture of DMEM/F12 and Neurobasal medium supplemented with 10 ng/mL epidermal growth factor (EGF), 20 ng/mL fibroblast growth factor (FGF), N-2 supplement, B-27 supplement, Penicillin/Streptomycin, Sodium-Pyruvate, GlutaMAX, MEM-NEAA, Heparin, HEPES, 2-mercaptoethanol and BSA (Schmitz et al, 2011;Schütze et al, 2023). NSCs are passaged using trypsin-EDTA and soybean trypsin inhibitor and kept in standard conditions at 37°C and 5% CO2.…”

Section: Mouse Nsc Culturementioning

confidence: 99%

“…Human and gorilla cortical organoids were generated according to the sliced neocortical organoid (SNO) method, as previously described in detail (Qian et al, 2020;Schütze et al, 2023). Briefly, hiPSC colonies of about 1.5 mm in size were detached with collagenase and transferred to ultra-low attachment 6-well plates (Corning, 3471), containing forebrain medium 1, to form 3D aggregates.…”

Section: Generation Of Human and Gorilla Organoidsmentioning

confidence: 99%

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Primate-expressed EPIREGULIN promotes basal progenitor proliferation in the developing neocortex

Cubillos,

Ditzer,

Kolodziejczyk

et al. 2023

Preprint

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Neocortex expansion during evolution is linked to higher numbers of neurons thought to result from increased proliferative capacity and neurogenic potential of basal progenitor cells (BPs) during development. Here we show that EREG, encoding the growth factor EPIREGULIN, is expressed in the human developing neocortex and in gorilla organoids, but not in the mouse neocortex. Addition of EPIREGULIN to the mouse neocortex increases proliferation of BPs via EGFR-mediated signaling, whereas ablation of EREG in human cortical organoids reduces BP proliferation. Addition of EPIREGULIN to cortical organoids promotes a further increase in proliferation of gorilla but not human BPs. Finally, we identify putative cis-regulatory elements that may contribute to inter-species differences in EREG expression. Overall, our results suggest that species-specific expression of EPIREGULIN may contribute to increased neocortex size in primates by providing a pro-proliferative signal to BPs in the subventricular zone progenitor niche.

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Section: Addition Of Epiregulin Does Not Induce Major Gene Expression...mentioning

confidence: 99%

Section: Micementioning

confidence: 99%

Section: Human and Primate Induced Pluripotent Stem Cell Linesmentioning

confidence: 99%

Section: Mouse Nsc Culturementioning

confidence: 99%

Section: Generation Of Human and Gorilla Organoidsmentioning

confidence: 99%

See 3 more Smart Citations

Primate-expressed EPIREGULIN promotes basal progenitor proliferation in the developing neocortex

Cubillos,

Ditzer,

Kolodziejczyk

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

The growth factor EPIREGULIN promotes basal progenitor cell proliferation in the developing neocortex

Cubillos,

Ditzer,

Kolodziejczyk

et al. 2024

EMBO J

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Neocortex expansion during evolution is linked to higher numbers of neurons, which are thought to result from increased proliferative capacity and neurogenic potential of basal progenitor cells during development. Here, we show that EREG, encoding the growth factor EPIREGULIN, is expressed in the human developing neocortex and in gorilla cerebral organoids, but not in the mouse neocortex. Addition of EPIREGULIN to the mouse neocortex increases proliferation of basal progenitor cells, whereas EREG ablation in human cortical organoids reduces proliferation in the subventricular zone. Treatment of cortical organoids with EPIREGULIN promotes a further increase in proliferation of gorilla but not of human basal progenitor cells. EPIREGULIN competes with the epidermal growth factor (EGF) to promote proliferation, and inhibition of the EGF receptor abrogates the EPIREGULIN-mediated increase in basal progenitor cells. Finally, we identify putative cis-regulatory elements that may contribute to the observed inter-species differences in EREG expression. Our findings suggest that species-specific regulation of EPIREGULIN expression may contribute to the increased neocortex size of primates by providing a tunable pro-proliferative signal to basal progenitor cells in the subventricular zone.

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Canonical and non-canonical PRC1 differentially contribute to the regulation of neural stem cell fate

Hoffmann,

Schütze,

Kolodziejczyk

et al. 2024

Preprint

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Neocortex development is characterized by sequential phases of neural progenitor cell (NPC) expansion, neurogenesis and gliogenesis. Polycomb-mediated epigenetic mechanisms are known to play important roles in regulating the lineage potential of NPCs during development. The composition of Polycomb Repressive Complex 1 (PRC1) is highly diverse in mammals and was hypothesized to contribute to context-specific regulation of cell fate. Here, we have performed side-by-side comparison of the role of canonical PRC1.2/1.4 and non-canonical PRC1.3/1.5, all of which are expressed in the developing neocortex, in NSC proliferation and differentiation. We found that the deletion ofPcgf2/4in NSCs led to a strong reduction in proliferation and to altered lineage fate, both during the neurogenic and gliogenic phase, whereasPcgf3/5played a minor role. Mechanistically, genes encoding stem cell and neurogenic factors were bound by PRC1 and differentially expressed uponPcgf2/4deletion. Thus, rather than different PRC1 sub-complexes contributing to different phases of neural development, we found that canonical PRC1 played a more significant role in NSC regulation during proliferative, neurogenic and gliogenic phases compared to non-canonical PRC1.

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Profiling Cell Type-Specific Gene Regulatory Regions in Human Cortical Organoids

Cited by 4 publications

References 42 publications

Primate-expressed EPIREGULIN promotes basal progenitor proliferation in the developing neocortex

Primate-expressed EPIREGULIN promotes basal progenitor proliferation in the developing neocortex

The growth factor EPIREGULIN promotes basal progenitor cell proliferation in the developing neocortex

Canonical and non-canonical PRC1 differentially contribute to the regulation of neural stem cell fate

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