Cell-type specific defects in PTEN-mutant cortical organoids converge on abnormal circuit activity
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Updated: Jul 25, 2025
Electroporation of Sliced Human Cortical Organoids for Studies of Gene Function
Published on: November 29, 2024
Martina Pigoni1,2, Ana Uzquiano1,2, Bruna Paulsen1,2
1Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA.
Abstract:
De novo heterozygous loss-of-function mutations in phosphatase and tensin homolog (PTEN) are strongly associated with autism spectrum disorders; however, it is unclear how heterozygous mutations in this gene affect different cell types during human brain development and how these effects vary across individuals. Here, we used human cortical organoids from different donors to identify cell-type specific developmental events that are affected by heterozygous mutations in PTEN. We profiled individual organoids by single-cell RNA-seq, proteomics and spatial transcriptomics and revealed abnormalities in developmental timing in human outer radial glia progenitors and deep-layer cortical projection neurons, which varied with the donor genetic background. Calcium imaging in intact organoids showed that both accelerated and delayed neuronal development phenotypes resulted in similar abnormal activity of local circuits, irrespective of genetic background. The work reveals donor-dependent, cell-type specific developmental phenotypes of PTEN heterozygosity that later converge on disrupted neuronal activity.
Insights
Mutations in phosphatase and tensin homolog (PTEN) disrupt human brain development. These PTEN gene mutations cause cell-type specific timing abnormalities, leading to altered neuronal circuit activity in autism spectrum disorders.
Area of Science:
- Neuroscience
- Developmental Biology
- Genetics
Background:
- Heterozygous loss-of-function mutations in phosphatase and tensin homolog (PTEN) are linked to autism spectrum disorders.
- The precise impact of these PTEN mutations on distinct cell types during human brain development and individual variability remains unclear.
Purpose of the Study:
- To investigate cell-type specific developmental effects of heterozygous PTEN mutations in human cortical organoids.
- To understand how donor genetic background influences these developmental abnormalities.
- To examine the consequences of these mutations on neuronal circuit activity.
Main Methods:
- Utilized human cortical organoids from multiple donors.
- Employed single-cell RNA-sequencing, proteomics, and spatial transcriptomics for comprehensive profiling.
- Conducted calcium imaging in intact organoids to assess neuronal circuit function.
Main Results:
- Identified abnormalities in developmental timing affecting outer radial glia progenitors and deep-layer cortical projection neurons.
- Observed that these developmental phenotypes varied depending on the donor's genetic background.
- Found that both accelerated and delayed neuronal development led to similar disruptions in local circuit activity, irrespective of genetic background.
Conclusions:
- Heterozygous PTEN mutations induce donor-dependent, cell-type specific developmental phenotypes during human brain development.
- These distinct cellular effects converge to disrupt neuronal activity in the developing brain.
- This study provides insights into the cellular and circuit-level mechanisms underlying PTEN-associated neurodevelopmental disorders.
