# Investigating the impact of inflammation on brain development using human cerebral organoids.

> **NIH NIH R21** · ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI · 2020 · $466,125

## Abstract

Schizophrenia (SCZ) and autism spectrum disorder (ASD) are severe neurodevelopmental disorders that are
caused by the interaction between genetic and environmental risk factors. Large-scale epidemiological have
consistently shown that inflammation during pregnancy is associated with an increased risk of SCZ and ASD in
offspring. A role for inflammation in the pathogenesis of SCZ and ASD is also supported by animal studies, as
well as genetic and post-mortem brain studies. How inflammation during pregnancy contributes to ASD and
SCZ at the molecular and cellular level is still largely unknown. The long-term goal of this project is to deepen
our insight into the role of inflammatory pathways in ASD and SCZ. To reach this goal we propose to use
cerebral organoids derived from human induced pluripotent stem cells (hiPSCs). Cerebral organoid cultures
have been shown to closely mimic molecular and cellular processes during the first two trimesters of in utero
human brain development. However, protocols up to now were lacking development of microglia, the immune
cells of the brain. We have developed an organoid protocol in which microglia develop innately. These
organoid-grown microglia resemble primary microglia at the phenotypic and functional level. In addition, we are
setting up a novel co-culture model for hiPSC-derived microglia and forebrain organoids. These models
provide novel possibilities to study the impact of inflammation on human brain development. The overall
objective of this project is to further develop these in vitro models to study the acute and long-lasting cellular
changes of the developing human brain induced by maternal inflammation. We will expose microglia-
containing organoids to the inflammatory triggers LPS or IFN-γ at a developmental stage reflecting the end of
the first trimester of gestation in humans. We will harvest the organoids at two early and two later timepoints. In
Aim 1. we will characterize the impact of immune activation on microglia number and phenotype, as well as the
expression of key inflammatory markers. We will analyze the microglia using immunostainings and the
expression of inflammatory markers using qPCR and western blots. Microglia will be sorted and transcriptomic
and protein expression changes analyzed using RNA sequencing and CyTOF. In Aim 2. we will investigate the
response of other neural cell types to LPS or IFN-γ stimulation. We will analyze organoid size and total cell
number, as well as the size and number of progenitor zones. In addition, we will analyze the expression of cell-
type specific markers in the organoids at different time point after stimulation. On a subsample of organoids,
we will perform single cell RNA-seq to determine changes in cell-type composition and cell-type specific
changes in gene expression. In the transcriptomic datasets we will analyze whether differential expressed
genes are enriched for ASD and SCZ-associated genes. We expect that this project will provide an in...

## Key facts

- **NIH application ID:** 9979244
- **Project number:** 1R21MH120581-01A1
- **Recipient organization:** ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
- **Principal Investigator:** Lotje Dorothee de Witte
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $466,125
- **Award type:** 1
- **Project period:** 2020-04-01 → 2023-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9979244

## Citation

> US National Institutes of Health, RePORTER application 9979244, Investigating the impact of inflammation on brain development using human cerebral organoids. (1R21MH120581-01A1). Retrieved via AI Analytics 2026-06-12 from https://api.ai-analytics.org/grant/nih/9979244. Licensed CC0.

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