Developing Human Pluripotent Stem Cell-Based Cerebral Organoids with a Controllable Microglia Ratio for Modeling Brain Development and Pathology

Xu, Renfeng; Boreland, Andrew J.; Li, Xiaoxi; Erickson, Caroline; Jin, Mengmeng; Atkins, Colm; Pang, Zhiping P.; Daniels, Brian P.; Jiang, Peng

doi:10.1101/2020.10.09.331710

Cited by 11 publications

(22 citation statements)

References 78 publications

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“…However, in the cerebral organoids, microglia extend fewer processes and do not show morphology as highly ramified as they do in vivo (Benito-Kwiecinski and Lancaster, 2020; Ormel et al, 2018; Xu et al, 2021). Consequently, we did not observe any significant morphological differences between DS vs. Cont microglia in cell size, as indicated by the quantification of microglia cell volume (Fig.…”

Section: Resultsmentioning

confidence: 87%

“…We recently developed a novel microglia-containing cerebral organoid model that provides a convenient tool to study human microglial functions in neurodevelopmental disorders (Xu et al, 2021). As shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…2A, here we used PMPs derived from the three pairs of Cont and DS hiPSCs and primitive neural progenitor cells (pNPCs) derived from Cont hiPSC cells (Xu et al, 2019). Then, hiPSC-derived pNPCs and PMPs were mixed to develop organoids, as described in our recent study (Xu et al, 2021). By week 4, both Cont and DS organoids contained cells that expressed CD45, which is a marker for all nucleated cells of the hemopoietic lineage (Carson et al, 1998) and labels microglia in the organoids (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Each brain organoid was generated from a total of 10,000 cells (7,000 pNPCs and 3,000 PMPs), in each well of ultra-low-attachment 96-well plates in the presence of ROCK inhibitor Y-27632 (10 mM) (Xu et al, 2021). The culture medium was composed of a 1:1 mixture of PMP medium and NPC medium (1:1 mixture of Neurobasal and DMEM/F12, supplemented with 1x N2, 1x B27-RA, and FGF2 (20 ng/ml, Peprotech) for three days (day 3).…”

Section: Brain Organoid Culturementioning

confidence: 99%

“…Recent advances in stem cell technology have led to the efficient generation of microglia from human induced pluripotent stem cells (hiPSCs) (Abud et al, 2017; Brownjohn et al, 2018; Douvaras et al, 2017; Haenseler et al, 2017; Jiang et al, 2020; Muffat et al, 2016; Pandya et al, 2017), providing an unlimited source of human microglia to study their pathophysiology. Very recently, we have developed new hiPSC-based microglia-containing cerebral organoid and chimeric mouse brain models (Jiang et al, 2020; Xu et al, 2021; Xu et al, 2020), in which hiPSC-derived microglia undergo maturation and develop appropriate functions. With DS hiPSC established in our previous studies (Chen et al, 2014; Xu et al, 2019), these new models provide unprecedented opportunities to investigate the role of human microglia in DS brain development and DSAD.…”

Section: Introductionmentioning

confidence: 99%

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Type I Interferon Signaling Drives Microglial Dysfunction and Senescence in Human iPSC Models of Down Syndrome and Alzheimer’s Disease

Jin

Alam

et al. 2021

Preprint

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Microglia are critical for brain development and play a central role in Alzheimers disease (AD) etiology. Down syndrome (DS), also known as trisomy 21, is the most common genetic origin of intellectual disability and the most common risk factor for AD. Surprisingly, little information is available on the impact of trisomy of human chromosome 21 (Hsa21) on microglia in DS brain development and AD in DS (DSAD). Using our new induced pluripotent stem cell (iPSC)-based human microglia-containing cerebral organoid and chimeric mouse brain models, here we report that DS microglia exhibit enhanced synaptic pruning function during brain development. Consequently, electrophysiological recordings demonstrate that DS microglial mouse chimeras show impaired synaptic neurotransmission, as compared to control microglial chimeras. Upon being exposed to human brain tissue-derived soluble pathological tau, DS microglia display dystrophic phenotypes in chimeric mouse brains, recapitulating microglial responses seen in human AD and DSAD brain tissues. Further flow cytometry, single-cell RNA-sequencing, and immunohistological analyses of chimeric mouse brains demonstrate that DS microglia undergo cellular senescence and exhibit elevated type I interferon signaling after being challenged by pathological tau. Mechanistically, we find that shRNA-mediated knockdown of Hsa21encoded type I interferon receptor genes, IFNARs, rescues the defective DS microglial phenotypes both during brain development and in response to pathological tau. Our findings provide first in vivo evidence supporting a paradigm shifting theory that human microglia respond to pathological tau by exhibiting accelerated senescence and dystrophic phenotypes. Our results further suggest that targeting IFNARs may improve microglial functions during DS brain development and prevent human microglial senescence in DS individuals with AD.

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Section: Resultsmentioning

confidence: 87%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Brain Organoid Culturementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Type I Interferon Signaling Drives Microglial Dysfunction and Senescence in Human iPSC Models of Down Syndrome and Alzheimer’s Disease

Jin

Alam

et al. 2021

Preprint

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Microglia innate immune response contributes to the antiviral defense and blood–CSF barrier function in human choroid plexus organoids during HSV‐1 infection

Qiao

Chiu

Liang

et al. 2023

Journal of Medical Virology

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The choroid plexus (ChP) is the source of cerebrospinal fluid (CSF). The ChP-CSF system not only provides the necessary cushion for the brain but also works as a sink for waste clearance. During sepsis, pathogens and host immune cells can weaken the ChP barrier and enter the brain, causing cerebral dysfunctions known as sepsisassociated encephalophagy. Here, we used human ChP organoid (ChPO) to model herpes simplex virus type 1 (HSV-1) infection and found ChP epithelial cells were highly susceptible to HSV-1. Since the current ChPO model lacks a functional innate immune component, particularly microglia, we next developed a new microgliacontaining ChPO model, and found microglia could effectively limit HSV-1 infection and protect epithelial barrier in ChPOs. Furthermore, we found the innate immune cyclic GMP-AMP synthase (cGAS)-STING pathway and its downstream interferon response were essential, as cGAS inhibitor RU.512 or STING inhibitor H-151 abolished microglia antiviral function and worsened ChP barrier in organoids. These results together indicated that cGAS-STING pathway coordinates antiviral response in ChP and contributes to treating sepsis or related neurological conditions.

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Toward the next generation of vascularized human neural organoids

Gao

Zhao

et al. 2022

Medicinal Research Reviews

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Thanks to progress in the development of threedimensional (3D) culture technologies, human central nervous system (CNS) development and diseases have been gradually deciphered by using organoids derived from human embryonic stem cells (hESCs) or human induced pluripotent stem cells (hiPSCs). Selforganized neural organoids (NOs) have been used to mimic morphogenesis and functions of specific organs in vitro. Many NOs have been reproduced in vitro, such as those mimicking the human brain, retina, and spinal cord. However, NOs fail to capitulate to the maturation and complexity of in vivo neural tissues. The persistent issues with current NO cultivation protocols are inadequate oxygen supply and nutrient diffusion due to the absence of vascular networks.In vivo, the developing CNS is interpenetrated by vasculature that not only supplies oxygen and nutrients but also provides a structural template for neuronal growth.To address these deficiencies, recent studies have begun to couple NO culture with bioengineering techniques and methodologies, including genetic engineering, coculture, multidifferentiation, microfluidics and 3D bioprinting, and transplantation, which might promote NO maturation and create more functional NOs. These cutting-edge methods could generate an ever more reliable NO model in vitro for deciphering the codes of human CNS development, disease progression, and translational application. In this review, we

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Developing Human Pluripotent Stem Cell-Based Cerebral Organoids with a Controllable Microglia Ratio for Modeling Brain Development and Pathology

Cited by 11 publications

References 78 publications

Type I Interferon Signaling Drives Microglial Dysfunction and Senescence in Human iPSC Models of Down Syndrome and Alzheimer’s Disease

Type I Interferon Signaling Drives Microglial Dysfunction and Senescence in Human iPSC Models of Down Syndrome and Alzheimer’s Disease

Microglia innate immune response contributes to the antiviral defense and blood–CSF barrier function in human choroid plexus organoids during HSV‐1 infection

Toward the next generation of vascularized human neural organoids

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