Altered neuronal migratory trajectories in human cerebral organoids derived from individuals with neuronal heterotopia

Klaus, Johannes; Kanton, Sabina; Kyrousi, Christina; Ayo-Martin, Ane Cristina; Giaimo, Rossella Di; Riesenberg, Stephan; O'Neill, Adam C; Camp, J. Gray; Tocco, Chiara; Santel, Malgorzata; Rusha, Ejona; Drukker, Micha; Schroêder, M.; Götz, Magdalena; Robertson, Stephen; Treutlein, Barbara; Cappello, Silvia

doi:10.1038/s41591-019-0371-0

Cited by 143 publications

(159 citation statements)

References 37 publications

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“…Single cell RNA-seq can identify discrete cell types in cultured cells and in complex tissues, based on each individual cell’s transcriptome. The ability to interrogate the transcriptomes of large numbers of individual cells has led to the discovery of previously unrecognized cell types [8, 9] and has detected changes in cellular diversity in response to genetic or environmental perturbation[37-39]. Recovering individual cells from skeletal tissue is more challenging than from cultured cells or from other tissues whose cells can be easily separated by brief digestion and/or physical disruption.…”

Section: Discussionmentioning

confidence: 99%

Single cell RNA sequencing of calvarial and long bone endocortical cells

Ayturk

Scollan

Vesprey

et al. 2019

Preprint

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Single cell RNA-seq (scRNA-seq) is emerging as a powerful technology to examine transcriptomes of individual cells. We determined whether scRNA-seq could be used to detect the effect of environmental and pharmacologic perturbations on osteoblasts. We began with a commonly used in vitro system in which freshly isolated neonatal mouse calvarial cells are expanded and induced to produce a mineralized matrix. We used scRNA-seq to compare the relative cell type abundances and the transcriptomes of freshly isolated cells to those that had been cultured for 12 days in vitro. We observed that the percentage of macrophage-like cells increased from 6% in freshly isolated calvarial cells to 34% in cultured cells. We also found that Bglap transcripts were abundant in freshly isolated osteoblasts but nearly undetectable in the cultured calvarial cells. Thus, scRNA-seq revealed significant differences between heterogeneity of cells in vivo and in vitro. We next performed scRNA-seq on freshly recovered long bone endocortical cells from mice that received either vehicle or Sclerostin-neutralizing antibody for 1 week. Bone anabolism-associated transcripts were also not significantly increased in immature and mature osteoblasts recovered from Sclerostin-neutralizing antibody treated mice; this is likely a consequence of being underpowered to detect modest changes in gene expression, since only 7% of the sequenced endocortical cells were osteoblasts, and a limited portion of their transcriptomes were sampled. We conclude that scRNA-seq can detect changes in cell abundance, identity, and gene expression in skeletally derived cells. In order to detect modest changes in osteoblast gene expression at the single cell level in the appendicular skeleton, larger numbers of osteoblasts from endocortical bone are required.

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Section: Discussionmentioning

confidence: 99%

Single cell RNA sequencing of calvarial and long bone endocortical cells

Ayturk

Scollan

Vesprey

et al. 2019

Preprint

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“…Similarly, by preserving the 3D structure and by using markers to track cells within the organoids, live imaging techniques can be used to follow on tissue and directly on live cells to study diverse dynamic cell behaviors. For example, cell division of the progenitor cells or cell migration can be studied in a human‐specific system and help unravel the mechanisms controlling these behaviors in humans(Bershteyn et al, ; Klaus et al, ; Lancaster et al, ; Xiang et al, ) (Figure c). Since brain organoids are a key human‐specific brain model, importance should be given to the investigation of the maturation process, the functional aspects and physiological properties of the human neurons when found in a 3D structure.…”

Section: How Can We Use Brain Organoids To Study Human Neurodevelopmentmentioning

confidence: 99%

“…Another important advantage of this method is the possibility of tracking the electroporated cells with a fluorescent marker (Bershteyn et al, 2017;Klaus et al, 2019;O'Neill, Kyrousi, Einsiedler, et al, 2018;O'Neill, Kyrousi, Klaus, et al, 2018;Lancaster et al, 2013;Renner et al, 2017). The electroporation of cerebral organoids is also very useful for live imaging of the cells as discussed in the next paragraph (Bershteyn et al, 2017;Klaus et al, 2019;Lancaster et al, 2017;Xiang et al, 2017). Through electroporation, gene regulation can be performed using CRISPR-Cas9-based genome engineering constructs (Cárdenas et al, 2018).…”

Section: Acute Genome Alterationsmentioning

confidence: 99%

“…In order to overcome these difficulties, some scientists have turned to brain organoids to model brain disorders. There is a big effort being made to model neurodevelopmental disorders such as microcephaly (Gabriel et al, 2016;Lancaster et al, 2013), macrocephaly (Li, Muffat, et al, 2017), lissencephaly (Bershteyn et al, 2017;Iefremova et al, 2017;Li, Muffat, et al, 2017), periventricular heterotopia (Klaus et al, 2019;O'Neill, Kyrousi, Einsiedler, et al, 2018;O'Neill, Kyrousi, Klaus, et al, 2018) and autism spectrum disorder (Mariani et al, 2015). Other types of diseases, which were already modeled using brain organoids are genetic, mental and neurodegenerative disorders such as schizophrenia (Ye et al, 2017), Sandhoff disease (Allende et al, 2018), Alzheimer (Raja et al, 2016), frontotemporal dementia (Seo et al, 2017) and Rett-syndrome (Mellios et al, 2018).…”

Section: Diseasementioning

confidence: 99%

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Using brain organoids to study human neurodevelopment, evolution and disease

Kyrousi

Cappello

2019

WIREs Developmental Biology

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The brain is one of the most complex organs, responsible for the advanced intellectual and cognitive ability of humans. Although primates are to some extent capable of performing cognitive tasks, their abilities are less evolved. One of the reasons for this is the vast differences in the brain of humans compared to other mammals, in terms of shape, size and complexity. Such differences make the study of human brain development fascinating. Interestingly, the cerebral cortex is by far the most complex brain region resulting from its selective evolution within mammals over millions of years. Unraveling the molecular and cellular mechanisms regulating brain development, as well as the evolutionary differences seen across species and the need to understand human brain disorders, are some of the reasons why scientists are interested in improving their current knowledge on human corticogenesis. Toward this end, several animal models including primates have been used, however, these models are limited in their extent to recapitulate human‐specific features. Recent technological achievements in the field of stem cell research, which have enabled the generation of human models of corticogenesis, called brain or cerebral organoids, are of great importance. This review focuses on the main cellular and molecular features of human corticogenesis and the use of brain organoids to study it. We will discuss the key differences between cortical development in human and nonhuman mammals, the technological applications of brain organoids and the different aspects of cortical development in normal and pathological conditions, which can be modeled using brain organoids. This article is categorized under: Comparative Development and Evolution > Regulation of Organ Diversity Nervous System Development > Vertebrates: General Principles

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“…However, this challenge has motivated researchers to develop in vitro culturing technologies that generate functional and physiologically relevant 3D human brain organoids from pluripotent stem cells as an important first step . While brain organoids have demonstrated their ability to recapitulate neurogenesis, migration, positioning and layering of cortices, they still fail to meet certain critical aspects of in vivo, especially when 3D brain organoids are used as models of brain development. For example, organoids have limited growth potential due to inadequate oxygen and nutrient supply, leading to necrosis at the inner core of the organoid.…”

Section: Introductionmentioning

confidence: 99%

The Emergence of Stem Cell‐Based Brain Organoids: Trends and Challenges

Gopalakrishnan

2019

BioEssays

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Recent developments in 3D cultures exploiting the self‐organization ability of pluripotent stem cells have enabled the generation of powerful in vitro systems termed brain organoids. These 3D tissues recapitulate many aspects of human brain development and disorders occurring in vivo. When combined with improved differentiation methods, these in vitro systems allow the generation of more complex “assembloids,” which are able to reveal cell diversities, microcircuits, and cell–cell interactions within their 3D organization. Here, the ways in which human brain organoids have contributed to demystifying the complexities of brain development and modeling of developmental disorders is reviewed and discussed. Furthermore, challenging questions that are yet to be addressed by emerging brain organoid research are discussed.

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Altered neuronal migratory trajectories in human cerebral organoids derived from individuals with neuronal heterotopia

Cited by 143 publications

References 37 publications

Single cell RNA sequencing of calvarial and long bone endocortical cells

Single cell RNA sequencing of calvarial and long bone endocortical cells

Using brain organoids to study human neurodevelopment, evolution and disease

The Emergence of Stem Cell‐Based Brain Organoids: Trends and Challenges

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