cortical organoids podcasts

Cross-site reproducibility of human cortical organoids reveals consistent cell type composition and architecture

Play Episode Listen Later Jul 29, 2023

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.07.28.550873v1?rss=1 Authors: Glass, M. R., Waxman, E. A., Yamashita, S., Lafferty, M., Beltran, A., Farah, T., Patel, N. K., Matoba, N., Ahmed, S., Srivastava, M., Drake, E., Davis, L. T., Yeturi, M., Sun, K., Love, M. I., Hashimoto-Torii, K., French, D. L., Stein, J. L. Abstract: Background: Reproducibility of hCO phenotypes remains a concern for modeling neurodevelopmental disorders. While guided human cortical organoid (hCO) protocols reproducibly generate cortical cell types in multiple cell lines at one site, variability across sites using a harmonized protocol has not yet been evaluated. We present an hCO cross-site reproducibility study examining multiple phenotypes. Methods: Three independent research groups generated hCOs from one induced pluripotent stem cell (iPSC) line using a harmonized miniaturized spinning bioreactor protocol. scRNA-seq, 3D fluorescent imaging, phase contrast imaging, qPCR, and flow cytometry were used to characterize the 3 month differentiations across sites. Results: In all sites, hCOs were mostly cortical progenitor and neuronal cell types in reproducible proportions with moderate to high fidelity to the in vivo brain that were consistently organized in cortical wall-like buds. Cross-site differences were detected in hCO size and morphology. Differential gene expression showed differences in metabolism and cellular stress across sites. Although iPSC culture conditions were consistent and iPSCs remained undifferentiated, primed stem cell marker expression prior to differentiation correlated with cell type proportions in hCOs. Conclusions: We identified hCO phenotypes that are reproducible across sites using a harmonized differentiation protocol. Previously described limitations of hCO models were also reproduced including off-target differentiations, necrotic cores, and cellular stress. Improving our understanding of how stem cell states influence early hCO cell types may increase reliability of hCO differentiations. Cross-site reproducibility of hCO cell type proportions and organization lays the foundation for future collaborative prospective meta-analytic studies modeling neurodevelopmental disorders in hCOs. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

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Improved Protocol for Reproducible Human Cortical Organoids Reveals Early Alterations in Metabolism with MAPT Mutations

PaperPlayer biorxiv neuroscience

Play Episode Listen Later Jul 11, 2023

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.07.11.548571v1?rss=1 Authors: Bertucci, T., Bowles, K. R., Lotz, S., Qi, L., Stevens, K., Goderie, S. K., Borden, S., Oja, L., Lane, K., Lotz, R., Lotz, H., Chowdhury, R., Joy, S., Arduini, B. L., Butler, D. C., Miller, M., Baron, H., Sandhof, C. A., Silva, M. C., Haggarty, S. J., Karch, C. M., Geschwind, D. H., Goate, A. M., Temple, S. Abstract: Cerebral cortical-enriched organoids derived from human pluripotent stem cells (hPSCs) are valuable models for studying neurodevelopment, disease mechanisms, and therapeutic development. However, recognized limitations include the high variability of organoids across hPSC donor lines and experimental replicates. We report a 96-slitwell method for efficient, scalable, reproducible cortical organoid production. When hPSCs were cultured with controlled-release FGF2 and an SB431542 concentration appropriate for their TGFBR1/ALK5 expression level, organoid cortical patterning and reproducibility were significantly improved. Well-patterned organoids included 16 neuronal and glial subtypes by single cell RNA sequencing (scRNA-seq), frequent neural progenitor rosettes and robust BCL11B+ and TBR1+ deep layer cortical neurons at 2 months by immunohistochemistry. In contrast, poorly-patterned organoids contain mesendoderm-related cells, identifiable by negative QC markers including COL1A2. Using this improved protocol, we demonstrate increased sensitivity to study the impact of different MAPT mutations from patients with frontotemporal dementia (FTD), revealing early changes in key metabolic pathways. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

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Cell specificity of adeno-associated virus (AAV) serotypes in human cortical organoids

PaperPlayer biorxiv neuroscience

Play Episode Listen Later Apr 13, 2023

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.04.13.536491v1?rss=1 Authors: Stanton, M. M., Hariani, H. N., Sorokin, J., Taylor, P. M., Modan, S., Rash, B. G., Rao, S. B., Enriquez, L., Quang, D., Hsu, P.-K., Paek, J., Owango, D., Castrillo, C., Nicola, J., Ramkumar, P., Lash, A., Flanzer, D., Shah, K., Kato, S., Skibinski, G. Abstract: Human-derived cortical organoids (hCOs) recapitulate cell diversity and 3D structure found in the human brain and offer a promising model for discovery of new gene therapies targeting neurological disorders. Adeno-associated viruses (AAVs) are the most promising vehicles for non-invasive gene delivery to the central nervous system (CNS), but reliable and reproducible in vitro models to assess their clinical potential are lacking. hCOs can take on these issues as they are a physiologically relevant model to assess AAV transduction efficiency, cellular tropism, and biodistribution within the tissue parenchyma, all of which could significantly modulate therapeutic efficacy. Here, we examine a variety of naturally occurring AAV serotypes and measure their ability to transduce neurons and glia in hCOs from multiple donors. We demonstrate cell tropism driven by AAV serotype and hCO donor and quantify fractions of neurons and astrocytes transduced with GFP as well as overall hCO health. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

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Human Cortical Organoids

Daily Discoveries

Play Episode Listen Later Feb 16, 2023 21:09

This week Sanjana, Oskar, and Hayden discuss a study where an imaging technology was created in order to observe a mouse brain implant over time without disturbing the mouse or the implant.

oskar cortical organoids

Cell-type specific developmental defects in PTEN-mutant cortical organoids converge on abnormal circuit activity

PaperPlayer biorxiv neuroscience

Play Episode Listen Later Nov 16, 2022

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2022.11.15.516664v1?rss=1 Authors: Pigoni, M., Uzquiano, A., Paulsen, B., Kedaigle, A. J., Yang, S. M., Symvoulidis, P., Adiconis, X., Velasco, S., Sartore, R., Kim, K., Tucewicz, A., Tsafou, K., Jin, X., Barrett, L., Chen, F., Boyden, E., Regev, A., Levin, J. Z., Arlotta, P. Abstract: De novo heterozygous loss-of-function mutations in PTEN are strongly associated with Autism spectrum disorders (ASD); however, it is unclear how heterozygous mutations in this gene affects 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. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

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Methadone alters transcriptional programs associated with synapse formation in human cortical organoids

PaperPlayer biorxiv neuroscience

Play Episode Listen Later Nov 5, 2022

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2022.11.04.515240v1?rss=1 Authors: Dwivedi, I., Zhou, D., Caldwell, A. B., Subramaniam, S., Haddad, G. G. Abstract: Opioid use disorder (OUD) among pregnant women has become an epidemic in the United States. Pharmacological interventions for OUD involve methadone, a synthetic opioid analgesic that attenuates withdrawal symptoms and behaviors linked with maternal drug addiction. However, methadone's ability to readily accumulate in neural tissue, and cause long-term neurocognitive sequelae, has led to concerns regarding its effect on prenatal brain development. We took advantage of human cortical organoid (hCO) technology to probe how this drug impacts the earliest mechanisms giving rise to the cerebral cortex. To this end, we conducted bulk mRNA sequencing of 2-month-old hCOs derived from two cell lines that were chronically treated with a clinically relevant dose of 1M methadone for 50 days. Differential expression and gene ontology analyses revealed a robust transcriptional response to methadone associated with functional components of the synapse, the underlying extracellular matrix (ECM), and cilia. Further unsupervised co-expression network and predictive protein-protein interaction analyses demonstrated that these changes occurred in concert, centered around a regulatory axis consisting of growth factors, developmental signaling pathways, and matricellular proteins. Our results demonstrate that exposure to methadone during early cortico-genesis fundamentally alters transcriptional programs associated with synapse formation, and that these changes arise by modulating extra-synaptic molecular mechanisms in the ECM and cilia. These findings provide novel insight into methadone's putative effect on cognitive and behavioral development and a basis for improving interventions for maternal opioid addiction. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

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Human cortical organoids forge functional circuits in rat brains

Spectrum Autism Research

Play Episode Listen Later Oct 26, 2022 5:32

The transplanted cells integrate into living animals' neural circuitry and influence behavior. The post Human cortical organoids forge functional circuits in rat brains appeared first on Spectrum | Autism Research News.

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Human cortical organoids forge functional circuits in rat brains

Spectrum Autism Research

Play Episode Listen Later Oct 12, 2022 5:32

The transplanted cells integrate into living animals' neural circuitry and influence behavior.

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Self-Organizing 3D Human Choroid Plexus-Ventricle-Cortical Organoids

PaperPlayer biorxiv neuroscience

Play Episode Listen Later Oct 1, 2020

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.09.30.321554v1?rss=1 Authors: Shaker, M. R., Cooper-White, J., Wolvetang, E. J. Abstract: Both the choroid plexus (CP) and the cortex are derived from the rostral neural tube during early embryonic development. In addition to producing CSF, the CP secretes essential factors that orchestrate cortical development and later neurogenesis. Previous brain modeling efforts with human pluripotent stem cells (hPSCs) generated either cortical or CP tissues in 3D culture. Here, we used hPSC-derived neuroectodermal cells, the building blocks of the anterior body, to simultaneously generate CP that forms ventricles and cortical cells in organoids (CVCOs), which can be maintained as 3D organoid cultures. Large scale culture revealed reproducibility of the protocol independent of cell lines, clones or batches. CVCOs contain mature and functional CP that projects multiple cilia into the ventricle-like fluid filled cysts and is in direct contact with appropriately patterned cortical cells. CVCOs thus recapitulate key features of developing forebrain structures observed in in vivo and constitute a useful for dissecting the role of CP in human forebrain development in health and disease. Copy rights belong to original authors. Visit the link for more info

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Cortical Organoids Model Early Brain Development Disrupted by 16p11.2 Copy Number Variants in Autism

PaperPlayer biorxiv neuroscience

Play Episode Listen Later Jun 27, 2020

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.06.25.172262v1?rss=1 Authors: Urresti, J., Zhang, P., Moran-Losada, P., Yu, N.-K., Negraes, P. D., Trujillo, C. A., Antaki, D., Amar, M., Chau, K., Pramod, A. B., Diedrich, J., Tejwani, L., Romero, S., Sebat, J., Yates, J. R., Muotri, A. R., Iakoucheva, L. M. Abstract: Reciprocal deletion and duplication of 16p11.2 is the most common copy number variation (CNV) associated with Autism Spectrum Disorders, and has significant effect on brain size. We generated cortical organoids to investigate neurodevelopmental pathways dysregulated by dosage changes of 16p11.2 CNV. We show that organoids recapitulate patients' macrocephaly and microcephaly phenotypes. Deletions and duplications have "mirror" effects on cell proliferation, neuronal maturation and synapse number, consistent with "mirror" effects on brain development in humans. Excess neuron number along with depletion of neural progenitors in deletions, and "mirror" phenotypes in duplications, demonstrate dosage-dependent impact of 16p11.2 CNV on early neurogenesis. Transcriptomic and proteomic profiling revealed synaptic defects and neuron migration as key drivers of 16p11.2 functional effect. Treatment with the RhoA inhibitor Rhosin rescued neuron migration. We implicate upregulation of small GTPase RhoA as one of the pathways impacted by the 16p11.2 CNV. This study identifies pathways dysregulated by the 16p11.2 CNV during early neocortical development. Copy rights belong to original authors. Visit the link for more info