Modeling autism-associated SHANK3 deficiency using human cortico-striatal organoids generated from single neural rosettes

Abstract

Our understanding of the human brain is limited by the lack of experimental models to mechanistically probe the properties of brain cells at different developmental stages under normal and pathological conditions. We developed a new method for generating human cortico-striatal organoids from stem cell-derived single neural rosettes (SNRs) and used it to investigate cortico-striatal development and deficits caused by the deficiency of an autism- and intellectual disability-associated gene SHANK3. We show that SNR-derived organoids consist of different cortico-striatal cells, including pallial and subpallial progenitors, primary cortical and striatal neurons, interneurons, as well as macroglial and mural cells. We also demonstrate that neurons in SNR-derived organoids are predictably organized, functionally mature, and capable of establishing functional neural networks. Interestingly, we found that the cellular and electrophysiological deficits in SHANK3-deficient SNR-derived organoids are dependent on the level of SHANK3 expression and that organoids with complete hemizygous SHANK3 deletion have disrupted expression of several clustered protocadherins and multiple primate-specific zinc-finger genes. Together, this study describes a new method for using SNRs to generate organoids, provides new insights into the cell lineages associated with human cortico-striatal development, and identifies specific molecular pathways disrupted by hemizygous SHANK3 deletion, which is the most common genetic abnormality detected in patients with 22q13 deletion syndrome.

Neurite outgrowth deficits caused by rare PLXNB1 mutation in pediatric bipolar disorder

“…2). Patient and control telencephalic organoids were produced from iPSC-derived single neural rosettes (SNR; Fig 2A-B(46). We previously demonstrated that 1-month-old SNR-derived organoids consist of telencephalic neural progenitors and cortical excitatory and lateral ganglionic eminence-derived inhibitory neurons (46).…”

The Application of Brain Organoids in Assessing Neural Toxicity

“…Brain organoids typically contain dozens of neural tubes, and the number of neural tubes in each organoid is different, which leads to heterogeneity in the size of brain organoids. Therefore, generating brain organoids containing a single neural tube might ensure the production of uniform brain organoids (Wang et al, 2021). However, current methods are based on manual FIGURE 3 | Application of brain organoids in neurotoxicity assessment.…”

iPSC toolbox for understanding and repairing disrupted brain circuits in autism

“…The molecular mechanisms disrupted by PMDS-related hemizygous SHANK3 deletion were investigated in cortico-striatal organoids generated from stem cell-derived single neural rosettes (SNRs) [ 113 ]. This research confirmed the presence of synaptic and intrinsic excitability deficits in SHANK3-deficient organoids and identified impaired expression of several clustered protocadherins as potentially responsible.…”

“…One limitation of organoids for connectivity studies is the unpredictable organization of cells and germinal zones, which may create conflicting gradients of secreted morphogens, affecting neuronal migration and maturation [ 126 ]. To overcome this limitation, we developed a method for generating telencephalic organoids from SNRs [ 113 ]. Each SNR is a neural tube-like structure containing neuroepithelial cells radially organized around the lumen [ 127 ].…”

“…To a certain extent, this mimics neural tissue development from a singular neural tube. Indeed, NPs and neurons in SNR-derived organoids are predictably organized around the lumen [ 113 ]. However, the potential advantages for studying connectivity remain unknown.…”