Characterization of human iPSC-derived astrocytes with potential for disease modeling and drug discovery

SNCA, the first gene associated with Parkinson’s disease, encodes the α-synuclein protein, the predominant component within pathological inclusions termed Lewy bodies. The presence of Lewy bodies is one of the classical hallmarks found in the brain of patients with Parkinson’s disease, and Lewy bodies have also been observed in patients with other synucleinopathies. However, the study of α-synuclein pathology in cells has relied largely on two-dimensional culture models, which typically lack the cellular diversity and complex spatial environment found in the brain. Here, to address this gap, we use 3D midbrain organoids, differentiated from human induced pluripotent stem cells derived from patients carrying a triplication of the SNCA gene and from CRISPR/Cas9 corrected isogenic control iPSCs. These human midbrain organoids recapitulate key features of α-synuclein pathology observed in the brains of patients with synucleinopathies. In particular, we find that SNCA triplication human midbrain organoids express elevated levels of α-synuclein and exhibit an age-dependent increase in α-synuclein aggregation, manifested by the presence of both oligomeric and phosphorylated forms of α-synuclein. These phosphorylated α-synuclein aggregates were found in both neurons and glial cells and their time-dependent accumulation correlated with a selective reduction in dopaminergic neuron numbers. Thus, human midbrain organoids from patients carrying SNCA gene multiplication can reliably model key pathological features of Parkinson’s disease and provide a powerful system to study the pathogenesis of synucleinopathies.

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“…1G ). Further double staining for GFAP and S100β 53 confirmed the presence of astrocytes in the hMOs ( Supplementary Fig. 2D ).…”

Section: Resultsmentioning

confidence: 55%

Midbrain organoids with anSNCAgene triplication model key features of synucleinopathy

Mohamed

Sirois

Ramamurthy

et al. 2021

Brain Communications

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“…The NF‐κB pathway plays an essential role in MES transition and acquired tumor treatment resistance in GBM. [ 49–51 ] Interestingly, a significant increase in mRNA expression of NF‐κB target genes (TNFAIP3, ICAM1) was detected 24 h after post‐treatment with MES EVs in different PN GSC lines (PN157, MGG6, MGG8, BT07; Figure S2B, Supporting Information), and this increase appears to be dose‐dependent, with the most significant effect after addition of 2–4 × 10 10 EVs mL −1 (Figure S2C,D, Supporting Information). These results were confirmed by an increased expression of NF‐κB reporter luciferase activity over time in response to EVs from different MES donor GSCs (MES83, MES326) and in different PN recipient GSCs (PN157, BT07, MGG8; Figure 4F and Figure S2E,F, Supporting Information).…”

Section: Resultsmentioning

confidence: 99%

Extracellular Vesicles Induce Mesenchymal Transition and Therapeutic Resistance in Glioblastomas through NF‐κB/STAT3 Signaling

Schweiger

Giovanazzi

et al. 2020

Adv. Biosys.

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Gliomas account for about 80% of all malignant CNS tumors, of which glioblastoma (GBM) is the most common and malignant form. [1] Despite a multimodal treatment approach including surgery, radiation, and temozolomide (TMZ) chemotherapy, the prognosis remains dismal. The Central Brain Tumor Registry reports that the five-year survival rate of adult patients with GBM is 4.3%. Although tumors may initially respond to this treatment armamentarium, recurrence remains inevitable. [2-5] Large-scale genomic, epigenomic, and transcriptomic characterization of different subtypes has provided valuable insights into the heterogeneous landscape of GBM. [6-9] Notably, comprehensive longitudinal Glioblastoma (GBM) is the most common primary malignant brain tumor and despite optimal treatment, long-term survival remains uncommon. GBM can be roughly divided into three different molecular subtypes, each varying in aggressiveness and treatment resistance. Recent evidence shows plasticity between these subtypes in which the proneural (PN) glioma stemlike cells (GSCs) undergo transition into the more aggressive mesenchymal (MES) subtype, leading to therapeutic resistance. Extracellular vesicles (EVs) are membranous structures secreted by nearly every cell and are shown to play a key role in GBM progression by acting as multifunctional signaling complexes. Here, it is shown that EVs derived from MES cells educate PN cells to increase stemness, invasiveness, cell proliferation, migration potential, aggressiveness, and therapeutic resistance by inducing mesenchymal transition through nuclear factor-κB/signal transducer and activator of transcription 3 signaling. The findings could potentially help explore new treatment strategies for GBM and indicate that EVs may also play a role in mesenchymal transition of different tumor types.

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“…Although only exogenous A53T α-syn gave statistically significant neuroinhibition with rat primary astrocytes, human astrocytes did show potent activation and upregulation of the secretory pathway with WT α-syn, so we cannot exclude the possibility that human astrocytes may respond more potently than rat astrocytes to the human α-syn protein in terms of neuroinhibition. Future studies could overcome these limitations by utilizing differentiated human induced pluripotent stem cell (iPSC) derived cell types [ 51 ].…”

Section: Discussionmentioning

confidence: 99%

Extracellular Alpha-Synuclein Promotes a Neuroinhibitory Secretory Phenotype in Astrocytes

Vieira

Radford

Hayashi

et al. 2020

Life

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Multiple system atrophy (MSA) and dementia with Lewy bodies (DLB) are α-synucleinopathies that exhibit widespread astrogliosis as a component of the neuroinflammatory response. Munc18, a protein critical to vesicle exocytosis, was previously found to strongly mark morphologically activated astrocytes in brain tissue of MSA patients. Immunofluorescence of MSA, DLB and normal brain tissue sections was combined with cell culture and co-culture experiments to investigate the relationship between extracellular α-synuclein and the transition to a secretory astrocyte phenotype. Increased Munc18-positive vesicles were resolved in activated astrocytes in MSA and DLB tissue compared to controls, and they were also significantly upregulated in the human 1321N1 astrocytoma cell line upon treatment with α-synuclein, with parallel increases in GFAP expression and IL-6 secretion. In co-culture experiments, rat primary astrocytes pretreated with α-synuclein inhibited the growth of neurites of co-cultured primary rat neurons and upregulated chondroitin sulphate proteoglycan. Taken together, these results indicate that the secretory machinery is significantly upregulated in the astrocyte response to extracellular α-synuclein and may participate in the release of neuroinhibitory and proinflammatory factors in α-synucleinopathies.

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Characterization of human iPSC-derived astrocytes with potential for disease modeling and drug discovery

Cited by 10 publications

References 40 publications

Midbrain organoids with anSNCAgene triplication model key features of synucleinopathy

Midbrain organoids with anSNCAgene triplication model key features of synucleinopathy

Extracellular Vesicles Induce Mesenchymal Transition and Therapeutic Resistance in Glioblastomas through NF‐κB/STAT3 Signaling

Extracellular Alpha-Synuclein Promotes a Neuroinhibitory Secretory Phenotype in Astrocytes

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