Granulin loss of function in human mature brain organoids implicates astrocytes in TDP-43 pathology

Majo, Martina de; Koontz, Mark; Marsan, Elise; Salinas, Nir; Ramsey, Marina; Kuo, Yien–Ming; Seo, Kyounghee; Li, Huinan; Dräger, Nina; Leng, Kun; Gonzales, Santiago L; Kurnellas, Michael; Miyaoka, Yuichiro; Klim, Joseph R.; Kampmann, Martin; Ward, Michael E.; Huang, Eric J.; Ullian, Erik M.

doi:10.1016/j.stemcr.2023.01.012

Cited by 21 publications

(12 citation statements)

References 60 publications

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Section: Discussionmentioning

confidence: 99%

“…The absence of overt MN loss may reflect the immaturity of MNs at the timepoints examined. In future studies, overcoming the challenges associated with long-term culture of MNs in monolayers (i.e., cell clumping and detachment) or use of three-dimensional culture models (i.e., spheroids, organoids) (Castellanos-Montiel et al, 2023; de Majo et al, 2023; Pereira et al, 2021) may enable neurodegeneration to be observed after several months of culture, without the need for exogenous treatments with stressors to elicit a phenotype. Alternatively, transcription factor-mediated transdifferentiation towards specific neurons, avoiding intermediate proliferative pluripotent stem cell stage, may help retaining specific age-related and epigenetic features involved in neurodegenerative disorders (Mollinari et al, 2018).…”

Section: Discussionmentioning

confidence: 99%

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Homozygous ALS-linked mutations in TARDBP/TDP-43 lead to hypoactivity and synaptic abnormalities in human iPSC-derived motor neurons

Lépine

Nauleau-Javaudin

Deneault

et al. 2023

Preprint

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Cytoplasmic mislocalization and aggregation of the RNA-binding protein TDP-43 is a pathological hallmark of the motor neuron (MN) disease amyotrophic lateral sclerosis (ALS). Furthermore, while mutations in the TARDBP gene (encoding TDP-43) have been associated with ALS, the pathogenic consequences of these mutations remain poorly understood. Using CRISPR/Cas9, we engineered two homozygous knock-in iPSC lines carrying mutations in TARDBP encoding TDP-43A382Tand TDP-43G348C, two common yet understudied ALS TDP-43 variants. MNs differentiated from knock-in iPSCs had normal viability and displayed no significant changes in TDP-43 subcellular localization, phosphorylation, solubility, or aggregation compared with isogenic control MNs. However, our results highlight synaptic impairments in both TDP-43A382Tand TDP-43G348CMN cultures, as reflected in synapse abnormalities and alterations in spontaneous neuronal activity. Collectively, our findings argue that MN dysfunction precedes the occurrence of TDP-43 pathology and neurodegeneration in ALS, and further implicates synaptic and excitability defects in the pathobiology of this disease.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Homozygous ALS-linked mutations in TARDBP/TDP-43 lead to hypoactivity and synaptic abnormalities in human iPSC-derived motor neurons

Lépine

Nauleau-Javaudin

Deneault

et al. 2023

Preprint

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“…These processes could also play a significant role in inhibiting proinflammatory pathways, including TNF-alpha production and the NF-κB inflammatory process [62,63]. Furthermore, granulin mutations may result in abnormal astrocyte and microglia-related mechanisms, such as their development and activation [64,65]. PSEN1 could also impact inflammatory pathways.…”

Section: Discussionmentioning

confidence: 99%

Patient with PSEN1 Glu318Gly and Other Possible Disease Risk Mutations, Diagnosed with Early Onset Alzheimer’s Disease

Yang,

Bagyinszky,

2023

IJMS

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In this manuscript, we introduced a French EOAD patient in Korea who carried the presenilin-1 (PSEN1) Glu318Gly mutations with four possible risk variants, including sortilin-related receptor 1 (SORL1) Glu270Lys, ATP-binding cassette subfamily A member 7 (ABCA7) Val1946Met, translocase of outer mitochondrial membrane 40 (TOMM40) Arg239Trp, and granulin (GRN) Ala505Gly. The patient started to present memory decline and behavioral dysfunction in his early 60s. His brain imaging presented amyloid deposits by positron emission tomography (PET-CT). The multimer detection system (MDS) screening test for plasma for amyloid oligomers was also positive, which supported the AD diagnosis. It was verified that PSEN1 Glu318Gly itself may not impact amyloid production. However, additional variants were found in other AD and non-AD risk genes, as follows: SORL1 Glu270Lys was suggested as a risk mutation for AD and could increase amyloid peptide production and impair endosome functions. ABCA7 Val1946Met was a novel variant that was predicted to be damaging. The GRN Ala505Gly was a variant with uncertain significance; however, it may reduce the granulin levels in the plasma of dementia patients. Pathway analysis revealed that PSEN1 Glu318Gly may work as a risk factor along with the SORL1 and ABCA7 variants since pathway analysis revealed that PSEN1 could directly interact with them through amyloid-related and lipid metabolism pathways. TOMM40 and PSEN1 could have common mechanisms through mitochondrial dysfunction. It may be possible that PSEN1 Glu318Gly and GRN Ala505Gly would impact disease by impairing immune-related pathways, including microglia and astrocyte development, or NFkB-related pathways. Taken together, the five risk factors may contribute to disease-related pathways, including amyloid and lipid metabolism, or impair immune mechanisms.

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“…Astrocytes were generated as described previously 16,61,62 . WTC11 iPSCs were cultured and expanded in six-well plates using Essential 8 media (Gibco/Thermo Fisher Scientific, Cat.…”

Section: Serum Free Protocolmentioning

confidence: 99%

CRISPRi-based screens in iAssembloids to elucidate neuron-glia interactions

Benitez

Boggess

et al. 2023

Preprint

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The sheer complexity of the brain has complicated our ability to understand its cellular mechanisms in health and disease. Genome-wide association studies have uncovered genetic variants associated with specific neurological phenotypes and diseases. In addition, single-cell transcriptomics have provided molecular descriptions of specific brain cell types and the changes they undergo during disease. Although these approaches provide a giant leap forward towards understanding how genetic variation can lead to functional changes in the brain, they do not establish molecular mechanisms. To address this need, we developed a 3D co-culture system termed iAssembloids (induced multi-lineage assembloids) that enables the rapid generation of homogenous neuron-glia spheroids. We characterize these iAssembloids with immunohistochemistry and single-cell transcriptomics and combine them with large-scale CRISPRi-based screens. In our first application, we ask how glial and neuronal cells interact to control neuronal death and survival. Our CRISPRi-based screens identified that GSK3β inhibits the protective NRF2-mediated oxidative stress response in the presence of reactive oxygen species elicited by high neuronal activity, which was not previously found in 2D monoculture neuron screens. We also apply the platform to investigate the role of APOE-ϵ4, a risk variant for Alzheimer's Disease, in its effect on neuronal survival. This platform expands the toolbox for the unbiased identification of mechanisms of cell-cell interactions in brain health and disease.

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Granulin loss of function in human mature brain organoids implicates astrocytes in TDP-43 pathology

Cited by 21 publications

References 60 publications

Homozygous ALS-linked mutations in TARDBP/TDP-43 lead to hypoactivity and synaptic abnormalities in human iPSC-derived motor neurons

Homozygous ALS-linked mutations in TARDBP/TDP-43 lead to hypoactivity and synaptic abnormalities in human iPSC-derived motor neurons

Patient with PSEN1 Glu318Gly and Other Possible Disease Risk Mutations, Diagnosed with Early Onset Alzheimer’s Disease

CRISPRi-based screens in iAssembloids to elucidate neuron-glia interactions

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