Abstract:Up-regulation of Mirta22/Emc10 is a major transcriptional effect of the 22q11.2-associated microRNA dysregulation and underlies key cellular as well as behavioral deficits. EMC10 is a component of the ER membrane complex, which promotes membrane insertion of a subset of polytopic and tail-anchored membrane proteins. Here we show that EMC10 expression is elevated in hiPSC-derived neurons from 22q11.2 deletion carriers and that reduction of EMC10 levels restores defects in neurite outgrowth and calcium signaling… Show more
“…We also tested whether there are alterations in neuronal activity in patient compared to control organoids using as a proxy system-wide glutamate-induced Ca 2+ transients. Comparisons of three pairs at day 259 identified a significant decrease in Ca 2+ peak amplitude in case compared to control cells ( P = 5.78×10 -12 , average number of cells per organoid: n=89 patient, n=124 control, Fig 1c,d,e ) consistent with previous findings in monolayer cultures of both mouse and patient neurons 12,13,14 . Thus, despite the lack of overt effects in growth, patient organoids display different functional properties, validating their usefulness as a preclinical model for studying the disease mechanisms underlying 22q11.2 deletions.…”
Section: Resultssupporting
confidence: 89%
“…We first examined the dysregulation of microRNAs (miRNAs) as a potential mechanism for the asynchronous expression of maturation-associated genes. We have previously shown that 22q11.2 deletion results in brain-enriched miRNA downregulation and an upregulation in target gene expression due to (i) hemizygosity of DGCR8 27,28 , a component of the “microprocessor” complex that is essential for miRNA production and (ii) hemizygosity of miRNA genes residing within the deletion 12,28, 29 . To assess the contribution of miRNA dysregulation to our observed cellular and RNA-seq phenotypes we performed bulk miRNA sequencing on suspension from 3 pairs of 8-10 DIV70 organoids.…”
Section: Resultsmentioning
confidence: 99%
“…We identified 3 main patterns of trajectory divergence between patients and controls (Fig 6d ). First, genes that are upregulated in patient progenitors and remain upregulated over time as exemplified by EMC10 (a regulator of membrane protein trafficking whose upregulation impedes the acquisition of full neuronal functionality in patient neurons and animal models 12,29,31 ) and PEG3 (a cell cycle regulator that is typically highly expressed in progenitor cells but not in differentiated neurons 32 ). Second, genes that show upregulation in progenitors but the changes dissipate over time as exemplified by TCF3 (a WNT signaling antagonist that increases neural progenitor cell self-renewal and represses neuronal differentiation 33,34 ).…”
Section: Resultsmentioning
confidence: 99%
“…Another noteworthy target is the EMC10 gene, which shows altered temporal expression trajectory along the entire EN lineage at DIV70 and significant upregulation in patient RG ( P = 1.56×10 - 6 ), IPC-nEN ( P = 5.3×10 - 5 ) and EN ( P = 7.3×10 - 6 ) but not in astrocytes, choroid or interneuron lineage cells ( Supplemental Table S2 ). Indeed effective knockdown to near WT levels of EMC10 in patient iPSC-derived neurons leads to restoration of key morphological and functional EN alterations emerging due to 22q11.2 deletions 12 . Furthermore, normalization of Emc10 levels in either developing or adult mouse brain rescues several 22q11.2 deletion associated behavioral deficits 12, 31 , establishing a clear link between miRNA regulatory mechanisms and aspects of the behavioral manifestations observed in 22q11.2 deletion carriers.…”
Section: Discussionmentioning
confidence: 99%
“…Indeed effective knockdown to near WT levels of EMC10 in patient iPSC-derived neurons leads to restoration of key morphological and functional EN alterations emerging due to 22q11.2 deletions 12 . Furthermore, normalization of Emc10 levels in either developing or adult mouse brain rescues several 22q11.2 deletion associated behavioral deficits 12, 31 , establishing a clear link between miRNA regulatory mechanisms and aspects of the behavioral manifestations observed in 22q11.2 deletion carriers.…”
Adults and children afflicted with the 22q11.2 deletion syndrome (22q11.2DS) exhibit cognitive, social, and emotional impairments, and are at significantly heightened risk for schizophrenia (SCZ). The impact of this deletion on early human brain development, however, has remained unclear. Here we harness organoid models of the developing human cerebral cortex, cultivated from subjects with 22q11.2DS and SCZ, as well as unaffected control samples, to identify cell-type-specific developmental abnormalities arising from this genomic lesion. Leveraging single-cell RNA-sequencing in conjunction with experimental validation, we find that the loss of genes within the 22q11.2 locus leads to a delayed development of cortical neurons. This compromised development was reflected in an elevated proportion of actively proliferating neural progenitor cells, coupled with a decreased fraction of more mature neurons. Furthermore, we identify perturbed molecular imprints linked to neuronal maturation, observe the presence of sparser neurites, and note a blunted amplitude in glutamate-induced Ca2+ transients. The aberrant transcription program underlying impaired development contains molecular signatures significantly enriched in neuropsychiatric genetic liability. MicroRNA profiling and target gene investigation suggest that microRNA dysregulation may drive perturbations of genes governing the pace at which maturation unfolds. Using protein-protein interaction network analysis we define complementary effects stemming from additional genes residing within the deleted locus. Our study uncovers reproducible neurodevelopmental and molecular alterations due to 22q11.2 deletions. These findings have the potential to facilitate disease modeling and promote the pursuit of therapeutic interventions.
“…We also tested whether there are alterations in neuronal activity in patient compared to control organoids using as a proxy system-wide glutamate-induced Ca 2+ transients. Comparisons of three pairs at day 259 identified a significant decrease in Ca 2+ peak amplitude in case compared to control cells ( P = 5.78×10 -12 , average number of cells per organoid: n=89 patient, n=124 control, Fig 1c,d,e ) consistent with previous findings in monolayer cultures of both mouse and patient neurons 12,13,14 . Thus, despite the lack of overt effects in growth, patient organoids display different functional properties, validating their usefulness as a preclinical model for studying the disease mechanisms underlying 22q11.2 deletions.…”
Section: Resultssupporting
confidence: 89%
“…We first examined the dysregulation of microRNAs (miRNAs) as a potential mechanism for the asynchronous expression of maturation-associated genes. We have previously shown that 22q11.2 deletion results in brain-enriched miRNA downregulation and an upregulation in target gene expression due to (i) hemizygosity of DGCR8 27,28 , a component of the “microprocessor” complex that is essential for miRNA production and (ii) hemizygosity of miRNA genes residing within the deletion 12,28, 29 . To assess the contribution of miRNA dysregulation to our observed cellular and RNA-seq phenotypes we performed bulk miRNA sequencing on suspension from 3 pairs of 8-10 DIV70 organoids.…”
Section: Resultsmentioning
confidence: 99%
“…We identified 3 main patterns of trajectory divergence between patients and controls (Fig 6d ). First, genes that are upregulated in patient progenitors and remain upregulated over time as exemplified by EMC10 (a regulator of membrane protein trafficking whose upregulation impedes the acquisition of full neuronal functionality in patient neurons and animal models 12,29,31 ) and PEG3 (a cell cycle regulator that is typically highly expressed in progenitor cells but not in differentiated neurons 32 ). Second, genes that show upregulation in progenitors but the changes dissipate over time as exemplified by TCF3 (a WNT signaling antagonist that increases neural progenitor cell self-renewal and represses neuronal differentiation 33,34 ).…”
Section: Resultsmentioning
confidence: 99%
“…Another noteworthy target is the EMC10 gene, which shows altered temporal expression trajectory along the entire EN lineage at DIV70 and significant upregulation in patient RG ( P = 1.56×10 - 6 ), IPC-nEN ( P = 5.3×10 - 5 ) and EN ( P = 7.3×10 - 6 ) but not in astrocytes, choroid or interneuron lineage cells ( Supplemental Table S2 ). Indeed effective knockdown to near WT levels of EMC10 in patient iPSC-derived neurons leads to restoration of key morphological and functional EN alterations emerging due to 22q11.2 deletions 12 . Furthermore, normalization of Emc10 levels in either developing or adult mouse brain rescues several 22q11.2 deletion associated behavioral deficits 12, 31 , establishing a clear link between miRNA regulatory mechanisms and aspects of the behavioral manifestations observed in 22q11.2 deletion carriers.…”
Section: Discussionmentioning
confidence: 99%
“…Indeed effective knockdown to near WT levels of EMC10 in patient iPSC-derived neurons leads to restoration of key morphological and functional EN alterations emerging due to 22q11.2 deletions 12 . Furthermore, normalization of Emc10 levels in either developing or adult mouse brain rescues several 22q11.2 deletion associated behavioral deficits 12, 31 , establishing a clear link between miRNA regulatory mechanisms and aspects of the behavioral manifestations observed in 22q11.2 deletion carriers.…”
Adults and children afflicted with the 22q11.2 deletion syndrome (22q11.2DS) exhibit cognitive, social, and emotional impairments, and are at significantly heightened risk for schizophrenia (SCZ). The impact of this deletion on early human brain development, however, has remained unclear. Here we harness organoid models of the developing human cerebral cortex, cultivated from subjects with 22q11.2DS and SCZ, as well as unaffected control samples, to identify cell-type-specific developmental abnormalities arising from this genomic lesion. Leveraging single-cell RNA-sequencing in conjunction with experimental validation, we find that the loss of genes within the 22q11.2 locus leads to a delayed development of cortical neurons. This compromised development was reflected in an elevated proportion of actively proliferating neural progenitor cells, coupled with a decreased fraction of more mature neurons. Furthermore, we identify perturbed molecular imprints linked to neuronal maturation, observe the presence of sparser neurites, and note a blunted amplitude in glutamate-induced Ca2+ transients. The aberrant transcription program underlying impaired development contains molecular signatures significantly enriched in neuropsychiatric genetic liability. MicroRNA profiling and target gene investigation suggest that microRNA dysregulation may drive perturbations of genes governing the pace at which maturation unfolds. Using protein-protein interaction network analysis we define complementary effects stemming from additional genes residing within the deleted locus. Our study uncovers reproducible neurodevelopmental and molecular alterations due to 22q11.2 deletions. These findings have the potential to facilitate disease modeling and promote the pursuit of therapeutic interventions.
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