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Freude, K. K.

Publications and source records attributed to Freude, K. K..

8 recordsLinked to original sources

Protein-state dysregulation and sex-specific neurodevelopmental signatures in schizophrenia forebrain organoids

Schizophrenia is highly heritable, yet the molecular mechanisms linking genetic risk to abnormal human brain development remain poorly understood. To address this, we generated dorsal forebrain organoids from 17 individuals with idiopathic schizophrenia and 17 age- and sex-matched controls and profiled them across multiple molecular layers, including single-nucleus transcriptomics, quantitative proteomics, metabolomics and deep post-translational modification (PTM) analysis. The organoids reproducibly modelled early cortical development and showed largely similar cellular composition between schizophrenia and control groups. Surprisingly, transcriptomic differences were relatively limited, with the strongest cell-type-specific changes observed in Cajal-Retzius neurons. In contrast, proteomic and particularly PTM-level analyses revealed widespread molecular disruption affecting pathways involved in neuronal migration, neurite development, synaptic function, protein kinase signalling, extracellular matrix organisation and lipid metabolism. Many of the earliest disease-associated changes emerged at the level of protein phosphorylation, consistent with altered neuronal maturation and neurite dynamics. At later developmental stages, schizophrenia organoids showed reduced abundance of synaptic proteins, fewer synaptic puncta and evidence of dysregulated retinoic acid and YAP1 signalling. Notably, most disease-associated alterations occurred independently of changes in transcript or protein abundance, indicating that key aspects of schizophrenia biology are encoded in protein state rather than expression level. These findings identify sex-specific dysregulation of protein state as a major molecular feature of schizophrenia and demonstrate the value of multi-layer proteomic approaches for uncovering disease mechanisms missed by transcriptomics alone.

neuroscience↗

Dysregulation of miRNAs Drives Premature GABAergic Maturation and Early Neurodevelopmental Defects in Schizophrenia

BackgroundSchizophrenia (SCZ) is a severe neurodevelopmental disorder with numerous genetic risk loci. However, little is known about the molecular alterations that occur during brain development in SCZ, particularly regarding the role of microRNA (miRNA) mediated regulatory mechanisms. This gap in knowledge is largely due to the limited availability of developing human brain tissue. Patient-derived brain organoids offer a promising alternative model. Here we use 3D dorsal forebrain organoids (DFOs) to investigate miRNA dysregulation in SCZ. MethodsDFOs were generated from human induced pluripotent stem cells (hiPSCs) derived from six SCZ patients and five matched controls and cultured for 120 days. Multi-omics analyses, immunohistochemistry, and in situ hybridization were employed to characterize molecular and spatial features. ResultsDFOs recapitulated key molecular hallmarks of human cortical development. Nineteen miRNAs were differentially expressed in SCZ: nine associated with neural progenitor proliferation were downregulated and ten linked to neuronal differentiation and synaptic maturation were upregulated, reflecting a compressed developmental timeline. Among 77 dysregulated mRNAs, 55 were predicted miRNA targets. SCZ DFOs exhibited significant upregulation of GABAergic pathway genes accompanied by altered expression of their regulatory miRNAs, indicating premature GABAergic lineage specification. The disrupted miRNA-mRNA network converged on glutamatergic and dopaminergic development, synaptic organization, and extracellular matrix remodeling. ConclusionDysregulated miRNAs in SCZ DFOs disrupt neuronal differentiation, excitatory-inhibitory balance, and early circuit formation, implicating miRNA-mediated post-transcriptional regulation as a key mechanism linking molecular alterations to cellular and network-level deficits in SCZ.

cell biology↗

Sex-specific remodeling of the tRNA epitranscriptome in Alzheimers disease

tRNA modifications are critical regulators of RNA stability, decoding fidelity, and cellular stress adaptation, yet their contribution to human neurodegenerative disease remains poorly understood. Beyond their established functions in translational control, emerging evidence shows that RNA modifications influence neurogenesis, neurodevelopment, neuronal function, brain-cell differentiation, and cellular plasticity. Consequently, dysregulation of these molecular processes is increasingly recognized as a mechanistic contributor to neurodegenerative disorders. Alzheimers disease (AD), characterized by amyloid pathology, synaptic dysfunction, and progressive neuronal loss, has recently been linked to disturbances in RNA metabolism, suggesting that alterations in the epitranscriptome may represent an underexplored dimension of AD pathophysiology. Here, we systematically profiled the tRNA epitranscriptome across cellular and animal models of AD, as well as in human postmortem brain tissue from non-demented controls and AD patients, using liquid chromatography-tandem mass spectrometry (LC-MS/MS). This method enables highly sensitive quantification of RNA modifications, with limits of detection in the low femtomole range. Across our models, we identified a conserved yet sex-specific remodeling of the tRNA modification landscape in AD. Because therapeutic options and early diagnostic tools for AD remain limited, we leveraged these findings to develop a tRNA-centered RNA-modification score that integrates both nucleobase-specific modification patterns and neuropathological disease severity into a quantitative metric. Together, our findings identify the tRNA epitranscriptome as a unifying molecular sex-specific signature of AD, linking disease pathology and sex to impaired RNA metabolism. This line of research opens a new path toward establishing early biomarkers or diagnostic tools for AD. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=77 SRC="FIGDIR/small/715648v1_ufig1.gif" ALT="Figure 1"> View larger version (25K): org.highwire.dtl.DTLVardef@174c203org.highwire.dtl.DTLVardef@6d610aorg.highwire.dtl.DTLVardef@fddeecorg.highwire.dtl.DTLVardef@136700e_HPS_FORMAT_FIGEXP M_FIG C_FIG

neuroscience↗

NINJ1 is activated by Mycobacterium tuberculosis ESX-1 secreted effector EsxA and mediates necrosis of infected human macrophages

Lytic cell death contributes to tissue damage and dissemination during Mycobacterium tuberculosis (Mtb) infection, yet the mechanisms governing plasma membrane rupture (PMR) remain incompletely defined. Here, we identify Ninjurin-1 (NINJ1), a mediator of PMR, as a central effector of macrophage lysis during Mtb infection. Mtb infection induced NINJ1 oligomerization, and genetic deletion or pharmacological inhibition of NINJ1 markedly reduced lactate dehydrogenase (LDH) release. Cytokine secretion was largely preserved in NINJ1-deficient macrophages, with the exception of CXCL10. Unexpectedly, inhibition of pyroptosis, apoptosis, necroptosis, and ferroptosis - individually or in combination - did not prevent NINJ1 activation or PMR, indicating that Mtb-induced membrane rupture proceeds independently of canonical regulated cell death pathways. However, NINJ1 oligomerization and PMR required the Mtb ESX-1 secretion system, identifying a bacterial virulence determinant as a critical upstream trigger. Although osmoprotective PEG partially reduced LDH release, neither calcium signaling nor cell swelling accounted for NINJ1-dependent PMR in Mtb-infected macrophages. Together, these findings establish NINJ1 as a key executioner of Mtb-induced lytic cell death and reveal an ESX-1-dependent pathway of PMR that is uncoupled from canonical host cell death programs. TeaserNINJ1 mediates plasma membrane rupture of macrophages infected with Mycobacterium tuberculosis

microbiology↗

Inferring systemic metabolic and oxidative stress susceptibility in normal-tension glaucoma through targeted skin fibroblast analysis

AimEnhance the understanding of intrinsic metabolic and oxidative stress vulnerability in normal-tension glaucoma (NTG) pathophysiology by targeted profiling of skin fibroblasts from NTG and control donors. BackgroundNTG is a primary open-angle glaucoma subtype characterised by glaucomatous neurodegeneration without elevated intraocular pressure. Increasing evidence links systemic metabolic and oxidative stress vulnerability to NTG pathophysiology, making non-ocular, somatic cells promising surrogate systems for assessing neurodegenerative predisposing mechanisms. MethodsSkin fibroblast cultures were obtained from four female NTG and four age- and gender-matched control donors. Targeted metabolic profiling of mitochondrial function, glycolytic capacity, and glucose and amino acid metabolism was performed using the Seahorse assay, gas chromatography-mass spectrometry, and high-performance liquid chromatography. Oxidative stress resilience to hydrogen peroxide was assessed employing the lactate dehydrogenase release assay. ResultsPure skin fibroblast cultures were obtained for all included donors. NTG and control fibroblast exhibited similar mitochondrial and glycolytic function. No group difference was demonstrated in relative glucose metabolism and absolute amino acid profile. Control and NTG fibroblasts exhibited similar susceptibility to oxidative stress. ConclusionNTG skin fibroblasts exhibit similar mitochondrial and glycolytic function, glucose metabolisation, amino acid profile, and oxidative stress resiliency compared to controls. Future studies should focus on mapping cell- and tissue-specific differences through combined genetic and transcriptomic profiling to guide and stratify the functional assessment of different endotypes within the NTG disease spectrum. Cell-specific dysregulation and the need for individual mechanistic grouping diminish the applicability of skin fibroblasts as a model system for exploiting NTG pathophysiology.

neuroscience↗

Human iPSC-derived pericyte-like cells carrying APP Swedish mutation overproduce beta-amyloid and induce cerebral amyloid angiopathy-like changes

Alzheimers disease (AD) patients often exhibit cerebral amyloid angiopathy (CAA), i.e beta-amyloid (A{beta}) accumulation within cerebral blood vessels causing cerebrovascular dysfunction. Pericytes wrap around vascular capillaries, thus regulating cerebral blood flow, angiogenesis, and vessel stability. Vascular dysfunction can promote the development and progression of neurodegenerative diseases, yet the specific contribution of pericytes to AD pathology remains unclear. Here we show that human induced pluripotent stem cell (iPSC)-derived pericyte-like cells (iPLCs) can generate A{beta} peptides, and that the cells carrying Swedish mutation in amyloid precursor protein (APPswe) secrete 10 times more A{beta}1-42 than the control cells. Additionally, APPswe iPLCs have an impaired capacity to support angiogenesis and barrier integrity, exhibit a prolonged contractile response, and produce increased levels of pro-inflammatory cytokines upon inflammatory stimulation. These functional alterations in APPswe iPLCs are accompanied by transcriptional upregulation of actin cytoskeleton and extracellular matrix organization-related genes. Therefore, the APPswe mutation in iPLCs recapitulates several features of CAA pathology in vitro. Our iPSC-based vascular cell model may thus serve as a platform for drug discovery targeting vascular dysfunction in AD.

cell biology↗

USP30 inhibition induces mitophagy and reduces oxidative stress in parkin-deficient and CCCP-stressed human iPSC-derived neurons

Ubiquitination of mitochondrial proteins plays an important role in the cellular regulation of mitophagy. The E3 ubiquitin ligase parkin (encoded by PARK2) and the ubiquitin-specific protease 30 (USP30) have both been reported to regulate ubiquitination of outer mitochondrial proteins and thereby mitophagy. Loss of E3 ligase activity is thought to be pathogenic in both sporadic and inherited Parkinsons disease (PD), with loss-of-function mutations in PARK2 being the most frequent cause of autosomal recessive PD. The aim of the present study was to evaluate whether mitophagy induced by USP30 inhibition provides a functional rescue in isogenic human induced pluripotent stem cell-derived dopaminergic neurons with and without PARK2 knockout (KO). Our data show that healthy neurons responded to CCCP-induced mitochondrial damage by clearing the impaired mitochondria and that this process was accelerated by USP30 inhibition. Parkin-deficient neurons showed an impaired mitophagic response to CCCP challenge, although mitochondrial ubiquitination was enhanced. USP30 inhibition promoted mitophagy in PARK2 KO neurons, independently of whether left in basal conditions or treated with CCCP. In PARK2 KO, as in control neurons, USP30 inhibition balanced oxidative stress levels by reducing excessive production of reactive oxygen species. Interestingly, non-dopaminergic neurons, were the main driver of the beneficial effects of USP30 inhibition. Our findings demonstrate that USP30 inhibition is a promising approach to boost mitophagy and improve cellular health, also in parkin-deficient cells, and support the potential relevance of USP30 inhibitors as a novel therapeutic approach in diseases with a need to combat neuronal stress mediated by impaired mitochondria.

neuroscience↗

APOE4 affects basal and NMDAR mediated protein synthesis in neurons by perturbing calcium homeostasis

Apolipoprotein E (APOE), one of the primary lipoproteins in the brain has three isoforms in humans - APOE2, APOE3, and APOE4. APOE4 is the most well-established risk factor increasing the pre-disposition for Alzheimers disease. The presence of the APOE4 allele alone is shown to cause synaptic defects in neurons and recent studies have identified multiple pathways directly influenced by APOE4. However, the mechanisms underlying APOE4 induced synaptic dysfunction remain elusive. Here, we report that the acute exposure of primary cortical neurons to APOE4 leads to a significant decrease in global protein synthesis. APOE4 treatment also abrogates the NMDA mediated translation response indicating an impairment of synaptic signaling. Importantly, we demonstrate that both APOE3 and APOE4 generate a distinct translation response which is closely linked to their respective calcium signature. Acute exposure to APOE3 causes a short burst of calcium through NMDARs in neurons leading to an initial decrease in protein synthesis which quickly recovers. Contrarily, APOE4 leads to a sustained increase in calcium levels by activating both NMDARs and L-VGCCs, thereby causing sustained translation inhibition through eEF2 phosphorylation, which in turn disrupts NMDAR response. Thus, we show that APOE4 affects basal and activity mediated protein synthesis response in neurons by affecting calcium homeostasis. We propose this as a possible mechanism to explain the synaptic dysfunction caused by APOE4. Highlights / SummaryO_LIAPOE3 treatment causes a short burst of calcium through NMDARs, leading to an acute increase in eEF2 phosphorylation which eventually recovers to basal levels. C_LIO_LIGlobal translation follows a similar temporal profile of initial inhibition followed by recovery in APOE3 treated neurons, thus unaffecting the NMDA mediated translation response. C_LIO_LIAPOE4 treatment activates both NMDARs and L-VGCCs leading to a marked elevation in calcium levels, thus causing sustained increase in eEF2 phosphorylation as well as global translation inhibition. C_LIO_LIHence, the NMDA mediated response is perturbed, potentially causing a stress-related phenotype in APOE4 treated neurons. C_LIO_LIThus, different calcium signatures and sources lead to distinct temporal profiles of translation. C_LI

neuroscience↗