A5072897813- Cellular transport and secretion
- Single-cell and spatial transcriptomics
- Pluripotent Stem Cells Research
- Metabolism, Diabetes, and Cancer
- MicroRNA in disease regulation
- CRISPR and Genetic Engineering
- Erythrocyte Function and Pathophysiology
- Genomics and Chromatin Dynamics
- Gene Regulatory Network Analysis
- RNA Research and Splicing
- Microtubule and mitosis dynamics
- Wnt/β-catenin signaling in development and cancer
- Renal and related cancers
- Renal cell carcinoma treatment
- Retinal Development and Disorders
- Legionella and Acanthamoeba research
- Angiogenesis and VEGF in Cancer
- Cancer-related gene regulation
- Tuberous Sclerosis Complex Research
- Cancer Genomics and Diagnostics
- Autophagy in Disease and Therapy
- Muscle Physiology and Disorders
- Renal and Vascular Pathologies
- Diabetes Treatment and Management
- Calcium signaling and nucleotide metabolism
Institute of Molecular Biotechnology
2016-2024
Universität Innsbruck
2023-2024
Vienna Biocenter
2016-2024
Austrian Academy of Sciences
2013-2022
University of California, San Francisco
2005-2014
San Francisco Foundation
2009-2013
Williams (United States)
2009
Medizinische Hochschule Hannover
1982-2005
Functional screen for microcephaly genes Genetic screens are widely used to identify regulators in biological processes. Human currently limited two-dimensional cell cultures, which lack the ability score tissue-dependent gene function. Esk et al. combined CRISPR-Cas9 screening with barcoded cellular lineage tracing enable loss-of-function three-dimensional human cerebral organoid tissue. By testing candidate genes, endoplasmic reticulum was found control extracellular matrix protein...
Abstract The development of the human brain involves unique processes (not observed in many other species) that can contribute to neurodevelopmental disorders 1–4 . Cerebral organoids enable study a context. We have developed CRISPR–human organoids–single-cell RNA sequencing (CHOOSE) system, which uses verified pairs guide RNAs, inducible CRISPR–Cas9-based genetic disruption and single-cell transcriptomics for pooled loss-of-function screening mosaic organoids. Here we show perturbation 36...
Organoids enable in vitro modeling of complex developmental processes and disease pathologies. Like most 3D cultures, organoids lack sufficient oxygen supply therefore experience cellular stress. These negative effects are particularly prominent models, such as brain organoids, can affect lineage commitment. Here, we analyze organoid fetal single-cell RNA sequencing (scRNAseq) data from published new datasets, totaling about 190,000 cells. We identify a unique stress signature the all...
During brain development, neural progenitors expand through symmetric divisions before giving rise to differentiating cell types via asymmetric divisions. Transition between those modes varies among individual stem cells, resulting in clones of different sizes. Imaging-based lineage tracing allows for analysis at high cellular resolution but systematic approaches analyse clonal behaviour entire tissues are currently lacking. Here we implement whole-tissue by genomic DNA barcoding 3D human...
The endocytic accessory clathrin assembly lymphoid myeloid leukemia protein (CALM) is the ubiquitously expressed homolog of neuron‐specific AP180 that has been implicated in retrieval synaptic vesicle. Here, we show CALM associates with α‐appendage domain AP2 adaptor via three peptide motifs 420 DPF, 375 DIF and 489 FESVF to a lesser extent amino‐terminal heavy chain. Reducing levels by RNA interference did not significantly affect localization, but depletion weakens its association plasma...
GLUT4, Clathrin, and Glucose In human muscle, the GLUT4 glucose transport pathway responds to insulin is responsible for 70 90% of clearance. basal metabolic state, sequestered away from cell surface released an intracellular membrane compartment in response insulin. This defective type II diabetes. Vassilopoulos et al. (p. 1192 ; see Perspective by Orme Bogan ) now describe a function CHC22 clathrin, second isoform clathrin that present humans not mice. participates biogenesis sequesters...
In the developing neocortex, progenitor cells expand through symmetric division before they generate cortical neurons multiple rounds of asymmetric cell division. Here, we show that orientation mitotic spindle plays a crucial role in regulating transition between those two modes. We demonstrate protein phosphatase PP4c regulates early cells. Upon removing PP4c, spindles fail to orient parallel neuroepithelial surface and progenitors divide with random orientation. As result, their divisions...
Article5 October 2016Open Access Source DataTransparent process MicroRNA-34/449 controls mitotic spindle orientation during mammalian cortex development Juan Pablo Fededa Corresponding Author [email protected] Institute of Molecular Biotechnology the Austrian Academy Sciences (IMBA), Vienna Biocenter (VBC), Vienna, Austria Search for more papers by this author Christopher Esk Beata Mierzwa Rugile Stanyte Shuiqiao Yuan Department Physiology and Cell Biology, University Nevada School Medicine,...
Clathrin heavy chain 22 (CHC22) is an isoform of the well-characterized CHC17 clathrin chain, a coat component vesicles that mediate endocytosis and organelle biogenesis. CHC22 has distinct role from in trafficking glucose transporter 4 (GLUT4) skeletal muscle fat, though its transfection into HEK293 cells suggests functional redundancy. Here, we show eightfold less abundant than muscle, other cell types have variably lower amounts CHC22, endogenous function independently nonmuscle cells....
The clathrin light chain (CLC) subunits participate in several membrane traffic pathways involving both and actin, through binding the actin-organizing huntingtin-interacting proteins (Hip). However, CLCs are dispensable for clathrin-mediated endocytosis of many cargoes. Here we observe that CLC depletion affects cell migration Hip reduces surface expression β1-integrin by interference with recycling following normal inactive β1-integrin. a modified also inhibit appearance gyrating...
Glucose transporter 4 (GLUT4) is sequestered inside muscle and fat then released by vesicle traffic to the cell surface in response postprandial insulin for blood glucose clearance. Here, we map biogenesis of this GLUT4 pathway humans, which involves clathrin isoform CHC22. We observe that transits through early secretory more slowly than constitutively secreted GLUT1 localize CHC22 ER-to-Golgi intermediate compartment (ERGIC). functions transport from ERGIC, as demonstrated an essential...
The HUSH (human silencing hub) complex contains the H3K9me3 binding protein M-phase phosphoprotein 8 (MPP8) and recruits histone methyltransferase SETDB1 as well Microrchidia CW-type zinc finger 2 (MORC2). Functional mechanistic studies of have hitherto been centered around while in vivo functions MPP8 MORC2 remain elusive. Here, we show that genetic inactivation Mphosph8 or Morc2a nervous system mice leads to increased brain size, altered architecture, behavioral changes. Mechanistically,...
Development of the human brain involves processes that are not seen in many other species, but can contribute to neurodevelopmental disorders (1–4). Cerebral organoids be used investigate a context limited by variability and low throughput. We have developed CRISPR-human organoids-scRNA-seq (CHOOSE) system utilizes verified pairs gRNAs, inducible CRISPR/Cas9-based genetic disruption, single-cell transcriptomics for pooled loss-of-function screening mosaic organoids. Genetic perturbations 36...
Mobilization of the GLUT4 glucose transporter from intracellular storage vesicles provides a mechanism for insulin-responsive import into skeletal muscle. In humans, clathrin isoform CHC22 participates in formation compartment muscle and fat. function is limited to retrograde endosomal sorting restricted its tissue expression species distribution compared conserved CHC17 that mediates endocytosis several other membrane traffic pathways. Previously, we noted was expressed at elevated levels...
Human brain organoids are powerful in vitro models for development and disease. However, their variability can complicate use biomedical research drug discovery. Both the specific protocol as well pluripotent starting cell line influence organoid result incomplete representation of types an experiment. Here, we systematically analyze cellular transcriptional landscape grown from multiple lines using four different protocols recapitulating dorsal ventral forebrain, midbrain, striatum. We...
Abstract Organoids enable disease modeling in complex and structured human tissue, vitro . Like most 3D models, they lack sufficient oxygen supply, leading to cellular stress. These negative effects are particularly prominent like brain organoids, where can prevent proper lineage commitment. Here, we analyze organoid fetal single cell RNA sequencing (scRNAseq) data from published new datasets totaling over 190,000 cells. We describe a unique stress signature found all samples, but not...
Neural organoids model the development of human brain and are an indispensable tool for studying neurodevelopment. Whole-organoid lineage tracing has revealed number progenies arising from each initial stem cell to be highly diverse, with sizes ranging one more than 20,000 cells. This high variability exceeds what can explained by existing stochastic models corticogenesis indicates existence additional source stochasticity. To explain this variability, we introduce SAN which distinguishes...
Abstract Background Single-cell RNA-seq suffers from unwanted technical variation between cells, caused by its complex experiments and shallow sequencing depths. Many conventional normalization methods try to remove this calculating the relative gene expression per cell. However, their choice of Maximum Likelihood estimator is not ideal for application. Results We present GTestimate , a new method based on Good-Turing estimator, which improves upon accounting unobserved genes. To validate we...
Abstract Glucose Transporter 4 (GLUT4) is sequestered inside muscle and fat, then released by vesicle traffic to the cell surface in response post-prandial insulin for blood glucose clearance. Here we map biogenesis of this GLUT4 pathway humans, which involves clathrin isoform CHC22. We observe that transits through early secretory more slowly than constitutively-secreted GLUT1 transporter localize CHC22 endoplasmic-reticulum-to-Golgi-intermediate compartment (ERGIC). functions transport...
The histotopographic localization of the nerves vas deferens was studied in dogs. are concentrated at two poles. A vasectomy with a nerve-sparing technique and conventional were performed groups 5 months later all dogs underwent vasovasostomy. intact nerve supply after controlled by electrophysical measurement. Sperm evaluation morphological findings compared advantages demonstrated. proliferative capacity epithelial layer has to be considered operations on vas.
Summary Cerebral organoids model the development of human brain and are an indispensable tool for studying neurodevelopment. Whole-organoid lineage tracing has revealed number progeny arising from each initial stem cell to be highly diverse, with sizes ranging one more than 20,000 cells. This exceeds what can explained by existing stochastic models corticogenesis, indicating existence additional source stochasticity. We propose explanation in terms SAN which this stochasticity is survival...