Biology subjects

Jungverdorben, J.

Publications and source records attributed to Jungverdorben, J..

5 recordsLinked to original sources

bioRxiv2026

Activated ADAM10 targeting antibody 1H5 suppresses NOTCH-driven glioma progenitors

Background: High-grade gliomas (HGGs) remain associated with high morbidity, mortality, and invariable recurrence after neurosurgical resection and chemoradiation. A major contributor to treatment failure is a population of stem-like glioma progenitor cells (GPCs) that depend on NOTCH-signaling for self-renewal and survival; clinical attempts to inhibit NOTCH with {gamma}-secretase inhibitors failed due to systemic toxicity. Methods: Here, we investigate the anti-HGG therapeutic potential of a fully-human monoclonal antibody, 1H5, that selectively targets activated-ADAM10, an -secretase required for NOTCH receptor processing, with high tumor specificity. We show that ADAM10 is markedly overexpressed and highly activated in primary and recurrent HGGs, using patient tissue, RNA-sequencing data, and early-passage HGG cell lines. Results: Across a panel of patient-derived HGG lines, sensitivity to 1H5 correlates with the cell-surface levels of activated-ADAM10, and shRNA-mediated ADAM10 knockdown recapitulates the growth-inhibitory effects of 1H5. ADAM10 inhibition suppresses NOTCH-pathway components and target genes, reduces expression of GPC markers, and impairs neurosphere formation while depleting Sox2- and Ki67-positive cells. We further find that 1H5 induces HGG cell death predominantly via necroptosis, characterized by survivin loss and increased phosphorylated MLKL and by transcriptional changes in necroptosis-related genes. In three-dimensional explants derived from primary HGG, 1H5 reduces growth and GPC marker expression while increasing TUNEL staining. Finally, convection-enhanced delivery of 1H5 in orthotopic xenograft models achieves high local intracranial concentrations, slows tumor growth, and prolongs survival, with regional pharmacodynamic effects consistent with NOTCH pathway suppression and necroptosis.

cancer biology↗

bioRxiv2026

Decoding subtype development and function in human pluripotent stem cell-derived midbrain dopaminergic neurons

Midbrain dopaminergic (mDA) neurons comprise molecularly and functionally distinct subtypes with differential vulnerability in neurodegenerative and psychiatric disorders. However, the mechanisms specifying subtype identity remain poorly understood, and protocols for the selective derivation of human mDA subtypes are lacking. Here we establish a strategy to derive substantia nigra (A9) and ventral tegmental area (A10) mDA neurons from human pluripotent stem cells (hPSCs). A9 identity is specified by dual-SMAD activation through Activin A and BMP7 at the midbrain floor-plate progenitor stage, whereas A10 identity is promoted by BMP inhibition. A9 mDA neurons are purified based on ALDH1A1 expression, and subtype identity is maintained by continued TGF-{beta} modulation and ESRRB activation in vitro and upon transplantation in vivo. Single-cell RNA sequencing and biochemical analyses demonstrate that hPSC-derived A9 neurons exhibit increased oxidative phosphorylation, neuromelanin-like pigmentation, elevated dopamine synthesis and release, and electrophysiological properties characteristic of A9 mDA neurons in vivo. Integration with human fetal midbrain datasets confirms strong transcriptional concordance between in vitro-derived and in vivo mDA subtypes. Neuromelanin-like structures produced by hPSC-derived A9 neurons trigger pro-inflammatory cytokine secretion from hPSC-derived microglia, and A9 neurons are primed to upregulate MHC-I genes in response to interferon-{gamma}, features which may contribute to the selective vulnerability of A9 neurons. Together, these results establish a robust in vitro platform to interrogate human mDA subtype development, function, and selective vulnerability, enabling mechanistic studies relevant to Parkinson's disease and the development of cell-based therapies.

developmental biology↗

bioRxiv2026

Rapid and efficient oligodendrocyte differentiation from human pluripotent stem cells via dual inhibition of BMP and Notch signaling

The protracted timing required for oligodendrocyte differentiation from human pluripotent stem cells (hPSCs) has limited their use in disease modeling, drug screening, and cell therapy. In particular, the signals that drive oligodendrocyte specification and maturation after neural induction and ventral patterning remain poorly understood. Here, we present a protocol to derive human oligodendrocytes from hPSCs that is based solely on extrinsic cues, and we identify dual inhibition of BMP and Notch signaling as critical drivers of oligodendrocyte commitment and maturation. By day 42 of differentiation, up to 70% of the cells are positive for the oligodendrocyte marker O4, with minimal astrocyte contamination, and show robust expression of mature myelin markers including MBP, MOG, and MAG. These hPSC-derived oligodendrocytes closely match the molecular identity of primary fetal human oligodendrocytes as assessed by single-cell RNA sequencing and are functional as shown by in vitro myelination assays. In addition to the rapid generation of myelinating oligodendrocytes, the new protocol can be modularly adapted for the efficient production of PDGFR+ oligodendrocyte precursors or mixed glial populations containing AQP4+ astrocytes, thereby providing a cellular toolbox for the study of human glial lineages in translational applications.

developmental biology↗

bioRxiv2024

Three-dimensional regulatory hubs support oncogenic programs in glioblastoma

Dysregulation of enhancer-promoter communication in the context of the three-dimensional (3D) nucleus is increasingly recognized as a potential driver of oncogenic programs. Here, we profiled the 3D enhancer-promoter networks of primary patient-derived glioblastoma stem cells (GSCs) in comparison with neuronal stem cells (NSCs) to identify potential central nodes and vulnerabilities in the regulatory logic of this devastating cancer. Specifically, we focused on hyperconnected 3D regulatory hubs and demonstrated that hub-interacting genes exhibit high and coordinated expression at the single-cell level and strong association with oncogenic programs that distinguish IDH-wt glioblastoma patients from low-grade glioma. Epigenetic silencing of a recurrent 3D enhancer hub--with an uncharacterized role in glioblastoma--was sufficient to cause concordant downregulation of multiple hub-connected genes along with significant shifts in transcriptional states and reduced clonogenicity. By integrating published datasets from other cancer types, we also identified both universal and cancer type-specific 3D regulatory hubs which enrich for varying oncogenic programs and nominate specific factors associated with worse outcomes. Genetic alterations, such as focal duplications, could explain only a small fraction of the detected hyperconnected hubs and their increased activity. Overall, our study provides computational and experimental support for the potential central role of 3D regulatory hubs in controlling oncogenic programs and properties. HIGHLIGHTS- 3D regulatory "hubs" in glioblastoma enrich for highly coregulated genes at a single-cell level and expand oncogenic regulatory networks. - Targeted perturbation of a highly recurrent 3D regulatory hub in GSCs results in altered transcriptional states and cellular properties. - 3D regulatory hubs across cancer types associate with tumor-specific and universal oncogenic programs and worse outcomes. - The majority of hyperconnected hubs do not overlap with structural variants, suggesting epigenetic mechanisms. eTOCHere we profile the 3D enhancer connectomes of primary patient-derived human glioblastoma stem cells (GSCs), identify hyperconnected 3D regulatory "hubs", and examine the impact of 3D hub perturbation on the transcriptional program and oncogenic properties.

genomics↗

bioRxiv2023

Generation of human cerebral organoids with a structured outer subventricular zone

Mammalian outer radial glia (oRG) emerge as cortical progenitor cells that directly support the development of an enlarged outer subventricular zone (oSVZ) and, in turn, the expansion of the neocortex. The in vitro generation of oRG is essential to model and investigate the underlying mechanisms of human neocortical development and expansion. By activating the STAT3 pathway using LIF, which is not produced in guided cortical organoids, we developed a cerebral organoid differentiation method from human pluripotent stem cells (hPSCs) that recapitulates the expansion of a progenitor pool into the oSVZ. The structured oSVZ is composed of progenitor cells expressing specific oRG markers such as GFAP, LIFR, HOPX, which closely matches human oRG in vivo. In this microenvironment, cortical neurons showed faster maturation with enhanced metabolic and functional activity. Incorporation of hPSC-derived brain vascular LIF- producing pericytes in cerebral organoids mimicked the effects of LIF treatment. These data indicate that the cellular complexity of the cortical microenvironment, including cell-types of the brain vasculature, favors the appearance of oRG and provides a platform to routinely study oRG in hPSC-derived brain organoids.

developmental biology↗