Biology subjects

Chneiweiss, H.

Publications and source records attributed to Chneiweiss, H..

6 recordsLinked to original sources

bioRxiv2025

Human cortico-vascular assembloids reveal a CELF2-AHNAK-dependent switch from neuronal to endothelial tropism in glioblastoma cells

Glioblastoma (GB) remains one of the most aggressive human cancers, driven by profound cellular plasticity and dynamic interactions with the neurovascular niche. Yet, existing preclinical models fail to reproduce the human brain-vascular interface, limiting studies of tumor-host crosstalk and invasion. To overcome this gap, we developed a human induced pluripotent stem cell (hiPSC)-derived cortico-endothelial (CO+EO) assembloid model by fusing cortical and endothelial organoids from the same genetic background. These assembloids spontaneously form branched vascular networks enriched in tight junction proteins (CLDN5, OCLN, ZO-1), thereby recapitulating a key blood-brain barrier (BBB)-like property and providing a physiologically relevant human platform for exploring glioblastoma-neurovascular interactions. Using this system, we uncover a previously unrecognized CELF2-dependent glioblastoma stem cell (GSC) tropism. CELF2-expressing GSCs preferentially infiltrate neural regions, where they trigger neuronal apoptosis and disrupt endothelial tight junction integrity, supporting an aggressive phenotype. Conversely, CELF2-deficient GSCs lose neurotropism, acquire mesenchymal features, and are redirected toward vascular compartments. This endothelial affinity requires the scaffold protein AHNAK, strongly expressed at the plasma membrane of CELF2-deficient cells and enriched at tumor-endothelial interfaces. AHNAK knockdown abolished endothelial infiltration, demonstrating its critical role in vascular tropism. Analysis of patient GB samples confirmed that CELF2-positive tumor cells are enriched in poorly vascularized, mitotically active areas and excluded from vessel-rich zones, closely paralleling assembloid findings. Transcriptomic profiling further revealed that CELF2 promotes a neuronal progenitor-like program while repressing mesenchymal and vascular-associated gene expression, thereby shaping tumor identity, invasive behavior, and tissue preference. Collectively, our study introduces CO+EO assembloids as an original human model of glioblastoma plasticity at the neurovascular interface. We identify CELF2 as a master regulator of GSC tropism and AHNAK as a mediator of vascular affinity, unveiling a novel molecular axis that governs glioblastoma invasion and highlighting new therapeutic opportunities. Key points- Developed cortico-endothelial assembloids mimicking the brain microenvironment. - Identified CELF2 level as a determinant of glioblastoma cell tropism toward neuronal or endothelial niches. - Showed CELF2-positive GB cells disrupt endothelial tight junctions, unlike CELF2-negative cells. Importance of the studyThis study presents a novel, physiologically relevant in vitro model of glioblastoma (GB) that faithfully recapitulates the complex human brain microenvironment, including both vascular and neuronal components. By combining cortical and endothelial organoids derived from human iPSCs into cortico-endothelial assembloids (CO+EOs), the model enables detailed analysis of GB cell behavior, including migration, tissue tropism, and impact on the neurovascular interface. The work identifies CELF2 as a key molecular determinant of GB cell tropism and neurovascular interaction. The discovery that CELF2-expressing GB cells preferentially infiltrate neuronal tissue and disrupt endothelial tight junctions, whereas CELF2-deficient cells favor endothelial regions without compromising tight junction integrity, offers critical insights into tumor heterogeneity and mechanisms of invasiveness. These findings not only enhance our understanding of how GB cells interact with distinct microenvironmental niches but also provide a powerful platform for testing therapeutic strategies targeting tumor-microenvironment interactions. Graphical AbstractSchematic representation of CO+EO assembloids comprising an endothelial (yellow) and a cortical (red) domain. On the left, control CELF2-expressing GB cells preferentialy infiltrate the neuronal compartment, where they trigger neuronal apoptosis and disrupt BBB-like tight junctions upon contact with endothelial cells. On the right, CELF2-deprived GB cells exhibit reduced neural invasion and increased tropism for endothelial cells, consistent with a mesenchymal signature shift partially driven by elevated AHNAK expression. These cells exert limited impact on neuronal viability and BBB integrity, and display partial loss of GSC identity, as indicated by reduced OLIG2 expression. The CO+EO assembloid model thus recapitulates key features of cortico-endothelial human tissue heterogeneity and provides a reliable platform to dissect the molecular mechanisms underlying GSC tropism and their tissue-specific interactions O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=145 SRC="FIGDIR/small/683634v1_ufig1.gif" ALT="Figure 1"> View larger version (52K): org.highwire.dtl.DTLVardef@26440org.highwire.dtl.DTLVardef@103b93eorg.highwire.dtl.DTLVardef@176ca48org.highwire.dtl.DTLVardef@1da502f_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

bioRxiv2025

Immune Cells Infiltration of Patient Derived Glioblastoma Cells spheroids in Acoustic Levitation in Bulk Acoustic Wave devices

We describe an acoustofluidic device that allows scaffold-free structuration and culture of multi-cellular tumoroids composed of patient-derived glioblastoma cells only or in combination with non-cancerous cell. A PDMS chip of controlled height was created to allow acoustic levitation of cells using a 2 MHz transducer held on top of the chip. Cells are introduced into the chip through a dedicated inlet upstream of the resonant cavity. The specific design of the cavity together with the acoustic field allow the formation of tumoroids of cells in a precise and controlled manner within the levitation chamber. The acoustic and fluidic environment of the device was determined through experiments confronted with numerical simulations. The control of the flow within the chip was optimized to allow long-term culture of tumoroids and injection of cell culture media without disturbing the tumoroids in levitation. The tumoroids can be also structured, with sequential injections of the different cell types. Using microglia, we show that the acoustofluidic device allows the formation and culture in acoustic levitation of tumoroids mixing cancer cells with other cells populating the tumor as well as immune cell infiltration within the tumoroids. These results demonstrate the suitability of acoustofluidic levitation as an original 3D culture method adapted to the exploration of cancer growth at multiple levels.

bioengineering↗

bioRxiv2024

A unique malignant cell type per patient tumor is encoded in each cancer cell transcriptome.

Deciphering shared features between patients through unsupervised analyses of tumor single-cell transcriptomes is hindered by the predominant clustering of malignant cells based on the patients tumor of origin. In contrast, cancer-associated non-malignant cells within tumors cluster according to their somatic cell type (e.g. macrophage, fibroblast, astrocyte), regardless of the individual patient. We investigated the origin of this contrasting clustering behavior using computational analyses and novel data sampling techniques across 160 tumors representing 9 distinct cancer types. This led to the unexpected disclosing of tumor-specific malignant cell types. We demonstrate that tumor-driven malignant cell clustering is independent from technical or computational biases, nor is it reducible to defined gene sets with tumor-specific expression across all examined cancers. On the opposite, we unveil redundant information dispersed across the transcriptome that encodes a unique identity shared by the malignant cells within each tumor. Additionally, we found a similar genomic encoding for the identity of normal cell types. Finally, we demonstrate that malignant cell identities are maintained across space and over time, as are the identities of normal somatic cells types. These findings suggest the establishment of a distinct type of malignant cells within each tumor, robustly and diffusively encoded across the entire transcribed genome.

cancer biology↗

bioRxiv2024

Nicotinamide Metabolism Reprogramming Drives Reversible Senescence of Glioblastoma Cells

Recent studies show that metabolites, beyond their metabolic roles, can induce significant changes in cell behavior. Herein, we investigate the non-canonical role of nicotinamide (vitamin B3) on glioblastoma (GB) cell behavior. Nicotinamide induced senescence in GB cells, characterized by reduced proliferation, chromatin reorganization, increased DNA damage, enhanced beta-galactosidase activity, and decreased Lamin B1 expression. Nicotinamide-induced senescence was accompanied by an unexpected reprogramming of its metabolism, marked by simultaneous downregulated transcription of NNMT (nicotinamide N-methyltransferase) and NAMPT (nicotinamide phosphoribosyl-transferase). Nicotinamide effects on GB cells were mediated by decreased levels of SOX2. Consistently, analyses of patients single cell transcriptome datasets showed that GB cells with low NNMT and NAMPT expression levels were enriched in gene modules related to senescence. Remarkably, senescent GB cells retained tumor-forming ability in vivo, albeit to a lesser extent compared to control cells. Further experiments at the single-cell level and transcriptomic analyses demonstrated that nicotinamide-induced senescence in GB cells is fully reversible. Overall, our findings identify a novel reversible senescent state in GB tumors and highlight the non-canonical role of NAM as a key driver of cancer cell plasticity.

cancer biology↗

bioRxiv2022

Human glioblastoma cell motility depends on the activity of the cysteine metabolism enzyme 3-Mercaptopyruvate sulfurtransferase

Cancer cells in similar functional states are found in all glioblastoma, despite the genomic heterogeneity observed between and within these brain tumors. Metabolism being downstream of all signaling pathways regulating cell behaviors, we looked for metabolic weaknesses in link with motility, a key functional state for glioblastoma aggressiveness. A signature-driven data reduction approach highlighted motile cells present in thirty tumors from four independent single-cell transcriptomic datasets. Analyses integrating trajectory modeling disclosed, as characteristic of motile cells, enhanced oxidative stress coupled with mobilization of the cysteine metabolism enzyme 3-Mercaptopyruvate sulfurtransferase (MPST). The soundness of this prediction was verified using migration and invasion assays with patient-derived cells and tissue organoids. Pharmacological and genetic manipulations showed that enhanced ROS production and MPST activity are required for the cells motility. Biochemical assays indicated that MPST acts by protecting protein cysteine residues from dismal hyperoxidation. In vivo, MPST knockdown translated in reduced tumor burden, and a robust increase in mice survival. These results show that enhanced oxidative stress coupled with MPST mobilization plays a key role in glioblastoma cell motility.

cancer biology↗

bioRxiv2019

Capture at the single cell level of metabolic modules distinguishing aggressive and indolent glioblastoma cells

Glioblastoma cell ability to adapt their functioning to microenvironment changes is a source of the extensive intra-tumor heterogeneity characteristic of this devastating malignant brain tumor. A systemic view of the metabolic pathways underlying glioblastoma cell functioning states is lacking. We analyzed public single cell RNA-sequencing data from glioblastoma surgical resections, which offer the closest available view of tumor cell heterogeneity as encountered at the time of patients diagnosis. Unsupervised analyses revealed that information dispersed throughout the cell transcript repertoires encoded the identity of each tumor and masked information related to cell functioning states. Data reduction based on an experimentally-defined signature of transcription factors overcame this hurdle. It allowed cell grouping according to their tumorigenic potential, regardless of their tumor of origin. The approach relevance was validated using an independent dataset of glioblastoma tissue transcriptomes, patient-derived cell lines and orthotopic xenografts. Overexpression of genes coding for amino acid and lipid metabolism enzymes involved in anti-oxidative, energetic and cell membrane processes characterized cells with high tumorigenic potential. Modeling of their expression network highlighted the very long chain polyunsaturated fatty acid synthesis pathway at the core of the network. Expression of its most downstream enzymatic component, ELOVL2, was associated with worsened patient survival, and required for cell tumorigenic properties in vivo. Our results demonstrate the power of signature-driven analyses of single cell transcriptomes to obtain an integrated view of metabolic pathways at play within the heterogeneous cell landscape of patient tumors.

cancer biology↗