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Advanced understanding of the neurotoxicity of brominated flame retardants: Integrating human neural organoids with multi-omics.

The developing nervous system is susceptible to environmental chemical exposure. Polybrominated diphenyl ethers (PBDEs) are brominated flame retardants that can induce neurodevelopmental impairments and neural dysfunction in animals. To investigate the toxicity of PBDEs to the human developing nervous system, neural organoid models, such as human cortical organoids (hCOs) and human retinal organoids (hROs), were established and exposed to BDE-47, BDE-209, and BDE-MIX (BDE-47/-209). Exposure to PBDEs caused morphological changes in both hCOs and hROs, as evidenced by decreased organoid size and…

Systematic production of human kidney organoids for transplantation in porcine kidneys during ex vivo machine perfusion.

Organoids derived from human pluripotent stem (hPS) cells hold promise for therapeutic purposes. However, technological advances to overcome their massive production while ensuring differentiation fidelity are still lacking. Here we report a procedure sustaining the derivation of kidney organoids from hPS cells (hPSC-kidney organoids) using a scalable, reproducible and affordable approach that allows hPSC-kidney organoid differentiation into different renal cell types. Using single-cell RNA sequencing, confocal image analysis, metabolic assays and CRISPR-Cas9 engineering for generation of fluo…

Origin and cell type specificity of mitochondrial DNA mutations in C9ORF72 ALS-FTLD human brain organoids.

Amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) are primarily genetic in ~20% of patients. Mutations in C9ORF72 are the most frequent cause, but it is not understood why there is notable regional pathology. An increased burden of mitochondrial DNA (mtDNA) mutations in ALS-FTLD brains implicates mitochondrial mechanisms; however, it remains unclear how and when these mutations arise. To address this, we generated cerebral organoids derived from human-induced pluripotent stem cells (hiPSCs) of patients with ALS-FTLD harboring the C9ORF72 hexanucleotide repeat exp…

Apoptosis regulators of the Bcl-2 family play a key role in chemoresistance of cholangiocarcinoma organoids.

Cholangiocarcinoma (CCA) is a rare but devastating liver cancer which is commonly diagnosed at a late stage and often resistant to chemotherapy. Bcl-2 family members, which control apoptotic cell death, are known to be involved in the chemoresistance of some cancer types. This study investigated the role of Bcl-2 family members in the chemoresistance of cholangiocarcinoma organoids (CCAOs) in both undifferentiated and matured branching phenotypes (BRCCAOs). Patient-derived CCAOs and BRCCAOs were cultured to assess chemoresistance to an FDA-approved anticancer drug panel by testing cell viabili…

Patient-derived medullary thyroid cancer organoids: a potential model for mechanistic studies on diagnostics and therapy.

OBJECTIVE: Medullary thyroid carcinoma (MTC) is a rare neuroendocrine thyroid tumor, with only 30 new patients annually in the Netherlands. … ene and protein expression were characterized by qPCR and immunofluorescent staining. Moreover, MTC-organoids (MTOs) were exposed to TKIs and PET tracers in proof-of-principle experiments. RESULTS: Ten MTC biopsies were processed and successfully cultured as MTOs. MTC-derived cells showed self-renewal potency for several passages, indicating the presence o …

Development of Sheep Duodenum Intestinal Organoids and Implementation of High-Throughput Screening Platform for Veterinary Applications.

New therapeutic molecules for farm animals are needed to address worldwide problems in the food industry, like the rise of resistance among ruminant parasites and pathogenic microbes. Since in vivo testing would involve an excessive number of animals, with consequent ethical and economic issues, the generation of sheep intestinal organoids represents a promising close-to-reality in vitro model for veterinary drug development; however, the characterization and application of such organoids remain limited. In this study, ovine intestinal organoids were generated from adult LGR5+ stem cells from …

Association of cytochrome c oxidase dysfunction with amyloidosis in Alzheimer's disease and patient-derived cerebral organoids.

Patients with Alzheimer's disease (AD) demonstrate brain mitochondrial dysfunction and energy deficiency that are closely associated with co … ition of CCO promoted amyloid production and elevated beta-secretase 1 (BACE1) activity in cerebral organoids derived from human induced pluripotent stem cells (hiPSCs) from one nonAD and one AD subject. The simplest interpretation of the results is that CCO dysfunction in the frontal lobe is a phenotypic mitochondrial change accompanying AD, which may cont …

Synergistic enhancement of AAV gene delivery in 2D cells and 3D organoids using polybrene and hydroxychloroquine.

Recent advances in three-dimensional (3D) culture platforms have enabled organoids to serve as physiologically relevant models for recapitulating human biology and assessing therapeutic efficacy and toxicity. Despite their promise, their complex architecture presents significant challenges for efficient gene delivery, thereby limiting their broader application in drug discovery and translational research. To overcome this challenge, we developed a sequential treatment strategy that combines polybrene (PB), which facilitates viral entry, and hydroxychloroquine (HCQ), which modulates endosomal p…

Lycii radicis cortex alleviates fibrosis in hiPSC-derived multilineage hepatic organoids via the cAMP-PKA pathway.

BACKGROUND: Liver fibrosis, driven by excessive extracellular matrix (ECM) deposition and activation of hepatic stellate cells (HSCs), still lacks effective therapies, partly due to the absence of human-relevant models. Lycii Radicis Cortex (LRC), a traditional Chinese medicine, exhibits reported anti-inflammatory and antioxidant activities, yet its anti-fibrotic potential has not been validated in human organoid-based systems. METHODS: We established hiPSC-derived multilineage hepatobiliary organoids (mHBOs) containing mesoderm-derived HSCs and implemented a TGF-β-induced fibrosis model with…

Celastrol reduces cisplatin-induced nephrotoxicity by downregulating SNORD3A level in kidney organoids derived from human iPSCs.

BACKGROUND: Celastrol, an active ingredient derived from Tripterygium wilfordii Hook F, has shown therapeutic potential for various kidney r … S: We investigated the ability of celastrol to reduce cisplatin-induced nephrotoxicity using kidney organoids. Kidney organoids were cultured and characterized, exhibiting renal tubular and glomerular structures and expressing specific kidney markers such as NPHS1, CD31, LTL, and SLC12A1. Data were obtained from in vitro experiments in which kidney organoid …

Quantitative Imaging Analysis of Internal Structural Complexity in Mouse Heart Organoids; Comparison to Mouse Embryonic Hearts.

We have established mouse heart organoids (mHOs) that are characterized by the presence of atrium- and ventricle-like structures that mimic entire embryonic hearts. However, maturation was not uniform, and little is known about their trabeculation status. Given the essential role of the trabeculae in cardiac morphogenesis, a new method that combines imaging analysis with a machine learning model was developed for quantifying the internal complexity in developing mHOs in a timely manner. We applied this method to screen for modified culture conditions and identified optimal treatment with valpr…

Connecting cilium, stress response, and proteostasis abnormalities inform variant and therapy assessment in RPGRIP1 retinal organoids.

RPGRIP1 encodes a connecting cilium (CC) protein essential for normal photoreceptor cell development and maintenance. Damaging variants in R … d other management options. Here, we use human induced pluripotent stem cell (iPSC)-derived retinal organoids to model RPGRIP1-associated IRD, detecting biomarkers of disease including CC interactome dysfunction, stress response, and proteostasis abnormalities. In parallel, utilizing these novel disease biomarkers, we demonstrate the pathogenicity of a miss …

Human fallopian tube epithelial organoids with TP53 mutation recapitulate features of serous tubal intraepithelial carcinoma (STIC).

OBJECTIVE: Serous tubal intraepithelial carcinoma (STIC) is the immediate precursor lesion for high-grade serous ovarian carcinoma (HGSOC) and harbors universal TP53 mutations. The lack of an appropriate in vitro model for STIC presents a major challenge in studying its pathogenesis. We aimed to develop a human in vitro model that mimics STIC lesions. METHODS: Using CRISPR-Cas9 gene editing, we generated human fallopian tube epithelial organoids with TP53 loss-of-function mutations (TP53-/- FTOs). We characterized TP53-/- FTOs on a cellular and molecular level using immunofluorescence confoca…

Enhancing pre-sexual and sexual differentiation of Toxoplasma gondii using retinal epithelial cells and intestinal organoids.

Toxoplasmosis, caused by Toxoplasma gondii, is a major global health concern due to its high prevalence, zoonotic transmission, and economic … re-sexual and sexual stages by combining a human retinal epithelial cell line and murine intestinal organoids with FELIX medium, which mimics feline intestinal biochemistry, and conditional MORC depletion. This system increases stage-specific gene expression and marker detection, demonstrating synergistic effects of host environment and genetic regulation. …

Macrophage-augmented organoids recapitulate the complex pathophysiology of viral diseases and enable development of multitarget therapeutics.

The pathophysiology of acute viral diseases is complex. It is characterized by strong inflammatory responses driven by immune cells, leading to tissue damage. Currently available in vitro models mainly recapitulate the viral life cycle but fail to model immune cell-mediated pathogenesis. Here we build macrophage-augmented organoids (MaugOs) by integrating macrophages into primary organoids that are cultured from human liver tissues. We test the infections of two RNA viruses, hepatitis E virus and SARS-CoV-2, and one DNA virus, monkeypox virus, which either primarily or secondarily affect the h…

Lung Organoids from hiPSCs Can Be Efficiently Transduced by Recombinant Adeno-Associated Viral and Adenoviral Vectors.

Background: Organoids are a valuable model for studying hereditary diseases such as cystic fibrosis (CF). Recombinant adenoviral (rAdV) and adeno-associated viral (rAAV) vectors are promising tools for CF gene therapy and genome editing. Objective: This study aims to determine the most efficient viral vector (rAdV5, rAAV serotypes 5, 6 and 9) and transduction protocol for delivering transgenes to lung organoids (LOs), providing a foundation for future CF gene therapy development. Methods: Three transduction protocols were used taking into account the specificities of LOs' cultivation in specif…

Modeling Mycobacterium tuberculosis pathogenesis in lung epithelial organoids reveals strain-specific host responses and intercellular crosstalk.

Emerging studies have identified alveolar epithelial cells as a conducive niche for Mycobacterium tuberculosis (Mtb) replication and spread … genesis. Transcriptomic analyses revealed that Mtb infection markedly alters cell death patterns in organoids and modulates signal transduction pathways in peripheral blood mononuclear cells (PBMCs). Western blot indicates the H37Rv strain induced ferroptosis, autophagy, and apoptosis while suppressing necroptosis in organoids. In contrast, BCG predominantl …

Человеческие миелионированные органоиды мозга с интегрированной микроглией в качестве модели для миелинового ремонта и ремиелионирующей терапии.

Олигодендроциты, миелионирующие клетки центральной нервной системы (ЦНС), необходимы для формирования миелиновых ножниц и имеют решающее значение для поддержания аксонной целостности и проводимости. Разрушение этих клеток и миелинных оболочек, которые они производят, является отличительной чертой таких демиелинизирующих условий, как множественный склероз или те, которые являются результатом некоторых побочных эффектов наркотиков, что приводит к глубоким неврологическим расстройствам. В этом исследовании мы создали органоид человеческого мозга, состоящий из нейронов, астроцитов и милинирующих о

Совпадение в аннотации оригинала
Oligodendrocytes, the myelinating cells of the central nervous system (CNS), are essential for the formation of myelin sheaths and pivotal f … ntegrating induced pluripotent stem cell-derived microglia, we endowed these myelinated human brain organoids (MHBOs) with immune characteristics. MHBOs with microglia (MHBOs +MG) enabled the investigation of demyelination and remyelination-a process in which myelin sheaths are regenerated-in a human context. After toxin-induced demyelination, we observed a …

Bacterial attachment and junctional transport function in induced apical-out polarized and differentiated canine intestinal organoids.

Dogs are increasingly recognized as valuable large animal models for understanding human intestinal diseases, as they naturally develop conditions similar to those in humans, such as Enterohemorrhagic E. coli, Clostridium difficile infection, inflammatory bowel disease, and ulcerative colitis. Given the similarity in gut flora between dogs and humans, canine in vitro intestinal models are ideal for translational research. However, conventional extracellular matrix-embedded organoids present challenges in accessing the lumen, which is critical for gut function. This study aimed to investigate t…

Optimization of culture conditions to generate vascularized multi-lineage liver organoids with structural complexity and functionality.

Hepatic organoids (HOs), primarily composed of hepatobiliary cells, do not represent the pathogenesis of liver diseases due to the lack of non-parenchymal cells. Multi-lineage liver organoids (mLOs) containing various cell types found in the liver offer a promising in vitro disease model. However, their structural complexity remains challenging to achieve due to the difficulty in optimizing culture conditions that meet the growth need of all component cell types. Here, we demonstrate that cystic HOs generated from hPSCs can be expanded long-term and serve as a continuous source for generating …