Search Results (589)

The development of patient-derived organoids (PDOs) has substantially advanced the study of gastrointestinal diseases by providing three-dimensional human models that faithfully recapitulate the structural, molecular, and functional characteristics of native tissues. Unlike conventional two-dimensional cultures and animal models, intestinal organoids preserve epithelial architecture, cellular heterogeneity, and patient-specific genetic features, enabling more physiologically relevant investigations of gastrointestinal physiology and disease. Recent technological advances, including co-culture systems, organoid-derived monolayers, and organ-on-chip platforms, have further expanded their ability to model epithelial interactions with immune cells, stromal components, and the gut microbiota. These developments have facilitated mechanistic studies of epithelial barrier function, host–microbiota communication, microbial metabolites, and endocrine signaling, while also supporting translational applications in inflammatory bowel disease, infectious disorders, inherited gastrointestinal diseases, and gastrointestinal cancers. Moreover, patient-derived organoids have emerged as promising platforms for drug screening, biomarker discovery, precision medicine, and regenerative therapies. Despite these advances, several challenges remain, including limited representation of the native tissue microenvironment, lack of standardized culture protocols, scalability, and regulatory issues that currently restrict routine clinical implementation. This review summarizes recent progress in gastrointestinal organoid technology, highlighting current applications, emerging experimental platforms, and future perspectives for integrating organoid-based models into translational research and personalized medicine. Full article

Glioblastoma (GBM) is a highly aggressive malignant brain tumor accounting for 15% of all brain tumors and 50% of all gliomas. The exact cause of GBM is not fully understood but risk factors include age, genetic mutations, exposure to ionizing radiation, and certain genetic disorders. Symptoms of GBM include headaches, seizures, cognitive impairment, and weaknesses on one side of the body. Myeloid cells account for 30–50% of the tumor mass and are instrumental in shaping the complex tumor microenvironment (TME). Inflammation in the TME is an important driver of tumor growth and invasion; however, as the environment evolves, the immunosuppressive TME poses a significant hurdle as it hinders the immune-mediated killing of tumor cells. Our work utilizes neuroimmune organoids containing neurons, astrocytes, microglia, and vascular-like cells, to which we add patient-derived GBM cells and/or iPSC-derived macrophages to model the GBM TME. Model characterization was performed using single-cell RNA sequencing and supernatant proteomics to determine cell-specific changes during coculturing. Our findings are consistent with this 7-day coculture model recapitulating key aspects of GBM early tumor establishment and immune activation, with transcriptomic and secretome signatures suggestive of an emerging immune evasion phenotype. Full article

Theranostic nanoparticles, which integrate diagnostic imaging and therapeutic delivery within a single nanoplatform, represent a transformative paradigm in oncological nanomedicine. Despite substantial preclinical progress, the field faces persistent gaps in rational nanoparticle design informed by tumor biology, preclinical model fidelity, and clinical translation. This review critically synthesizes theranostic nanoparticle research across three underexplored domains. First, we examine tumor microenvironment features—reactive oxygen species dynamics, glutathione gradients, hypoxia, and proteasomal dysregulation—as mechanistic drivers of nanoparticle responsiveness. Second, we evaluate redox-responsive and proteasome-targeted nanoplatforms that exploit these cues for stimuli-triggered drug release and simultaneous imaging readout. Third, we address the unmet need for three-dimensional organoid and microfluidic tumor models as predictive preclinical testing environments, given the well-documented limitations of conventional two-dimensional cultures. Cancer subtype-specific applications are discussed for breast cancer, HPV-associated malignancies, colorectal cancer, and prostate cancer. Clinical translation barriers—including pharmacokinetic constraints, protein corona formation, immune clearance, anti-PEG antibodies, complement activation-related pseudoallergy, and FDA/EMA regulatory pathways—are addressed from a clinical oncology perspective. The review concludes with a research roadmap integrating proteomics-guided nanoparticle engineering, patient-derived organoid biobanks, and artificial intelligence-assisted design as priority areas for next-generation oncological theranostics. Full article

Breast cancer is a highly heterogeneous disease characterized by distinct molecular subtypes, dynamic tumor–microenvironment interactions, and variable therapeutic responses. Despite the availability of multiple preclinical platforms, a major challenge remains the lack of a coherent multiscale framework capable of integrating biological complexity across experimental systems. This limitation reduces the predictive power of individual models and highlights the need for complementary strategies that reproduce disease progression across multiple biological scales. In this context, advanced imaging technologies have emerged as essential tools for linking preclinical platforms and enhancing their translational relevance. This review examines how multimodal imaging supports the integration of in vitro, ex vivo, and in vivo breast cancer models. We discuss how optical imaging, high-frequency ultrasound, magnetic resonance imaging, positron emission tomography/computed tomography, and intravital microscopy provide complementary molecular, functional, anatomical, and cellular information for the longitudinal assessment of tumor growth, metastatic dissemination, microenvironment remodeling, and therapeutic response. Particular attention is given to emerging translational workflows that combine patient-derived models with advanced imaging to investigate drug sensitivity, treatment resistance, and tumor progression within a precision oncology perspective. We also highlight the role of multimodal imaging in biomarker validation across platforms and in the development of clinically relevant preclinical pipelines. Overall, advanced imaging represents a critical translational bridge across breast cancer model systems, improving the predictive value of preclinical studies and supporting imaging-guided precision oncology from bench to bedside. Full article

The human placenta is a temporary organ structured to optimize exchange between the maternal and fetal circulatory systems. Its fetal component consists of highly branched chorionic villi, which are anchored to the maternal uterine wall and project into the intervillous space. The outer surface of these villi is lined by a multinucleated, continuous layer called the syncytiotrophoblast, which is supported by an underlying layer of proliferative cytotrophoblast cells and the invasive extravillous trophoblast (EVT). This cellular bilayer forms a selective barrier that directly bathes in maternal blood, allowing for the efficient transfer of oxygen and nutrients while structurally preventing the direct mixing of maternal and fetal blood cells. Human placental studies have been stymied by ethical and accessibility constraints. Stem cell biology has now revolutionized the capacity to model human placental development, in particular with the derivation of human trophoblast stem cells (hTSCs) and organoids. Authentic, self-renewing human trophoblast stem cells (hTSCs) were first derived not from pluripotent stem cells but from primary tissue—first-trimester villous cytotrophoblasts and blastocysts. Derivation from human pluripotent stem cells (PSCs) followed only subsequently, along two principal routes: conversion of naive PSCs, which retain extraembryonic competence, and induction from primed PSCs, as well as by direct reprogramming of somatic cells to induced hTSCs. An important advance underlying these improvements is the mapping of a global reprogramming roadmap. Multi-omic and lineage-tracing experiments have mapped the stepwise transcriptional and epigenetic conversions of fibroblasts to hTSCs, including sequential chromatin reconfiguration, trophoblast gene network activation, and repression of somatic signatures. These results identify major regulatory bottlenecks and intermediate states, improving reprogramming fidelity. The derivation of stem-cell-based trophoblast organoids now enables complex modeling of placental architecture, function, and disease susceptibility in vitro. These organoids accurately recapitulate placental barrier functions and immunological features, allowing for examinations of maternal–fetal health, pregnancy disorders, and placental infection response to viruses like cytomegalovirus and SARS-CoV-2. Looking ahead, the integration of reprogramming and organoid technologies will propel patient-specific and tailor-made models for personalized diagnostics, drug screening, and mechanism studies. As we unravel the molecular ballet of trophoblast induction, such discoveries have the potential to bridge basic translational gaps in reproductive biology and maternal–fetal medicine. Full article

Background: Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal solid malignancies, with poor survival driven by late presentation, aggressive tumor biology, and limited responsiveness to conventional systemic therapy. Advances in molecular profiling have expanded opportunities for biomarker-guided and targeted therapeutic approaches. Methods: A literature review was conducted using PubMed and the Cochrane Library through July 2026, supplemented by abstracts and proceedings from major international oncology conferences. Results: Pancreatic cancer is driven mainly by somatic changes in KRAS, TP53, CDKN2A, and SMAD4. Established precision approaches include maintenance olaparib for selected platinum-sensitive tumors with germline BRCA1 or BRCA2 pathogenic variants, immune checkpoint inhibition for mismatch repair-deficient or microsatellite instability-high tumors, and tropomyosin receptor kinase inhibition for cancers with neurotrophic tyrosine receptor kinase gene fusions. Direct inhibition of KRAS and RAS represents a major therapeutic breakthrough. KRAS G12C inhibitors established proof of concept, while agents targeting the more common KRAS G12D mutation are showing encouraging early activity. In the randomized phase III RASolute 302 trial, the multiselective RAS inhibitor daraxonrasib improved survival compared with chemotherapy in previously treated metastatic disease with oncogenic RAS mutations. Early studies of zoldonrasib combinations have extended this progress to KRAS G12D-mutant disease, although confirmation is required. Molecular profiling, next-generation sequencing, patient-derived organoids, and circulating tumor DNA may further improve treatment selection and monitoring. Conclusions: Precision oncology is becoming clinically relevant in pancreatic ductal adenocarcinoma. KRAS- and RAS-directed therapies are central advances, but resistance, toxicity, limited durability, and access to comprehensive testing remain important challenges. Full article

Cancer metabolism is a dynamic and context-dependent process shaped by both tumor-intrinsic programs and microenvironmental cues. Capturing this complexity remains a major challenge, which limits the translation of metabolic insights into clinically actionable strategies. Patient-derived tumor organoids, together with emerging engineered platforms such as organ-on-chip systems, vascularized assembloids, and bioprinted tumor models, have opened new avenues for investigating tumor metabolism in physiologically relevant settings. These models enable the study of metabolic heterogeneity across tumor types, disease stages, and treatment conditions while preserving clinically relevant tumor features. Importantly, they provide functional platforms for ex vivo metabolic profiling, identification of metabolic vulnerabilities, and prediction of therapeutic responses. In this review, we discuss recent advances in the use of patient-derived and engineered 3D tumor models to characterize context-dependent metabolic states and treatment-induced metabolic rewiring. We first review technologies currently available to interrogate metabolism in these systems, including optical metabolic imaging, spatial metabolomics, isotope tracing, and bioenergetic profiling. We then discuss how 3D tumor models are used to investigate metabolic interactions within the tumor microenvironment (TME), including stromal and immune crosstalk, acidosis, nutrient availability, and circadian regulation. Finally, we critically examine current limitations, particularly the insufficient physiological relevance of standard organoid culture conditions for metabolic studies, and discuss how advanced engineering approaches and computational modeling may contribute to metabolism-driven functional precision oncology. Full article

Background/Objectives: Sacituzumab govitecan (SG, IMMU-132) is a TROP2-directed antibody–drug conjugate carrying SN-38. Patients with colorectal liver metastasis (CRLM) still have limited treatment options after first-line systemic therapy, and the preclinical pharmacological value of TROP2-directed SN-38 delivery in CRLM remains insufficiently defined. Methods: Public single-cell RNA-sequencing data were reanalyzed to explore the distribution of TACSTD2/TROP2-positive epithelial-associated cells in adjacent normal tissues, primary colorectal cancer, and CRLM. The prognostic relevance of TACSTD2 was evaluated using the Kaplan–Meier Plotter database. TROP2-knockdown and TROP2-overexpressing colorectal cancer models were used to assess IMMU-132 response in vitro and in vivo. Pharmacological antitumor activity was further evaluated using subcutaneous xenografts, syngeneic intrasplenic liver metastasis models, and CRLM patient-derived organoids (PDOs). Transcriptomic profiling and γ-H2AX immunofluorescence were used to explore treatment-associated molecular changes. Results: At the patient/sample level, TACSTD2/TROP2-positive epithelial-associated cells showed numerically higher proportions in primary colorectal tumors and liver metastases than in adjacent normal tissues, and high TACSTD2 expression was associated with inferior overall survival in a public survival database. TROP2 knockdown attenuated IMMU-132-induced growth inhibition, apoptosis, and suppression of colony formation. In vivo, IMMU-132 suppressed colorectal tumor growth and reduced liver metastatic burden, with more evident activity in human TROP2-overexpressing models. In a limited exploratory CRLM PDO cohort established after first-line therapy, liver metastasis-derived PDOs showed lower normalized AUC values than primary tumor-derived PDOs. Transcriptomic analysis and representative γ-H2AX immunofluorescence suggested that IMMU-132 treatment was associated with changes in adhesion/cytoskeletal-related pathways, Wnt/cancer-associated transcriptional programs, and DNA damage-associated signals. Conclusions: These in vitro, in vivo, and PDO-based findings support further preclinical pharmacological evaluation of TROP2-directed SN-38 delivery by IMMU-132 in biomarker-annotated CRLM models, particularly in the post-first-line systemic therapy setting. Full article

Background: Patient-derived functional models have emerged as promising translational platforms capable of reproducing tumour-specific treatment sensitivity patterns, which could be used to personalise neoadjuvant treatment for patients with rectal cancer. Herein, we aimed to summarise the current comparative evidence in a meta-analytical framework on radiotherapy response between preclinical platforms and matched patient data. Methods: A systematic review was performed according to PRISMA principles to identify studies evaluating patient-derived functional models for the prediction of radiotherapy or chemoradiotherapy response in rectal cancer. Study characteristics, experimental protocols, predictive performance and clinical correlations were extracted. An exploratory hierarchical summary receiver operating characteristic (HSROC) meta-analysis was performed using studies providing sufficient data. Results: Eight studies involving patient-derived organoids and zebrafish patient-derived xenograft models were included. Most studies evaluated locally advanced rectal cancer treated with neoadjuvant chemoradiotherapy. The included studies demonstrated concordance rates ranging from 78% to 100% between ex vivo functional responses and matched clinical treatment outcomes. Reported predictive performance was favourable, with Yao et al. demonstrating 85.0% concordance, 78.0% sensitivity and 92.0% specificity, while Hsu et al. reported 87.5% sensitivity and 100% specificity using radiobiological modelling. Exploratory HSROC analysis demonstrated overall favourable discriminatory performance for prediction of treatment resistance and poor response. Conclusions: Patient-derived functional models, particularly PDOs, demonstrate promising potential as predictive biomarkers for radiotherapy and chemoradiotherapy response in rectal cancer. Although the current evidence remains exploratory and is limited by methodological heterogeneity and small cohorts, these platforms represent a promising translational strategy in precision radiation oncology, warranting prospective multicentre validation. Full article

Preclinical bladder cancer models are often judged by tumor take, tumor growth or treatment inhibition, yet these endpoints do not reveal which bladder-specific constraints a given model preserves or bypasses. The bladder is shaped by a specialized urothelial barrier, urine exposure, cyclic filling and emptying, inflammatory injury, metabolic stress and intravesical treatment pressure. In this review, we use susceptibility engineering as an organizing framework for model selection and validation. We define susceptibility engineering as the deliberate definition, perturbation and reporting of model states that alter tumor initiation, adhesion, colonization, survival or therapeutic exposure. This framework groups cell lines, patient-derived organoids, cell-line-derived xenograft (CDX) and patient-derived xenograft (PDX) models, orthotopic transplantation, N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN)-induced tumors, genetically engineered mouse models and large-animal platforms according to the biological constraints they test. We focus on three linked dimensions: urothelial barrier integrity and uroplakin-related tools; local colonization thresholds under bladder-specific selection; metabolic susceptibility involving peroxisome proliferator-activated receptor gamma (PPARG)-associated differentiation programs and candidate solute carrier family 25 (SLC25)-linked mitochondrial stress nodes. We further distinguish large-animal systems as platforms for local delivery, imaging, device testing and procedural scale rather than universal substitutes for mouse models. A susceptibility-based validation framework could improve model selection, explain divergent responses across systems and support tiered platforms that connect patient-derived biology, mechanistic mouse studies and clinically realistic intravesical evaluation. Full article

Colon cancer is one of the leading causes of death, requiring advanced therapies that need models for developing new drugs. Conventional cell culture models do not accurately and precisely reproduce the complexity of the tumor microenvironment, limiting their usefulness in research and therapy development. To address this weakness, patient-derived organoids have emerged as promising in vitro models. The implementation of these organoid-based models into more physiologically relevant systems is expected to improve their clinical relevance. Thus, integrating these organoids into microfluidic chips acting as bioreactors will likely improve the predictive response of therapies in personalized medicine. In this article, we report the development of a new colon cancer organoid-on-a-chip model that enables the in vitro culture of patient-derived colon cancer organoids under continuous culture media flow. We demonstrate how the developed organoids were derived from tumor biopsies of patients with colorectal cancer, expanded in standard three-dimensional (3D) culture, and cultured inside the microfluidic chips. The microfluidic chip chambers are designed to house organoids in a controlled environment, allowing the injection of therapies and monitoring by optical microscopy in real time. The in vitro therapies tested were a combination of drugs based on 5-fluorouracil and oxaliplatin at different concentrations. As a result, we demonstrate for the first time that this model proves the capability of this technology for in vitro testing colon cancer therapies. Full article

Reactive oxygen species (ROS)-regulated antioxidant defense is closely linked to non-small cell lung cancer (NSCLC) progression and therapy resistance. Wogonin (WGN), a flavonoid from Scutellaria baicalensis, has antitumor activity, but whether it is associated with ROS-dependent ferroptotic and mitochondrial stress in NSCLC remains incompletely defined. A549 and BEAS-2B cells, male BALB/c nude mouse A549 xenografts, patient-derived NSCLC organoids, and public transcriptomic cohorts were analyzed using viability, colony formation, migration/invasion, DCFH-DA ROS, JC-1, Annexin V/PI, Fe²⁺ and lipid ROS probes, RT-qPCR, Western blotting, immunofluorescence, inhibitor rescue, and c-Myc gain- and loss-of-function assays. WGN suppressed A549 growth and motility with weaker effects on BEAS-2B cells. WGN markedly increased intracellular ROS, Fe²⁺ accumulation and lipid peroxidation, decreased mitochondrial membrane potential, promoted Caspase-related apoptosis, reduced c-Myc/GPX4 and SLC7A11, and increased ACSL4. N-acetylcysteine, Z-VAD-FMK and Ferrostatin-1 partially rescued WGN-induced injury. c-Myc overexpression partially restored GPX4 and reduced lipid ROS/Fe²⁺ accumulation, whereas c-Myc knockdown decreased GPX4. Xenografts and organoids reproduced tumor inhibition and selected redox-associated molecular changes. Collectively, WGN suppresses A549-associated NSCLC phenotypes in association with ROS accumulation, ferroptosis-related lipid injury, mitochondrial dysfunction-associated apoptosis, and c-Myc/GPX4 downregulation. Full article

Current inflammatory bowel disease (IBD) therapies suppress immune pathways, yet epithelial recovery can remain incomplete. This review evaluates whether intestinal chloride and bicarbonate transport can support a distinct, adjunctive pharmacological strategy. Rather than cataloguing transport proteins, we compare the cystic fibrosis transmembrane conductance regulator (CFTR), SLC26A3/down-regulated in adenoma (DRA), and TMEM16A/ANO1 against an evidence hierarchy of human disease relevance, causal epithelial biology, pharmacological tractability, target engagement, functional rescue, and developability. CFTR and DRA form the most coherent module linking bicarbonate availability to mucin expansion, epithelial surface pH, fluid balance, and barrier organization, but their liabilities differ. CFTR is structurally and clinically druggable, yet its modulators are genotype-directed, and broad activation may worsen diarrhea. DRA has stronger evidence for a colonic barrier role and emerging support from human organoids, but no validated activator or stabilizer. TMEM16A has abundant chemical tools, yet uncertain selectivity, wide extra-epithelial expression, and no established disease-modifying role in IBD. No intervention has achieved mucosal healing through anion-transport rescue in IBD. We therefore define the decisive experiments required before translation: confirmation of persistent functional defects in human tissue, selective exposure-linked rescue in patient-derived epithelium, direct target engagement, and protection against hypersecretion or electrolyte imbalance. The evidence supports focused, mechanism-based evaluation of the CFTR-DRA axis rather than empirical repurposing. Full article

Alzheimer’s Disease (AD) is a multifaceted progressive neurodegenerative disease characterized by memory deficits and cognitive impairment. The disease is clinically diagnosed by the presence of β-amyloid (Aβ), hyperphosphorylated tau, and neurodegeneration. Animal models serve as an indispensable tool to understanding AD pathogenesis and evaluating potential therapeutic approaches. However, despite the development of more than 200 rodent models, species-specific differences limit the translational relevance. Brain organoids generated from induced pluripotent stem cells (iPSCs) have the potential to bridge the gap between transgenic mouse models and clinical trials in human patients. Herein, we describe a robust stimulus-based neuroimmune organoid model that demonstrates neuroinflammation, neurodegeneration, and lipid dysregulation with AD-relevant pathological markers. Using planar organoids containing neurons, astrocytes, microglia, and vascular cells, we performed in-depth characterization of the induced AD-like phenotype using supernatant proteomic analysis, immunofluorescence staining, NfL and GFAP release, scRNAseq, bulk RNAseq, and pathway analysis both acutely (24 h) and chronically (7 days). Furthermore, we show that the induced neuroinflammation, lipid dysregulation, and neurodegeneration can be ameliorated using small molecules. This defined inducible model system presents an opportunity for drug discovery and development using a complex multicellular brain microenvironment derived from human iPSCs. Full article

Chemotherapy resistance remains a major challenge in colorectal cancer (CRC) treatment, necessitating novel adjuvant strategies. This study employed an integrated analytical strategy combining network pharmacology, single-cell RNA sequencing (scRNA-seq) of patient-derived organoids (PDOs) and molecular dynamics simulations to identify bioactive compounds from Ganoderma lucidum and elucidate their chemo-sensitizing mechanisms. Network pharmacology identified five bioactive components of G. lucidum, corresponding to 267 potential targets. Integration with transcriptomic data, weighted gene co-expression network analysis (WGCNA), and known CRC genes refined these to 19 core targets. Cross-referencing with scRNA-seq data from irinotecan-treated PDOs pinpointed cyclin-dependent kinase 2 (CDK2) and Aurora kinase A (AURKA) as pivotal targets. Molecular dynamics simulations confirmed stable binding of the key component cerevisterol to both CDK2 and AURKA proteins, with binding free energies of −120.67 kJ/mol and −134.47 kJ/mol, respectively. In vitro cell viability assays across multiple CRC cell lines (HCT116, RKO, and HT-29) and PDOs demonstrated that cerevisterol significantly sensitized CRC cells to irinotecan (SN38). Notably, we observed that CDK2 was preferentially enriched in MSI-H tumors, whereas AURKA was enriched in MSS tumors, suggesting the potential of MSI status as a biomarker for patient stratification. Collectively, these findings identify cerevisterol as a dual-targeting natural product that modulates CDK2 and AURKA to overcome chemotherapy resistance, providing a quantitative analytical framework for discovering bioactive compounds and their molecular targets from medicinal fungi. Full article