中文导读
药物发现成本高、周期长。本文综述类器官与器官芯片如何提升筛选的生理相关性与预测性，加速药物发现。
以下为英文全文（English full text）
1. Introduction
Drug discovery remains one of the most challenging and expensive endeavors in modern biomedical research. Despite advances in molecular biology and computational chemistry, the translation of preclinical findings into clinical success continues to face significant hurdles. Traditional two-dimensional (2D) cell cultures and animal models, while valuable, often fail to recapitulate the complex physiology of human tissues, leading to high attrition rates in clinical trials. Organoids—self-organizing three-dimensional (3D) tissue cultures derived from stem cells—and organ-on-chip (OoC) microphysiological systems represent a paradigm shift in preclinical drug screening, offering human-relevant models that bridge the gap between simple cell cultures and animal studies.
This article provides a comprehensive overview of how organoid and organ-on-chip technologies are transforming drug screening across the pharmaceutical pipeline, from early-stage high-throughput compound screening to personalized medicine applications.
2. The Evolution of Drug Screening Models: From 2D to 3D to Organ-on-Chip
2.1 Two-Dimensional Cell Culture Era
For decades, immortalized cell lines grown on flat plastic surfaces have served as the workhorse of in vitro pharmacology. Models such as HeLa, HEK293, and HepG2 enabled rapid, scalable screening of thousands of compounds. However, 2D cultures fundamentally alter cellular morphology, polarity, and signaling pathways. Cells in monolayers lack the complex cell-cell and cell-matrix interactions that define native tissue architecture, leading to altered gene expression profiles and drug response phenotypes that poorly correlate with human in vivo behavior.
2.2 The Emergence of Three-Dimensional Organoid Culture
The development of organoid technology, pioneered by the seminal work of Clevers and colleagues on intestinal stem cell-derived organoids, marked a transformative leap in tissue modeling. Organoids self-assemble into structures that recapitulate key architectural features of their tissue of origin, including luminal organization, stem cell niches, and differentiated cell types. Unlike spheroids—which are typically aggregates of cells without inherent tissue architecture—organoids exhibit active self-organization driven by endogenous developmental programs.
Patient-derived organoids (PDOs) established from tumor biopsies or resected tissues have proven particularly powerful for drug screening. Driehuis et al. established a platform of 27 patient-derived pancreatic cancer organoids and screened them against a panel of 76 therapeutic agents, demonstrating that PDO responses could stratify patients by therapeutic sensitivity. Similarly, de Witte et al. showed that PDOs derived from ovarian cancer patients closely mimicked clinical responses and exhibited heterogeneous inter- and intrapatient drug responses, underscoring the value of patient-specific models in predicting treatment outcomes.
2.3 Organ-on-Chip: Microphysiological Systems for Dynamic Tissue Modeling
While organoids capture tissue architecture, they traditionally lack the dynamic physiological cues present in vivo—such as fluid shear stress, tissue-tissue interfaces, and vascular perfusion. Organ-on-chip technology addresses these limitations by integrating living cells within microfluidic devices that mimic the physical and biochemical microenvironment of native tissues. First developed by Ingber and colleagues at the Wyss Institute, organ chips use flexible, porous membranes to create tissue-tissue interfaces (e.g., epithelium-endothelium), with controlled media flow enabling nutrient delivery, waste removal, and physiologically relevant mechanical cues.
McAleer et al. demonstrated that a multi-organ chip integrating liver and heart tissues could recapitulate drug metabolism-dependent toxicity, revealing cardiotoxic effects of metabolites that remained undetected in static cultures. Shroff et al. (2022) provided a comprehensive review of multi-organ chips for metabolism studies, noting that these systems enable real-time assessment of organ cross-talk during drug exposure.
3. Advantages of Organoids in Drug Screening
3.1 Patient Specificity and Genetic Fidelity
Organoids established from patient tissues retain the genetic and epigenetic landscape of the donor. Tiriac et al. and Gahesh et al. demonstrated that PDOs preserve the mutational profiles and tissue identity of the original tumor, making them ideal platforms for precision medicine. Ooft et al. and Pasch et al. independently validated that PDO drug responses mirror clinical patient responses within clinically meaningful timeframes.
3.2 Preservation of Tumor Heterogeneity
Tumors are inherently heterogeneous, with subclonal populations exhibiting differential drug sensitivities. Traditional cell lines, often derived from a single clone, fail to capture this complexity. PDOs, by contrast, maintain intratumoral heterogeneity. de Witte et al. demonstrated that PDOs established from different regions of the same ovarian cancer patient exhibited distinct drug response profiles, highlighting the importance of multi-region sampling for comprehensive therapeutic assessment.
3.3 Physiological Relevance
Organoids recapitulate tissue-specific functions that are absent or diminished in 2D cultures. Liver organoids express cytochrome P450 (CYP) enzymes at levels approaching primary human hepatocytes, enabling physiologically relevant drug metabolism studies. Kidney organoids develop proximal tubule-like structures with functional transporter expression, critical for nephrotoxicity assessment. Cardiac organoids derived from human induced pluripotent stem cells (iPSCs) exhibit spontaneous contractility and electrophysiological properties suitable for cardiotoxicity screening.
4. Organ-on-Chip Applications in Drug Screening
4.1 ADMET Studies
Absorption, Distribution, Metabolism, Excretion, and Toxicity (ADMET) properties determine the fate of a drug in the human body and are major contributors to clinical failure. Organ-on-chip platforms offer unprecedented opportunities for integrated ADMET assessment. Shuler and colleagues constructed a co-culture model of Caco-2 intestinal cells and hepatocytes that successfully matched 22 of 24 clinically relevant oral bioavailability predictions. Shroff et al. (2022) reviewed multi-organ chips specifically designed for ADMET studies, including gut-liver, liver-kidney, and blood-brain barrier systems.
4.2 Efficacy Screening
Beyond ADMET, organ chips enable efficacy screening in physiologically relevant contexts. Tumor-on-chip systems incorporating vascularized microenvironments allow researchers to study drug penetration, tumor-stroma interactions, and immune cell infiltration. Griep et al. and Sung et al. demonstrated that multi-organ chips could be configured to model systemic drug distribution and organ-specific responses, enabling simultaneous assessment of target efficacy and off-target effects.
4.3 Toxicity Assessment
Organ chips are increasingly used for organ-specific toxicity profiling. A kidney-on-chip developed by Emulate Bio integrated tissue-embedded microsensors for oxygen, glucose, lactate, and glutamine, providing real-time assessment of nephrotoxicity. The system uncovered a previously unknown mechanism of cisplatin-induced injury involving glucose transport and predicted the protective effects of SGLT2 inhibitors—subsequently validated through retrospective clinical analysis of 247 patients.
5. High-Throughput Screening Platforms and Technologies
5.1 Automation and Miniaturization
The scalability of organoid-based screening has been significantly enhanced by advances in automation. Ma et al. (2025) described automated organoid platforms integrated with microfluidic technologies that enable parallel screening of hundreds to thousands of compounds. Schuster et al. highlighted the importance of microfluidic organoid chips and arrayed seeding approaches that miniaturize culture into multiwell plate formats, enabling high-throughput processing. Platforms now range from approximately 20 to 240 organoids per chip, with some systems compatible with 384-well formats for robotic screening.
5.2 High-Content Imaging and Analysis
Integration of automated liquid handling with high-content imaging and computational phenotyping has accelerated organoid-based screening. CellTiter-Glo 3D assays quantify ATP levels in PDO cultures as a measure of viability, while advanced imaging platforms enable multidimensional readouts including cell survival, mitochondrial function, and neural network dynamics. Phan et al. (2019) demonstrated a simple high-throughput approach using PDOs to identify actionable drug sensitivities, emphasizing the feasibility of scaling PDO screening for clinical applications.
6. Combined Organoid and Organ-on-Chip Screening Strategies
The integration of organoid biology with microfluidic engineering yields hybrid models that combine the tissue authenticity of organoids with the physiological control of organ chips. Organoid-on-chip platforms use micropillars or microwells to position organoids within perfusable microfluidic chambers, enabling continuous feeding, real-time imaging, and controlled exposure to drugs, metabolites, and immune cells. Zhu et al. and Wang et al. developed higher-throughput organoid-on-chip platforms that support complete differentiation workflows from embryoid body aggregation to maturation, eliminating manual transfer steps and improving reproducibility.
Novak et al. (2024) introduced a dynamic Microphysiological System Chip Platform (MSCP) with customizable 4-organ integration, demonstrating an intestine-liver-heart-lung cancer microphysiological system for multidimensional oral anti-lung cancer drug evaluation. This system evaluates drug efficacy and side effects by simulating physiological communication through fluid-based transport between organs.
7. Personalized Medicine and Companion Diagnostics
7.1 Patient-Derived Organoids as Companion Diagnostics
The predictive accuracy of PDOs in mirroring patient responses has positioned them as potential companion diagnostics. Verstegen et al. (2025), published in Nature Medicine, reviewed clinical applications of human organoids, noting that as of October 2024, ClinicalTrials.gov listed 36 trials using organoids for personalized medicine across multiple cancer types. PDOs are being used to test responses to standard-of-care chemotherapy, targeted therapies, and immunotherapies including CAR-T cell therapy.
Jacob et al. demonstrated that glioblastoma PDOs could predict responses to standard-of-care therapy and CAR-T immunotherapy within clinically relevant timelines. Weeber et al., Fujii et al., Pauli et al., and Vlachogiannis et al. collectively established that metastatic cancer site-derived PDOs effectively evaluate drug response and recapitulate patient outcomes.
7.2 Organoid Biobanks for Population-Scale Screening
Organoid biobanks—living repositories of patient-derived tumor organoids—enable population-scale drug screening and biomarker discovery. Calandrini et al. and van de Wetering et al. described biobanks that preserve the molecular diversity of patient cohorts, facilitating retrospective and prospective drug response studies. These biobanks are now being integrated with genomic and clinical databases to develop predictive algorithms for treatment selection.
8. From Preclinical to Clinical: Translation and Regulatory Considerations
Despite remarkable advances, the clinical translation of organoid and organ-on-chip technologies requires rigorous validation and regulatory acceptance. Zhou et al. (2025) emphasized that prospective studies demonstrating reproducible correlations between organoid responses and patient outcomes are essential before organoid testing can be recognized as a companion diagnostic modality. Regulators are developing guidance on how organoid-derived evidence should be generated, validated, and interpreted.
The FDA Modernization Act 2.0, which removed the mandate for animal testing in drug development, has accelerated interest in human-relevant alternative models. Organoids and organ chips are increasingly recognized as viable components of preclinical evidence packages, particularly when integrated with computational modeling and retrospective clinical validation.
9. Conclusion
Organoid and organ-on-chip technologies represent a fundamental evolution in drug screening, offering human-relevant models that address the limitations of traditional 2D cultures and animal models. From high-throughput compound screening to patient-specific treatment optimization, these platforms are reshaping the pharmaceutical pipeline. As automation, standardization, and regulatory frameworks mature, organoid and organ-on-chip systems will become indispensable tools for accelerating drug discovery, improving safety assessment, and delivering personalized therapies.