SCIENCE CHINA PRESS
REVIEW
Endocrine cancer organoids in basic and translational medical research
Dong Chen11, Zhulan Chen11, Huihui Yang21, Lei Zhang3, Chenchen Hu1, Zhuo Yang1, Peng Li1, Xi Su1, Xiaoling Liu1*, Wei Wei1* & Yongsheng Zhao1*
1Department of Thyroid and Breast Surgery and Department of Nuclear Medicine, Peking University Shenzhen Hospital, Shenzhen 518036, China
2Department of Pharmacy, The Eighth Affiliated Hospital, Sun Yat-sen University, Shenzhen 518033, China 3Department of Radiotherapy, Peking University Shenzhen Hospital, Shenzhen 518036, China
¡Contributed equally to this work *Corresponding authors (Xiaoling Liu, email: liuxiaoling97@aliyun.com; Wei Wei, email: rxwei1123@163.com; Yongsheng Zhao, email: zhaoys69@126.com)
Received 21 November 2024; Accepted 26 February 2025; Published online 5 June 2025
Endocrine cancers are a heterogeneous group of malignancies that originate from cells capable of secreting hormones. Examples include but are not limited to thyroid cancer, adrenocortical carcinoma, prostate cancer, and pancreatic cancer. Our limited understanding of endo- crine cancers is partially due to constraints related to model systems, which cannot accurately replicate the pathogenesis of these tumors. Patient-derived organoids (PDOs) are clusters of multiple cell types that grow in a three-dimensional environment. They have become innovative models that faithfully reproduce genotype and phenotype of the tissues from which they originated, facilitating the prediction of patient treatment responses and guiding the development of precision medicine. This article provides a comprehensive review of the establishment of endocrine cancer PDOs and their applications in cancer research, drug screening, and personalized therapy. These excellent preclinical models have the potential to advance our understanding of endocrine cancers in basic research and clinical practice. In addition, we discuss the challenges related to current organoid technologies and provide future perspectives on the applications of orga- noids in precision medicine to improve the management of endocrine cancers.
patient-derived organoids | endocrine cancers | precision medicine | drug screening | tumor microenvironment
Introduction
The endocrine system plays a critical role in regulating human physiology through the production and secretion of various hormones. Endocrine cells are situated within specialized glands or spread throughout nonendocrine organs, including cells that produce steroids (such as those in the adrenal cortex and ovary), thyroid follicular cells (responsible for synthesizing thyroid hormones and thyroglobulin), and neuroendocrine cells. The interactions between endocrine glands (such as the thyroid gland, thymus, pancreas, adrenal glands, prostate, ovaries, and testes) are highly coordinated (Figure 1). Endocrine cancers are a heterogeneous collection of malignancies that originate from cells capable of producing hormones (Latteyer et al., 2016). Common endocrine tumors include those in the endocrine glands and neuroendocrine tumors. Endocrine cancers often result in abnormal hormone production and affect many important physiological functions, such as growth, development, metabo- lism, sexual function, and reproduction. Although many endocrine tumors are benign, prostate, pancreatic, and ovarian cancers are leading causes of cancer-related death globally (Dasari et al., 2017; He et al., 2024). Therefore, novel treatment strategies and therapeutic targets are greatly needed for these
endocrine cancers. However, few reliable preclinical models exist for the study of potential therapeutic interventions.
Two-dimensional (2D) cell cultures, patient-derived xenografts (PDXs), and genetically engineered animal models are still commonly used in cancer research. However, these models cannot accurately recapitulate the complex biological character- istics of natural human tumors. Therefore, the establishment of high-fidelity preclinical tumor models is crucial for investigating cancer-related mechanisms and for the development of persona- lized anticancer therapies in the clinic.
Over the last ten years, three-dimensional (3D) cultured organoids have been increasingly utilized in preclinical studies to investigate tumor pathophysiology and responses to cancer therapy. Organoids are self-organizing multicellular ex vivo organ models derived from stem cells or primary tissues in specific 3D environments (Fatehullah et al., 2016). Organoids mimic the characteristics of real organs in vivo and can be stably expanded. Patient-derived organoids (PDOs) are usually formed from tumor tissues obtained via surgery or biopsy, and they structurally and functionally resemble the original cancer tissues. In addition, PDOs have been successfully developed using peripheral blood (Gao et al., 2014), ascites (Kopper et al., 2019), and malignant pleural effusion (Chen et al., 2020) samples from patients with
Citation: Chen, D., Chen, Z., Yang, H., Zhang, L., Hu, C., Yang, Z., Li, P., Su, X., Liu, X., Wei, W., et al. (2025). Endocrine cancer organoids in basic and translational medical research. Sci China Life Sci 68, 2842-2866. https://doi.org/10.1007/s11427-024-2888-8
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Pituitary
Thyroid & Parathyroid glands
Breast
Pancreas
Adrenal glands
Prostate
Ovaries
Male
Female
tumors. PDOs can be used to expand small tumor samples and analyze cancer at any stage of cultivation, greatly expanding the number of tumor types that can be propagated and studied in the laboratory (Tuveson and Clevers, 2019). These PDOs accurately recapitulate the pathological phenotype and heterogeneity of parental tissues, as genetic stability and disease-state phenotype are maintained throughout long-term passaging. Notably, PDOs have higher fidelity to primary tumors than traditional cell lines and PDX models (Drost and Clevers, 2018; Veninga and Voest, 2021). PDOs have shown their unique value and potential in recapitulating cancer phenotypes, genetic profiles, and drug sensitivity, rendering them powerful tools for predicting patient treatment responses (Figure 2). The development of organoid models not only helps improve the quality of cancer diagnosis and treatment but also opens new avenues for an in-depth understanding of disease progression and promotes the develop- ment of personalized therapy.
To date, PDOs have been successfully established from various endocrine cancers. However, there is still a lack of systematic studies on organoid technology in endocrine cancers. In this review, we present the latest advancements in PDO models of endocrine cancers, from basic mechanisms to clinical applica- tions, with a particular focus on innovative strategies for cancer modeling, drug assessment, and personalized medicine. We also discuss the limitations and future development prospects of organoid technology.
Establishment of PDOs from endocrine cancers
Submerged Matrigel or basement membrane culture is a classic method for cultivating organoids and can be used for endocrine cancer PDOs. This process includes dissociating original tumor
tissues into a dispersed cell suspension via enzymatic and mechanical methods. The cells are subsequently embedded in a matrix and placed in culture medium to promote the growth of PDOs (Figure 2A). Organoid culture medium must contain some components to maintain heterogeneity in PDOs, including essential components, various pathway inhibitors and growth factors. The common key cytokines required for PDO generation usually include epidermal growth factor (EGF), R-spondin (WNT pathway inducer), and noggin (a BMP antagonist), which collectively promote stem cell growth, differentiation, and self- renewal (Dutta et al., 2017; Sato et al., 2009). Representative culture media for endocrine cancer PDOs are shown in Table 1. R-spondin and noggin are critical components of organoid culture medium (Boj et al., 2015). EGF is a member of a growth factor family that stimulates the proliferation of cells in various organoids, including prostate cancer (Gao et al., 2014), breast cancer (Sachs et al., 2018), and pancreatic cancer (Boj et al., 2015) organoids. B27 supplements comprise a variety of nutrients that promote cell survival, facilitate organoid forma- tion, and prevent adhesion during cultivation. A83-01 is a transforming growth factor ß (TGFß)/activin receptor-like kinase 5 (ALK5) inhibitor that prevents epithelial-to-mesenchymal transition (EMT) in cancer organoids by inhibiting TGF-B expression (Fujii et al., 2016). Y-27632, a Rho-associated coiled-coil protein kinase (ROCK) inhibitor, has been reported to promote organoid growth and prevent anoikis (Watanabe et al., 2007). Additionally, other factors, such as N-acetylcysteine, nicotinamide, and neuregulin 1, which induce differentiation and promote cell survival, play distinct roles in organoid culture (Ma et al., 2022). Notably, the additives and culture conditions for PDOs are not fixed and should be customized depending on cancer types and PDO attributes. Currently, there are no
A Cancer modeling
Surgery or biopsy tissues, ascites, etc.
Dissociation
3D organoid culture
Living biobanks
B Identification and evaluation
MIN m ny
Histopathology
Multi-omics studies
TME interaction
Gene editing
Intra- & Inter-patient heterogenity
C Applications
Drug
Carcinogenesis research
Drug screening & discovery
Drug toxicology & safety
Immunotherapy
Personalized medicine
standardized culture media or protocols for PDO generation. The successful establishment of PDOs can be validated and evaluated by H&E staining, immunohistochemistry, and gene sequencing (Figure 2B). The purpose of these evaluations is to confirm the consistency of PDO features with those of the corresponding tumors, which is a prerequisite for follow-up research.
Pituitary tumor
The pituitary gland is the hub of the human endocrine system, and regulates various physiological processes, such as growth, metabolism, reproduction, and lactation. Pituitary tumors are the third most prevalent type of intracranial tumors, accounting for 10%-15% of all brain tumors (Daly and Beckers, 2020; Melmed et al., 2022). Although most pituitary tumors are benign, symptomatic pituitary adenomas can increase the morbidity and mortality due to hypo- or hypersecretion of hormones. Most pituitary tumors originate from the neuroendo- crine epithelium and are well-differentiated and nonmetastatic; these tumors are called pituitary neuroendocrine tumors (PitNETs) (Asa et al., 2022). Adrenocorticotropic hormone (ACTH)-secreting PitNETs, which stimulate the adrenal glands to produce excess cortisol, lead to Cushing’s disease, a severe endocrine disorder (Cushing, 1933; Loriaux, 2017). The treat-
ment modality for pituitary tumors includes surgical resection, pharmacotherapy, and radiation therapy (Varlamov et al., 2019). Owing to the lack of preclinical models to mimic the complexity of the PitNET microenvironment, we are unable to advance the knowledge of clinical care by implementing tumor- specific therapies that are more efficacious and better tolerated for patients with PitNETs.
Multiple in vitro 2D cell cultures and animal models have been established to reveal the pathobiology of pituitary tumors. Nevertheless, these models fail to depict the full spectrum of tumor histology. While some pituitary spheroid/aggregate/ organoid models have been reported, these cultures are composed of poorly differentiated cells with high replication potential, which may influence drug responses and yield results that are difficult to translate into clinical settings (Tsukada et al., 2013; Zhang et al., 2021a). However, organoids can be used to overcome several of the aforementioned drawbacks.
Several publications on pituitary tumor organoids have appeared in the last few years. Zhang et al. (2021a) developed organoids from corticotrope tumors. Tumor cells were embedded in Matrigel and cultured in optimized medium containing 10% fetal bovine serum and typical organoid growth factors including insulin-like growth factor-1 (IGF-1), EGF, fibroblast growth factor 8 (FGF8), triiodothyronine (T3), thyrotropin
| Pituitary adenoma | Thyroid cancer | Parathyroid hyperplasia | Adrenocortical carcinoma | Pancreatic cancer | Prostate cancer | Ovarian cancer | Breast cancer | |
|---|---|---|---|---|---|---|---|---|
| References | Nys et al., 2022 | Chen et al., 2021a | Noltes et al., 2022 | Arakawa et al., 2024 | Boj et al., 2015 | Gao et al., 2014 | Kopper et al., 2019 | Sachs et al., 2018 |
| Basic medium | Advanced DMEM/ F12 medium | Advanced DMEM/ F12 medium | DMEM/F12 medium | Advanced DMEM/F12 medium | Advanced DMEM/ F12 medium | Advanced DMEM/ F12 medium | Advanced DMEM/ F12 medium | Advanced DMEM/ F12 medium |
| Glutamax | 1% | 1% | 1× | 1× | 1× | 2 mmol L-1 | 1× | 1× |
| HEPES | – | 10 mmol L-1 | 10 mmol L-1 | – | 1× | 10 mmol L-1 | 10 mmol L-1 | 10 mmol L-1 |
| Antibiotic- antimycotic | – | 1% | – | – | – | – | – | – |
| Penicillin/ Streptomycin | – | – | 1× | – | 1× | – | 1× | 1× |
| Primocin | – | – | – | – | 1 mg mL-1 | 1% | 0.2% | 50 µg mL-1 |
| B27 supplement | 2% | 1× | 0.5x | 1x | 1× | 1× | 2% | 1× |
| N-acetylcysteine (mmol L-1) | 1.25 | 1.25 | – | – | 1 | 1.25 | 1.25 | 1.25 |
| N2 | 1% | – | – | 1× | – | – | – | – |
| Nicotinamide | – | 10 mmol L-1 | 100 umol L-1 | – | 10 mmol L-1 | 10 mmol L-1 | 10 mmol L-1 | 5 mmol L-1 |
| Wnt 3a | – | – | 50% conditioned medium | – | 50% conditioned medium | – | – | – |
| R-spondin1 | 200 ng mL-1 | 500 ng mL-1 | 10% conditioned medium | – | 10% conditioned medium | 5% conditioned medium | 10% conditioned medium | 10% conditioned medium |
| Noggin | 100 ng mL-1 | 100 ng mL-1 | 25 ng mL-1 | – | 10% conditioned medium or 0.1 µg mL-1 | 10% conditioned medium | 1% conditioned medium | 100 ng mL-1 |
| EGF (ng mL-1) | 50 | 50 | 20 | 50 | 50 | 50 | 5 | 5 |
| FGF2 (ng mL-1) | 20 | – | – | 20 | – | 1 | – | – |
| FGF7 (ng mL-1) | – | 5 | – | – | – | – | – | 5 |
| FGF8 (ng mL-1) | 200 | – | – | – | – | – | – | – |
| FGF10 (ng mL-1) | 100 | 10 | 50 | – | 100 | 10 | 10 | 20 |
| A83-01 | 0.25 umol L-1 | 500 nmol L-1 | 5 umol L-1 | – | 0.5 umol L-1 | 0.5 umol L-1 | 500 nmol L-1 | 500 nmol L-1 |
| SB202190 | 100 nmol L-1 | 10 pmol L-1 | – | – | – | 10 pmol L-1 | – | 500 nmol L-1 |
| Y-27632 (umol L-1) | – | 10 | 10 | 10 | 10 | 10 | 5 | 5 |
| Special reagents | HGF: 20 ng mL-1 SHH: 100 ng mL-1 Cholera Toxin: 100 ng mL-1 IL-6: 20 ng mL-1 IGF-1: 40 ng mL-1 | – | VEGF-121: 10 ng mL-1 Angiotensin: 0.1 nmol mL-1 | IGF-2: 20 ng mL-1 | Gastrin: 10 nmol L-1 | DHT: 0.1 or 1 nmol L-1 | ß-Estradiol: 100 nmol L-1 Hydrocortisone: 0.5 µg mL-1 Forskolin: 10 umol L-1 Heregulinß-1: 37.5 ng mL-1 | Neuregulin 1: 5 nmol L-1 |
a) DHT, dihydrotestosterone; EGF, epidermal growth factor; FGF, fibroblast growth factor; HGF, hepatocyte growth factor; IGF, insulin-like growth factor; IL-6, interleukin-6; SHH, sonic hedgehog; VEGF-121, vascular endothelial growth factor 121.
releasing hormone (TRH), and bovine hypothalamus extract. It was observed that the tumor organoids contained corticotrope (ACTH+), stem/progenitor (SOX2+), and intermediate (PITX1+, TBX19+) cell types after 5 weeks of culture. In the 2D culture system derived from primary tumor tissue, ACTH secretion was rapidly lost, whereas in the 3D organoid culture system, hormone secretion continued for up to 18 weeks. Nys et al. (2022) successfully developed organoids from different types of pituitary tumors, such as hormone-producing/functioning and non-hormonal/non-functioning tumors. The isolated cells were embedded in a matrix and cultured in optimized medium supplemented with hepatocyte growth factor (HGF) and IL-6. These organoids recapitulated the histological features and stem cell phenotypes of the original tumor samples. Unfortunately, these organoids could not be expanded, which was consistent with the observed upregulation of pro-apoptotic and hypoxia- induced genes, as well as the downregulation of pro-survival genes in the organoids compared with the parental tissues. Using
a slightly adapted protocol, PitNET organoids were generated from 35 tumor samples of Cushing’s disease patients; the differentiated cell lines and stem/progenitor cells exhibited multicellular properties closely resembling the patients’ adeno- mas in terms of the pathology and complexity (Chakrabarti et al., 2022). The organoids consisted of multiple cell types, including stem cells from functional tumors and organoids, which successfully secreted ACTH in vitro. Whole-exome sequencing revealed that these organoids harbored the same genetic changes as the original tumor. Cui et al. (2025) developed organoids from different PitNET types, with an overall success rate of 87.5%. Compared with functional PitNETs (6/9, 66.7%), organoids derived from clinically nonfunctional Pit- NETs (22/23, 95.7%) were more likely to be successfully generated. The cellular structure, mutational characteristics, subtype-specific neuroendocrine profiles, and tumor microenvir- onment (TME) heterogeneity of PitNET samples were preserved in PDOs.
Thyroid cancer
Thyroid cancer is the most prevalent endocrine cancer with a rising incidence. In the thyroid gland, papillary thyroid cancer (PTC), follicular thyroid cancer (FTC), and anaplastic thyroid cancer (ATC) originate from follicular cells, while medullary thyroid cancer (MTC) arises from parafollicular cells. Differen- tiated thyroid cancers (DTCs), including PTCs and FTCs, account for more than 90% of thyroid malignancies, with PTCs being the most common. The treatment of DTCs typically involves surgical thyroid resection followed by complete ablation with radioactive iodine. PTC generally has a good prognosis, and the 5-year survival rate ranges from 95% to 97%. However, approximately 20% of patients present with tumor recurrence, metastasis, and radioactive iodine-refractory disease (RAIRD) within 10 years. Furthermore, ATC and MTC are very rare and aggressive types of thyroid cancer, suggesting the need for systemic treatment (Saini et al., 2018; Sampson et al., 2007). The emergence of organoid models has provided a powerful preclinical tool for the treatment of thyroid cancer. Research on thyroid cancer organoids is limited and has focused mostly on surgically excised thyroid cancer tissues. In 2021, our group successfully established organoid lines derived from PTC patients (Chen et al., 2021a). Cells were isolated and collected from freshly resected PTC (n=14) and nodular thyroid goiter (n=4) samples, and organoids were generated from 10 out of 14 PTC samples and 3 out of 4 NTG samples. Interestingly, the histopathological characteristics, genetic muta- tions, and somatic copy number variations of the parental tumors were retained in the PTC organoids. In 2023, we presented a detailed, step-by-step procedure for the generation of PTC organoids from clinical specimens. The success rate of developing PTC organoids from clinical samples was 77.6% (Yang et al., 2023). Vilgelm et al. (2020) described a simple fine-needle biopsy technique that can be employed to establish PDOs from various cancer types, including thyroid cancer. They then outlined the potential downstream applications of this technology, such as immune cell characterization, high-throughput chemotherapy screening and in vivo PDO xenograft establishment.
PTC organoids derived from patient specimens may be a potential diagnostic tool for radioactive iodine-resistant patients. Sondorp et al. (2020) generated PDOs from 13 patients with PTC and 3 patients with RAIRD. The PTC and RAIRD organoids and their corresponding source tissues had similar expression profiles for thyroid-specific and transporter/receptor markers. Substan- tial differences in gene and protein expression were observed between PTC and RAIRD organoids, implying potential biomar- kers that could enable the early identification of radioactive iodine-resistant patients before treatment. Pecce et al. (2022) reported a method for the development and long-term main- tenance of PDOs from PTC, lymph node metastasis of PTC, and ATC. These organoids retained the phenotypic and genetic characteristics of their original tissues, including key mutations in the mature thyroid cancer driver genes BRAF, N-K-H-RAS, PTEN, and TP53. MTC-derived organoids have also been established, but they only secreted calcitonin before 3-4 passages, indicating a potential limitation in short-term experi- ments utilizing these MTC organoids (Baregamian et al., 2023). Thyroid cancer-derived organoids can exhibit histopathological properties similar to those of primary tumors. This property can be utilized to elucidate the mechanisms of tumorigenesis and progression and to identify effective targets for intervention.
Additionally, these organoids can be used for the personalized diagnosis and treatment of thyroid cancer patients.
Parathyroidoma
The parathyroid glands are important endocrine organs that secrete parathyroid hormone to regulate calcium homeostasis (Abate and Clarke, 2017; Khundmiri et al., 2016). The parathyroid glands typically include four delicate structures that are 3-4 mm in size and are located on the side of and behind the thyroid gland. The parathyroid glands are composed of master cells that express calcium-sensing receptors (CaSRs) and regulate the blood calcium concentration by mediating the secretion of parathyroid hormone (PTH) (Ritter et al., 2012). Primary hyperparathyroidism (PHPT) is the predominant type of para- thyroid dysfunction, with parathyroid adenomas accounting for the vast majority of PHPT cases. As the main regulator of calcium metabolism, the parathyroid glands play a vital role in the metabolism of the entire body, and hyperfunctional parathyroid glands disrupt major metabolic pathways.
Parathyroid organoids provide a viable option for the study of parathyroid disease, and are new and efficient modeling tools for in vitro and in vivo experiments. Noltes et al. (2022) isolated human parathyroid stem cells from original tissues and studied their potential for in vitro expansion and formation of parathyroid organoids to generate functional parathyroid tissues. Further- more, parathyroid organoids were evaluated for use in physio- logical studies and their potential for use in the identification of new therapeutic targets, drug screening, and imaging tracer testing. Sekhar et al. (2023) performed global, non-targeted metabolomics and live-cell functional metabolism studies using parathyroid organoids. Under semi-solid culture conditions, PDOs were developed from human PHPT tissues (n=6) using a fine-needle aspiration technique. The organoids accurately reproduce the metabolic profile of PHPT tissues, providing a detailed representation of the metabolic microenvironment complexity of parathyroid tissues. Non-targeted metabolomics revealed high fidelity of metabolites between PDOs and the corresponding tissues, indicating a lack of significant variation between PDOs and tissue cohort metabolites, with only some PDOs or tissue-specific metabolite signatures present. The differences in parathyroid-specific metabolites and metabolic pathway changes observed in PDOs are probably related to the metabolic adaptations required for the formation, proliferation, equilibrium, and survival of parathyroid cells under 3D in vitro conditions. Likewise, a minor fraction of parathyroid tissue- specific metabolites that were not detected in PDOs may be complementary to the reproduction of the overall metabolic profile of PHPT tissues, but may also be unrelated to parathyroid metabolism under in vitro conditions (Sekhar et al., 2023). Baregamian et al. (2023) successfully developed parathyroid tumor organoids. Transmission electron microscopy images of parathyroid adenoma PDOs under higher magnification of the cell body revealed membrane-bound granular structures and a greater abundance of elongated mitochondria. Importantly, these PDOs retained intact CaSR expression and function in regulating PTH secretion in response to calcium fluctuations, offering a vital opportunity for future research to focus on targeting CaSRs to manage permanent hypoparathyroidism or persistent hyperparathyroidism in patients with benign or malignant parathyroid disease (Baregamian et al., 2023).
Adrenocortical cancer
The adrenal gland contains two embryologically different areas: the cortex and the medulla. In general, tumors of the adrenal cortex can be classified into adenomas and carcinomas. Adrenocortical adenoma (ACA) is a benign tumor that originates from the secretory cells of the adrenal gland, with an incidence of 425/100,000, representing 33% to 96% of aberrant adrenal incidentalomas (Terzolo et al., 2011). In contrast to the relatively high incidence of ACA, adrenocortical carcinoma (ACC) is a very rare malignant endocrine tumor with a poor prognosis, where the median 5-year overall survival rate is only 22%-44%, varying on the basis of staging and surgical selection (Ayala- Ramirez et al., 2013). In addition, ACC is related to cancer predisposition syndromes, including Li Fraumeni syndrome, Beckwith-Weidemann syndrome, Lynch syndrome, Werner syndrome, and congenital adrenal hyperplasia (Batalini et al., 2019; Else et al., 2014; Lapunzina, 2005). Owing to its rarity, clinical data are scarce, and currently, only a limited number of cell lines and PDXs have been developed in the last three decades. Therefore, the development of novel preclinical models for ACC is essential in mechanistic research and drug development.
Over the past few years, limited ACC organoid or 3D tumor platform studies have been published on either cell lines or patient-derived tissues. Before 2022, 3D models of ACC were composed only of spheroids generated from H295R and SW-13 cells, mainly employed in drug-screening strategies (Cerquetti et al., 2021; Nilubol et al., 2012). Haider et al. (2020) utilized 3D ACC cell line spheroids to assess drug delivery efficiency of mitotane versus mitotane delivered in the form of micelles. Another study reported successful generation of Matrigel-based organoids from ACCs and adrenal neoplasms in vitro. Transmis- sion electron microscopy of ACC PDOs showed abundant secretory granules. These PDOs formed within a timeframe of 1-3 weeks and sustained cortisol secretion for only 2-3 passages (Baregamian et al., 2023). Arakawa et al. (2024) generated six ACC organoids from surgical specimens and examined their response to an identified compound, TAK-243. Although these organoids could not be passaged for a long time, the authors were able to conduct drug testing within the initial 2-3 passages. They also established organoids from ACC PDX models. Immunostain- ing of the PDX-derived organoids was positive for ACC-specific molecules, such as ß-catenin, inhibin-a, SF-1, and INSM-1. Dedhia et al. (2023) utilized matrix metalloproteinase experi- ments to study the metastasis of ACC in organoids and microfluidic models, and identified key differences between tumor cells that can and cannot metastasize through RNA sequencing to identify new intervention targets.
Unlike ACCs, pheochromocytomas (PCCs) and paragangliomas (PGLs), collectively known as PPGLs, arise from adrenal medullary chromaffin cells or the sympathetic or parasympa- thetic paraganglia. Approximately 10%-15% of PCCs and 35%- 40% of PGLs are metastatic (Amar et al., 2005; Patel et al., 2020). PPGLs generally have slow growth rates, with an estimated doubling time of 4-7 years, which makes it difficult to model cell lines of the adrenal medulla. Patient-derived spheroids from four PPGL patients demonstrated differential drug sensitivity (Wang et al., 2022a). At the 2023 AACR conference, Calucho et al. (2023) reported that they established 10 PPGL organoids with conserved secretory profiles and drug respon- siveness similar to those of the original tumors. The generation of
additional cell lines or PDOs and pairing such models with metastasis-on-a-chip platforms can enhance our knowledge of these diseases including drivers, biomarkers of malignancy, and targeted therapies.
Pancreatic cancer
The pancreas contains both exocrine and endocrine components. The exocrine tissue makes up more than 95% of the pancreas, whereas 1%-2% of the tissue performs endocrine functions (Arutyunyan et al., 2020). The endocrine pancreas is composed of the islets of Langerhans, which are made up of various types of cells that produce hormones to regulate whole-body energy balance and the function of the exocrine pancreas. Pancreatic cancer is one of the most aggressive and deadliest human malignancies, with pancreatic ductal adenocarcinoma (PDAC) accounting for 85%-90% of all pancreatic tumors, while pancreatic neuroendocrine tumors (NETs) accounting for less than 5% (Ho et al., 2020; Werner et al., 2023). PDAC is often characterized by rapid progression, poor prognosis, high hetero- geneity, and intricate heterocellular systems (Siegel et al., 2022). Despite advancements in diagnostic tools and treatment approaches for PDAC, the 5-year overall survival rate for patients remains as low as 10% (Park et al., 2021). Heterogeneity and individual variability significantly hinder the effectiveness of standardized medical therapies, resulting in variable patient responses to chemotherapy (Connor and Gallinger, 2022). Clinicians often rely on their clinical experience to select drug treatments for individual patients. Therefore, a high-fidelity preclinical model that can mimic the genetic heterogeneity and complex microenvironment of pancreatic cancer for personalized detection and therapy is urgently needed.
In recent years, rapid advancements have been made in the application of PDOs in preclinical research on PDAC, with researchers focusing on optimizing the growth conditions and characterizing PDOs to assess their suitability as representations of patient tumors. Several groups have established a biobank of living PDAC organoids derived from primary tumors, surround- ing healthy tissue, and metastatic lesions, with a success rate of 70%-85%. PDAC organoids have been shown to accurately reflect the genomic, molecular, and structural characteristics of primary tumors, making them valuable tools for studying cancer in patients with local, late-stage, and metastatic diseases (Boj et al., 2015; Driehuis et al., 2019; Huang et al., 2015; Romero- Calvo et al., 2019; Tiriac et al., 2018). Pancreatic cancer organoids have been established and can be used to study tumor biology and facilitate clinical applications.
Boj et al. (2015) described the development of tumor organoids from human PDAC tissues and biopsies. There PDOs could be passaged continuously and cryopreserved, displayed ductal features, and closely resembled original tumors in terms of disease stage, tumor organization, and pathology. Upon ortho- topic transplantation into mice, these organoids formed invasive malignant tumors with poorly defined glandular structures and a noticeable desmoplastic reaction. These PDOs were applied to identify new genes upregulated in PDAC, indicating that they can be used to study pathways involved in pancreatic cancer development. These findings will help in the development of new therapeutic and diagnostic strategies. In a similar study in 2015, Huang et al. (2015) successfully generated PDAC PDOs from primary tumors, with these PDOs maintaining the tumor
differentiation status, heterogeneity, histoarchitecture, and patient-specific physiologic characteristics. PDAC organoids also formed tumors in xenograft models within 4-7 weeks when a small number of cells were injected into NSG mice. Seino et al. (2018) established PDAC organoid models from 39 patients and identified 3 functional subtypes according to their stem cell niche factor reliance on R-spondin and Wnt. Transcriptional analysis and CRISPR-Cas9-based genome editing of PDAC driver genes demonstrated the functional heterogeneity of Wnt niche independence in PDAC, which occurred through non-genetic tumor progression. Tiriac et al. (2018) developed pancreatic cancer PDOs that closely mimicked the mutational spectrum, transcriptional subtypes, and therapeutic response of primary tumors, providing a platform for prospective therapeutic selec- tion.
Driehuis et al. (2019) generated 30 PDOs from pancreatic and distal bile duct tumors. These PDOs recapitulated the histological and genetic alterations of pancreatic cancers. Whole-genome sequencing of these organoids revealed that the expression and mutation status of specific genes, such as SMAD4, can impact the growth of organoids in specific media. As a result, the authors suggested examining samples to identify the suitable medium components for organoid culture, as specific growth factors may be necessary to support the growth of PDOs (Driehuis et al., 2019). PDAC PDOs can also be generated from endoscopic ultrasound-guided fine-needle aspiration biopsies, allowing for molecular analysis and drug testing. This method can be efficiently implemented in clinical practice, with extensive significance (Dantes et al., 2020). Single-cell RNA sequencing of PDOs from primary PDAC and the matched liver metastases demonstrated a high level of concordance in subtype assign- ments with the original tumors, suggesting that differences among PDAC patients are maintained in these models. Addi- tionally, PDOs displayed cell state diversity and exhibited a conserved hierarchy of differentiation akin to that observed in primary PDAC tissues (Krieger et al., 2021).
Prostate cancer
Prostate cancer (PCa) is the most prevalent malignant tumor and the second leading cause of cancer-related death in men worldwide (Sandhu et al., 2021; Sung et al., 2021). The vast majority of PCa cases are adenocarcinomas of a luminal phenotype that express the transcription factor androgen receptor (AR) and rely heavily on AR signaling. While localized PCa can be effectively treated with surgery or radiotherapy, approximately 20% of men eventually experience lethal recur- rence. Treatment of recurrent disease with AR signaling inhibitors frequently leads to metastatic castration-resistant prostate cancer (mCRPC).
Generating human PCa organoids is a challenging task, with a success rate of 15%-20%. Gao et al. (2014) reported a biobank of organoids derived from six PCa biopsies and one sample from circulating tumor cells (CTCs), with a success rate of 20%. These PDOs were demonstrated to capture the genetic and molecular diversity of PCa and exhibit similar histopathological features. Furthermore, PDOs represent various subtypes of CRPCs, including AR-dependent adenocarcinoma, AR-negative adeno- carcinoma, and neuroendocrine carcinoma. Drost et al. (2016) described a method for generating prostate organoids from healthy mouse and human prostate cells, metastatic PCa lesions
and CTCs using a fully defined serum-free conditioned medium. Full-grown organoids can usually be obtained in about two weeks after the plating of digested tissue material. In another study, four organoid lines from three needle biopsies were successfully developed and maintained in vitro for 10 weeks (Beshiri et al., 2018). Servant et al. (2021) reported a biobank consisting of PCa organoids derived from 81 primary and metastatic PCa samples, 9% of which can be maintained long- term. Karkampouna et al. (2021) developed PCa PDOs featuring specific biological and genetic profiles that can be utilized to study tumor growth, metastasis, drug resistance in the early stages of the disease, and the response to chemotherapy. Heninger et al. (2021) established PDOs from locally advanced PCa that maintained the complexity of the TME as seen in the original tumor. Orthogonal flow cytometry analysis revealed that these PDOs maintained a distinct subpopulation of epithelial cells and preserved the expression profiles of AR and AR-related genes observed in the parental PCa cells. Servant et al. (2021) improved the culture conditions for PDOs derived from 81 samples of primary and metastatic prostate cancer, with a 69% success rate for long-term maintenance. Furthermore, by employing mouse prostate organoids, Grbesa et al. (2021) revealed that mutations in SPOP, the tumor suppressor gene most commonly mutated in human primary PCa, influence AR accessibility and binding patterns that are similar to those in primary PCa.
Neuroendocrine prostate cancer (NEPC) organoids were generated from needle biopsies of metastatic lesions with a success rate of only 16% (4/25). The authors demonstrated genomic, transcriptomic, and epigenomic similarities between organoids and patient tumors (Puca et al., 2018). Another group investigated the impact of the extracellular matrix (ECM) on organoid growth in CRPC and NEPC samples (Mosquera et al., 2022). They analyzed tumor ECMs through transcriptomics, proteomics, and immunohistochemistry and assessed the com- position of various variants of synthetic hydrogels. The authors reported that the growth, epigenetics, and gene expression of organoids were influenced by the type and stiffness of the hydrogel.
Ovarian cancer
The ovaries are the endocrine glands responsible for producing estrogen and progesterone as well as regulating female reproduc- tion. Ovarian cancer (OC) is a highly aggressive form of cancer that is often diagnosed at a later stage, making treatment extremely difficult. Despite improvements in cancer treatment, OC remains a lethal disease in women, with an unfavorable prognosis and a poor 5-year overall survival (Torre et al., 2018). Moreover, OC is a highly heterogeneous disease, with epithelial ovarian cancer (EOC) accounting for more than 90% of cases. EOC can be categorized into two distinct types: type I EOC, which includes endometrioid, clear cell, seromucinous, mucinous, low- grade serous carcinomas, and malignant Brenner tumors; and type II EOC, which consists of high-grade serous ovarian carcinomas (HGSOCs), carcinosarcomas, and undifferentiated carcinomas (Kurman and Shih, 2016). HGSOC accounts for 70%-80% of mortality, with a high recurrence rate and chemoresistance posing significant challenges in clinical man- agement. PDOs have provided a unique platform to investigate OC, conduct drug screening, and advance personalized medicine.
PDOs have been established from primary and metastatic
tumors of various OC subtypes, with a success rate of 65%-90%. Hill et al. (2018) performed a functional analysis of DNA repair in short-term HGSOC PDOs. They tested 33 PDOs from 22 HGSOC patients to detect defects in homologous recombination and replication fork protection. Whole-genome sequencing and histopathological assessments demonstrated that PDOs exhibited genomic and histological characteristics similar to those of the parental tumors. Based on human fallopian tube organoid culture medium, Kopper et al. (2019) developed 56 PDOs from samples of 32 cases, covering a wide range of OC subtypes, such as serous borderline tumors, mucinous borderline tumors, low- grade serous borderline tumors, mucinous carcinomas, endome- trioid carcinomas, clear cell carcinomas, and high-grade serous subtypes. The authors created two culture conditions by adding forskolin, hydrocortisone, and heregulin-ß-1 to the medium, with or without the addition of WNT-conditioned medium. The overall success rate was 65% for all histological types, and the success rate was 55% for HGSOC. OC organoids exhibited stemness and self-renewal properties, faithfully captured the genomic and pathophysiological characteristics of the tumors from which they originate, and demonstrated inter- and intra-patient heteroge- neity. Moreover, the organoid platform allowed for long-term expansion and manipulation, thereby providing a valuable tool for drug screening of different OC subtypes (Kopper et al., 2019). Senkowski et al. (2023) developed two specialized culture media for HGSOC PDOs using a method similar to that of Kopper, emphasizing the need for stricter criteria in organoid cultivation with a success rate of 53%. The criteria for successful establish- ment were as follows: stable passaging over 10 generations, no growth stalling, and confirmed genomic stability.
de Witte et al. (2020) reported 36 whole-genome characterized PDOs derived from 23 OC patients with documented clinical backgrounds. These PDOs retained the genomic characteristics of the primary tumor lesions. Hoffmann et al. (2020) established 15 HGSOC organoid cultures that closely match the phenotype of primary tumors. During prolonged culture periods, they noted unavoidable growth arrest, emphasizing the advantage of a low- Wnt culture medium for the formation and development of HGSOC organoids. Activating the Wnt pathway inhibits PDO growth, while activating BMP signaling facilitates PDO forma- tion. Maenhoudt et al. (2020) comprehensively assessed various components and recognized neuregulin-1 as a crucial factor in enhancing the growth and development of OC PDOs, despite an overall derivation efficiency of 44% for all OC patients. Nanki et al. (2020) examined the differential genomic profiles of parent tumors and PDOs and showed that they share key DNA variants, including BRCA1 and 2, MLH1, TP53, and PIK3CA. In addition, there was a 59.5% overlap in gene variations between organoids derived from HGSOC, clear cell, endometroid, and parental tumors.
Malignant effusion is a special form of tumor metastasis. OC PDOs are derived not only from primary or metastatic tumors but also from body fluids containing OC cells. Chen et al. (2020) established organoids from the ascites and pleural effusion fluid of HGSOC patients. Despite the rarity of tumor cells in ascites and effusions, this approach is appropriate for rapid drug screening in short-term organoid cultures when the tumor progresses rapidly. In another study, Bi et al. (2021) established 21 PDOs from 28 OC tissue or ascites samples, with a success rate of 75%. These findings indicate that PDOs can serve as efficient tools for developing personalized cancer treatments for patients with OC.
In summary, these studies highlight that OC PDOs exhibit patient tumor-dependent characteristics, and retain the marker expression and mutational landscape of OC, irrespective of the various preparation methods used in different studies.
Breast cancer
The development of the breast is predominantly controlled by ovarian steroids and pituitary hormones. Breast cancer (BC) is one of the most frequently diagnosed malignancies among women worldwide (Teng et al., 2024; Trapani et al., 2022). BC is a heterogeneous tumor, usually classified based on the hormone receptor status of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2). Tumors lacking the expression of these receptors are referred to as triple-negative breast cancer (TNBC), which is considered the most aggressive type of BC. Treatment of BC mainly includes surgery, radiotherapy, endocrine therapy, systemic chemother- apy, and anti-human HER2-targeted therapy. The variability in gene mutations and individual differences contribute to the failure of BC therapy. The emergence of organoid technology has opened up new possibilities for investigating BC samples.
To date, numerous studies have been published on the development of PDOs from human BC tissues. Nguyen-Ngoc et al. (2012) attempted to generate BC organoids to investigate the role of the ECM in tumor cell invasion and metastasis. Sachs et al. (2018) successfully established 95 BC organoids from 155 surgical primary and metastatic tumors, achieving a success rate of more than 80%. The authors found that neuregulin 1, a key complementary factor and ligand for HER tyrosine kinases 3 and 4, is associated with breast development and tumor formation. The addition of neuregulin 1 to culture medium promoted the effective generation of BC organoids and allowed for long-term expansion. Morphological and histopathological analysis re- vealed that BC organoids closely resembled the original tumor tissues. More specifically, ductal carcinoma produced solid organoids, while lobular carcinoma produced loose organoids. The expression of ER, PR, and HER2 in BC tissues was accurately retained in the corresponding organoids. Transcriptome and genomic characteristics of BC organoids were also similar to those of the parental tumors. These organoids are valuable preclinical models for BC and have broad applications in academic, clinical, and pharmaceutical research (Sachs et al., 2018).
Goldhammer et al. (2019) reported that human BC organoids best represented the tissue of origin. Mazzucchelli et al. (2019) introduced a novel technique for generating BC organoids from surgical and biopsy specimens, and proposed a new enzymatic digestion protocol to increase the efficiency of organoid forma- tion. Chen et al. (2021b) developed 99 PDOs from 132 BC samples and assessed ER, PR, HER2, and Ki-67 expression in tumor tissues and paired organoids through immunohistochem- ical analysis. They found that the expression profiles of these BC markers were well maintained in the organoids, regardless of whether the organoids had undergone systemic anticancer treatment. Bhatia et al. (2022) established a repository of organoids from 83 BC tissues. These authors not only demon- strated that PDOs accurately recapitulated the characteristics of patient tumors but also observed that when TNBC organoids were transplanted into NOD/SCID mice, the tumors produced were morphologically similar to the original tumors. Saeki et al.
(2023) generated organoids from 10 individuals with BC and found that the PDOs presented varying growth rates and patterns. Single-cell RNA sequencing analysis showed that each PDO consisted of multiple subpopulations with distinct cellular states and cycles, estrogen responsiveness, and EMT-like gene expression programs. These findings indicate that specific cell clusters within organoids correspond to distinct PDO character- istics. Furthermore, RNA-seq analysis revealed that although the expression profiles of the two tumor tissues and the paired organoids were not fully consistent, the PDOs maintained a certain level of tumor heterogeneity observed in the primary tissue after continuous passaging.
Organoids for cancer pathogenesis research
The pathogenesis of cancers is a complex process, and studying the mechanism of cancer pathogenesis is an important topic in the field of oncology. PDOs can simulate the growth process of cancers in the human body, thus furthering our understanding of the pathogenesis of cancers, and highlighting their value as tools for exploring cancer pathogenesis. PDOs can be used to investigate the proliferation, metastasis, drug resistance, and other characteristics of cancer cells, thereby providing crucial insights for cancer prevention, diagnosis, and treatment. In addition, tumors can vary greatly in terms of their stage, genetic background, and molecular behavior. PDOs can preserve the phenotypic heterogeneity of primary tumors, providing a suitable alternative in vitro model for research on tumor development and therapy.
Due to the heterogeneity of tumors, identifying driver mutations in cancers is difficult. Therefore, PCa organoids with heterogeneity can offer a unique opportunity for the study of PCa (Chai et al., 2023; Kretzschmar, 2021; Tuveson and Clevers, 2019). Organoids derived from both normal and tumor tissues from the same patient can be used to compare gene expression profiles and detect differences in the activation of specific oncogenic pathways in PCa. Moreover, organoids derived from healthy tissues are relatively more genetically stable and may be employed as models to assess the functional roles of specific tumor mutations in driving PCa development (Pietrzak et al., 2020). Compared with traditional cell lines, PDOs have the advantages of retaining tumor heterogeneity and preserving a higher level of TME. PDOs can provide a more in-depth understanding of the molecular and cellular mechanisms involved in BC progression. By using a 3D microfluidic system and computational model, Hwang et al. (2023) found that during the collective migration of heterogeneous tumor and tumor stromal cell clusters, the main migrating cells segregate into a specific subgroup characterized by their collective migratory potential, especially the keratin-14 and calponin-3- positive subgroups. This process enables migrating cells to regulate their protrusion dynamics through the local producing laminin, emphasizing how these cells guide collective migration by interacting with the microenvironment.
The occurrence of tumors is not yet fully understood. PDOs allow the modeling of human cancer development in vitro and thus represent an excellent approach for studying cancer progression. Research on the tumorigenesis of pituitary adeno- mas has been conducted primarily through animal tumor cell lines and animal models. However, studies on human pituitary adenomas are challenging due to notable interspecies differences
and the lack of dependable cell lines. Nys et al. (2022) revealed and highlighted the activation phenotype of stem cells in pituitary tumors, identified the activation of stem cells in pituitary tumorigenesis, and extended this finding to the derived organoids, which may be involved in currently unknown pathogenic processes. Pancreatic cancer PDX-derived organoids have been proven to preserve complex glycosylation variants (Huang et al., 2020b), promoting tumor progression through dysregulated protein levels, stability, and localization (Pinho and Reis, 2015). In pancreatic organoid models, KRAS can regulate epithelial-macrophage crosstalk and promote pancreatic carci- nogenesis (Bishehsari et al., 2018). Huang et al. (2020a) and Beato et al. (2021) developed an intraductal papillary mucinous neoplasia (IPMN) organoid biobank to study the genetic and biological mechanisms of IPMN. Naruse et al. (2020) utilized four genotoxic substances, ethylmethanesulfonate, acrylamide, diethylnitrosamine, and 7,12-dimethylbenz[a]anthracene, to investigate their chemical carcinogenic effects. The 7,12- dimethylbenz[a]anthracene-treated mammary tissue-derived or- ganoids with a heterozygous Trp53 knockout exhibited tumor- igenicity, while those with wild-type Trp53 did not. Furthermore, PDOs have also been employed to study pathways involved in tumorigenesis.
Cancer metastasis refers to the process by which cancer cells spread from their original site to other organs and is the primary cause of death in patients with cancer (Ganesan et al., 2024; Steeg, 2016). Cancer metastasis is a complex molecular process, and the maintenance of cancer pathological and physiological features by PDOs renders them a suitable research platform. Organoids have been widely used to elucidate the mechanisms that drive cancer metastasis. By using PDOs, Huang et al. (2020c) identified SMAD4 as the transcription factor that drives EMT and coordinates invasion program in PDAC. SOX4 has been demonstrated to support the maintenance of an undifferentiated and proliferative state in BC organoids (Roukens et al., 2021). Compared with SOX4-positive BC organoids, SOX4-knockdown BC organoids contained more differentiated cells expressing intraluminal or basal genes, lower levels of cell cycle genes, and impaired tumor growth and metastasis capability (Roukens et al., 2021). Another application of organoids in BC is the study of pathways involved in tumorigenesis and metastasis. For exam- ple, the roles of EGF receptors and integrins (Furuta and Bissell, 2016), MALAT1 long non-coding RNA (Arun et al., 2016), and FZD6 (Corda et al., 2017) have been studied in BC organoids. The roles of RANK ligands (Nolan et al., 2016) and JNK (Cellurale et al., 2012) in mammary and cancer development, as well as the roles of the transcription factors Sox9 and Slug in controlling mammary stem cell fate, have been investigated in pre-tumor tissue-derived organoid cultures (Guo et al., 2012). Sachs et al. (2018) generated organoids from primary and metastatic BC tissues and faithfully reproduced the tissue histology, hormone receptor and HER2 status, and DNA copy number alterations of the original tissues. These findings indicate that organoids can effectively replicate native and metastatic tumors and can be studied to expand our understanding of cancer biology. Furthermore, Dedhia et al. (2023) used a single-chip metastasis platform to characterize ACC metastasis and performed RNA sequencing to identify key differences between tumor cells that can metastasize and those that cannot, aiming to identify new intervention targets. In short, PDOs offer a reliable tumor model for studying the mechanisms that promote and inhibit tumor
invasion.
Genome editing technology has the potential to be applied in establishing PDOs for studying the molecular mechanisms of cancers (Driehuis and Clevers, 2017; Feng et al., 2021; Tiroille et al., 2023). Chua et al. (2014) developed PCa organoids from castration-resistant Nkx3.1-expressing cells that contained basal and luminal cells. Through genetic manipulation involving the deletion of Pten and the activation of KrasG12D, the AR signaling pathway was elevated in PCa organoids derived from castration- resistant Nkx3.1-expressing cells. This discovery emphasized the key roles of Pten deletion and activated Kras mutation in the development of PCa. CRISPR/Cas9-based genome editing and ectopic expression of oncogenes/tumor suppressor genes can be used to investigate the molecular mechanisms of PCa develop- ment (Koo et al., 2011; Nie and Hashino, 2017; Teriyapirom et al., 2021). Karkampouna et al. (2021) developed PCa models with specific biological and genetic characteristics for studying tumor progression, metastasis, and resistance to drugs at the initial phases of cancer, as well as for evaluating the response to chemotherapy. The combination of CRISPR/Cas9 technology with PDOs can be utilized to investigate the pathogenesis of BC (Dekkers et al., 2020). In a study by Jardé’s team, Wnt1 was created by activating the HER2/neu and Wnt pathways in a less- studied type of BC (Jardé et al., 2016). PDOs subjected to gene editing by the CRISPR system can be used as valuable tools for identifying interactions between drugs and genes (Hirt et al., 2022; Lo et al., 2021). Hirt et al. (2022) reported the relationship between specific missense mutations in AT-rich interaction domain 1A (ARID1A) and drug responsiveness in PDAC organoids through the use of CRISPR/Cas9 technology.
PDOs have been proven to be valuable tools for epigenomic research, including the exploration of DNA methylation changes in PCa. Lu et al. (2019) and Stepper et al. (2017) demonstrated that the fusion of DNA methyltransferase genes (DNMT1 and DNMT3) to nuclease-inactivated CRISPR/Cas9 led to reduced DNA methylation of target genes by blocking the activity of DNMT1 and DNMT3. Similarly, changes in histone modifications have been observed in the development and progression of PCa (Pomerantz et al., 2020). It has been proposed that dysregulation of histone demethylases and histone methyltransferases is closely related to the aggressiveness and drug resistance of PCa (Duan et al., 2019; Wang et al., 2023). Interestingly, the application of dCas9 fused to genes encoding histone modification enzymes is considered an effective method for investigating histone mod- ifications. The use of this CRISPR-dCas9-based technology to study epigenomic alterations in PDOs can provide molecular insights into the mechanisms of epigenetic/epigenomic altera- tions in PCa (Driehuis and Clevers, 2017; Elbadawy et al., 2020; Hilton et al., 2015).
Organoids for drug screening and personalized medicine
Precision medicine or personalized therapy is a rapidly develop- ing method that enables clinicians to choose treatments with greatly increased effectiveness to improve patient quality of life. PDOs replicate the phenotype and genotype of the original tumor, thus providing new avenues for the development of cancer models that better mimic real pathophysiological states. As in vitro personalized preclinical models, PDOs can be used to rapidly identify the most suitable drugs, develop optimal and efficient
drug treatment plans, and lower the risk of side effects, drug resistance, and tumor recurrence, thereby achieving optimal treatment outcomes. Over the past decade, various PDO biobanks have been established for drug testing and toxicology assess- ments. PDOs can also serve as vital models for drug discovery, potentially shortening the preclinical trial period, reducing development costs and risks, enhancing the success rate, and promoting drug discovery. PDO biobanks can be used to reveal new treatment options for subsequent evaluation in clinical trials. Many research institutions have performed single- or multi-center clinical trials with PDOs to evaluate their potential in predicting drug responses and guiding drug selection (Drost and Clevers, 2018). Furthermore, some clinical trials are investigating whether PDO-guided therapy can improve cancer prognosis (Drost and Clevers, 2018). For refractory tumors, PDOs can be used to evaluate the efficacy of different combination therapy strategies, enabling the selection of the most effective approach (Amodio et al., 2020). Therefore, PDOs represent a promising advanced functional testing platform in the field of precision medicine.
Pituitary tumor
Surgical resection is the first-line treatment for TSH-, GH-, and ACTH-producing adenomas. However, complete resection is technically challenging, and many patients require persistent drug treatment after surgery. Organoids that mimic pituitary adenomas can be derived from induced pluripotent stem cells (iPSCs). Due to the lack of specificity, poor tolerability, and low efficacy of second-line drug treatments, Cushing’s disease presents a therapeutic challenge. Chakrabarti et al. (2022) developed organoids from iPSCs harboring familial mutations in MEN1 and CDH23, as well as PitNET tissues of Cushing’s disease patients.
Drug screening with small-molecule compounds revealed that PDOs from each patient exhibited patient-dependent responses, reflecting individual genetic and molecular differences. Further elucidation of the mode of action of the identified active drugs can unveil novel treatment targets for patients with Cushing’s disease (Chakrabarti et al., 2022). In a subsequent study, the CRISPR/ Cas9 system was used to introduce somatic mutations in genes implicated in corticotropinoma tumorigenesis (i.e., USP8 and USP48) in iPSCs derived from healthy donors (Mallick et al., 2023; Sbiera et al., 2019). The genetically engineered organoids (iPSCUSP8/iPSCUSP48) were used to study the impacts of the glucocorticoid receptor antagonist mifepristone and the gluco- corticoid receptor modulator relacorilant (Mallick et al., 2023). Organoids responded differently to these modulators in terms of somatostatin receptor expression, ACTH secretion, and cell apoptosis/proliferation. This research revealed the potential efficacy of relacorilant in combination with somatostatin analogs and demonstrated that relacorilant is superior to mifepristone, supporting its further development for the therapy of Cushing’s disease (Mallick et al., 2023). Cui et al. (2025) validated that PitNET organoids recapitulated the histological and genomic features and microenvironmental heterogeneity of parental tumors, and demonstrated the predictive value of PDOs in personalized drug testing. They observed distinctive drug response profiles in PDOs obtained from patients with different PitNET subtypes, indicating that PDOs can serve as a platform for personalized drug screening for PitNET patients. Therefore,
organoids may provide important translational insights for drug discovery and mechanism identification. These models will promote drug discovery and expand the understanding of pathophysiology, thereby contributing to breakthroughs in the clinical management of patients with pituitary adenomas.
Thyroid cancer
Clinical treatment of thyroid cancer mainly includes surgical treatment and radioactive iodine therapy. About 15%-33% of PTC patients have difficulty in receiving radiation therapy due to that their thyroid follicles are unable to absorb radioiodine (Durante et al., 2006; Pacini et al., 2012). Therefore, radioactive iodine-refractory (RAIR) thyroid tumors are a therapeutic challenge. In addition, patients with MTC and ATC have a worse prognosis, and currently, there are no better treatment options. PDOs can serve as potential diagnostic and therapeutic tools for iodine-refractory and highly invasive thyroid cancers.
In 2021, our team screened 24 targeted drugs and chemother- apeutic agents in PTC organoids and demonstrated that these organoids derived from different patient sources exhibited varying sensitivities to drugs. We also found that the response of PTC organoids to drugs correlated strongly with the results of patient genomic testing, suggesting the high value of combining the results of genomic sequencing with drug susceptibility testing of PTC organoids (Chen et al., 2021a). Clinical and epidemiologic reports indicate that thyroid cancer is 3-4 times more prevalent in women than in men, suggesting a role for estrogen in thyroid cancer. We examined whether estrogen affects the proliferation of PTC organoids and the types of ERs involved in this process. We indicated that ERa is the key player in the estrogen-induced proliferation of PTC organoids, demonstrating that the inhibition of ERa may be a promising therapeutic strategy for the treatment of female thyroid cancer (Chen et al., 2021a). BRAFV600E is the most common driver mutation in PTC. Several clinical trials have demonstrated limited benefits from BRAF-targeted therapy in patients with thyroid cancer, suggesting that combination therapy based on BRAF inhibitors may be a promising treatment strategy. In 2023, we reported the development of both wild-type and BRAFV600E-mutant PTC organoids to assess the efficacy of targeted therapeutic drugs. We assessed the effectiveness of BRAF inhibitors (dabrafenib and vemurafenib), both individually and in combination with MEK inhibitors (selumetinib and trametinib), receptor tyrosine kinase inhibitors (sorafenib, cabozantinib, lenvatinib, vandetanib, and sunitinib), or chemotherapy drugs (vincristine, doxorubicin, paclitaxel, and cisplatin), and found that these combinations were more effective than single treatment (Chen et al., 2023). These PTC organoids have shown promise in assessing drug responses in individual patients, indicating their potential for personalized treatment of thyroid cancer. Moreover, Sondorp et al. (2020) developed organoid models from PTC and RAIR-PTC patients with the potential for pre-treatment diagnosis of [131-resistant patients. These organoid models retained tumor features and the phenotype of RAIR disease, and have the potential to predict patient response to radioiodine with the promise of early identification of [131_ resistant patients to avoid ineffective treatments and side effects.
Adrenocortical cancer
Currently, the treatment of metastatic ACC typically involves the
use of mitotane and cytotoxic drugs, but the efficacy is limited. Arakawa et al. (2024) explored the mechanism of the clinical ubiquitin-activating enzyme inhibitor TAK-243, its synergistic effects with ACC treatment drugs, and its effectiveness in ACC PDOs and mouse xenografts. The results revealed that TAK-243 alone or in combination with the BCL2 inhibitor venetoclax was effective against ACC cell lines as well as PDOs. Wang et al. (2022a) created human PPGL primary cultures from tumor specimens of surgical patients for personalized drug testing and correlated the outcomes with germline or somatic driver gene mutations in the respective tumors.
Pancreatic cancer
PDAC is a severe disease and has the lowest 5-year survival rate. The standard therapy for PDAC comprises surgery, chemother- apy, and radiation therapy. Nevertheless, because of the difficulties linked to early detection and therapy, only a small percentage of patients are diagnosed in the initial stage and eligible for surgical resection. The standard chemotherapy approach involves the combination of various cytotoxic antic- ancer drugs, including the FOLFIRINOX protocol (consisting of oxaliplatin, irinotecan, leucovorin, and 5-fluorouracil) and gemcitabine in combination with albumin-bound paclitaxel (Park et al., 2021; van Eijck et al., 2024). However, individual patients exhibit varying responses to similar drugs. Additionally, both adjuvant and neoadjuvant chemotherapy have limited efficacy and promote drug resistance, thereby limiting survival outcomes.
Many studies have shown that pancreatic cancer PDOs can mimic all phases of pancreatic cancer and related genetic changes. The drug response of pancreatic cancer PDOs corre- sponds closely to clinical outcomes in patients, which makes pancreatic cancer organoids useful for assessing chemotherapy sensitivity and synchronous tumor metastasis over time. Currently, PDOs are used as model systems to develop novel precision medicine strategies, serving as a predictive platform for selecting treatment options for patients with PDAC. Huang et al. (2015) examined the response of PDOs from five patient tumors to the standard PDAC medication gemcitabine, and found that all PDOs displayed poor responses to the drug, with an average growth inhibition rate of 30%. The authors continued to utilize the same five personalized PDOs in a drug-screening approach to evaluate five different epigenetic inhibitors. The authors also reported that the PDOs preserved patient-specific characteristics, such as the inhibition of epigenetic markers, oxygen consump- tion, and EZH2 dependency, highlighting the utility of their system in further drug screening. Walsh et al. (2016) used their developed PDOs to assess the response to gemcitabine and AZD1480 (a new ATP-competitive JAK2 inhibitor) individually or in combination. While gemcitabine and the combination led to a notable decrease in the OMI index (optical metabolic imaging, used to assess the effect on PDO proliferation), AZD1480 alone did not cause a significant reduction. Romero-Calvo et al. (2019) reported on dose-dependent and drug-specific response testing of PDOs. This study investigated the response of PDOs from two patients to gemcitabine and the combination of gemcitabine with abraxane. The main discovery of this research was the identification of two clones from the organoids, highlighting the promise of these models in providing important insights into clonal populations. However, the response of PDOs to treatment
varies and may not always align with patient response due to rapid advancement of the disease. Gout et al. (2021) validated a quadruple-targeting strategy (PARP, ATR, ATM, and DNA-PKcs inhibition) for PDOs and observed a significant induction of cell death even in non-homologous recombination deficiency-mu- tant PDOs by artificially inducing an HRD-deficient status. In another study, patient-derived PDAC organoids were used as a validation tool for a NOXA-dependent RUNX1 inhibitor, which was initially identified in a large unbiased drug screening experiment (Doffo et al., 2022). Furthermore, April-Monn et al. (2021) reported that patient-derived pancreatic NET organoids showed variable sensitivities to sunitinib, everolimus, and temozolomide. These 3D models provide unique insights into NET biology and may become a routine part of preclinical assessment of therapeutic targets for NETs.
Some recent studies indicate that the response of PDOs to drugs is consistent with the clinical response of patients with pancreatic cancer (Driehuis et al., 2019; Farshadi et al., 2021; Grossman et al., 2022; Tiriac et al., 2018). Tiriac et al. (2018) developed a panel of PDOs from 66 PDAC patients to be used for genetic profiling and drug screening, and demonstrated that drug testing in PDOs could aid in therapy selection for patients within a clinically relevant period. They discovered new common genetic alterations in these PDOs that act as driver oncogenes in PDAC. These PDOs exhibited patient-like responses to standard-of-care chemotherapeutics, such as 5-fluorouracil, paclitaxel, gemcita- bine, irinotecan, and oxaliplatin. Furthermore, they identified genetic and transcriptome characteristics of organoids that may predict chemotherapy sensitivity. The application of PDO pharmacotyping in clinical practice will greatly improve preci- sion medicine and ultimately patient outcomes. Another group also pharmacotyped PDO models from PDAC patients’ endo- scopic biopsies and surgical specimens to assess chemotherapy sensitivity, indicating the practicality of this strategy for adjuvant therapy selection within a clinically relevant timeframe (Seppälä et al., 2020). Indeed, pharmacotype-guided chemotherapeutic selection showed benefit in identifying effective drugs for patients, indicating that PDO-driven drug screening results serve as a reliable biomarker for predicting the response to chemotherapy in individuals with PDAC. Recently, this group prepared and analyzed PDAC PDOs using next-generation sequencing and pharmacotyping. Through a randomized controlled clinical trial, the PDO-specific pharmacotype was prospectively evaluated as a predictor of clinical treatment response. This study revealed a significant timeframe for collecting data from PDOs that can help promote precise treatment, highlighting that the sensitivity of PDAC PDOs can predict the response to chemotherapy based on biomarker assessments (Seppälä et al., 2022).
Additionally, Driehuis et al. (2019) conducted extensive drug screening of 76 chemotherapeutic agents in 24 PDO models, aiming to evaluate the drug sensitivity of PDOs and correlate it with the clinical outcomes of patients. Notably, the authors uncovered synergistic effects when the HER2/EGFR inhibitor lapatinib was combined with gemcitabine. Nevertheless, only the clinical records of four patients, all of whom received gemcitabine treatment were available. Hennig et al. (2022) investigated the potential use of PDOs in tailoring poly-chemotherapy regimens, such as neoadjuvant and adjuvant chemotherapy, for treating PDAC. Pharmacotyping of chemotherapy-naïve patients and neoadjuvant-treated PDOs revealed the potential for PDOs to help develop personalized poly-chemotherapy plans. Demyan et al.
(2022) reported a large single-institution PDO biobank of PDAC and indicated the possibility of rapid PDO drug screening and obtaining results within one week of tissue resection. The ability to develop PDOs from chemotherapy-naïve and post-neoadjuvant tissue allows longitudinal PDO generation to evaluate dynamic sensitivity profiling to chemotherapy. Grossman et al. (2022) reported a strong correlation between the drug sensitivity test results of PDOs and the clinical response of individual PDAC patients. For example, a patient’s PDOs demonstrated the greatest sensitivity to irinotecan, followed by trametinib, and in accordance with these findings, the patient exhibited stable disease in response to FOLFIRINOX (contains irinotecan) and a partial response to trametinib/lapatinib. Nevertheless, these studies have been conducted in relatively small cohorts, emphasizing the importance of carefully designed, broader co- clinical trials in larger patient cohorts.
Prostate cancer
The treatment for PCa typically involves androgen deprivation therapy (ADT), surgical intervention, and pharmaceutical management (Sandhu et al., 2021). Androgen receptor signaling plays a key role in the development and proliferation of PCa, making ADT the mainstay of standard-of-care for PCa. However, many patients treated with ADT develop CRPC, which is diagnosed by an increased level of prostate-specific antigen despite castration (Nuhn et al., 2019). Despite current treatment options for CRPC including hormonal therapy, chemotherapy, immunotherapy, radionuclide therapy, and targeted therapy, metastatic CRPC remains a deadly disease with poor survival rates (Nuhn et al., 2019).
PCa organoids are valuable models for evaluating the efficacy of anticancer drugs due to their retention of genetic features and tumor heterogeneity. PCa organoids generated from metastatic CRPC patient biopsies were used to study the efficacy of targeted drugs such as BET domain inhibitors (Welti et al., 2018). For instance, a study showed that the combination of CX-5461 (an RNA polymerase I inhibitor) and CX-6258 (a pan-PIM kinase inhibitor) exerted potent anti-tumor effects on PCa organoids derived from PDXs (Lawrence et al., 2018). Additionally, CRPC organoids have been employed to assess the efficacy of BKM-120 (a PI3K inhibitor) (Gao et al., 2014; Triscott et al., 2023), everolimus (a mTOR inhibitor) (Elbadawy et al., 2020; La Manna et al., 2020), and enzalutamide (an AR antagonist) (Risbridger et al., 2018; Van Hemelryk et al., 2021). Moreover, Wu et al. (2023) demonstrated that Z15, an AR antagonist and selective AR degrader, effectively inhibits AR/AR-V7 activity in CRPC cell lines and organoids.
PDOs from NEPC patients were established and used as new models for investigating the pathogenesis of NEPC, resulting in the discovery of ALK as a promising target for drug therapy (Carneiro et al., 2018; Patel et al., 2022). Four CRPC- neuroendocrine (CRPC-NE) organoids and two CRPC-adenocar- cinoma (CRPC-Aden) organoids were developed to conduct high- throughput screening of 129 chemotherapy and targeted therapy drugs. According to the findings, AR antagonists (such as enzalutamide) and other chemotherapy drugs (such as cabazitaxel and docetaxel) had higher efficacy in organoids derived from CRPC patients (Puca et al., 2018). Drug candidates, such as vandetanib and pozotinib, have also been evaluated in PCa organoids. Both drugs demonstrated notable inhibitory
effects on CRPC-NE organoids (Puca et al., 2018). Jansson et al. (2018) conducted high-throughput screening of more than 100 drugs in CRPC-LuCaP PDX-derived organoids. Among these drugs, heat shock protein-90 (HSP90) inhibitors have shown significant anti-tumor activity in CRPC-LuCaP PDX-derived organoids. Ganetespib, an HSP90 inhibitor, demonstrated the most potent inhibitory effect on CRPC-LuCaP PDX models by targeting the AR and PI3K signaling pathways. Furthermore, Beshiri et al. (2018) demonstrated that LuCaP-derived organoids deficient in BRCA2 were sensitive to olaparib in a clinical trial. Some preclinical models derived from CRPC samples have been used as in vitro systems to test the efficacy of PARP inhibitors (e.g., olaparib) in combination with cisplatin (Elsesy et al., 2023). In general, these findings indicate that PDOs can serve as excellent models for drug screening, potentially advancing personalized treatment.
Several studies have emphasized the potential application opportunities of PCa PDOs in personalized medicine. PCa PDOs are considered promising in vitro models for assessing drug efficacy, with the results serving as indicators of clinical outcomes (Karkampouna et al., 2021; Pauli et al., 2017). Indeed, PDOs have been utilized to assess drug efficacy in several clinical trials for PCa. For example, Puca et al. (2018) established PCa organoids from CRPC-NE patient samples and conducted a clinical trial using these models. A small subset of single drugs and drug combinations identified via high-throughput drug screening revealed new possibilities for treating this disease. These organoids were also employed in a clinical study to evaluate the efficacy of the aurora kinase inhibitor alisertib. This finding aligns with the outcomes of a clinical trial (NCT01799278) in which alisertib showed significant effects in a subset of patients with CEPC-NE (Beltran et al., 2019). These studies suggest that PCa organoid models can serve as powerful tools for assessing drug efficacy in clinical studies and developing new treatment strategies.
Ovarian cancer
Over the past 25 years, standardized treatment for OC patients has consisted of cytoreductive surgery and platinum/taxane- based adjuvant chemotherapy. In some cases, local radiotherapy can also be used for the treatment of OC. While OC patients respond well to the primary chemotherapy regimen, approxi- mately 70%-80% of patients eventually experience tumor recurrence and develop increased chemoresistance (Pignata et al., 2017). The ability to predict the response to platinum-based chemotherapy and PARP inhibitors is limited by certain genetic features (Pujade-Lauraine et al., 2017; Swisher et al., 2017). It also remains unclear whether maintenance-targeted therapy increases sensitivity to chemotherapy. This highlights the need for accurate and real-time predictive models to help guide personalized and precise treatment strategies.
OC is a heterogeneous disease frequently detected in its advanced stages. PDOs accurately represent the heterogeneity and disease markers of OC, mirroring their genomic character- istics. Therefore, PDOs can provide a better understanding of cancer pathobiology, drug screening and discovery. PDOs have been proven to be promising tools for new drug development and preclinical high-throughput drug screening, enabling the pre- diction of clinical responses before treatment and providing a basis for personalized medicine. This strategy helps to identify
candidate drugs that are highly likely to be effective for individual patients, thus decreasing the reliance on animal experiments, while simultaneously increasing the amount of supporting data and accelerating the drug development process.
Many groups have attempted to develop PDO models for OC to identify possible underlying mechanisms and discover new and effective treatment options. Jabs et al. (2017) compared PDOs and 2D cell cultures obtained from tumor tissues, ascites, and pleural effusions of nine patients with ovarian serous adenocar- cinoma. The results indicated that PDOs displayed drug responses that were closely linked to patient genotypes, implying that PDOs are more effective in replicating clinical responses. Approxi- mately 50% of HGSOC cases involve DNA repair defects that can be targeted by blocking the nuclear enzyme poly (ADP-ribose) polymerase. Hill et al. (2018) established a platform to analyze DNA repair function in short-term HGSOC PDOs and discovered that homologous recombination defects were positively corre- lated with sensitivity to PARP inhibitors and that replication fork protection defects were associated with sensitivity to carboplatin, and Checkpoint Kinase 1 and Ataxia telangiectasia and Rad3- related protein inhibitors. In combination with genomic screen- ing, functional testing of PDOs is an effective tool for identifying targetable defects in DNA damage repair. HGSOC PDOs were also developed to assess the effect of DNA repair on OC phenotypes and predict the clinical response to DNA repair inhibitors (Hill et al., 2018). PDOs from different patients have varying sensitivities to chemotherapy drugs. Kopper et al. (2019) captured the responses of different tumor subtypes of OC PDOs to the gold standard of platinum-based chemotherapy. PDOs are used to screen various anticancer drugs and the mechanisms of chemotherapy resistance, demonstrating their potential in treatment development and personalized medicine. Using the “mini-rings” seeding technique, Phan et al. (2019) screened 240 kinase inhibitors in four OC PDOs for tailored treatment response prediction. Shigeta et al. (2021) established three PDO cultures from tumor tissues and ascites of two ovarian clear cell carcinoma patients. They then conducted high-throughput drug screening of 42 FDA-approved or late-stage developmental tumor drugs using two ascites-derived organoids. However, due to the small sample size, the homogeneity of organoids derived from ascites and tumor tissue from the same patient could not be effectively proven, leading to controversy over the promotion of this screening strategy. Vias et al. (2023) demonstrated that PDOs replicated clinically relevant chromosomal instability characteristics observed in HGSOC patients, including copy number-driven gene expression signatures, and that their drug responses matched those of the original tumors. Using single-cell DNA sequencing, they identified clonal populations with distinct copy number characteristics. The developed models reproduced the heterogeneity of chromosomal instability in HGSOC, indicat- ing their potential value in treatment selection. A major obstacle in developing treatments is incomplete genetic information and a lack of immunocompetent models. To assess the impact of certain mutations and their combinations in HGSOC, Zhang et al. (2021b) generated organoids from fallopian tubes that address both cell-autonomous and cell-nonautonomous effects. They proposed an effective combination therapy using FDA-approved drugs for CCNE1-amplified HGSOC. These syngeneic organoid models provide a platform for developing innovative approaches for identifying genomic subgroups of this complex disease.
An increasing number of preclinical studies have confirmed
the viability and practicality of using OC PDOs for personalized drug response evaluation at the individual patient level. de Witte et al. (2020) evaluated the ability of PDOs to predict clinical drug responses and tumor heterogeneity in OC patients and showed that at least one drug yields high responsiveness in 88% of patients. They observed a strong correlation between the sensitivity of PDOs to neoadjuvant carboplatin/paclitaxel che- motherapy and patients’ clinical response. This research indicated that PDOs serve as valuable preclinical platforms offering insights into individualized treatment for OC patients. Gorski et al. (2021) examined the sensitivity of HGSOC PDOs to drugs by comparing their EC50 values with clinically achievable Cmax values and characterized patient responses based on progression-free survival (PFS) assessed using the RECIST criteria. The authors found that one PDO had an EC50 value that was above the clinically achievable plasma Cmax, corre- sponding to a significantly lower PFS than that of the other patients. This study was the first to use PFS as an indicator to show the ability of PDOs to predict clinical response in OC patients. Gray et al. (2023) documented a case of a patient with platinum-resistant advanced LGSOC. A drug sensitivity assay conducted on the PDOs provided guidance for treatment planning. After treatment, the patient’s condition quickly stabilized. Notably, the drug sensitivity results were inconsistent with the results of genetic testing, highlighting the potential of PDO drug sensitivity as a valuable supplement to treatment when genomics analyses lack accuracy.
Breast cancer
BC PDOs are mainly used for drug testing and drug discovery. PDOs have the capability to assess the safety and effectiveness of drugs or their metabolism in organs. Walsh et al. (2014) xenotransplanted original tumors and organoids into mice to conduct drug sensitivity tests, revealing that the drug sensitivity of the organoids closely mirrored that of the original tumors to a certain degree. A panel of drugs evaluated in BC organoids confirmed the heterogeneity of organoid responses to drugs. Sachs et al. (2018) demonstrated the effectiveness of six EGFR/ AKT/mTORC inhibitors targeting the HER signaling pathway in HER2-overexpressing BC organoids. Similarly, BC organoids with high BRCA1/2 signatures displayed sensitivity to the PARP inhibitors olaparib and niraparib, while those with lower BRCA1/2 signatures did not. Campaner et al. (2020) observed that BC organoids accurately reflect the characteristics of the original tumor tissues, enabling the assessment of standard treatment efficacy and the identification of resistant cell populations within the tumor. Guillen et al. (2022) found that the drug responses of PDX-derived organoids were in line with the responses observed in vivo. Wu et al. (2022) reported that the type I protein arginine methyltransferase (PRMT) inhibitor MS023, which is currently in the clinical development stage, exhibited antitumor effects on TNBC cell lines. Nevertheless, PDOs of TNBC showed varying responses to MS023, with only half of the PDOs showing sensitivity to this inhibitor, while the rest displayed resistance. This research offers valuable knowledge on the pharmacological mechanism of MS023, aiding in the development of targeted therapies for TNBC, and presents potential avenues for cancer immunotherapy. Ma et al. (2022) conducted high-throughput screening of a large number of tumor suppressor (LATS) kinase inhibitors and identified an
effective LATS inhibitor, VT02956, from approximately 17,000 compounds. They found that VT02956 represses the expression of ESR1 by targeting the Hippo pathway, leading to the inhibition of proliferation in ER+ BC cells and PDOs, with minimal cytotoxicity to other cells. Therefore, LATS represents a promis- ing target for cancer therapy, especially in cases of endocrine- resistant BC, providing a novel approach for the treatment of ER + BC. To explore new drugs for the treatment of BC patients and bone metastasis, Sun et al. (2022) tested approximately 120,000 compounds from a small-molecule chemical library. They discovered that the compound S6 significantly inhibited the proliferation of SCP2 cells in a dose- and time-dependent manner, and effectively suppressed the growth of BC PDOs. Therefore, they recognized S6 as a potential candidate compound for future development as a new cancer therapeutic drug. Soleimani et al. (2022) reported that JNK-IN-8 (a c-Jun N-terminal kinase inhibitor) significantly inhibited TNBC cells, which exhibited translucent cytoplasmic vacuoles with lysosomal characteristics. Moreover, JNK-IN-8 effectively hindered the growth of TNBC organoids, which displayed rupture and disintegration pheno- types. In addition to inhibiting JNK, JNK-IN-8 also triggers lysosomal biogenesis and autophagy by targeting TFEB/TFE3. These findings suggest that JNK-IN-8 could be a new and promising therapy for TNBC. BC PDOs can also be applied in screening natural compounds. Our group evaluated the ther- apeutic efficiency of oxypalmatine and artemisitene on BC PDOs, supporting them as potential drug candidates for the treatment of BC (Chen et al., 2024; Lin et al., 2023). We also found that liensinine diperchlorate and artemisitene can synergistically attenuate the growth of BC PDOs (Lin et al., 2024).
BC organoids allow for high-throughput screening of different antitumor drugs and select more suitable treatment options for patients receiving existing or emerging therapies, demonstrating the value of these organoids in clinical applications. Using PDOs generated from metastatic BC biopsies, Sachs et al. (2018) reported that in vitro responses of PDOs to tamoxifen mimic those of the corresponding patients, suggesting that PDOs may serve as surrogates for BC to predict drug efficacy in vitro. Deng’s team collected 132 BC tissues from 125 patients, with 77 samples from treatment-naïve patients, and 55 samples from patients who had received systemic anticancer therapy. They established 99 individualized organoid models from these tumor tissues. By conducting large-scale drug testing on PDOs, they identified the most effective drugs for individual patients and successfully developed personalized treatment strategies (Chen et al., 2021b). Shu et al. (2022) assessed the sensitivity of PDOs derived from BC biopsies to different neoadjuvant chemotherapy drugs and found that these PDOs exhibited varied responses to these drugs. They subsequently compared the clinical outcomes of patients with those of in vitro tests and found that PDOs produced consistent responses to drugs, as reflected in the clinical responses of patients. Therefore, BC PDOs have the potential to predict patient responses to both monotherapy and combination drug therapies, facilitating the development of personalized treatment.
Organoids as tools for drug resistance research
Gene mutation, chromosomal amplification and rearrangement are the main factors leading to drug resistance in tumor cells (Fojo, 2007). PDOs have many advantages for investigating drug resistance mechanisms and can simulate the progression of drug
resistance. For example, by employing an increasing concentra- tion gradient method to transform sensitive PDOs into resistant PDOs, it is possible to obtain sensitive or resistant PDOs from the same sample, facilitating the comparison of their gene expression profiles. The outcomes achieved with this method reduce the impact of other interfering variables while maintaining the strong representativeness of each patient, thus revealing the high value of PDOs in drug resistance modeling.
The mechanism of resistance to dopamine agonists (DAs) in prolactinomas is not clear, and no feasible alternative drug treatments are currently available. Through transcriptome sequencing and single-cell sequencing analysis, Cheng et al. (2024) revealed that resistance to DAs in prolactinomas is linked to the upregulation of the focal adhesion signaling pathway. Using DA-resistant prolactinoma organoid models, 180 small molecule compounds were screened, and genistein was identified as a promising treatment option for DA-resistant prolactinomas. Further research revealed that genistein can suppress tumor growth by blocking the FA pathway, and in vivo tests also demonstrated its ability to prevent the formation of subcutaneous tumors.
The aggressiveness of PDAC and its primary and acquired drug resistance are important factors leading to poor prognosis (Kamposioras et al., 2019). The progression and drug resistance of pancreatic cancer are complex processes involving multiple mechanisms, including genetic and epigenetic changes, tumor heterogeneity, metabolic reprogramming, TME, and immune evasion. Common treatment options for PDAC include the gemcitabine+nab-paclitaxel or FOLFIRINOX (5-fluorouracil, leucovorin, irinotecan, and oxaliplatin). These chemotherapy treatments inevitably lead to toxicity and tumor resistance and only prolong the median survival for about one year. To determine the mechanism of chemoresistance in PDAC, orga- noids also represent a reliable approach. Ponz-Sarvise et al. (2019) reported that over-activation of ERBB is a key parameter in the resistance of PDAC to combined inhibition of MAPK and PI3K. They further showed that the combined inhibition of MEK and ERBB using the MEK inhibitor selumetinib and the pan-ERBB inhibitor neratinib resulted in tumor regression in human organoid xenografts in a short-term intervention study. Farshadi et al. (2021) developed 10 primary and FOLFIRINOX-treated PDOs and clarified that the histological, transcriptional, and genetic features of the PDOs reproduce those of the original tissues. Subsequent treatment of PDOs with either individual drugs or the FOLFIRINOX combination regimen demonstrated significant drug resistance in tumors after FOLFIRINOX treat- ment. Peschke et al. (2022) proposed a longitudinal precision oncology platform using functional model systems, including the identification of chemotherapy-induced vulnerabilities using PDOs. Previously, tumor resistance to chemotherapy was often characterized as the selection of resistant populations of tumor cells. However, the authors showed that PDAC cells appeared to develop resistance not only through genetic alterations but also through induced cellular plasticity, leading to the emergence of new drug-sensitive resistant phenotypes. Therefore, this study supports the use of PDOs to identify novel therapeutic targets after standard-of-care treatments in patients with PDAC.
At least three major mechanisms contribute to drug resistance in CRPC, including mutations in the AR, TP53, ETS, and PTEN genes; chromosomal amplification and chromosomal rearrange- ment; and the activation of bypass signaling pathways, such as
the glucocorticoid receptor pathway, which compensates for the loss of AR signaling (Arora et al., 2013; Ku et al., 2017; Mu et al., 2017; Robinson et al., 2015). PCa organoids are also being used to explore the molecular mechanisms of drug resistance. Pappas et al. (2019) utilized PCa organoids to examine the efficacy of anti-androgen drugs on p53- and Pten-deficient PCa. Dhimolea et al. (2021) investigated drug-resistant tumor cells in xenografts, organoids, and cancer patients. This research demonstrated that suppressing BRD4 (a MYC co-activator) led to decreased drug cytotoxicity and enhanced resistance. Lee et al. (2022) developed organoids from patient-derived xenograft models of bone meta- static PCa. Organoids resistant to ADT can be produced by cultivating PCa cells in the absence of androgens or in the presence of anti-androgens such as enzalutamide. Androgen deprivation resulted in the development of reversibly dormant basal-luminal-like hybrid cells, underscoring the potential contribution to disease progression by some cancer treatments, suggesting the need for a reevaluation of treatment strategies (Lee et al., 2022). A synthetic ECM-based hydrogel organoid model was used to study the impact of EZH2 and DRD2 inhibitors on CRPC-NEPC. The results from RNA sequencing, proteomics, special-omics, and functional analyses indicated that epigenetic inhibitors followed by DRD2 treatment can counteract drug- resistance ECM conditions in CRPC-NEPCs (Mosquera et al., 2022).
Chemoresistance often occurs in OC, resulting in a lack of response or recurrence (Patch et al., 2015). PDOs can be utilized to identify new targets that contribute to chemoresistance and sensitivity and help determine appropriate clinical treatment strategies. Sun et al. (2020) explored the mechanism of cisplatin chemoresistance by using OC organoids. The authors reported that Aurora-A regulates cell senescence and glucose metabolism to develop cisplatin resistance through the SOX8/FOXK1 signaling pathway in OC. McCorkle et al. (2021) analyzed the differences in gene expression between paclitaxel-sensitive and resistant OC organoids through RNA sequencing and identified significant upregulation of the ABCB1 gene in resistant organoids. To assess sensitivity to first-line treatment and genomic drivers related to carboplatin resistance, Gorski et al. (2021) developed HGSOC PDOs and assessed their sensitivity to different doses of drugs. By comparing the results of mutation analysis between resistant and sensitive PDOs, they identified TEME178B as a novel gene related to chemotherapy resistance. Wang et al. (2022b) established cisplatin-resistant OC organoids by increasing the concentration of cisplatin. RNA sequencing of cisplatin-sensitive and resistant PDOs revealed that the Fibrillin- 1/VEGFR2/STAT2 signaling pathway regulates glucose metabo- lism and angiogenic processes, promoting chemoresistance. Another approach involves obtaining paired samples before and after recurrence from the same patient and performing short- term organoid culture in vitro. Pietilä et al. (2021) used this method to culture PDOs from the ascites of an HGSOC patient before chemotherapy and after relapse. They proposed that collagen-6 adhesion was upregulated by cisplatin and that collagen-6 increased resistance to cisplatin-induced cytotoxicity, particularly in recurrent HGSOC organoids.
Organoid modeling of the tumor microenvironment
The TME consists of the ECM and stromal cells, which include immune cells, endothelial cells (ECs), cancer-associated fibro-
blasts (CAFs), adipocytes, pericytes, and so on (Figure 3A). Stromal cells interact with one another and tumor cells by releasing growth factors, chemokines, and regulatory molecules, and participate in tumorigenesis, progression, metastasis, and drug resistance (Carmona-Fontaine et al., 2017; Rodrigues et al., 2021). Although PDOs have exhibited promise in drug assess- ment, it is undeniable that these PDOs are composed mainly of epithelial cells and lack stromal components present in the native TME. Many drug responses to tumors are related to the TME and are ultimately strongly influenced by it. Therefore, the develop- ment of PDO models containing tumor stromal components is crucial for recapitulating the parental TME and for conducting preclinical drug testing. To establish advanced models that can mimic tumor structure, heterogeneity, and assumed cell-cell interactions, efforts have been made to create co-culture models of endocrine tumor organoids with other cell types (Figure 3A). Currently, three innovative methods have been reported to model the TME in a more physiologically significant way (Figure 3B). (i) Tumor cells and their environment can be embedded in gels inside transwell culture dishes for air-liquid interface (ALI) culture; (ii) bioprinting can be used to develop organoids containing TME cells, providing a more authentic structure for examining tumor behavior; and (iii) organoids can be cultured in microfluidic devices containing collagen, thereby maintaining tumor cells and endogenous immune cells, and these systems can be utilized on their own or in combination with bioprinting, to replicate dynamic elements such as blood flow or targeted drug administration.
An ALI method was reported for the propagation of PDOs from primary clinical tumors. This method can preserve the multi- component features of the TME with various components of the tumor parenchyma and stroma, including functional tumor- specific infiltrating lymphocytes. This method ensures the retention and expansion of the endogenous stroma and immune cells present in organoids and the corresponding tumor tissues. Using the ALI method, PDOs were extracted from more than 100 human biopsies or mouse tumors in syngeneic immunocompe- tent hosts, where tumor epithelia were co-cultured with native embedded immune cells such as T cells, B cells, NK cells, and macrophages. Importantly, PDOs effectively simulated immune checkpoint blockade with anti-PD-1 and/or anti-PD-L1 agents, expanding and activating tumor antigen-specific TILs and leading to tumor cell cytotoxicity, highlighting the potential of this model for developing personalized immunotherapies for lymphomas (Neal et al., 2018). A large-scale study involving 100 excised tumor samples, including PDAC samples, revealed that ALI PDOs highly preserved the integrated stromal CAFs and tumor architecture (Grönholm et al., 2021).
CAFs are tumor-specific stromal components of the TME that can enhance tumor growth by synthesizing and reshaping the ECM as well as secreting growth factors that promote angiogen- esis. Moreover, they can impact the treatment response by negatively affecting the entry of drugs into the tumor. Co-culture techniques have been used to supplement PDOs with CAFs. Seino et al. (2018) reported that co-culturing human PDAC organoids with CAFs that exhibit immunosuppressive characteristics with- in the TME can stimulate the growth of organoids representing WNT-nonproducing PDAC subtypes through CAF-produced WNT. On the other hand, TGFß and IL-1 ligands that are secreted by PDOs can increase the heterogeneity of CAFs and trigger different subtypes of myofibroblast and inflammatory
CAFs, respectively (Biffi et al., 2019). Tsai et al. (2018) reported a platform of pancreatic cancer organoids incorporating patient- derived CAFs and peripheral blood lymphocytes. The activation of myofibroblast-like CAFs and tumor-dependent lymphocyte infiltration were identified in these complex models. This in vitro model is suitable for studying tumor-stroma interactions and evaluating immunotherapy drugs. Compared with PDAC orga- noids cultured alone, organoids co-cultured with CAFs exhibited increased resistance to commonly employed PDAC drugs. Single- cell RNA sequencing showed differences in gene expression in PDAC organoids stimulated with CAFs, leading to the induction of EMT in cancer cells. This study indicated that EMT induction plays a role in supporting the chemoresistance observed in their co-culture system (Schuth et al., 2022). Davaadelger et al. (2019) introduced fibroblasts into organoids to explore the impact of BRCA1 mutations on progesterone in breast cells. A BC organoid-fibroblast co-culture model revealed a shared pro- tumorigenic paracrine signaling mechanism between normal breast fibroblasts and CAFs (Chatterjee et al., 2019). This mechanism involves the release of CCL7, IL-6, and IL-8 by normal breast fibroblasts or CAFs, resulting in the release of platelet-derived growth factor BB from ER-positive BC cells.
The vascularization of organoids is another aspect that should be considered. Organ-on-chip technology is a new combination of microfabrication techniques and biomimetics. With the microfluidics system regulating fluid flow, a 3D microsystem of human organs-on-a-chip can be developed (Zimmermann, 2021). The vascularization of organoids using microfluidic platforms can replicate the transport characteristics of the TME in vivo (Sontheimer-Phelps et al., 2019). For example, Shirure et al. (2018) established an organoid chip for BC featuring three interconnected microfluidic devices supporting endothelial cell vasculature to produce a 3D network of self-assembled perfusable vessels, allowing the growth of a BC organoid structure. Swaminathan et al. (2019) used bioprinting technology to print 3D spheres of breast epithelial cells and co-cultured them with vascular endothelial cells, leading to the successful development of tissue models suitable for drug efficacy research. Choi et al. (2021) established PDAC PDOs with a vascular niche, and demonstrated that those PDOs retained cancer-initiating cells by interacting with endothelial cells through the Wnt and Notch signaling pathways.
However, the preservation of various cell populations, as well as the extended conservation of stromal elements and immune cells in organoid cultures, remains under investigation. The culture medium should be optimized to support various cell types without favoring clone selection. Raghavan et al. (2021) emphasized that the in vitro TME and culture medium formula may not only non-genetically drive the state of pancreatic cancer cells cultured as PDOs, but also significantly affect drug responses.
Organoids for cancer immunotherapy
The application of immunotherapy has become increasingly prominent in the field of cancer treatment. It stimulates or restores the immune system of patients, enhances antitumor responses in the TME, and regulates the elimination of tumor cells (Rosenberg and Restifo, 2015). Nevertheless, due to the complexity of the human immune system and the immune escape mediated by the TME, the efficacy of in vitro immunother-
A
Tumor cell
B cell
T cell
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NK cell
Dendritic cell
Adipocyte
Pericyte
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Cancer-associated fibroblast
Tumor tissue
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Myeloid-derived suppressor cell
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apy drug models is often inconsistent with clinical responses in many patients (Prasetyanti and Medema, 2017). Hence, the obstacles associated with immune tumor models are significant, and there is a pressing need for in vitro models that can be used to evaluate individual effectiveness in a clinical setting. PDOs provide a new opportunity for tumor immunotherapy. Co- culture systems of PDOs and immune cells that can simulate the cellular and molecular complexity of the TME, encompassing the interaction between tumor cells and immune cells, allowing for a more precise representation of resistance mechanisms to immunotherapy. Utilizing PDOs derived from specific patients facilitates the prediction of individual responses to various immune-based therapeutic strategies. Additionally, PDOs provide essential perspectives for the advancement of new immunothera- pies (Grönholm et al., 2021; Ye et al., 2020).
Wan et al. (2021) employed a 3D matrix to co-culture PDOs with immune cells, replicating the TME of HGSOC. The bispecific anti-PD-1/PD-L1 antibody treatment led to the activation of NK and T cells, resulting in more active and cytotoxic phenotypes via the suppression of bromodomain-containing protein 1 (BRD1). Therefore, inhibiting BRD1 could be a novel treatment option for OC patients to overcome immune cell evasion. Tumor-infiltrating mast cells (TIMs) can act as either promoters or suppressors of tumors, and are linked to resistance to anti-PD-1 therapy. Compared with PDOs from HGSOC patients with high stromal TIMs, those with low stromal TIMs showed enhanced sensitivity
to anti-PD-1 therapy, indicating that the accumulation of stromal TIMs could contribute to resistance against PD-1 blockade and serve as a novel biomarker for immunotherapy resistance (Cao et al., 2021). Zhang et al. (2021b) showed different responses of tumorigenic organoids to HGSOC che- motherapeutics. By neutralizing the cytokines/chemokines produced by organoids, the distinct immune microenvironments induced can be modulated. These results may be applied to increase the efficiency of immunotherapy and chemotherapy for patients with OC.
Chimeric antigen receptor (CAR)-T cell therapy has shown encouraging outcomes in certain hematologic cancers, but its efficacy in solid tumors is still being explored. Compared with other preclinical models commonly employed in CAR-T cell therapy research, PDOs provide unique advantages as they preserve the biological characteristics of original tumors, which are crucial for studying CAR-T cell therapy in early-stage solid tumors. Many research teams have developed co-culture models of organoids with lymphocytes, particularly T cells and NK cells, to model immunotherapies such as CAR-T cells and immune checkpoint blockade. Researchers co-cultured CAR-T cells with PDOs to verify the anti-tumor effects of novel targeted CAR-T cells, modified CAR-T cells, and CAR-T combination therapies. For example, Schnalzger et al. (2019) employed CAR-NK-92 cells that target organoids expressing EGFRvIII to assess tumor antigen-specific cytotoxicity and examine the efficacy of CAR-
and tumor-specific therapy. These findings revealed that CAR- NK-92 cells can directly target the ubiquitous epithelial antigen EPCAM and effectively target multiple class I organoids. Co- culturing of CAR-derived cells with PDOs can capture the molecular and cellular processes of immunotherapy, demonstrat- ing great promise in predicting treatment efficacy and cytotoxi- city.
Oncolytic viruses have replicative ability and tumor-targeting specificity, and they effectively infect and destroy tumor cells, which induces anti-tumor immune responses in the host (Kaufman et al., 2016). Pancreatic cancer (Raimondi et al., 2020) and breast cancer (Carter et al., 2022) organoids have been employed as screening platforms for oncolytic virus treatment. According to research conducted by Raimondi et al. (2020), oncolytic adenovirus exhibited good replication selectiv- ity in PDAC organoids but not in normal pancreatic tissue organoids. Notably, PDOs exhibit individualized responses, confirming their reliability as in vitro models for testing preclinical responses to oncolytic viruses. Although some studies have suggested the potential of PDOs for investigating the infectivity and cytotoxicity of oncolytic viruses, to the best of our knowledge, the immune responses induced by oncolytic viruses have not been studied in complex immune organoids.
Research on immunotherapy has driven the development of new cancer treatments, whether used independently or in combination with other treatments, which target immune regulatory pathways to control cancer cells. PDOs combined with TME characteristics simulate immunotherapy responses. Additionally, co-culturing with immune cells can be used to study immunotherapy efficiency and the mechanism of drug resistance. In the future, organoid methods will significantly promote basic scientific research and the translation of immuno- oncology, accelerating the progress of personalized immunother- apy for human cancers.
Limitations and perspectives
As a promising technology, PDO models have become important tools for basic, preclinical, and clinical research. However, several limitations remain that warrant further development in the following aspects (Figure 4).
The limited availability of patient materials is one challenge. Typically, there are few opportunities to collect materials from each patient, and if palliative aspiration of ascites or secondary debulking surgery is performed, there are rarely any additional chances. Methods used for collecting tumor samples, such as biopsies and surgical resection, also affect the efficiency of organoid formation. Standardized culture procedures are crucial and need to be developed and optimized. There are also variations in PDO production due to the absence of a universally accepted standardized approach. Unstandardized sampling procedures can reduce the activity of tumor tissues, thus affecting the success rate of PDOs. Compared with healthy tissues, tumor-derived organoids generally have a slower growth rate, leading to contamination by healthy tissues (Dijkstra et al., 2020; Karthaus et al., 2014). The medium components for PDO cultures vary by tumor type. The addition of some growth factors or pathway inhibitors may result in the unintentional selection of specific clones within the PDOs. There are advantages and disadvantages to the long-term and short-term expansion of PDO cultures. Long-term expansion allows for multiple tests to improve
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Ethical issues
Limited patient materials
Incomplete microenvironment components
No standardized protocols
LIMITATIONS
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Contamination from healthy tissues
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Heterogeneity within PDOs
reproducibility but increases the risk of clonal selection during prolonged culture, leading to a loss of heterogeneity. Further- more, the production of PDOs is currently expensive, and the technology is still developing and challenging to integrate into healthcare systems. The ethical implications of PDO biobanking also require further consideration.
Current organoid technology cannot easily maintain the complexity of the patient-specific immune microenvironment. Due to the absence of tumor matrix, blood vessels, and immune cells, PDO models fail to fully replicate the tumor’s characteristics (Jabs et al., 2017; Ma et al., 2021). PDOs lacking immune and vascular systems also cannot be used to evaluate immune-related drugs and anti-angiogenic drugs. In recent years, “organ-on-a- chip” technology, which involves the integration of microfluidic chips and organoids, has effectively addressed some of these issues. This novel technology can overcome some limitations in the application of organoids by controlling the behavior of stem cells and the cellular microenvironment through further design improvements. This technology has been investigated in the study of cancers including breast cancer (Azimian Zavareh et al., 2022) and pancreatic cancer (Haque et al., 2021; Schuster et al., 2020), providing a quick and reliable platform for drug testing. Tumor organoids can mimic the TME either by cultivating tumor spheroids in microfluidic devices to maintain autologous myeloid and lymphoid cell populations, or by incorporating immune components such as CAFs to control organoid vascularization and perfusion to retain endogenous stromal components (Wang et al., 2016). Microfluidic chip technology can also accurately replicate the characteristics of blood vessels and flow, addressing the shortcomings of the vascular system in PDO models (Dijkstra et al., 2018). For example, Shirure et al. (2018) employed this technology and organoid integration to simulate perfused vessels to study the progression and response of PDOs to chemotherapy and anti-angiogenic therapy. Additionally, the fluid in micro- fluidic devices can link organoids from varied tissue sources. This enables the system to mimic the exchange of nutrients and
substances and the generation of vascular networks between tumors, thereby better modeling the behaviors of tumor metastasis and spread (Neufeld et al., 2022; Sharifi et al., 2021).
Many studies have revealed that organoids effectively reflect inter-patient and intra-tumor heterogeneity. Nevertheless, the variability in drug responses across organoid lines derived from distinct regions of the same primary tumor has raised concerns about the representativeness of organoids in personalized drug screening. The lack of standardized analysis techniques hinders high-throughput screening using organoids as models, necessi- tating automation for reliable high-throughput applications. In addition, there is currently a lack of unified standards for defining patient clinical response and drug effectiveness for PDOs. Models with greater clinical relevance are necessary to establish criteria for interpreting data for predicting drug responses. To verify and advance the use of PDOs in forecasting patient responses, extensive randomized controlled trials must be conducted for validation, and uniform standards should be established.
Despite the remaining challenges, PDOs represent a rapidly developing technology with tremendous potential in advancing personalized medicine. Organoids have proven beneficial in the study of endocrine tumors, which will continue to bring revolutionary changes to preclinical research, paving the way for the development of new, effective, and appropriate treatment modalities. In the future, the integration of multiple research models is the optimal approach to further advancing medical research. By continuously optimizing the culture conditions of in vitro models, a more reliable platform can be established for the study of endocrine tumors and drug screening, leading to a broader clinical application of this new technology.
Conclusion
PDO models have been proven to be powerful platforms for both basic and translational cancer research. We reviewed the establishment, application, and future prospects of endocrine tumor organoids. These models are promising tools for cancer research as they mimic the structural and histological char- acteristics of tumors and exhibit specific gene phenotypes of patients. Therefore, they can serve as tools for studying tumorigenesis, progression, metastasis, and drug resistance. Organoid technology is not only applicable in basic research but also plays an important role in clinical studies. Currently, PDOs show great potential as models for preclinical high- throughput drug screening and novel drug development and can provide evidence for clinical trials. In addition, some complex co-culture models may serve as tools for personalized immu- notherapy validation and toxicity examination in early clinical trials. From in-depth investigations of relevant mechanisms to the application of personalized medicine, organoid platforms map a hopeful landscape for cancer research and serve as the cornerstone for more precise oncology strategies.
Compliance and ethics
The authors declare that they have no conflict of interest.
Acknowledgement
This work was supported by the Guangdong Basic and Applied Basic Research Foundation (2023A1515220165), the Shenzhen Science and Technology Program (KCXFZ20211020163407011, JCYJ20230807095802005, JCYJ20230807095803006, JCYJ20230807115159050, RCBS20221008093330066), the National Natural Science Foundation of China (32401178), and the Bethune Charitable Foundation (JKM2022-A03).
Open Access
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