Precision Medicine in Urothelial Carcinoma Using Patient-Derived Organoids and Microfluidic Chips

Urothelial carcinoma is a common cancer of the urinary system, and a major challenge is that patients often respond differently to the same chemotherapy, making personalized treatment difficult. Taiwan is famous for its excellence in both medical care and chip manufacturing. Therefore, we aim to integrate microfluidic chip technology to optimize treatment strategies. We successfully established patient-derived tumor organoids from three urothelial carcinoma cases. These organoids closely resemble the original tumors in both morphology and protein expression. Drug testing revealed significant variability among patients, highlighting their potential in predicting drug sensitivity. By combining organoids with microfluidic chip systems to mimic blood flow, we provide a promising platform for advancing precision cancer therapy.

Problem:

Cancer remains a major global health challenge and is one of the leading causes of death worldwide. Recent global estimates reported nearly 20 million new cancer cases and 9.7 million cancer deaths in 2022, highlighting its substantial burden on patients and healthcare systems [1,2] (Figure 1). Urothelial carcinoma is a common cancer in the urinary system (Figure 2). However, current cancer therapies still face several limitations [3-5] (Figure 3):

Patients often have different responses to chemotherapy.

It is difficult to choose the most effective treatment for each patient.

Traditional models may not accurately reflect the original tumor structure and molecular features.

Current drug screening methods may not provide fast, personalized, and clinically useful results.

Solution:

Taiwan is famous for its excellence in both medical care and chip manufacturing. Therefore, we aim to integrate microfluidic chip technology to optimize treatment strategies. (Figure 4)

Establish patient-derived urothelial carcinoma organoids that highly retain the tumor’s original structure and molecular characteristics [6].

Test chemotherapy drugs on organoids to evaluate individual drug sensitivity and support precision treatment selection [7].

Integrate microfluidic chips with tumor organoids to enhance drug screening efficiency for improving clinical decision-making and treatment outcomes [8].

Therefore, developing advanced tumor models that combine organoids with microfluidic technology may help identify effective treatments earlier, reduce trial-and-error therapy, lower medical costs, and advance personalized cancer care.

Finding better ways to treat urothelial carcinoma inspired us to build a more realistic and patient-specific tumor model. We asked ourselves a simple question:

“Can we predict which chemotherapy drug works best for each patient before treatment?”

To answer that, we established patient-derived urothelial carcinoma organoids and explored their integration with microfluidic chip technology (Figure 5). First, tumor tissues obtained from patients with urothelial carcinoma are processed to isolate cancer cells, which are then embedded in Matrigel to establish three-dimensional organoid cultures (Figure 6-9). We observed that these organoids preserve key structural and molecular features of the original tumors by hematoxylin and eosin (H&E) staining and immunohistochemical staining [9].

Next, commonly used chemotherapeutic drugs in urothelial carcinoma—including cisplatin, epirubicin, gemcitabine, and paclitaxel—are applied to the organoids at different concentrations. Cell viability is measured using the CellTiter-Glo® 3D reagent to evaluate drug responses. This allows comparison of drug sensitivity across different patient-derived samples.

Furthermore, we incorporate a microfluidic chip system that enables dynamic perfusion of culture medium using a peristaltic pump. This system mimics physiological conditions such as fluid flow and nutrient exchange, providing a more realistic tumor microenvironment compared to static culture.

Through this integrated approach, we aim to build a platform that more accurately predicts patient-specific drug responses.

In this study, we successfully established patient-derived urothelial carcinoma tumor organoids from three individuals (Figure 6-7) and systematically evaluated their biological characteristics and drug responses. Morphologically, the organoids exhibited diverse shapes and sizes both between different patients and within the same patient, reflecting the intrinsic heterogeneity of tumors (Figure 8-9).

Histological analyses, including hematoxylin and eosin staining and immunohistochemical staining for Ki-67 and FGFR3, demonstrated that the organoids closely resembled their corresponding original tumor tissues in both structural organization and molecular expression. These findings confirm that the organoid model faithfully preserves tumor-specific characteristics (Figure 10).

Drug response experiments revealed significant variability among organoids derived from different patients using the CellTiter-Glo® 3D cell viability assay. When treated with four commonly used chemotherapeutic agents, each organoid displayed a unique sensitivity profile (Figure 11). Some drugs were more effective in certain organoids, while others showed limited effects. This highlights the presence of patient-specific differences in drug responsiveness (Figure 12).

In addition, the study demonstrated the feasibility of culturing tumor organoids within a microfluidic chip system (Figure 13). Under dynamic perfusion conditions, organoids maintained stable growth and preserved their three-dimensional structures. Microscopic observations confirmed that organoids could adhere, grow, and form spheroid or clustered structures within the chip environment.

Overall, the results indicate that tumor organoids not only retain key biological features of original tumors but also serve as a reliable platform for evaluating individualized drug responses. The successful integration with microfluidic technology further enhances the physiological relevance of the model.

The results of our study demonstrate that patient-derived urothelial carcinoma organoids are a highly promising platform for personalized drug testing and precision cancer treatment (Figure 14). By preserving key structural and molecular features of the original tumor, these organoids provided a more realistic model than conventional cell culture systems. This makes them especially valuable for evaluating how individual patients may respond to chemotherapy.

Our findings showed that tumor organoids could be successfully established from urothelial carcinoma tissues and that they retained important pathological characteristics of the original tumors. In addition, drug testing revealed clear patient-to-patient differences in chemotherapy sensitivity, highlighting the importance of individualized treatment selection rather than relying on a one-size-fits-all approach.

The integration of microfluidic chip technology further strengthened this platform by creating a more dynamic and physiologically relevant tumor environment. This combination improves the potential for more accurate drug screening, better simulation of in vivo conditions, and more efficient evaluation of treatment response.

Together, these results support the clinical potential of our organoid-based platform. It may help doctors identify effective treatments earlier, reduce ineffective therapy, lower the risk of recurrence, and minimize unnecessary side effects. This could ultimately improve treatment outcomes while reducing the physical, emotional, and economic burden on patients.

Overall, our study highlights the potential of combining patient-derived organoids with microfluidic technology to advance urothelial carcinoma research and move cancer treatment closer to the goal of truly personalized medicine.

Long-term goal is to develop a more efficient organoid-on-a-chip platform that can be translated into real-world applications for precision medicine in urothelial carcinoma (Figure 15) . Future work will focus on validating the correlation between organoid drug responses and actual patient clinical outcomes through longitudinal follow-up. In addition, further optimization of organoid-on-a-chip systems will aim to incorporate more components of the tumor microenvironment, such as immune cells and stromal cells. These advances may enable the development of high-throughput, clinically applicable platforms for personalized therapy and drug discovery.

First, I want to thank all the teachers at the Taiwan International Science Fair for making it such an enjoyable and memorable experience. I would also like to thank my lab mentor, Dr. Hao-Lun Luo, for generously providing laboratory equipment, patiently answering my research questions, and helping me quickly master experimental procedures. Special thanks to Professor Kwang-Huei Lin for his unwavering encouragement, guidance, and helpful suggestions. Finally, I’m really thankful to my parents for always supporting me. They spent so much time driving me to labs and school, helping me follow what I love in science.

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