"Nature" sub-issue: Tumor organoids + chip, realizing rapid anti-cancer drug sensitivity detection!

Recently, Researcher Liu Peng's research team from the Department of Biomedical Engineering, Tsinghua University School of Medicine, together with the team of Academician Wang Jun from Peking University People's Hospital, and Chen Xiaofang's research team from Beijing University of Aeronautics and Astronautics, published new research results on the combination of organoids and chips in the academic journal Nature Communications "Patient-derived organoids analyzed on a superhydrophobic microwell array for predicting drug response of lung cancer patients within a week"
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The research results combine the two cutting-edge technologies of microfluidic chips and tumor organoids, significantly improving the efficiency and timeliness of predicting the clinical efficacy of anti-cancer drugs for tumor patients
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In the era of precision treatment, in vitro drug susceptibility prediction of tumors is an important research direction for individualized treatment
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In recent years, important progress has been made in tumor organoid technology, and in vitro models for predicting the efficacy of anti-cancer drugs for patients have been developed in many cancer types
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However, in the field of lung cancer, the construction and application of tumor organoid models are limited by issues such as efficiency and time-consuming, which are more difficult than other cancer types
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The research team combined microfluidic chip and tumor organoid technology, carried out a cross-field, clinical-oriented joint research, and developed a new integrated superhydrophobic microporous array chip (InSMAR-chip), which can obtain tumor organoids within one week The results of drug sensitivity prediction have great prospects for clinical transformation
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The research team first improved the processing method of tumor samples, using mechanical processing methods to cultivate a large number of lung cancer organoids (LCOs) from fresh tumor tissues resected and biopsy, confirming that LCOs retain the histology and genetics of parental tumors Scientific characteristics, and has the potential for unlimited passage and expansion
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At the same time, the team developed InSMAR-chip and used it for the high-throughput three-dimensional culture and analysis of LCOs
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Because the micropore volume on the chip is in the order of nanoliters, the sample consumption and culture time are greatly reduced, and the drug reaction test is completed in only one week, and the drug sensitivity result is obtained
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Subsequent experiments fully proved that the test results of these drugs are highly consistent with the patient-derived xenografts, tumor gene mutations and clinical results
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The combination of LCOs model and microfluidic chip provides an effective and reliable technical means for predicting patient-specific drug response in clinical practice
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Research results 1.
The LCOs culture team from the patient’s cancer tissues used mechanical methods to process 103 surgically resected lung tumor samples, including 71 adenocarcinoma (AC), 23 squamous cell carcinoma (SCC), and 4 For small cell lung cancer (SCLC) and 5 other types of lung cancer, the final organoid culture success rate was 79% (55/71 AC, 18/23 SCC, 4/4 SCLC, 4/5 other types of lung cancer)
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Figure 1.
The establishment of LCOs from the patient’s tumor for subsequent long-term culture and a one-week drug sensitivity test on InSMAR-chip.
2.
The LCOs established and cultured by the characteristic team of LCOs retain the histology and genetics of the original tumor tissue It can be kept stable after long-term in vitro culture and passage.
The results in the figure below show that the 0th generation LCOs have the main characteristics of tumor organoids
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Figure 2.
H&E and immunohistochemical staining images of lung cancer tissue and derived tumor organoids
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H&E staining results show that LCOs have a 3D structure, which is consistent with tumors in vivo
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LCOs derived from adenocarcinoma maintain the acinar or solid structure, and the expression patterns of thyroid transcription factor 1 (TTF-1) and cytokeratin 7 (CK7) are also retained in the organoid structure
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Squamous cell carcinoma organoid cells LC97-O showed poor differentiation, high proliferation, p40 positive expression and partial expression of CK5/6, which reproduced the characteristics of the original tumor tissue
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Figure 3.
Heat map of lung cancer tissue and LCOs to genome-wide copy number variation (CNV)
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The increase (red) and decrease (blue) of the DNA copy number found in the original lung cancer tissue are preserved in the corresponding tumor organoids
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Compared with the original tumor tissue (T: tissue, O: organoid), all LCOs can see signal amplification
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3.
Integrated superhydrophobic micropore array chip drug sensitivity test verification.
The integrated superhydrophobic micropore array chip has the characteristics of high throughput, small size, saving samples and reagents, and real-time tracking and addressing.
The surface of the chip has the characteristics of high throughput, small size, and real-time tracking and addressing.
With the super-hydrophobic coating, the micropores are physically isolated, ensuring an independent liquid environment for each micropore, which is convenient for the operation of sample filling and liquid exchange.
In addition, the chip micropore spacing is consistent with the 1536-well plate, which is more suitable Equipped with commercial imaging and testing equipment
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Figure 4.
Image of InSMAR-chip chip with droplet array in the microwell
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Due to the repulsion of the superhydrophobic surface with a contact angle greater than 160°, when excess medium is sucked out of the chip, a uniform medium droplet array can be spontaneously formed in the microwell array
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Figure 5.
Comparison of LCOs on-chip and off-chip culture
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On the 7th day, the comparison of the growth rate (g) and viability (h) of the four LCOs showed that there was no significant difference between the LCOs cultured on and off the chip (n = 20, the results are shown as mean + SD )
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Figure 6.
Flow chart of a week-long drug susceptibility test on InSMAR-chip
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AB-1: Use alamaBlueTM for cell viability test before drug treatment, AB-2: Cell viability test after drug treatment
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Figure 7.
Comparison process of drug sensitivity test using PDX-derived organoids (PDXO) and PDX mouse model.
Figure 4.
One-week drug sensitivity test based on LCOs to predict the efficacy of lung cancer targeted drugs on the chip The prediction of the efficacy of LCOs on lung cancer targeted drugs can be obtained earlier than the NGS test, and the results are consistent with the results of gene mutations
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Figure 8.
The fitted dose-response curves of EGFR mutant LC132O and EGFR wild-type LC133-O indicate the difference in the sensitivity of the two tumors to targeted drugs, which is very consistent with the clinical results
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5.
The one-week drug sensitivity test represents the heterogeneity of tumor response to chemotherapy.
Figure 9.
The heterogeneous response of lung adenocarcinoma organoids to cisplatin chemotherapy
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The fitted dose-response curve shows the response of AC organoids to pemetrexed + cisplatin (PC) and gemcitabine + cisplatin (GC)
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Each data point represents the average of 3 replicates
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The heat map on the right is an example of two organoids sensitive to PC and two other organoids sensitive to GC
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R indicates that IC50 is not obtained by fitting
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Figure 10.
The drug response curve on the left shows the results of on-chip drug response testing using LC96-O for three chemotherapy regimens (PC, GC, and paclitaxel + cisplatin)
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The line graph on the right represents the tumor volume of PDX with the same chemotherapy regimen used for LCOs
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Of these three cisplatin-based chemotherapy regimens, none of them can effectively reduce the survival rate of organoids
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The same chemotherapy drugs given to PDX mice failed to effectively inhibit the growth of xenograft tumors, which is consistent with the results of LCOs-based testing
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Although effective drugs have not been successfully identified, these data indicate the feasibility of LCOs-based drug screening on a chip to accurately select anticancer drugs for individual patients
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In summary, the research team has established a set of effective lung cancer sample processing and analysis methods, and successfully shortened the drug susceptibility test to one week.
Compared with the PDO-based drug susceptibility test, the detection time is several weeks or even months.
The new method The advantage of timeliness is outstanding, and the detection time meets the requirements of clinical practice
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The results of LCOs drug sensitivity testing using InSMAR-chip, through comparison with PDX models, gene mutations, and actual clinical efficacy, have proved that it can predict the response of tumors in vivo to anticancer drugs and has the potential for drug screening
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It is believed that this cutting-edge technology combining chips and organoids will bring new breakthroughs for tumor organoids to predict the clinical response of drugs.
The research team will continue to work together and look forward to developing more application scenarios and transforming this technology in the future.
In clinical practice, we have opened up the last mile of precision medicine
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Dr.
Hu Yawei from Tsinghua University School of Medicine, Sui Xichao, deputy chief physician of Thoracic Surgery, Peking University People’s Hospital, are the co-first authors of this article.
Researcher Liu Peng, Academician Wang Jun and Associate Professor Chen Xiaofang are the corresponding authors of this article
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Link to the paper: https:// The article is transferred from: Tsinghua University School of Medicine