Table 1.
Relationship between patient’s clinicopathological features and the culture success rate of gastric cancer organoids.
Fig 1.
Study Workflow and histopathological characterization of GC PDOs.
(A) Schematic overview of GC PDOs experimental workflow: tissue isolation, organoid culture, histopathological evaluation, genomic analysis, in vitro drug sensitivity testing, and prediction of clinical response. (B) Representative bright-field images of GC-010 PDOs from day 0 to day 9 post-seeding in Matrigel. Scale bars, 100 µm. Similar growth patterns were observed across all 17 established PDO lines. (C) Bright-field images highlighting the morphological diversity among PDOs derived from SRCC (GC-014, GC-027) and non-SRCC (GC-010, GC-011). Scale bars, 100 µm. (D) Representative H&E staining images of GC PDOs and their corresponding parental tumors. GC-007 is SRCC and GC-025 is non-SRCC. Black arrow indicates signet ring cells; blue arrow indicates solid or cystic structures. Scale bars, 50 µm. (E) IHC staining of Ki67, CK7, CK20 and CDX2 in GC PDOs and their parental tumors from GC-007 and GC-025. GC PDOs faithfully recapitulated histologic features of their parental tumors. Scale bars, 50 µm. P, passage. D, day. GC, gastric cancer. PDOs, patient-derived organoids. SRCC, signet-ring cell carcinoma. Non-SRCC, non-signet-ring cell carcinoma.
Fig 2.
Genetic features of GC PDOs and their parental tumors.
(A) Heatmap depicting CNVs in GC PDOs and their matched parental tumors. Columns represent individual samples, and rows represent genomic positions across chromosomes 1-22. The color scale indicates DNA copy number gains (red) and losses (blue). (B) Genome-wide CNVs profiles for two representative GC PDOs and their parental tumors (GC-010 and GC-014). The upper panels show genome-wide CNV profiles (chromosomes 1-22) with copy number gains (red), losses (blue), and regions with no change (green). The lower panels display B-allele frequency (BAF) distribution, showing balanced heterozygosity (orange), allelic imbalance (blue), and LOH events (BAF = 0/1). (C) Heatmap showing genetic alterations in the commonly mutated genes in GC, with alterations color-coded by type (e.g., missense, nonsense). (D) Distribution of base substitution types detected in GC PDOs and their corresponding parental tumor tissues (GC-007, GC-010, GC-014, GC-025). GC, gastric cancer. PDOs, patient-derived organoids.
Fig 3.
Response of GC PDOs to chemotherapeutic drugs.
(A-E) Dose-response curves illustrating the sensitivity of PDOs to (A) 5-FU, (B) oxaliplatin, (C) SN-38, (D) paclitaxel, and (E) epirubicin. Data are presented as the mean ± SD from three biological replicates. Experiments were performed using 13 PDO lines. (F-G) Representative bright-field images of GC PDOs (GC-012, non-SRCC; GC-014, SRCC) treated with the indicated chemotherapeutic drugs at various concentrations. The tested concentrations were as follows: 5-FU (C1 = 0.002 µM, C2 = 0.02 µM, C3 = 0.2 µM, C4 = 2 µM, C5 = 20 µM, C6 = 200 µM), Oxaliplatin (C1 = 0.01 µM, C2 = 0.1 µM, C3 = 1 µM, C4 = 10 µM, C5 = 100 µM, C6 = 1000 µM), SN-38 (C1 = 0.03 nM, C2 = 0.3 nM, C3 = 3 nM, C4 = 30 nM, C5 = 300 nM, C6 = 3000 nM), Paclitaxel (C1 = 0.08 nM, C2 = 0.8 nM, C3 = 8 nM, C4 = 80 nM, C5 = 800 nM), Epirubicin (C1 = 0.002 µM, C2 = 0.02 µM, C3 = 0.2 µM, C4 = 2 µM, C5 = 20 µM). Scale bars, 100 µm. C, concentration. GC, gastric cancer. PDOs, patient-derived organoids. SRCC, signet-ring cell carcinoma. Non-SRCC, non-signet-ring cell carcinoma.
Fig 4.
Sensitivity evaluation of GC PDOs to chemotherapeutic drugs.
(A) Heatmap of tumor inhibition rates (IR) for 13 PDOs treated with five chemotherapeutic agents at steady-state plasma concentrations. (B-C) Comparison of the mean tumor IR among different chemotherapeutic drugs at steady-state plasma concentrations in (B) SRCC (n = 4) and (C) non-SRCC (n = 9) organoids. Data are presented as the mean ± SD. Statistical significance was determined using one-way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001. (D-H) Comparison of the mean tumor IR between PDOs from SRCC (n = 4) and non-SRCC (n = 9) for each chemotherapeutic drug. Data are presented as the mean ± SD. Statistical significance was determined using Welch’s t-test in (D) and Student’s t-test in (E-H). (I) Average sensitivity ranking and 95% CI of different chemotherapeutic drugs in SRCC (n = 4) and non-SRCC (n = 9). Sensitivity ranking was determined by the ratio of the IC50 values of PDOs to the drug’s steady-state plasma concentration, with lower values indicating higher sensitivity. (J) P values for the comparison of sensitivity rankings among chemotherapeutic drugs in SRCC and non-SRCC PDOs. Bold green font indicates that the average sensitivity ranking of drugs in column a is significantly lower than that in column b. Bold red font indicates that the average sensitivity ranking in column a is significantly higher than that in column b. Statistical significance was determined using one-way ANOVA. (K) Differences in the average sensitivity ranking and 95% CI between PDOs from SRCC and non-SRCC for each chemotherapeutic drug. Statistical significance was determined using Student’s t-test. P < 0.05 indicates a statistically significant difference. n, number. GC, gastric cancer. PDOs, patient-derived organoids. IR, inhibition rate. SRCC, signet-ring cell carcinoma. Non-SRCC, non-signet-ring cell carcinoma. CI, confidence interval.
Fig 5.
GC PDOs predict clinical outcomes of patients treated with chemotherapeutic agents.
(A) Kaplan-Meier survival analysis showing DFS outcomes of the patients with matched PDOs. Based on the in vitro drug-sensitivity results of PDOs treated with the corresponding clinical adjuvant chemotherapy regimen, patients were divided into two groups: drug-sensitive (n = 6) and drug-resistant (n = 4). The P value was determined by log-rank test. (B) Kaplan-Meier survival analysis showing PFS outcomes of the patients with matched PDOs. Based on the in vitro drug-sensitivity results of PDOs treated with the corresponding clinical chemotherapy regimen, patients were divided into two groups: drug-sensitive (n = 1) and drug-resistant (n = 2). The P value was determined by log-rank test. (C) Heatmap summarizing the concordance between PDO drug screening results and patients’ clinical outcomes (13 patient cases). The heatmap displays patient outcomes and PDO responses to 5-FU and oxaliplatin. (D) Concordance analysis of drug sensitivity between clinical responses and matched GC PDOs samples. (E) Representative bright-field images of GC PDOs treated with vehicle, 5-FU and oxaliplatin in three selected cases. (F) Fitted dose-response curves of PDOs (GC-021, GC-026 and GC-012) representing the diverse patients’ responses to 5-FU and oxaliplatin. Data are presented as the mean ± SD from three biological replicates. (G) Clinical treatment timeline for patient GC-021 and corresponding drug screening results on matched PDOs. (H) CT scans of patient GC-021 at baseline and after postoperative adjuvant treatment. (I) Clinical treatment timeline for patient GC-026 and corresponding drug screening results on matched PDOs. (J) CT scans of patient GC-026 at baseline and at the metastasis stages. (K) Clinical treatment timeline for patient GC-012 and corresponding drug screening results on matched PDOs. (L) CT scans of patient GC-012 at baseline and after postoperative adjuvant treatment. Red arrows indicate tumors. GC, gastric cancer. PDOs, patient-derived organoids. DFS, disease-free survival. PFS, progression-free survival. NA, not reached. PD, progressive disease.