Article Figures & Data
Figures
- Figure 1
Establishment of patient-derived organoids as in vitro tumor models for colorectal cancer. Morphology of patient-derived organoids (PDOs) on days 1 and 7. After 7-day culture, the organoids were used to evaluate hyperthermia and chemotherapy. The figure shows a flowchart of hyperthermia and chemotherapy evaluation. PDOs are bifurcated to evaluate the following chemotherapy drugs at 37 °C and 43 °C: mitomycin, abraxane, 5-fluorouracil, oxaliplatin, raltitrexed, gemcitabine, and lobaplatin.
- Figure 2
Raltitrexed has the highest hyperthermia chemotherapy sensitization enhancement ratio score among the screened chemotherapy drugs. (A) The dose-response curves of organoids derived from patient 1 after treatment with different drugs for 90 min at 37 °C and 43 °C. Cell viability at 37 °C was measured with 10% CCK-8 after 2 days under drug-free conditions after drug treatment. The hyperthermia chemotherapy sensitization enhancement ratio (HCSER) score was obtained by calculating the ratio of the IC50 values between 37 °C and 43 °C for each drug. That is, HCSER = IC50 (37 °C)/IC50 (43 °C). (B) The HCSER scores of organoids derived from the other 10 patients. Raltitrexed had the highest HCSER score. (C) The highest HCSER scores of the 7 chemotherapy drugs in organoids from 22 patients. (D) Dose-response curves of organoids derived from DLD-1 cell lines, and the HCSER summary for all evaluated drugs. (E) The HCSER scores of the organoids derived from another 2 colorectal cancer cell lines. Raltitrexed showed the highest HCSER scores. (F) Counts of the highest HCSER scores of the 7 chemotherapy drugs in the 4 colorectal cancer cell lines.
- Figure 3
Gene expression profiles of raltitrexed hyperthermia sensitization. (A) Clustered heat map of differentially expressed genes. G1 denotes patients insensitive to raltitrexed (P15, P20, and P22). G2 denotes patients sensitive to raltitrexed (P5 and P10). (B) Enrichment in KEGG pathway terms for differentially expressed genes between the G1 and G2 groups. The horizontal axis denotes the enriched KEGG pathways, and the vertical axis displays the −log10 (P-value). The red boxes represent the 27 genes participating in these pathways, and the red arrows represent pathways reported to be associated with hyperthermia. (C) Western blot analysis was conducted to assay the downregulated expression of PI3K, p-PI3K, AKT, p-AKT, 4E-BP1, p-4E-BP1, Focal Adhesion Kinase, and p-Focal Adhesion Kinase in the raltitrexed high-HCSER group. The low-HCSER group contains patients 13, 18, and 21, and the high-HCSER group contains patients 2, 3, and 6.
- Figure 4
Patient-derived organoids reflect personal response to chemotherapy in combination with hyperthermic intraperitoneal chemotherapy. (A) Treatment and procedure timeline in patient 8. The values highlighted in blue indicate the administration of chemotherapy. The values highlighted in red indicate the efficacy of hyperthermic intraperitoneal chemotherapy. (B) Left: CT image of patient 8 before surgery. Right: CT image of patient 8 at 1 month after surgery. The small nodules and peritoneal fluid decreased, as compared with those in the left CT image. (C) Cell viability of organoids derived from patient 8 in response to raltitrexed, mitomycin, and oxaliplatin, and the HCSER for those drugs.
Tables
Drug name Drug brand Drug specifications Raltitrexed NANJING CHIA TAI TIANQING 2 mg Mitomycin HANHUI PHARMA 10 mg Oxaliplatin SANOFI 50 g 5-fluorouracil PUDEP HARMA 0.25 g Lobaplatin Hainan Changan International Pharmaceutical Co. Ltd. 50 mg Gemcitabine NANJING CHIA TAI TIANQING 0.2 g Abraxane CSPC 100 mg Drug name Recommended dose in clinically of single chemotherapy Intraperitoneal perfusion concentration Experimental setting Raltitrexed 3 mg/m2 2.8 μM 0/0.2/2/20/200/2,000 μM Mitomycin 6–8 mg 5.9 μM 0/0.05/0.5/5/50/500 μM Oxaliplatin 130 139 μM 0/0.1/1/10/100/1,000 μM 5-Fu 500–600 mg/m2 1.96 μM 0/0.02/0.2/2/20/200 μM Lobaplatin 50 mg/m2 53.5 μM 0/0.05/0.5/5/50/500 μM Gemcitabine 1,000/1,250 mg/m2 1.8 μM 0/0.02/0.2/2/200/2,000 μM Abraxane 260 mg/m2 122.9 μM 0/0.1/1/10/100/1,000 μM High-HCSER group Low-HCSER group Cases Average SD Cases Average SD 11 167.37 297.4 11 4.89 5.90 P 0.0019 The High-HCSER group contains patients 1–11, and the low-HCSER group contains patients 12–22. SD, standard deviation. P < 0.05 was considered statistically significant.
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