Construction of patient-derived glioblastoma organoid models and sensitivity analysis of temozolomide and lomustine as monotherapy and in combination

YANG Guangzhao; ZOU Cheng; LIU Xiao; QIN Haozhe; PAN Jiahao; CAO Zhengcong; ZHANG Qi; LI Juan; GU Jintao; HE Yalong; LIN Wei

doi:10.13276/j.issn.2097-1656.2026.07.002

Abstract：Objective To establish patient-derived organoid(PDO)models of glioblastoma(GBM)and evaluate their application value in predicting sensitivity to temozolomide and lomustine,both as monotherapies and in combination.Methods Tumor tissues were collected from five patients with pathologically confirmed GBM who had not received any prior systematic treatment.PDO models were constructed using a suspension culture method.The fidelity of the PDOs to the primary tumors was validated by HE staining,immunofluorescence(IF)staining,and whole-exome sequencing(WES)to assess histological morphology and genetic characteristics.Invasive behavioral traits were evaluated using a 3D invasion assay in vitro.Drug sensitivity of stably passaged PDOs was assessed using a lactate dehydrogenase release assay following drug administration.Dose-response curves were fitted to calculate the IC50·Synergistic effects of drug combinations were evaluated by calculating the combination index(CI)using the Chou-Talalay method.Results Three GBM PDO lines capable of stable subculture for over three passages were successfully established,with a culture success rate of 60％.HE and IF staining demonstrated high histological concordance between the PDO models and the primary tumors.WES analysis revealed that the paired PDOs retained up to 92％of the genomic mutation heterogeneity present in the parental tissues.Drug sensitivity testing using two common chemotherapeutic agents showed significant inter-individual variability.The IC50 values of the three PDO lines for temozolomide were 58.52 μmol/L[95％CI(52.17 μmol/L,65.52 μmol/L)],2 923 μmol/L[95％CI(1 784 μmol/L,6 133 μmol/L)],and 187.10 μmol/L[95％CI(159.80 μmol/L,220.60 μmol/L)],respectively.The IC50 values for lomustine were 12.65 μmol/L[95％CI(10.56 μmol/L,15.11 μmol/L)],105.90 μmol/L[95％CI(83.69 μmol/L,141.30 μmol/L)],and 19.64 μmol/L[95％CI(16.41 μmol/L,23.57 μmol/L)],respectively.In combination experiments,heterogeneous dose-effect relationships were observed following temozolomide and lomustine treatment across different GBM PDO models.A141 PDO exhibited antagonism at low doses,synergy at some moderate doses,and antagonism at high doses.The response patterns for A137 PDO and A140 PDO were more complex,transitioning through phases of low-dose synergy,moderate-dose antagonism followed by synergy,and reverting to antagonism at high doses.This indicated that the synergistic effect of this drug combination was strictly confined to a specific moderate dose range.Conclusion The GBM PDO models established in this study successfully recapitulate the morphological and genetic features of the primary tumors and retain their invasive behavioral characteristics.When integrated with drug sensitivity testing,these models effectively simulate the heterogeneous drug response profiles of patients' tumors,offering a promising rapid and reliable preclinical platform for guiding and optimizing personalized clinical treatment strategies.