目前，结直肠癌（colorectal cancer，CRC）是世界范围内常见的恶性肿瘤之一，具有发病率高、治疗难度大及发病年龄趋于年轻化等特点。当前临床治疗主要依赖于化学治疗和免疫治疗策略。但不同患者对化疗或免疫治疗的响应差异巨大，因此，亟需能探寻一种预测或评估个体化治疗效果的精准医疗模型进行临床前的药效评价。类器官作为新型肿瘤研究模型，具有与原始肿瘤特性一致，且操作简便的优势，其作为药效评价模型不仅能弥补肿瘤细胞系在传代过程中易发生基因漂变的特性，且显著弥补小鼠异种移植（patient-derived xenograft，PDX）模型构建周期长、成本高和不能反应肿瘤微环境的缺陷。因此，利用CRC类器官进行体外药效评价，有望为CRC的个体化治疗提供更加精准和高效的预判手段，推动精准医疗在CRC中的临床转化与应用。本研究旨在评估CRC类器官作为体外药效评价模型的可行性。为此，构建了由多例人源CRC组织衍生的类器官生物样本库，并系统开展了多种化疗药物及免疫治疗药物的体外敏感性检测，以验证其在预测患者个体治疗反应中的潜力和应用价值。

本研究成功建立了包含86例CRC类器官的生物样本库；并通过苏木精-伊红染色（hematoxylin-eosin staining，HE）、免疫组化染色（immunohistochemistry staining，IHC）以及全外显子（whole exome sequencing，WES）测序分析表明，CRC类器官保留了与原发肿瘤相似的组织学以及遗传学特征，其CK20、Ki67、CDX2和Villin蛋白的表达与原发肿瘤相似；化疗药物药敏性试验表明，不同患者来源的CRC类器官表现出对化疗药物敏感性的差异；转录组测序的初步分析发现，内质网应激反应相关基因（ERN1等）的表达上调，可能与5-氟尿嘧啶（5-Fluorouracil，5-FU）耐药相关；编码胆固醇转运蛋白基因NPC1L1表达上调，可能与奥沙利铂耐药相关。免疫治疗敏感性评估显示，患者自体来源T细胞对CRC类器官具有显著杀伤活性，但抗程序性死亡受体1（programmed death-ligand 1，PD-L1）单克隆抗体帕博利珠单抗（pembrolizumab）未能显著增强免疫细胞的肿瘤杀伤效应，提示免疫检查点抑制剂在当前模型中未展现预期药效。鉴于CRC的高度异质性及个体间生物学差异，后续的研究仍需通过扩大样本量来开展多中心、系统性研究，以进一步明确免疫检查点抑制剂在CRC类器官模型中的作用机制及临床预测价值。

综上所述，类器官作为一种具有前景的新型临床前模型，在药物药效评价以及药物筛选中展示巨大潜力。CRC类器官在遗传背景、分子表达谱及药物反应性等方面均高度保留了原发肿瘤的关键特征，使其能够有效反映个体差异并预测治疗响应。因此，基于CRC类器官的药效评价不仅为精准制定化疗和免疫治疗方案提供了重要参考，也为实现个体化医疗奠定了坚实基础。未来，一方面，有必要在更大规模的前瞻性临床试验中进一步验证CRC类器官在预测治疗反应及指导个体化治疗方面的临床应用价值；另一方面，应充分利用该模型深入解析化疗药物及免疫治疗产生耐药性的潜在机制，以推动新型治疗策略的开发与优化。通过持续拓展类器官平台的应用范围，有望加速CRC从基础研究向精准医学的转化进程。

Currently,colorectal cancer (CRC) ranks as one of the most common malignant tumors worldwide,characterized by high incidence,significant treatment challenges,and a trend toward younger onset.Current clinical management primarily relies on chemotherapy and immunotherapy strategies. However,there is substantial inter-patient variability in response to chemotherapy or immunotherapy,underscoring the urgent need for precision medicine models that can predict or evaluate individualized treatment efficacy in preclinical drug efficacy assessments.As a novel tumor research model,organoids offer the advantages of maintaining the characteristics of the original tumor and operational simplicity. As drug efficacy evaluation models,they not only effectively address the tendency of tumor cell lines to undergo genetic drift during passaging but also significantly surpass the limitations of mouse xenograft (Patient-Derived Xenograft, PDX) models,including long construction cycles,high costs, and the inability to recapitulate the tumor microenvironment. Therefore,the use of CRC organoids for in vitro drug efficacy evaluation holds promise as a more precise and efficient predictive tool for personalized CRC treatment,driving the clinical translation and application of precision medicine in CRC.The aim of this study was to assess the feasibility of CRC organoids as in vitro drug efficacy evaluation models.To this end, an organoid biobank derived from multiple human CRC tissues was established,and systematic in vitro sensitivity testing of various chemotherapeutic and immunotherapeutic agents was conducted to validate their potential and application value in predicting individual patient treatment responses.

Firstly,we successfully established a biobank comprising 86 CRC organoids. Analyses including hematoxylin-eosin (HE) staining, immunohistochemistry (IHC) staining, and whole exome sequencing (WES) demonstrated that CRC organoids retained histological and genetic characteristics similar to their parental tumors. The expression of CK20,Ki67,CDX2,and Villin proteins in the organoids closely mirrored that of the original tumors.Chemotherapy drug sensitivity assays revealed interpatient heterogeneity in responses among CRC organoids derived from different patients.Preliminary transcriptome sequencing analysis identified upregulated expression of endoplasmic reticulum stress response-related genes (e.g.,ERN1),potentially associated with 5-Fluorouracil (5-FU) resistance,and elevated expression of the cholesterol transporter gene NPC1L1,potentially linked to oxaliplatin resistance.Finally,immunotherapy sensitivity assessment showed that patient-derived autologous T cells exhibited significant cytotoxic activity against CRC organoids.However,the anti-Programmed Death-Ligand 1 (PD-L1) monoclonal antibody Pembrolizumab failed to significantly enhance the tumor-killing effect of immune cells,suggesting that immune checkpoint inhibitors did not demonstrate the expected efficacy in this model.Given the high heterogeneity of CRC and interindividual biological variations,subsequent research requires multicenter systematic studies with expanded sample sizes to further clarify the mechanisms of action and clinical predictive value of immune checkpoint inhibitors in the CRC organoid model.

In conclusion,as a promising new preclinical model,organoids show great potential in the evaluation of drug efficacy and drug screening.CRC organoids highly retain the key characteristics of the primary tumor in terms of genetic background,molecular expression profile, and drug reactivity,enabling them to effectively reflect individual differences and predict treatment responses.Therefore,the evaluation of drug efficacy based on CRC organoids not only provides an important reference for the precise formulation of chemotherapy and immunotherapy regimens but also lays a solid foundation for the realization of personalized medicine.In the future,on the one hand, it is necessary to further verify the clinical application value of CRC organoids in predicting treatment responses and guiding personalized treatment in larger-scale prospective clinical trials.On the other hand,this model should be fully utilized to deeply analyze the potential mechanisms of drug resistance generated by chemotherapeutic drugs and immunotherapy,so as to promote the development and optimization of new treatment strategies.By continuously expanding the application scope of the organoid platform,it is expected to accelerate the transformation process from basic research to precision medicine for colorectal cancer.