【Biomaterials】浙江大学吴宏伟等|48小时快速构建、100%植入率！新型细胞环类器官攻克肉瘤精准医疗核心瓶颈#

文章标题：Engineering cell ring organoids for efficient establishment of patient-derived orthotopic xenotransplantation (r-PDOX) model in sarcoma

通讯作者：Hongwei Wu, Hongwei Ouyang, Xiaowen Liang

文章链接：https://doi.org/10.1016/j.biomaterials.2026.124382

文章概要#

引言#

肉瘤（包括骨肉瘤和软组织肉瘤）具有极高的异质性和罕见性，导致临床上极难找到有效的精准靶向药物。尽管原位移植动物模型（PDOX）能较好地模拟肿瘤微环境，但传统方法存在细胞利用率低、建模周期漫长（需数月）以及基质环境缺失等致命缺陷，无法满足临床快速用药指导的需求。为此，研究团队开创性地开发了一种无支架细胞环类器官平台，利用独特的几何结构与基质细胞共培养技术，成功打破了肉瘤 preclinical 模型构建的时间与效率瓶颈。

Fig. 1. Cell ring organoids fabrication and characterization. (A) Schematic diagram of patient tumor cells mixed with human fibroblasts to form cell ring organoids on the cell rings. (B) Cell ring organoids from different angles of view, including the top view (i), side view (ii), on-chip (iii), and in tissue culture medium (iv). Scale bar = 1 mm. (C) The thickness of cell ring organoids formed from different initial cell numbers (0.6, 0.8, and 1 million). Scale bar = 1 mm. (D) H&E and IHC staining of patient-derived cell ring organoids and corresponding patient tumor tissues, including the expression of α-SMA, Ki-67, and SATB2. Scale bar = 50 μm. (Data are presented as mean ± SD, one-way ANOVA and Tukey’s HSD post hoc test were used).#

主要实验及结论#

研究人员将患者来源的原代肉瘤细胞与商业化人皮肤成纤维细胞（HSFs）按 1:1 的比例混合，在专用的微型肿瘤芯片中进行无支架自组装。实验表明，该平台在48小时内即可快速生成结构紧密、尺寸可控且无中心坏死的肿瘤环状类器官。转录组测序分析证实，这种共培养模式成功诱导了成纤维细胞向癌相关成纤维细胞（CAF）样活化，重建了高保真的肿瘤微环境，使类器官与患者原始肿瘤组织表现出高达 0.87 的基因表达强相关性，完整保留了患者特异性的分子特征。

Fig. 2. RNA sequencing analysis of patient-derived cell ring organoids and corresponding patient tumor tissues. (A) PCA analysis of patient-derived organoids OS0113, OS0114, and OS0812 and their corresponding patient tumor tissues S0113, S0114, and S0812. (B) Scatter plots of the correlation of average log2(FPKM) between the patient tumor tissues and patient-derived cell ring organoids from all three patients (Pearson’s correlation). (C) Heatmap of DEG expression in individual patient samples and their corresponding organoids (FC > 1.5 or < −1.5; false discovery rate [FDR] < 0.05). Log2(RPKM), log2(reads per kilobase of transcript per million reads mapped). (D) Venn diagram of DEGs in the organoids and corresponding tumor tissues analyzed by pairwise comparisons between different patients. (E) Representative GO of biological processes enriched in patient tumor tissues and corresponding organoids in each patient compared to the other two patients.#

Fig. 3. Cell ring organoid transplantation had higher tumor modeling efficiency than cell suspension injection. (A) Schematic diagram of orthotopic implantation of cell ring organoids and cell suspension injection in the femur of animals and the following assessments. (B) In vivo fluorescent imaging of tumor formation from cell suspension or cell ring organoids at the cell number of 0.6 million (B) or 1.2 million (D) and the quantification of tumor fluorescence at implantation cell number of 0.6 million (C) or 1.2 million (E). Micro-CT analysis (F) and bone destruction percentage (G) of the tumors formed by cell ring organoids compared to cell suspension formed tumors and normal tissues. (H). H&E staining imaging of normal tissue and the tumor tissues formed by cell ring organoids and cell suspension. Scale bar, Left: Scale bar = 3 mm; Right: Scale bar = 500 μm. (Data are presented as mean ± SD, unpaired Student’s t-test was used; ∗∗: p < 0.01; ∗∗∗: p < 0.001; ∗∗∗∗: p < 0.0001).#

在活体动物实验中，研究团队将自组装完成的细胞环整块原位移植至小鼠股骨缺损处，成功建立了新型环状原位异种移植（r-PDOX）模型。对比传统细胞悬液注射，新方法不仅简化了手术操作、杜绝了细胞泄漏，更在节省 50% 细胞用量的前提下，将模型建立时间缩短至 7 天以内，且实现了 100% 的肿瘤植入成功率。病理和显微 CT 评估显示，r-PDOX 模型完美重现了临床骨肉瘤典型的骨质破坏、骨膜新骨形成及丰富的血管新生等恶性进展特征。在此基础上，研究团队进行了初步的临床药物敏感性探索，发现该模型对临床一线化疗药物异环磷酰胺及靶向药安罗替尼的反应趋势，与相应患者的实际临床疗效展现出令人振奋的初步一致性。

Fig. 4. r-PDOX models exhibited similar therapeutic outcomes as in osteosarcoma patients. (A) Treatment regime and timeline of representative case in patients and r-PDOX models. (B) Imaging comparisons of three osteosarcoma patients before and after treatment. Scale bar = 5 cm. (C) Relative tumor size before and after treatment. (D) H&E staining of tumor tissue before and after treatment. (E) The proportion of cell necrosis before and after treatment. (F) Photographs of tumors treated with saline or IFO treatment in r-PDOX models. Scale bar = 1 cm. (G) The quantification of tumor size treated with saline or IFO of r-PDOX models. (H) The quantification of tumor size treated with saline or IFO of patients. (I) H&E staining of tumors from r-PDOX models. Scale bar = 100 μm. (J) The quantification of necrosis percentages in tumor tissues treated with saline or IFO of r-PDOX models. (Data are presented as mean ± SD, unpaired Student’s t-test was used; ∗: p < 0.05; ∗∗: p < 0.01; ∗∗∗∗: p < 0.0001).#

Fig. 5. r-PDOX models exhibited similar therapeutic outcomes as in metastatic soft tissue sarcoma patients. (A) Treatment regime and timeline of representative case in patients and r-PDOX models. (B-D) Comparison of pre-treatment and post-treatment pulmonary CT findings in patients with metastatic soft tissue sarcoma and lung metastases. (E) Photographs of tumors treated with saline or anlotinib treatment in r-PDOX models. Scale bar = 1 cm. (F) The quantification of tumor size treated with saline or anlotinib of r-PDOX models. (Data are presented as mean ± SD, unpaired Student’s t-test was used; ∗∗: p < 0.01; ∗∗∗: p < 0.001; ∗∗∗∗: p < 0.0001).#

总结及展望#

这项研究成功攻克了肉瘤精准医疗中 preclinical 模型构建周期长、成功率低的长期痛点。该细胞环类器官平台不仅能在极短的时间窗口内高保真地还原肿瘤微环境与基因表达谱，其独特的空心环状结构还被证实能让巨噬细胞等免疫细胞良好存活，为未来构建多细胞免疫微环境模型奠定了坚实基础。尽管其临床预测准确性仍需在更大规模的患者队列中进行验证，但这一创新平台无疑为肉瘤以及其他基质丰富型实体瘤的药物筛选、病理机制研究及个体化精准治疗开辟了一条极具临床转化前景的新路径。