类器官在肿瘤转化医学中的应用和进展

doi:10.19401/j.cnki.1007-3639.2022.11.009

摘要/Abstract

摘要：

随着精准医学概念的提出，肿瘤作为一类高度异质性的疾病，其个体化治疗已成为精准医学的一个关键领域也受到更多关注。近期提出的类器官模型为肿瘤的基础研究和个体化治疗带来了新突破。类器官指从干细胞或器官祖细胞来源，以类似体内细胞分化的方式组织成的器官特异性的细胞集合。肿瘤类器官指利用原代恶性细胞经过体外3D培养构建的恶性细胞团，在体外培养条件下可持续增殖，一定程度上保留了原代肿瘤的病理学形态特征、基因组与转录组特征、药物敏感性及恶性细胞间异质性，为体外肿瘤研究提供了新方法，尤其在预测患者药物敏感性、药物高通量筛选等方面有巨大潜力，为肿瘤个体化治疗作出了贡献。但该模型仍存在不足，如无法重现体内肿瘤微环境等，使其在临床等方面的应用受限。目前将肿瘤类器官与其他肿瘤成分共培养、与微流控设备和生物打印技术等联合应用将有望弥补其缺陷、突破肿瘤治疗领域的瓶颈。本文将常用肿瘤研究模型的特点进行对比，总结部分肿瘤类器官的培养方法，并描述了肿瘤类器官的临床相关应用，最后对其与其他技术联合应用进行了叙述，并对未来肿瘤类器官的发展方向予以展望。

关键词: 肿瘤, 类器官, 转化医学, 药物敏感性预测, 药物筛选

Abstract:

With the introduction of precision medicine, the individualized treatment of cancer has raised increasing interest in the field. Recently, the utilization of organoids as a powerful tool for precision medicine has achieved great breakthroughs for basic research and potential clinical usage. Organoids usually derived from stem/progenitor cells are organ-specific multi-cell clusters organized in a manner like the cell differentiation patterns of their original in vivo counterpart. While keeping the basic pathological/genetic/biological phenotypes of their parental primary cells/tissues, tumor organoids still have a certain potential for proliferative immortalization, which makes organoid-based research/clinical models hopeful tools for precision medicine, especially in predicting drug sensitivity for clinical patients or conducting high-throughput screening of new compounds/drugs for pharmaceutical development. However, obstacles still exist to the clinical application of organoid-based tools, such as the lack of in vivo tumor microenvironment. Currently, with the development of co-culture, microfluidic and bioprinting techniques, breakthroughs have been made for the practical clinical/industrial utilization of organoids. In the current review, we compared the characteristics of common ex vivo tumor models with organoid-based models, summarized the protocols for tumor organoid culturing, reviewed the clinical/industrial application of tumor organoids, and finally look forward to the new outlooks of tumor organoid-based tools in the future.

Key words: Tumor, Organoid, Translational medicine, Drug susceptibility prediction, Drug screening

中图分类号:

王若彤, 王欣, 沈波. 类器官在肿瘤转化医学中的应用和进展[J]. 中国癌症杂志, 2022, 32(11): 1105-1114.

WANG Ruotong, WANG Xin, SHEN Bo. Application and progress of organoids in tumor translational medicine[J]. China Oncology, 2022, 32(11): 1105-1114.

图/表 2

表1

表2

常见的类器官培养基成分概述"

Tumor	Author and reference	Purposes	Key resources	Buffers/media	Nultrition	Antibiotic	Supplements	Others
Lung cancer	Shi R,et al^[40]	Drug response including targeting drugs; Establishing a platform	Advanced DMEM/F12, glutamax, HEPES, antibiotic-antimycotic, B-27 supplement, N-acetylcysteine, rec. hEGF, rec. hFGF-10, rec. hFGF-4, rec. hNoggin, SAG (smoothened ligand), A83-01 (TGF-b receptor inhibitor), Y-27632, CHIR 99021(GSK-3 inhibitor)	Advanced DMEM/F12, HEPES	GlutaMAX, B-27 supplement, N-acetylcysteine	Antibiotic-antimycotic	Rec. hEGF, rec. hFGF-10, rec. hFGF-4, rec. hNoggin, SAG (smoothened ligand)	A83-01 (TGF-b receptor inhibitor), Y-27632, CHIR 99021 (GSK-3 inhibitor)
Liver cancer	Broutier L, et al^[22]	Prognosis indicators; drug susceptibility prediction; Identification of new targets	Advanced DMEM/F12, 1% penicillin/streptomycin, 1% glutamax, HEPES, B27 supplement (without vitamin A), N2 supplement, N-acetylcysteine, R-spondin1 (conditioned medium), nicotinamide, rec. h[Leu15]-gastrin Ⅰ, rec. hEGF, rec. hFGF10, rec. hHGF, forskolin (CAMP inhibitor), A8301, Y27632 (remove after passaging) and dexamethasone (remove after passaging)	Advanced DMEM/F12, HEPES	1% glutamax, B27 supplement (without vitamin A), N2 supplement, N-acetylcysteine, R-spondin1 (conditioned medium), nicotinamide	1% Penicillin/Streptomycin	Rec. h[Leu15]-gastrin Ⅰ, rec. hEGF, rec. hFGF10, rec. hHGF	Forskolin (CAMP inhibitor), A8301, Y27632 (remove after passaging) and dexamethasone (remove after passaging)
Rectal cancer	Ganesh K, et al^[25]	Establishing a platform, radiotherapy, chemotherapy and targeted drug response prediction	Advanced DMEM/F12 supplemented with antibiotic-antimycotic, B27, N2, glutamax, gastrin Ⅰ, HEPES, N-acetylcysteine, nicotinamide, 50% Wnt-3A (conditioned medium and remove after passaging), 20% R-spondin-1 (conditioned medium and remove after passaging), rec. mNoggin (remove after passaging), rec. hEGF, A83-01, SB202190	advanced DMEM/F12, HEPES	B27, N2, glutamax, N-acetylcysteine, nicotinamide	Antibiotic-antimycotic	Gastrin Ⅰ, 50% Wnt-3A (conditioned medium and remove after passaging), 20% R-spondin-1 (conditioned medium and remove after passaging), rec. mNoggin (and remove after passaging), rec. hEGF	A83-01, SB202190
Gastric cancer	Bartfeld S, et al^[41]	Culture protocol, modeling organogenesis	Advanced DMEM/F12 supplemented with penicillin/ streptomycin, HEPES, glutamax, B27, N-acetylcysteine, EGF, Noggin (conditioned medium) 10%, R-spondin1 (conditioned medium) 10%, Wnt (conditioned medium) 50%, FGF10, gastrin, TGF-βi (A-83-01). nicotinamide^, RHOKi (Y-27632), IGF^, p38 inhibitor (SB202190)^, GSK3β inhibitor (CHIR99021)^, PGE2^*	advanced DMEM/F12, HEPES	Glutamax, B27, N-Acetylcysteine	Penicillin/streptomycin	EGF, Noggin conditioned medium 10%, 10% R-spondin1 (conditioned medium), 50% Wnt (conditioned medium), FGF10, gastrin, IGF*	TGFβi (A-83-01), RHOKi (Y-27632), p38 inhibitor (SB202190), GSK3β inhibitor (CHIR99021), PGE2*
Gastric cancer	Yan H H N, et al^[31]	Tumor heterogeneity; drug susceptibility prediction	Modifying the medium of Bartfeld S by adding nutlin-3a (p53/MDM2 inhibitor)	advanced DMEM/F12, HEPES	Glutamax, B28, N-Acetylcysteine	Penicillin/streptomycin	Rec. hEGF, rec. hFGF10, Wnt3A (conditioned medium), R-spondin1 (conditioned medium), Noggin (conditioned medium)	Nutlin-3a (p53/MDM2 inhibitor)
Pancreatic cancer	Tiriac H,et al^[42]	Biomarker, genome character, prognosis, drug response prediction	Advanced DMEM/F12, HEPES, glutamax, A83-01, hEGF, mNoggin, hFGF10, hgastrin Ⅰ, N-acetylcysteine, nicotinamide, PGE2, B27 supplement, R-spondin1 (conditioned media), Afamin/Wnt3A (conditioned media)	Advanced DMEM/F12, HEPES	Glutamax, N-acety-lcysteine, nicotinamide, B27 supplement	NM	Rec. hEGF, Afamin/Wnt3A, rec. hFGF10, rec. hgastrin I, PGE2, R-spondin1 (conditioned media), Afamin/Wnt3A (conditioned media)	A83-01
Prostate cancer	Chua C W, et al^[43]	Modeling prostate organoid	Hepatocyte culture medium, EGF, Y-27632, glutamax, 5% activated carbon stripped FBS	Hepatocyte culture medium	5% activated carbon stripped FBS, glutamax	NM	EGF	Y-27632
Bladder cancer	Lee S H,et al^[34]	Establish PDO biobank and describe character	Hepatocyte culture medium, EGF, Y-27632, glutamax, 5% activated carbon stripped FBS	Hepatocyte culture medium	5% activated carbon stripped FBS, glutamax	NM	EGF	Y-27632

表2

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Item	Cancer cell line	PDX	Tumor spheroid	Organoid
Tumor microenvironment	No	Limited	No	No
Physiologic representation	Limited	Semiphysiologic	Limited	Semiphysiologic
Difficulty	Easy	Hard	Easy	Easy
Repopulation activity	Good	Medium	Poor	Good
Genome editing	Yes	No	Yes	Yes
High-throughput screening	Yes	No	Yes	Yes
Tumor heterogeneity	No	Yes	No	Yes
Library establishment	Yes	Limited	Yes	Yes