结直肠腺瘤（CA），尤其是高危CA（HRCA），是一种具有高发病率和复发率的癌前病变，在全球范围内占非遗传性结直肠癌约90%的发病例数。目前，复发性CA只能通过反复侵入性息肉切除治疗，而由于缺乏可靠的CA相关药物筛选体外模型，安全且有前景的药物研发策略仍然缺失。我们建立了一个大规模的高危结直肠腺瘤器官样（HRCA-PDO）样本库，该样本库包含了从33名患者中获得的37条PDO细胞系，并进行了一系列高通量和高内容的HRCA药物筛选。我们使用非WNT3a培养基建立了一种原代培养系统，该培养基显著提高了HRCA-PDO的纯度，同时保持了其生存能力。我们还证明了HRCA-PDO复制了原发性腺瘤的组织学特征、细胞多样性、基因突变和分子特征。特别是，我们确定了LGR5、c-Myc和OLFM4等失调的干细胞基因作为腺瘤的标志，这些基因在HRCA-PDO中得到了很好的保留。基于HRCA-PDO样本库，我们应用了一个定制的139种化合物库进行了药物筛选。其中包括二甲双胍、BMS754807、泊尼帕坦和AT9283等四种药物被筛选为潜在的具有一致抑制效果的药物。作为代表，我们发现二甲双胍通过限制干细胞特性的维持，在体内外抑制了HRCA-PDO的生长。本研究建立了一种有前景的HRCA-PDO样本库，并进行了首次的高通量和高内容的HRCA药物筛选，以期为结直肠癌的预防提供启示。© 2023. BioMed Central Ltd., part of Springer Nature.

Colorectal adenoma (CA), especially high-risk CA (HRCA), is a precancerous lesion with high prevalence and recurrence rate and accounts for about 90% incidence of sporadic colorectal cancer cases worldwide. Currently, recurrent CA can only be treated with repeated invasive polypectomies, while safe and promising pharmaceutical invention strategies are still missing due to the lack of reliable in vitro model for CA-related drug screening.We have established a large-scale patient-derived high-risk colorectal adenoma organoid (HRCA-PDO) biobank containing 37 PDO lines derived from 33 patients and then conducted a series of high-throughput and high-content HRCA drug screening.We established the primary culture system with the non-WNT3a medium which highly improved the purity while maintained the viability of HRCA-PDOs. We also proved that the HRCA-PDOs replicated the histological features, cellular diversity, genetic mutations, and molecular characteristics of the primary adenomas. Especially, we identified the dysregulated stem genes including LGR5, c-Myc, and OLFM4 as the markers of adenoma, which are well preserved in HRCA-PDOs. Based on the HRCA-PDO biobank, a customized 139 compound library was applied for drug screening. Four drugs including metformin, BMS754807, panobinostat and AT9283 were screened out as potential hits with generally consistent inhibitory efficacy on HRCA-PDOs. As a representative, metformin was discovered to hinder HRCA-PDO growth in vitro and in vivo by restricting the stemness maintenance.This study established a promising HRCA-PDO biobank and conducted the first high-throughput and high-content HRCA drug screening in order to shed light on the prevention of colorectal cancer.© 2023. BioMed Central Ltd., part of Springer Nature.