Knudsen Erik S, Balaji Uthra, Mannakee Brian, Vail Paris, Eslinger Cody, Moxom Christopher, Mansour John, Witkiewicz Agnieszka K
University of Arizona Department of Medicine, University of Arizona, Tucson, Arizona, USA.
University of Arizona Cancer Center, University of Arizona, Tucson, Arizona, USA.
Gut. 2018 Mar;67(3):508-520. doi: 10.1136/gutjnl-2016-313133. Epub 2017 Jan 10.
Pancreatic ductal adenocarcinoma (PDAC) is a therapy recalcitrant disease with the worst survival rate of common solid tumours. Preclinical models that accurately reflect the genetic and biological diversity of PDAC will be important for delineating features of tumour biology and therapeutic vulnerabilities.
27 primary PDAC tumours were employed for genetic analysis and development of tumour models. Tumour tissue was used for derivation of xenografts and cell lines. Exome sequencing was performed on the originating tumour and developed models. RNA sequencing, histological and functional analyses were employed to determine the relationship of the patient-derived models to clinical presentation of PDAC.
The cohort employed captured the genetic diversity of PDAC. From most cases, both cell lines and xenograft models were developed. Exome sequencing confirmed preservation of the primary tumour mutations in developed cell lines, which remained stable with extended passaging. The level of genetic conservation in the cell lines was comparable to that observed with patient-derived xenograft (PDX) models. Unlike historically established PDAC cancer cell lines, patient-derived models recapitulated the histological architecture of the primary tumour and exhibited metastatic spread similar to that observed clinically. Detailed genetic analyses of tumours and derived models revealed features of ex vivo evolution and the clonal architecture of PDAC. Functional analysis was used to elucidate therapeutic vulnerabilities of relevance to treatment of PDAC.
These data illustrate that with the appropriate methods it is possible to develop cell lines that maintain genetic features of PDAC. Such models serve as important substrates for analysing the significance of genetic variants and create a unique biorepository of annotated cell lines and xenografts that were established simultaneously from same primary tumour. These models can be used to infer genetic and empirically determined therapeutic sensitivities that would be germane to the patient.
胰腺导管腺癌（PDAC）是一种治疗棘手的疾病，在常见实体瘤中生存率最低。准确反映PDAC遗传和生物学多样性的临床前模型对于描绘肿瘤生物学特征和治疗易损性至关重要。
采用27个原发性PDAC肿瘤进行遗传分析和肿瘤模型开发。肿瘤组织用于异种移植和细胞系的衍生。对原发肿瘤和开发的模型进行外显子组测序。采用RNA测序、组织学和功能分析来确定患者来源模型与PDAC临床表现之间的关系。
所采用的队列捕捉到了PDAC的遗传多样性。在大多数病例中，都开发了细胞系和异种移植模型。外显子组测序证实了所开发细胞系中保留了原发性肿瘤突变，且随着传代次数增加这些突变保持稳定。细胞系中的遗传保守水平与患者来源异种移植（PDX）模型中观察到的相当。与历史上建立的PDAC癌细胞系不同，患者来源模型重现了原发性肿瘤的组织学结构，并表现出与临床观察到的相似的转移扩散。对肿瘤和衍生模型的详细遗传分析揭示了PDAC的体外进化特征和克隆结构。功能分析用于阐明与PDAC治疗相关的治疗易损性。
这些数据表明，通过适当的方法可以开发出维持PDAC遗传特征的细胞系。此类模型是分析遗传变异重要性的重要底物，并创建了一个独特的生物样本库，其中包含从同一原发性肿瘤同时建立的注释细胞系和异种移植。这些模型可用于推断与患者相关的遗传和经验性确定的治疗敏感性。
