用于生成胸膜间皮瘤的绒毛尿囊膜患者来源异种移植（CAM-PDX）模型以及进行临床前成像以促进癌症研究转化和药物筛选的方法。

Methodology for generating chorioallantoic membrane patient-derived xenograft (CAM-PDX) models of pleural mesothelioma and performing preclinical imaging for the translation of cancer studies and drug screening.

作者信息

Schulze Jan, Barnett Sarah, Shaw Liam, Herrmann Anne, Poptani Harish, Rassl Doris M, Haragan Alexander, Shackcloth Michael, Sacco Joseph J, Coulson Judy M

机构信息

Molecular and Clinical Cancer Medicine, University of Liverpool Institute of Systems Molecular and Integrative Biology, Liverpool, England, UK.

Egg Facility, Liverpool Shared Research Facilities, University of Liverpool Faculty of Health and Life Sciences, Liverpool, England, UK.

出版信息

F1000Res. 2025 May 7;14:481. doi: 10.12688/f1000research.163596.1. eCollection 2025.

DOI:10.12688/f1000research.163596.1

PMID:40568242

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12188182/

Abstract

BACKGROUND

Pleural mesothelioma is a cancer of the lung lining associated with asbestos exposure. Platinum/pemetrexed chemotherapy has been used for many years but provides little benefit and, despite recent immunotherapy advances, prognosis remains poor underpinning the need for development of novel therapeutics or drug repurposing. Fertilized hens' eggs provide a rapid and cost-effective alternative to murine models of pleural mesothelioma which are commonly used in preclinical studies, with chorioallantoic membrane (CAM) xenografts being a partial replacement for mouse flank xenografts. Here we describe methods to generate mesothelioma patient-derived xenografts on the CAM (CAM-PDX), and to subsequently assess these PDX nodules by preclinical imaging and histology.

METHODS

Fragments of surplus mesothelioma tissue obtained from patient biopsies were implanted onto the CAM on embryonic day 7 (E7), fresh or following cryopreservation, with the established PDX dissected on E14 and fixed for histological/immunohistochemical analysis. The optimal freezing method was determined by comparing tissue integrity and cellular content of cryopreserved tissue fragments with paired fresh samples via histological/immunohistochemical analyses. [ F]-fluorodeoxyglucose positron emission tomography/computed tomography (FDG-PET/CT) was used to assess viability of PDXs

RESULTS

Methodologies for processing, cryopreservation, re-animation, and engraftment of mesothelioma tissue fragments were established. Cryopreservation of biopsy samples and parallel processing of contiguous sections allows for assessment of mesothelioma cellularity. CAM-PDXs, generated from fresh or slow-frozen tissue, were well vascularized whilst maintaining the architecture and cellular composition of the patient tissue. Furthermore, uptake of [ F]-FDG following intravenous injection could be visualized and quantified.

CONCLUSIONS

The CAM is a rapid platform for engrafting patient-derived tissue, maintaining elements of the tumor microenvironment and recapitulating heterogeneity observed in mesothelioma. Combining the CAM-PDX model and FDG-PET/CT provides a quantitative platform for pre-screening of novel treatment strategies and drug combinations, with the potential for development of patient tumor avatars for predicting clinical response.

摘要

背景

胸膜间皮瘤是一种与接触石棉相关的肺膜癌症。铂类/培美曲塞化疗已使用多年，但益处不大，尽管近期免疫治疗取得进展，但预后仍然很差，这突出了开发新型疗法或药物重新利用的必要性。受精母鸡的鸡蛋为临床前研究中常用的胸膜间皮瘤小鼠模型提供了一种快速且经济高效的替代方案，绒毛尿囊膜（CAM）异种移植可部分替代小鼠侧腹异种移植。在此，我们描述了在CAM上生成间皮瘤患者来源异种移植瘤（CAM-PDX）的方法，并随后通过临床前成像和组织学评估这些PDX结节。

方法

从患者活检中获取的多余间皮瘤组织碎片在胚胎第7天（E7）植入到CAM上，新鲜组织或冷冻保存后植入，已建立的PDX在E14解剖并固定用于组织学/免疫组织化学分析。通过组织学/免疫组织化学分析将冷冻保存的组织碎片与配对的新鲜样本的组织完整性和细胞含量进行比较，以确定最佳冷冻方法。使用[F]-氟脱氧葡萄糖正电子发射断层扫描/计算机断层扫描（FDG-PET/CT）评估PDX的活力。