Head and Neck Squamous Cell Carcinoma Biopsies Maintained Ex Vivo on a Perfusion Device Show Gene Changes with Time and Clinically Relevant Doses of Irradiation.
Head and Neck Squamous Cell Carcinoma Biopsies Maintained Ex Vivo on a Perfusion Device Show Gene Changes with Time and Clinically Relevant Doses of Irradiation.
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DOI：
10.3390/cancers15184575
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发表时间：
2023-09-15
期刊：
影响因子：
5.2
通讯作者：
Greenman, John
中科院分区：
文献类型：
--
作者：
Green, Victoria;Baldwin, Lydia;England, James;Marshall, Gayle;Frost, Lucy;Moore, Craig;Greenman, John
减少在研究和药物测试中使用动物的愿望导致了三维模型开发的热潮，这些模型试图模仿人体的部分。然而，这些模型如何真正模仿人类的情况仍有待充分阐明。作者开发了一种技术（灌注装置），使小块人体组织能够保持在体外，从而能够研究各种治疗对患者自身组织的影响。目前的研究描述了这项技术如何用于研究组织中发生的基因变化，同时保持在灌注设备上，以及辐射的影响，从而更深入地了解患者组织从体内取出后的行为以及该模型是否适用于未来的治疗测试。在过去的二十年中，用于研究疾病的三维（3D）培养模型的进展显着增加，但充分了解这些模型在体内的表现仍需要进一步的研究。本研究调查了基线样本和在组织芯片灌注装置上维持长达96小时的样本之间的基因表达差异，有和没有临床相关剂量的照射，以区分模型和治疗效果。将来自7名头颈部鳞状细胞癌（HNSCC）患者的肿瘤组织样品细分，并在切除后立即固定或在有或没有2戈瑞（戈伊）或10戈伊照射的情况下在组织芯片装置中维持48和96小时。使用nCounter® PanCancer Progression Panel测量基因表达。使用nSolver软件（版本4.0）鉴定离体培养前后以及对照和辐照样品之间的差异表达基因。使用Proteome Profiler™平台分析来自芯片上肿瘤的分泌组中细胞因子的存在。显著数量的基因都增加（n = 6和64）和减少（n = 18和58）的表达在芯片上保持48和96小时，分别与新鲜组织相比，然而，所选择的照射时间表没有诱导基因表达或细胞因子分泌的显着变化。尽管离体维持的HNSCC组织显示大部分改变的基因减少，但25%和53%（48和96小时）确实显示表达增加，表明组织保持功能。肿瘤组织芯片的辐照需要进行更长的时间才能观察到特定的基因变化，但我们首次证明了使用这种灌注平台研究HNSCC组织活检的基因组反应的可行性。
The desire to reduce the use of animals in research and drug testing has led to an upsurge in the development of 3-Dimensional models, which try and mimic parts of the human body. However, how these models truly mimic the human situation remains to be fully elucidated. The authors have developed technology (perfusion device) that enables small pieces of human tissue to be maintained outside of the body, enabling the investigation of the effects of various treatments on the patient’s own tissue. The current study describes how this technology has been used to study the gene changes occurring in the tissue, whilst being maintained on the perfusion device, and also the effects of irradiation, providing a deeper understanding of how the patient tissue behaves once it is removed from the body and whether this model will be useful for future treatment testing. Advancements in 3-Dimensional (3D) culture models for studying disease have increased significantly over the last two decades, but fully understanding how these models represent in vivo still requires further investigation. The current study investigated differences in gene expression between a baseline sample and that maintained on a tissue-on-chip perfusion device for up to 96 h, with and without clinically-relevant doses of irradiation, to allow differentiation of model and treatment effects. Tumour tissue samples from 7 Head and Neck Squamous Cell Carcinomas (HNSCC) patients were sub-divided and either fixed immediately upon excision or maintained in a tissue-on-chip device for 48 and 96 h, with or without 2 Gray (Gy) or 10 Gy irradiation. Gene expression was measured using an nCounter® PanCancer Progression Panel. Differentially expressed genes between pre- and post-ex vivo culture, and control and irradiated samples were identified using nSolver software (version 4.0). The secretome from the tumour-on-chip was analysed for the presence of cytokines using a Proteome Profiler™ platform. Significant numbers of genes both increased (n = 6 and 64) and decreased (n = 18 and 58) in expression in the tissue maintained on-chip for 48 and 96 h, respectively, compared to fresh tissue; however, the irradiation schedule chosen did not induce significant changes in gene expression or cytokine secretion. Although HNSCC tissue maintained ex vivo shows a decrease in a large proportion of altered genes, 25% and 53% (48 and 96 h) do show increased expression, suggesting that the tissue remains functional. Irradiation of tumour tissue-on-chip needs to be conducted for longer time periods for specific gene changes to be observed, but we have shown, for the first time, the feasibility of using this perfusion platform for studying the genomic response of HNSCC tissue biopsies.
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DOI：
10.1002/hed.26004
发表时间：
2020-02
期刊：
Head & neck
影响因子：
--
作者：
Naghavi AO;Kim Y;Yang GQ;Ahmed KA;Caudell JJ
通讯作者：
Caudell JJ
影响因子：
4.7
作者：
Diehl V;Huber LS;Trebicka J;Wygrecka M;Iozzo RV;Schaefer L
通讯作者：
Schaefer L
影响因子：
5
作者：
Goel, Bharat;Tiwari, Anoop Kumar;Pandey, Rajeev Kumar;Singh, Akhand Pratap;Kumar, Sujeet;Sinha, Abhishek;Jain, Shreyans K.;Khattri, Arun
通讯作者：
Khattri, Arun
影响因子：
2.3
作者：
Kim SI;Kang JW;Noh JK;Jung HR;Lee YC;Lee JW;Kong M;Eun YG
通讯作者：
Eun YG
影响因子：
7.3
作者：
Manduca N;Maccafeo E;De Maria R;Sistigu A;Musella M
通讯作者：
Musella M