Translational Tumor Engineering Laboratory, Department of Biomedical Engineering, National University of Singapore, Singapore, 119276, Singapore.
The N.1 Institute for Health, National University of Singapore, Singapore, 117456, Singapore.
Adv Sci (Weinh). 2024 May;11(20):e2307129. doi: 10.1002/advs.202307129. Epub 2024 Mar 17.
Recently mapped transcriptomic landscapes reveal the extent of heterogeneity in cancer-associated fibroblasts (CAFs) beyond previously established single-gene markers. Functional analyses of individual CAF subsets within the tumor microenvironment are critical to develop more accurate CAF-targeting therapeutic strategies. However, there is a lack of robust preclinical models that reflect this heterogeneity in vitro. In this study, single-cell RNA sequencing datasets acquired from head and neck squamous cell carcinoma tissues to predict microenvironmental and cellular features governing individual CAF subsets are leveraged. Some of these features are then incorporated into a tunable hyaluronan-based hydrogel system to culture patient-derived CAFs. Control over hydrogel degradability and integrin adhesiveness enabled derivation of the predominant myofibroblastic and inflammatory CAF subsets, as shown through changes in cell morphology and transcriptomic profiles. Last, using these hydrogel-cultured CAFs, microtubule dynamics are identified, but not actomyosin contractility, as a key mediator of CAF plasticity. The recapitulation of CAF heterogeneity in vitro using defined hydrogels presents unique opportunities for advancing the understanding of CAF biology and evaluation of CAF-targeting therapeutics.
最近绘制的转录组图谱揭示了癌症相关成纤维细胞 (CAF) 的异质性超出了先前建立的单一基因标志物的程度。对肿瘤微环境中单个 CAF 亚群的功能分析对于开发更准确的 CAF 靶向治疗策略至关重要。然而，缺乏能够在体外反映这种异质性的强大临床前模型。在这项研究中，利用从头颈部鳞状细胞癌组织中获得的单细胞 RNA 测序数据集来预测控制单个 CAF 亚群的微环境和细胞特征。然后将其中一些特征纳入可调节的透明质酸基水凝胶系统中，以培养患者来源的 CAF。通过改变细胞形态和转录组谱，控制水凝胶的降解性和整合素粘附性，从而衍生出主要的肌成纤维细胞和炎症性 CAF 亚群。最后，使用这些水凝胶培养的 CAF，确定了微管动力学，但不是肌动球蛋白收缩性，作为 CAF 可塑性的关键介质。使用定义明确的水凝胶在体外再现 CAF 异质性为深入了解 CAF 生物学和评估 CAF 靶向治疗提供了独特的机会。
