[A049]模拟肿瘤微环境的三维微流控平台

A. Pavesi, S. Wong, R. Kamm, S. Lee, Giulia Adriani
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引用次数: 0

摘要

免疫治疗是目前癌症治疗的主要突破,但由于免疫抑制肿瘤微环境(immunosuppressive tumor microenvironment, TME)的挑战,实体瘤的治疗方法在临床场景中仍然存在局限性。具体来说,在招募到TME的免疫细胞中,单核细胞/巨噬细胞尤其丰富。体外巨噬细胞分为经典活化的M1巨噬细胞和交替活化的M2巨噬细胞。然而,巨噬细胞是高度可塑性的细胞,它们在体内TME中经常呈现混合表型。单核/巨噬细胞参与癌细胞增殖、细胞侵袭、细胞杀伤、血管生成、t细胞免疫抑制,并且在广泛的癌症中经常与不良预后相关。然而,破坏单核/巨噬细胞的肿瘤活性及其与TME中复杂细胞系统的相互作用仍然是一个挑战。因此,更好地了解单核/巨噬细胞表型的调节机制及其与肿瘤的相互作用可能会使它们成为相关的治疗策略。为了研究免疫抑制的TME,我们开发了基于微流体的集成平台,与肿瘤细胞和免疫细胞的三维(3D)共培养,以研究TME中的物理和分子线索如何调节细胞相互作用。我们的3D多细胞平台提供了相当大的好处,并且在之前的研究中已经证明,在模拟TME和筛选不同治疗方法方面,3D多细胞平台比传统的2D平台有明显的优势。我们之前的研究表明,免疫系统与癌细胞的相互作用在原发性肿瘤部位或继发性转移部位的转移中起着至关重要的作用。此外,我们研究了单核细胞和程序性死亡配体1 (PD-L1)免疫检查点对t细胞受体(TCR)工程t细胞的影响。我们现在开发了一个新的3D微流控平台来研究间质流(IF)的影响,IF是通过肿瘤基质的液体流动,它是TME的一个重要组成部分,可能有助于巨噬细胞向肿瘤表型的极化。微流控模型允许肿瘤细胞和单核/巨噬细胞共培养,用IF刺激它们,并在3D中量化细胞迁移。据我们所知,这项研究代表了第一个将IF和肿瘤细胞和免疫细胞结合在一起的微流控肿瘤模型。我们的初步结果证实,肿瘤细胞和肿瘤细胞分泌因子(TSF)的存在增加了巨噬细胞向癌细胞的迁移速度和方向性,可能有助于癌细胞的传播。有趣的是,基于if的机械提示(培养中没有肿瘤细胞)的存在导致巨噬细胞迁移的类似增加。此外,通过联合使用TSF和IF,我们观察到对巨噬细胞迁移没有协同作用或相加作用。在TSF和/或IF的刺激下,观察到巨噬细胞表型(M1和M2)的混合群体,而在没有TSF或IF的情况下,观察到M1和M2标记物的表达水平较低。重要的是,从这项研究中获得的理解可以帮助设计和功能测试免疫治疗策略,以调节TME中的巨噬细胞极化,达到抗肿瘤表型。引文格式:Andrea Pavesi, Siew Cheng Wong, Roger Kamm, Sharon Wei Ling Lee, Giulia Adriani。模拟肿瘤微环境的三维微流控平台[摘要]。第四届CRI-CIMT-EATI-AACR国际癌症免疫治疗会议:将科学转化为生存;2018年9月30日至10月3日;纽约,纽约。费城(PA): AACR;癌症免疫学杂志2019;7(2增刊):摘要nr A049。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Abstract A049: Three-dimensional microfluidic platform mimicking the tumor microenvironment
Immunotherapy is currently a main breakthrough in cancer treatment, but therapeutic approaches for solid tumors still present limitations in the clinical scenario due to the challenges posed by the immunosuppressive tumor microenvironment (TME). Specifically, among immune cells recruited to the TME, monocytes/macrophages are especially abundant. Macrophages in vitro have been classified as classically activated M1 macrophages and alternatively activated M2 macrophages. However, macrophages are highly plastic cells and they often present mixed phenotypes in the in vivo TME. Monocytes/macrophages are involved in cancer cell proliferation, cell invasion, cell killing, vascular angiogenesis, T-cell immunosuppression, and are often correlated with a poor outcome in an extended range of cancers. However, disrupting the protumor activity of monocytes/macrophages and their interactions with the complex cellular system in the TME remains a challenge. Thus, a better understanding of the mechanisms that modulate monocyte/macrophage phenotype and their interactions with tumors may lead to make them a relevant therapeutic strategy. To study the immunosuppressive TME, we developed microfluidic-based integrated platforms with a 3-dimensional (3D) co-culture of tumor cells and immune cells to investigate how physical and molecular cues in the TME regulate the cellular interplay. Our 3D multicellular platforms offer considerable benefits and have already demonstrated in previous studies a clear advantage over classical 2D platforms to model the TME and to screen for different therapeutic approaches. Our previous studies demonstrated the crucial role of immune system interactions with cancer cells in metastasis either at a primary tumor site or at a secondary metastatic site. Further, we investigated the impact of monocytes and Programmed Death Ligand 1 (PD-L1) immune checkpoint on T-cell receptor (TCR)-engineered T-cells. We have now developed a new 3D microfluidic platform to study the effects of interstitial flow (IF), the flow of fluid through tumor stroma, which is an important component of the TME that may contribute to the polarization of macrophages toward a protumor phenotype. The microfluidic model allows the co-culture of tumor cells and monocytes/macrophages, to stimulate them with IF and to quantify cell migration in 3D. To the best of our knowledge, this study represents the first microfluidic tumor model that incorporates IF and both tumor and immune cells. Our preliminary results confirmed that the presence of tumor cells and hence tumor-cell secreted factors (TSF) increased the migration speed and directedness of macrophages toward cancer cells, potentially contributing to cancer cell dissemination. Interestingly, the presence of the IF-based mechanical cue (without tumor cells in culture) resulted in a similar increase in macrophage migration. Further, by combining both TSF and IF we observed no synergistic or additive effect on macrophage migration. A mixed population of macrophage phenotypes (M1 and M2) was observed with either the stimulus of TSF and/or IF while lower expression levels of M1 and M2 markers were observed without TSF or IF. Importantly, the understanding gained from this investigation can help in the design and functional testing of immunotherapeutic strategies to modulate macrophage polarization in the TME towards an antitumor phenotype. Citation Format: Andrea Pavesi, Siew Cheng Wong, Roger Kamm, Sharon Wei Ling Lee, Giulia Adriani. Three-dimensional microfluidic platform mimicking the tumor microenvironment [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr A049.
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