Characterizing breast cancer invasion and proliferation when co-aggregated with adipocytes in multicellular spheroids created with a custom bioreactor to augment cell-cell connectivity.

项目摘要 乳腺癌是女性最常见的癌症形式，被认为是现代流行病， 特别是因为高度的异质性。多项研究表明，肿瘤 微环境促进了肿瘤的生长和化疗耐药。癌症-脂肪细胞的相互作用 随着它们之间的串扰被证明促进肿瘤增殖和 癌症转移增加。然而，以前的共培养方法的生理相关性是 有问题。过去的几项研究没有使用人类细胞，也没有区分白色和棕色 脂肪细胞，具有不同的表型。在这项建议中，我们将分析人类肿瘤的侵袭性 三阴性和腔A乳腺癌细胞与人棕色和白色脂肪细胞共培养。至 为此，我们建议使用3D多细胞肿瘤球体系统。肿瘤球体提供了一个生理性的 研究癌症生物学和药物发现的相关环境。研究表明，与2D范式相比， 3D模型更好地概括了在人类体内肿瘤中发现的化疗耐药性和肿瘤生长。在实验室里，我们 已经开发出一种生物反应器，它利用表面振动产生法拉第波，使细胞聚集在内部 一个液室。具有类似技术的设备已被用于组装无支架3D细胞构造 快速和可扩展的方式，不需要额外的化学物质或侵入性程序。在……里面 在这项资助中，我们还试图确定这项技术是否能产生具有更大细胞-细胞的多细胞球体 附着力，与标准的自组装相比。为了实现这笔赠款的科学和技术目标， 提出了两个具体目标：第一个目标是比较多细胞的生存能力和连通性 用我们的表面振动生物反应器形成的球体与标准的自组装球体相比。第二 AIM将测量用人类癌症制成的肿瘤球体中肿瘤侵袭力和增殖能力的变化 细胞和人类脂肪细胞。我们将比较两种不同类型的乳腺癌细胞：三阴性和 腔A和两种不同类型的脂肪细胞：白色和棕色脂肪细胞。这些组合将使我们能够 探讨不同肿瘤表型与特定脂肪细胞类型的相互作用。总体而言， 拟议的研究将产生重大的科学和技术影响，通过扩大我们对 乳腺癌-脂肪细胞相互作用和测试一种技术，该技术可以通过 以可扩展的方式改进了蜂窝内连接。这将对癌症生物学领域产生重大影响 和治疗，因为在许多肿瘤中观察到癌症和脂肪细胞的关联，包括 前列腺癌、卵巢癌和胰腺癌。

Project Summary Breast cancer is the most common form of cancer in women and is considered a modern-day epidemic, especially because of the high degree of heterogeneity. Multiple studies have shown that the tumor microenvironment contributes to cancer growth and chemoresistance. Cancer-adipocyte interactions have gained increased attention as the crosstalk between them has been shown to promote tumor proliferation and increased cancer migration. However, the physiological relevance of previous co-culture methods is questionable. Several past studies did not use human cells, or differentiate between white and brown adipocytes, which have different phenotypes. In this proposal we will analyze tumor invasiveness of human triple negative and luminal A breast cancer cells when co-cultured with human brown and white adipocytes. To do this, we propose to use a 3D multicellular tumor spheroid system. Tumor spheroids provide a physiological relevant environment to study cancer biology and drug discovery. Compared to 2D paradigms, research shows 3D models better recapitulate chemoresistance and tumor growth found in human in vivo tumors. In the lab, we have developed a bioreactor that uses surface vibrations to produce Faraday waves that aggregate cells inside a liquid chamber. Devices with similar technology have been used to assemble scaffold-free 3D cell constructs and spheroids in a rapid and scalable way, without the need for additional chemicals or intrusive procedures. In this grant, we also seek to determine if this technology generates multicellular spheroids with greater cell-cell adhesion, compared to standard self-assembly. To achieve the scientific and technological goals of this grant, two specific aims are proposed: The first aim will compare the viability and connectivity of multicellular spheroids formed with our surface vibration bioreactor versus standard self-assembly spheroids. The second aim will measure changes in tumor invasiveness and proliferation in tumor spheroids made with human cancer cells and human adipocytes. We will compare two different types of breast cancer cells: triple negative and luminal A and two different types of adipocytes: white and brown adipocytes. These combinations will allow us to investigate the interactions of different cancer phenotypes with specific adipocyte cell types. Overall, the proposed research will have a major scientific and technological impact by expanding our understanding of breast cancer-adipocyte interactions and testing a technology that could deliver multicellular spheroids with improved intracellular connectivity in a scalable manner. This will significantly impact the field of cancer biology and therapeutics, as cancer-adipocyte associations have been observed in a multitude of tumors, including prostate, ovarian, and pancreatic cancer.