Bioengineered Prostate-on Chip: Mechanisms of Stromal Dysregulation in Prostate Cancer

生物工程前列腺芯片：前列腺癌基质失调的机制

• 批准号: 10593937
• 负责人: Cynthia K Miranti
• 金额: $ 46.84万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-07 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10593937
• 关键词: AR gene; Acceleration; Address; Age; Androgen Receptor; Basal Cell; Basic Cancer Research; Biological Testing; Biology; Biomedical Engineering; Cell Differentiation process; Cell Line; Cell Survival; Cells; Coculture Techniques; Data; Dependence; Development; Diagnosis; Disease; Epithelial Cells; Epithelium; FGF10 gene; Fibroblasts; Genetic Transcription; Gland; Human; In Situ; Individual; MAP3K7 gene; Maintenance; Malignant Neoplasms; Malignant neoplasm of prostate; Measures; Mediating; Microfluidic Microchips; Modeling; NOTCH3 gene; Oncogenes; Oncogenic; Oranges; Patients; Phenotype; Physiological; Proliferating; Prostate; Receptor Signaling; Repression; Signal Induction; Signal Transduction; Stratified Epithelium; Stromal Cells; Structure; TNF gene; Testing; Tissues; Transforming Growth Factor beta; Tumor Cell Line; Tumor-Derived; cancer cell; carcinogenesis; gene repression; human model; in vivo; man; men; morphogens; neoplastic cell; new technology; overexpression; preservation; prostate cancer model; receptor expression; translational cancer research; tumor; tumorigenesis; tumorigenic

ABSTRACT It is well accepted that intrinsic action of the androgen receptor (AR) within the prostate epithelium drives prostate cancer proliferation and survival. Less appreciated is the fact that AR is also expressed in the stroma surrounding the epithelium. Stromal-expressed AR acts extrinsically to maintain the differentiated striated basal and luminal epithelium of the normal gland. During prostate cancer development, AR expression in the stroma is lost. How AR is lost from the stroma and how its loss promotes prostate cancer development is unknown. Our objectives are to define the mechanism that leads to stromal-specific AR loss and determine how AR loss in the stroma, in conjunction with epithelial oncogenesis, promotes prostate cancer development Based on our preliminary data, we hypothesize that tumor-derived TNFα/TGFβ1 transcriptionally suppresses AR expression in the stroma, causing loss of FGF10 and Wnt16 secretion, which are required to maintain the stratified epithelium through induction of luminal cells and maintenance of basal cells, respectively. To test this, we developed the first human Prostate-on-Chip model by culturing basal epithelial cells next to prostate stromal cells within a microfluidic device. Within this model, we can fully recapitulate the stromal AR-dependent induction of luminal epithelial cell differentiation. Furthermore, co-culturing normal stroma with tumor cells within this model leads to the induction of CAF phenotypes and reduced stromal AR expression, mimicking the tumor/host interactions seen in vivo. Models that recapitulate human glandular organization and its dysregulation during disease development are critical for our mechanistic understanding of how stroma and oncogenic epithelial interactions drive tumor development. We will test our hypothesis in three aims: 1) Determine the mechanism by which stromal AR maintains prostate epithelial cell differentiation. Our working hypothesis is that stromal AR signaling induces secretion of stromal FGF10 and Wnt16, which are required for induction of luminal epithelial cells and maintenance of basal epithelial cells, respectively. 2) Determine the mechanism by which AR expression is lost in the tumor stroma. Our working hypothesis is that tumor-secreted factors, TNFα and TGFβ1, acting through NF-κB signaling, suppress transcription of the stromal AR gene independent of CAF conversion. 3) Determine the functional consequence of tumor-induced stromal AR loss on prostate epithelial differentiation in a new de novo in situ human prostate cancer model. Our working hypothesis is that tumor-induced stromal loss of AR- dependent induction of Wnt16 and FGF10, via TNFα/TGFβ1, co-operates with epithelial oncogenes to accelerate tumor development and induce loss of basal epithelial cells. The proposed studies will be the first to demonstrate how TNFα/TGFβ-mediated suppression of stromal AR expression leads to the loss of Wnt16 and FGF10 to promote prostate cancer development. These studies will also provide the framework for further development of the first human Prostate-on-Chip model, which recapitulates human prostate biology, for basic and translation cancer research.

摘要 人们普遍认为，前列腺上皮内雄激素受体(AR)的内在作用驱动着前列腺 癌症的增殖和存活率。较少被认识到的事实是AR也在基质周围表达 上皮细胞。间质表达的AR外源性作用于维持分化的纹状基底和管腔 正常腺体的上皮组织。在前列腺癌的发展过程中，AR在间质中的表达会丢失。多么 AR从基质中丢失，它的丢失如何促进前列腺癌的发展尚不清楚。我们的目标 是定义导致间质特异性AR损失的机制，并确定AR在间质中的损失 根据我们的初步数据，与上皮肿瘤的发生相结合，促进前列腺癌的发展， 我们假设肿瘤来源的肿瘤坏死因子α/转化生长因子β1转录抑制间质中AR的表达， 导致FGF10和WNT16分泌的丧失，这是维持复层上皮所必需的 腔细胞的诱导和基底细胞的维持。为了测试这一点，我们开发了第一个人类 微流控培养前列腺基质细胞旁的基底层上皮细胞的前列腺片上模型 设备。在这个模型中，我们可以完全概括腔上皮细胞对间质AR依赖的诱导 差异化。此外，在该模型中，将正常间质与肿瘤细胞共培养可导致诱导 CAF表型和间质AR表达减少，模拟了体内肿瘤/宿主的相互作用。 概述了人类腺体组织及其在疾病发展过程中的失调的模型是 对于我们从机制上理解间质和致癌上皮相互作用如何驱动肿瘤至关重要 发展。我们将在三个目标上检验我们的假设：1)确定间质AR的机制 维持前列腺上皮细胞分化。我们的工作假设是间质AR信号诱导 分泌基质FGF10和WNT16，这是诱导腔上皮细胞和 分别维持基底上皮细胞。2)确定AR表达丢失的机制 在肿瘤间质中。我们的工作假设是肿瘤分泌因子，肿瘤坏死因子α和转化生长因子β1，通过 NF-κB信号，抑制基质AR基因的转录，而不依赖于CAF的转换。3)确定 肿瘤诱导的间质AR缺失对前列腺上皮细胞分化的功能影响 Novo原位人前列腺癌模型。我们的工作假设是肿瘤诱导的AR间质丢失- WNT16和FGF10通过肿瘤坏死因子α/转化生长因子β1的依赖诱导，与上皮癌基因协同加速 肿瘤的发生和导致基底上皮细胞的丧失。拟议的研究将是第一个证明 肿瘤坏死因子α/转化生长因子β介导的间质AR表达抑制导致WNT16和FGF10丢失的机制 促进前列腺癌的发展。这些研究还将为进一步发展 第一个人类前列腺片上模型，它概括了人类前列腺生物学，用于基础和翻译 癌症研究。