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: 10204253
• 负责人: Cynthia K Miranti
• 金额: $ 48.42万
• 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/10204253
• 关键词: AR gene; Address; Age; Ally; Androgen Receptor; Basal Cell; Biological Testing; Biology; Biomedical Engineering; Cell Differentiation process; Cell Line; Cell Maintenance; Cell Survival; Cells; Coculture Techniques; Data; Dependence; Development; Diagnosis; Disease; Duct (organ) structure; Epithelial; Epithelial Cells; FGF10 gene; Fibroblasts; Genetic Transcription; Gland; Human; In Situ; Individual; MAP3K7 gene; Malignant Neoplasms; Malignant neoplasm of prostate; Measures; Mediating; Microfluidic Microchips; Modeling; NOTCH3 gene; Oncogenes; Oncogenic; Oranges; Patients; Phenotype; Physiological; Prostate; Receptor Signaling; Repression; Signal Transduction; Stratified Epithelium; Stromal Cells; Structure; TNF gene; Testing; Tissues; Transforming Growth Factor beta; Translations; Tumor Cell Line; Tumor-Derived; anticancer research; base; cancer cell; carcinogenesis; host neoplasm interaction; human model; in vivo; man; men; morphogens; neoplastic cell; new technology; overexpression; preservation; prostate cancer model; receptor expression; 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.

抽象的 人们普遍认为前列腺上皮内雄激素受体 (AR) 的内在作用驱动前列腺 较少人认识到 AR 也在周围基质中表达。 基质表达的 AR 发挥外在作用以维持分化的基底层和管腔纹状体。 正常腺体上皮细胞在前列腺癌发展过程中，间质中的 AR 表达会丢失。 AR 从基质中丢失，其丢失如何促进前列腺癌的发展尚不清楚。 的目的是定义导致基质特异性 AR 损失的机制，并确定基质中 AR 损失的方式， 与上皮肿瘤发生结合，促进前列腺癌的发展 根据我们的初步数据， 我们发现肿瘤来源的 TNFα/TGFβ1 转录抑制基质中 AR 的表达， 导致 FGF10 和 Wnt16 分泌丧失，而 FGF10 和 Wnt16 是维持复层上皮所需的 分别诱导管腔细胞和维持基底细胞为了测试这一点，我们开发了第一个人类。 通过在微流体内培养前列腺基质细胞旁边的基底上皮细胞来建立前列腺芯片模型 在这个模型中，我们可以完全重现管腔上皮细胞的基质 AR 依赖性诱导。 此外，在该模型中将正常基质与肿瘤细胞共培养会导致诱导。 CAF 表型的改变和基质 AR 表达的减少，模拟了体内观察到的肿瘤/宿主相互作用。 概括人类腺体组织及其在疾病发展过程中失调的模型是 对于我们理解基质和致癌上皮相互作用如何驱动肿瘤的机制至关重要 我们将在三个目标上检验我们的假设：1）确定基质 AR 的机制。 我们的工作假设是基质 AR 信号传导诱导。 基质 FGF10 和 Wnt16 的分泌，这是诱导管腔上皮细胞和 分别维持基底上皮细胞2)确定AR表达丢失的机制。 我们的工作假设是肿瘤分泌因子 TNFα 和 TGFβ1 通过作用发挥作用。 NF-κB 信号传导，抑制基质 AR 基因的转录，与 CAF 转换无关 3) 确定。 肿瘤诱导的基质 AR 损失对前列腺上皮分化的功能影响 我们的工作假设是肿瘤诱导的 AR- 基质损失。 通过 TNFα/TGFβ1 依赖性诱导 Wnt16 和 FGF10，与上皮癌基因合作加速 拟议的研究将首次证明其会影响肿瘤的发展并诱导基底上皮细胞的损失。 TNFα/TGFβ 介导的基质 AR 表达抑制如何导致 Wnt16 和 FGF10 缺失 这些研究还将为前列腺的进一步发展提供框架。 第一个人类前列腺芯片模型，概括了人类前列腺生物学，用于基础和翻译 癌症研究。