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

摘要 众所周知，前列腺上皮内雄激素受体（AR）的内在作用驱动前列腺增生， 癌细胞增殖和存活。不太了解的是，AR也表达在周围的基质中， 上皮细胞间质表达的AR在细胞外起作用，维持分化的纹状体基底和管腔 正常腺体的上皮。在前列腺癌发展过程中，基质中的AR表达丢失。如何 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。我们的目标 是为了确定导致基质特异性AR损失的机制，并确定基质中的AR损失， 结合上皮肿瘤发生，促进前列腺癌的发展基于我们的初步数据， 我们假设肿瘤源性TNFα/TGFβ1在转录上抑制间质中的AR表达， 导致FGF 10和Wnt 16分泌的损失，所述FGF 10和Wnt 16分泌是维持复层上皮细胞所需的， 分别诱导腔细胞和维持基底细胞。为了验证这一点，我们开发了第一个人类 通过在微流体内培养邻近前列腺基质细胞的基底上皮细胞的前列腺芯片模型 设备.在这个模型中，我们可以完全概括间质AR依赖性的管腔上皮细胞诱导， 分化此外，在该模型中，将正常基质与肿瘤细胞共培养导致诱导 CAF表型和减少的基质AR表达，模拟体内观察到的肿瘤/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 发展我们将从三个方面检验我们的假设：1）确定基质AR 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FGF 10和Wnt 16，它们是诱导腔上皮细胞所需的， 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TNFα和TGFβ1通过 NF-κB信号传导抑制不依赖于CAF转化的基质AR基因的转录。3)确定 肿瘤诱导的间质AR缺失对前列腺上皮分化的功能影响 新的原位人前列腺癌模型。我们的工作假设是肿瘤诱导的AR- 通过TNFα/TGFβ1对Wnt 16和FGF 10的依赖性诱导，与上皮癌基因协同作用， 肿瘤发展并诱导基底上皮细胞的损失。拟议中的研究将首次证明 TNFα/TGFβ介导的间质AR表达抑制如何导致Wnt 16和FGF 10的丢失， 促进前列腺癌的发展。这些研究还将为进一步发展 第一个人类前列腺芯片模型，它概括了人类前列腺生物学，用于基础和翻译 癌症研究。

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.