23 research outputs found
Estrogen action on the prostate gland: a critical mix of endocrine and paracrine signaling
No full textA pro-tumourigenic loop at the human prostate tumour interface orchestrated by oestrogen, CXCL12 and mast cell recruitment
No full textArticle first published online: 9 JUL 2014Abstract not availableStuart J Ellem, Renea A Taylor, Luc Furic, Ola Larsson, Mark Frydenberg, David Pook, John Pedersen, Bree Cawsey, Andrew Trotta, Eleanor Need, Grant Buchanan and Gail P Risbridge
In vitro modeling of the prostate cancer microenvironment
No full textProstate cancer is the most commonly diagnosed malignancy in men and advanced disease is incurable. Model systems are a fundamental tool for research and many in vitro models of prostate cancer use cancer cell lines in monoculture. Although these have yielded significant insight they are inherently limited by virtue of their two-dimensional (2D) growth and inability to include the influence of tumour microenvironment. These major limitations can be overcome with the development of newer systems that more faithfully recreate and mimic the complex in vivo multi-cellular, three-dimensional (3D) microenvironment. This article presents the current state of in vitro models for prostate cancer, with particular emphasis on 3D systems and the challenges that remain before their potential to advance our understanding of prostate disease and aid in the development and testing of new therapeutic agents can be realised
Novel imaging of the prostate reveals spontaneous gland contraction and excretory duct quiescence together with different drug effects
No full textProstate carcinoma and benign prostate hyperplasia (BPH) with associated lower urinary tract symptoms (LUTS) are among the most prevalent and clinically relevant diseases in men. BPH is characterized by an enlargement of prostate tissue associated with increased tone of smooth muscle cells (SMCs) which surround the single glands composing the prostate. Secretions of the glands leave the prostate through local excretory ducts during the emission phase of ejaculation. Pharmacological treatment of BPH suggests different local drug targets based on reduction of prostate smooth muscle tone as the main effect and disturbed ejaculation as a common side effect. This highlights the need for detailed investigation of single prostate glands and ducts. We combined structural and functional imaging techniques—notably, clear lipid-exchanged, acrylamide-hybridized rigid imaging/immunostaining/in situ hybridization–compatible tissue-hydrogel (CLARITY) and time-lapse imaging—and defined glands and ducts as distinct SMC compartments in human and rat prostate tissue. The single glands of the prostate (comprising the secretory part) are characterized by spontaneous contractions mediated by the surrounding SMCs, whereas the ducts (excretory part) are quiescent. In both SMC compartments, phosphodiesterase (PDE)-5 is expressed. PDE5 inhibitors have recently emerged as alternative treatment options for BPH. We directly visualized that the PDE5 inhibitors sildenafil and tadalafil act by reducing spontaneous contractility of the glands, thereby reducing the muscle tone of the organ. In contrast, the ductal (excretory) system and thus the prostate’s contribution to ejaculation is unaffected by PDE5 inhibitors. Our differentiated imaging approach reveals new details about prostate function and local drug actions and thus may support clinical management of BPH.—Kügler, R., Mietens, A., Seidensticker, M., Tasch, S., Wagenlehner, F. M., Kaschtanow, A., Tjahjono, Y., Tomczyk, C. U., Beyer, D., Risbridger, G. P., Exintaris, B., Ellem, S. J., Middendorff, R. Novel imaging of the prostate reveals spontaneous gland contraction and excretory duct quiescence together with different drug effects.</p
Mimicking the tumour microenvironment (TME): Angiogenesis in tumour progression
No full textTwo-dimensional (2D) substrates cannot accurately mimic the\ud
complex matrix of native TME, whereas 3D models can recapitulate the natural\ud
tumour progression in vitro. As part of the tumour stroma, fibroblasts and\ud
endothelial cells (ECs) are well-known to not only support tumour growth but\ud
also to reduce the efficacy of anti-cancer drugs. Particularly, ECs are involved\ud
in the process of tumour vascularisation which represents a crucial step in the\ud
progression of cancer. Most of the previous studies are carried out in animal\ud
models or 2D cultures; hence, a detailed evaluation of experimental data is\ud
poor. To address this issue, we aim to develop a novel 3D in vitro approach, to\ud
mimic native tumour angiogenesis in 3D and to quantify the developed vascular\ud
network
3D bioengineered microtissues reveal key role of tumour microenvironment in early prostate carcinogenesis
No full textThe tumour microenvironment (TME) plays a fundamental role in prostate carcinogenesis. Classical tumour recombination experiments have shown that cancer‐associated stroma directs tumour formation in benign epithelia. Despite this, stromal components are often overlooked in human PCa models. Currently, there are no models of human PCa which adequately examines the combined contribution of cancer‐associated fibroblasts (CAF), their aberrant extracellular matrix (ECM) and a key resident immune population, mast cells (MC). Here we describe a three‐dimensional (3D) bioengineered TME microtissue in vitro model which interrogates the interaction and contribution of these components in early prostate carcinogenesis.
Melt‐electrospun scaffolds were formatted from medical grade poly(ε‐caprolactone). Patient‐derived primary CAFs or non‐malignant prostatic fibroblasts (NPFs) were incorporated into the scaffolds, forming a 3D microtissue. Once confluent, tagged benign epithelial cells (BPH‐1 or RWPE‐1) were co‐cultured on the microtissues ± MC (HMC‐1 or LAD2), MC‐conditioned media (CM) or recombinant tryptase. Subsequently, microtissues were fixed and tumourigenicity was assessed by analysing the 3D morphological transformation of epithelial cells. Additionally, live cell migration assays were performed to further quantify invasive potential.
Our data show that CAF and NPF proliferate and each deposit distinct ECM to form a 3D stromal network within the scaffolds. CAF, but not NPF, microtissues induce an invasive morphology in the benign epithelium. MCs cooperate with CAFs, potentiating CAF‐induced tumourigenic effects. These effects are mediated by MC secreted factors, specifically tryptase (a serine protease). Our data indicate that MC‐derived tryptase likely acts by remodelling the native aberrant ECM deposited by primary CAFs, conferring tumourigenicity on the benign epithelia.
Overall, this model demonstrates the cascade of interactions between CAFs, ECM and MCs, mediated by tryptase, to drive early epithelial transformation in the human prostate. Our data also highlight tryptase as a key mediator of these effects, which may be a novel therapeutic target to slow carcinogenesis.No Full Tex
Brief Report: A Bioassay to Identify Primary Human Prostate Cancer Repopulating Cells
No full textAbstract
Cancer cells are heterogeneous in both their phenotypes and ability to promote tumor growth and spread. Xenografting is used to identify the most highly capable cells of regenerating tumors, referred to as cancer repopulating cells. Because prostate cancers (PCa's) rarely grow as xenografts, indentifying PCa repopulating cells has not been possible. Here, we report improved methods to xenograft localized primary PCa tissues using chimeric grafts with neonatal mouse mesenchyme. Xenograft survival of tumor tissue was significantly increased by neonatal mesenchyme (six of six patients, 66% of grafts, versus four of six patients, 41% of grafts) and doubled the proliferation index of xenografted cancer cells. When applied to isolated PCa cells, neonatal mesenchyme effectively reconstituted PCa's and increased xenograft survival (four of nine patients; 32% of grafts with mesenchyme and 0% without), and supported active cancer cell proliferation. Using this assay, we showed that unfractionated α2β1integrinhi and α2β1integrinlo cells from primary localized PCa's demonstrated tumor formation at comparable rates, similar to previous reports using metastatic specimens. Thus, this new protocol efficiently established tumors and enabled proliferative expansion of both intact tumor tissue and fractionated cancer cells, providing a bioassay to identify and therapeutically target PCa repopulating cells.</jats:p
