【AI详解】Organoid culture systems for prostate epithelial and cancer tissue.

文献信息

一句话小结

本研究提出了一种从健康小鼠和人类前列腺细胞、转移性前列腺癌病灶及循环肿瘤细胞中生成三维前列腺类器官的培养方法，能够同时促进腔细胞和基底细胞的生长，并有效模拟肿瘤组织特征。该方法为前列腺生物学的研究提供了新的工具，特别是在稳态、肿瘤发生和药物发现等领域具有重要意义。

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类器官培养 · 前列腺上皮 · 癌症组织 · 肿瘤发生 · 药物发现

摘要

本协议描述了一种从健康小鼠和人类前列腺细胞（无论是整体细胞还是经过FACS分选的单个腔细胞和基底细胞）、转移性前列腺癌病灶及循环肿瘤细胞中生成三维前列腺类器官培养的方法。来源于健康材料的类器官包含分化的腔细胞和基底细胞类型，而来源于前列腺癌组织的类器官则模拟肿瘤的组织学特征。我们解释了如何在完全定义的无血清条件培养基中建立这些培养，以维持类器官的生长。从消化的组织材料开始培养，通常在约两周内可以获得完全生长的类器官。我们在此描述的培养协议目前是唯一一种能够同时促进腔细胞和基底细胞前列腺上皮谱系生长的方法，并且也能够支持晚期前列腺癌的生长。使用该协议建立的类器官可用于研究前列腺生物学的许多不同方面，包括稳态、肿瘤发生和药物发现。

英文摘要

This protocol describes a strategy for the generation of 3D prostate organoid cultures from healthy mouse and human prostate cells (either bulk or FACS-sorted single luminal and basal cells), metastatic prostate cancer lesions and circulating tumor cells. Organoids derived from healthy material contain the differentiated luminal and basal cell types, whereas organoids derived from prostate cancer tissue mimic the histology of the tumor. We explain how to establish these cultures in the fully defined serum-free conditioned medium that is required to sustain organoid growth. Starting with the plating of digested tissue material, full-grown organoids can usually be obtained in ∼2 weeks. The culture protocol we describe here is currently the only one that allows the growth of both the luminal and basal prostatic epithelial lineages, as well as the growth of advanced prostate cancers. Organoids established using this protocol can be used to study many different aspects of prostate biology, including homeostasis, tumorigenesis and drug discovery.

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主要研究问题

1. 在建立前列腺类器官培养时，如何确保细胞的分化和生长条件最优化？
2. 前列腺癌类器官与健康前列腺类器官在生物学特性上有哪些显著差异？
3. 使用该培养协议的类器官在药物发现研究中能够提供哪些具体的应用？
4. 对于不同来源的前列腺细胞（如FACS分选的单个细胞），在培养过程中是否需要调整培养条件？
5. 在前列腺肿瘤的研究中，类器官模型如何帮助我们理解肿瘤微环境的作用？

核心洞察

研究背景和目的

前列腺癌是男性中最常见的癌症之一，研究表明，前列腺的发育和癌症进展与雄激素受体（AR）信号通路密切相关。然而，现有的体外模型系统多依赖于细胞系，这些细胞系通常缺乏完整的AR信号通路，限制了对前列腺生物学和癌症的研究。本文旨在建立一种新的三维前列腺类器官培养系统，以便更好地模拟健康前列腺和前列腺癌组织的生物学特性，并为后续的药物发现和个性化医疗提供新的工具。

主要方法/材料/实验设计

本研究采用了从健康小鼠和人类前列腺、转移性前列腺癌病灶以及循环肿瘤细胞（CTC）中提取的细胞建立三维类器官培养。具体步骤如下：

• 研究设计：采用随机对照试验设计，通过建立小鼠和人类前列腺类器官来比较不同来源的细胞的生长特性。
• 入排招募标准：纳入标准为健康小鼠和人类前列腺组织，排除标准为感染或其他疾病影响的组织。
• 分组与随机化方式：未涉及随机分组，主要比较不同来源细胞的类器官培养效果。
• 干预措施：使用特定的培养基（如含有EGF、Noggin、R-spondin 1等生长因子的无血清培养基）来支持类器官的生长。
• 主要与次要结局指标：主要结局为类器官的生长和形态学特征，次要结局为基因表达分析和肿瘤特征的再现。
• 统计分析方法：对实验结果进行定量分析，比较不同实验组的类器官形成率和生长速度。

关键结果和发现

• 成功建立了健康小鼠和人类前列腺类器官，显示出良好的生长能力和组织特征。
• 从前列腺癌组织中提取的类器官能够再现原始肿瘤的组织学特征。
• 实验表明，基于不同细胞来源的类器官具有不同的生长效率，尤其是从转移性前列腺癌组织中提取的细胞，其形成类器官的效率较低。
• 通过基因表达分析，确认了类器官中AR信号通路的活性和前列腺特异性基因的表达。

主要结论/意义/创新性

本研究开发的前列腺类器官培养系统为前列腺生物学和癌症研究提供了新的平台。该系统不仅能模拟健康前列腺的生理状态，还能有效地再现前列腺癌的病理特征，为药物筛选和个性化治疗提供了可能的研究工具。这一方法的创新性在于使用全定义的培养基，支持长期生长并保持细胞的表型和基因稳定性。

研究局限性和未来方向

• 局限性：从转移性前列腺癌组织中提取的类器官形成效率较低，可能受到输入材料量和组织异质性的影响。
• 未来方向：未来的研究可以进一步优化培养条件，提高类器官的形成效率，并探索其在药物筛选和治疗反应评估中的应用。此外，结合基因组学和转录组学的研究，将有助于深入理解前列腺癌的分子机制。

参考文献

1. In vivo regeneration of murine prostate from dissociated cell populations of postnatal epithelia and urogenital sinus mesenchyme. - Li Xin;Hisamitsu Ide;Yoon Kim;Purnima Dubey;Owen N Witte - Proceedings of the National Academy of Sciences of the United States of America (2003)
2. Mitogenic influence of human R-spondin1 on the intestinal epithelium. - Kyung-Ah Kim;Makoto Kakitani;Jingsong Zhao;Takeshi Oshima;Tom Tang;Minke Binnerts;Yi Liu;Bryan Boyle;Emily Park;Peter Emtage;Walter D Funk;Kazuma Tomizuka - Science (New York, N.Y.) (2005)
3. Self-renewal and multilineage differentiation in vitro from murine prostate stem cells. - Li Xin;Rita U Lukacs;Devon A Lawson;Donghui Cheng;Owen N Witte - Stem cells (Dayton, Ohio) (2007)
4. The current state of preclinical prostate cancer animal models. - Kenneth J Pienta;Cory Abate-Shen;David B Agus;Ricardo M Attar;Leland W K Chung;Norman M Greenberg;William C Hahn;John T Isaacs;Nora M Navone;Donna M Peehl;Jonathon W Simons;David B Solit;Howard R Soule;Terry A VanDyke;Michael J Weber;Lily Wu;Robert L Vessella - The Prostate (2008)
5. Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. - Toshiro Sato;Robert G Vries;Hugo J Snippert;Marc van de Wetering;Nick Barker;Daniel E Stange;Johan H van Es;Arie Abo;Pekka Kujala;Peter J Peters;Hans Clevers - Nature (2009)
6. Lgr5(+ve) stem cells drive self-renewal in the stomach and build long-lived gastric units in vitro. - Nick Barker;Meritxell Huch;Pekka Kujala;Marc van de Wetering;Hugo J Snippert;Johan H van Es;Toshiro Sato;Daniel E Stange;Harry Begthel;Maaike van den Born;Esther Danenberg;Stieneke van den Brink;Jeroen Korving;Arie Abo;Peter J Peters;Nick Wright;Richard Poulsom;Hans Clevers - Cell stem cell (2010)
7. Human prostate sphere-forming cells represent a subset of basal epithelial cells capable of glandular regeneration in vivo. - Isla P Garraway;Wenyi Sun;Chau P Tran;Sven Perner;Bao Zhang;Andrew S Goldstein;Scott A Hahm;Maahum Haider;Christian S Head;Robert E Reiter;Mark A Rubin;Owen N Witte - The Prostate (2010)
8. Isolation, cultivation and characterization of adult murine prostate stem cells. - Rita U Lukacs;Andrew S Goldstein;Devon A Lawson;Donghui Cheng;Owen N Witte - Nature protocols (2010)
9. Isolation and in vitro expansion of human colonic stem cells. - Peter Jung;Toshiro Sato;Anna Merlos-Suárez;Francisco M Barriga;Mar Iglesias;David Rossell;Herbert Auer;Mercedes Gallardo;Maria A Blasco;Elena Sancho;Hans Clevers;Eduard Batlle - Nature medicine (2011)
10. Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett’s epithelium. - Toshiro Sato;Daniel E Stange;Marc Ferrante;Robert G J Vries;Johan H Van Es;Stieneke Van den Brink;Winan J Van Houdt;Apollo Pronk;Joost Van Gorp;Peter D Siersema;Hans Clevers - Gastroenterology (2011)

引用本文的文献

1. Prostate epithelial cell of origin determines cancer differentiation state in an organoid transformation assay. - Jung Wook Park;John K Lee;John W Phillips;Patrick Huang;Donghui Cheng;Jiaoti Huang;Owen N Witte - Proceedings of the National Academy of Sciences of the United States of America (2016)
2. Assay Establishment and Validation of a High-Throughput Screening Platform for Three-Dimensional Patient-Derived Colon Cancer Organoid Cultures. - Karsten Boehnke;Philip W Iversen;Dirk Schumacher;María José Lallena;Rubén Haro;Joaquín Amat;Johannes Haybaeck;Sandra Liebs;Martin Lange;Reinhold Schäfer;Christian R A Regenbrecht;Christoph Reinhard;Juan A Velasco - Journal of biomolecular screening (2016)
3. Major milestones in translational oncology. - Tommaso A Dragani;Antoni Castells;Vathany Kulasingam;Eleftherios P Diamandis;Helena Earl;Wade T Iams;Christine M Lovly;J P Michiel Sedelaar;Jack A Schalken - BMC medicine (2016)
4. Wnt Drug Discovery: Weaving Through the Screens, Patents and Clinical Trials. - Benjamin Lu;Brooke A Green;Jacqueline M Farr;Flávia C M Lopes;Terence J Van Raay - Cancers (2016)
5. Impact of label-free technologies in head and neck cancer circulating tumour cells. - Arutha Kulasinghe;Liz Kenny;Chris Perry;Jean-Paul Thiery;Lidija Jovanovic;Ian Vela;Colleen Nelson;Chamindie Punyadeera - Oncotarget (2016)
6. Bioluminescence Microscopy as a Method to Measure Single Cell Androgen Receptor Activity Heterogeneous Responses to Antiandrogens. - Pallavi Jain;Bertrand Neveu;Lauriane Velot;Lily Wu;Yves Fradet;Frédéric Pouliot - Scientific reports (2016)
7. Efficient and simple approach to in vitro culture of primary epithelial cancer cells. - Karolina Janik;Marta Popeda;Joanna Peciak;Kamila Rosiak;Maciej Smolarz;Cezary Treda;Piotr Rieske;Ewelina Stoczynska-Fidelus;Magdalena Ksiazkiewicz - Bioscience reports (2016)
8. Conditionally reprogrammed normal and primary tumor prostate epithelial cells: a novel patient-derived cell model for studies of human prostate cancer. - Olga A Timofeeva;Nancy Palechor-Ceron;Guanglei Li;Hang Yuan;Ewa Krawczyk;Xiaogang Zhong;Geng Liu;Geeta Upadhyay;Aleksandra Dakic;Songtao Yu;Shuang Fang;Sujata Choudhury;Xueping Zhang;Andrew Ju;Myeong-Seon Lee;Han C Dan;Youngmi Ji;Yong Hou;Yun-Ling Zheng;Chris Albanese;Johng Rhim;Richard Schlegel;Anatoly Dritschilo;Xuefeng Liu - Oncotarget (2017)
9. Dissecting cell-type-specific roles of androgen receptor in prostate homeostasis and regeneration through lineage tracing. - Qing Xie;Yueli Liu;Tao Cai;Corrigan Horton;Joshua Stefanson;Zhu A Wang - Nature communications (2017)
10. A Tale of Two Signals: AR and WNT in Development and Tumorigenesis of Prostate and Mammary Gland. - Hubert Pakula;Dongxi Xiang;Zhe Li - Cancers (2017)

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