PO.TB04.05 · 肿瘤生物学
来自原发性人类前列腺癌的类器官在肿瘤相关上皮细胞状态方面表现出异质性
Organoids from primary human prostate cancer exhibit heterogeneity in the states of tumor-associated epithelial cells
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
前列腺癌（PCa）是男性中最常被诊断的恶性肿瘤，也是全球癌症相关死亡的第二大原因。尽管局限性PCa的长期生存率较高，但转移性疾病即使在接受强化多模态治疗后仍基本无法治愈。我们对晚期PCa侵袭性行为的驱动因素以及导致其对各种药物治疗无反应的分子通路的了解有限，这凸显了在体外研究肿瘤以便评估更有效疗法的必要性。肿瘤来源类器官代表了研究癌症的一种先进体外模型，正日益被用于开发新的治疗策略，成为推进肿瘤学精准医学的宝贵工具。我们的目标是表征来自FMRP-USP前列腺样本队列的患者来源类器官，这些类器官能够准确模拟原始患者肿瘤。我们的方法是在Matrigel中从活检样本建立较长期的类器官培养物。为此，我们使用了9个新鲜前列腺切除组织样本，通过机械解剖和酶消化进行处理，然后过滤并在含补充剂的高级DMEM/F12培养基中培养（GlutaMAX、B27、N-乙酰半胱氨酸、重组EGF、FGF-10和FGF-2、A-83-01、SB202190、烟酰胺、DHT、PGE2、Y-27632、Noggin和R-spondin）。9名患者年龄在50至75岁之间，诊断为高危PCa，ISUP分级为4级或5级（伦理委员会编号88216025.3.0000.5440）。使用形态学三维成像和组织学（H&E染色、IHC和IF）方法对类器官进行表征，并与同一肿瘤的原发样本进行比较，以确认其癌症建模能力和适用于下游检测的适宜性。我们成功建立了长达2个月的长期类器官培养。通过表征我们证实类器官呈现出与其起源组织相似的形态和肿瘤免疫特征。总之，我们成功建立并表征了长期患者来源的PCa类器官，它们紧密重现了原始肿瘤，为未来的临床前研究和精准肿瘤学应用提供了一个稳健的平台。
查看英文原文 English abstract
Prostate cancer (PCa) is the most commonly diagnosed malignant tumour in men and the second leading cause of cancer-related mortality worldwide. Despite the high long-term survival rates in localised PCa, metastatic disease remains largely incurable even after intensive multimodal therapy. Our limited understanding of the drivers of aggressive behaviour in advanced PCa and the molecular pathways that underlie their failure to respond to various drug therapies underscores the need to study tumours in vitro so that more effective therapies can be evaluated. Tumour-derived organoids represent an advanced in vitro model for studying cancer and are increasingly being used in the development of new therapeutic strategies, serving as valuable tools for advancing precision medicine in oncology. Our goal was to characterise patient-derived organoids from our prostate samples cohort of the FMRP-USP, which accurately model the original patient tumours. Our approach involved establishing longer-term organoid cultures from biopsy samples in Matrigel. For this, we used 9 fresh prostatectomy tissue samples that were processed by mechanical dissection and enzymatic digestion, then filtered and cultured in advanced DMEM/F12 medium with supplements (GlutaMAX, B27, N-acetylcysteine, recombinant EGF, FGF-10 and FGF-2, A-83-01, SB202190, nicotinamide, DHT, PGE2, Y-27632, Noggin, and R-spondin). Nine patients were aged between 50 and 75 years and diagnosed with high-risk PCa with ISUP grade of 4 or 5 (Ethics Committee Number 88216025.3.0000.5440). The organoids were characterized using morphological 3D imaging and histological (H&E staining, IHC and IF) approaches and compared to the primary sample of the same tumor to confirm their cancer-modelling capacity and suitability for the downstream assays. We successfully established long-term organoid culture for up to 2 months. We verified through characterisation that the organoids presented a morphology and tumour immune profile similar to the tissues of origin. In summary, we have successfully established and characterised long-term patient-derived PCa organoids, which closely recapitulate the original tumours, providing a robust platform for future pre-clinical studies and precision oncology applications.
利益披露 Disclosure
N. J. Santos, None..
L. C. R. Caldeira da Costa, None..
F. O. Buono, None..
F. P. Saggioro, None..
A. A. Rodrigues Junior, None..
R. Neuppmann Feres, None..
R. Borges dos Reis, None..
J. A. Squire, None..
L. Fröhlich Archangelo, None.