Mesenchymal stroma drives axonogenesis and nerve-induced aggressiveness in osteosarcoma.

IF 12.8 1区 医学 Q1 ONCOLOGY
Gemma Di Pompo, Thimios A Mitsiadis, Pierfrancesco Pagella, Alessandro Pasquarelli, Giuliano Bettini, Silvia Sabattini, Alberto Righi, Sofia Avnet, Nicola Baldini
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引用次数: 0

Abstract

Background: Osteosarcoma (OS), the most common primary bone malignancy, is a leading cause of cancer-related mortality in children and adolescents. Besides genomic abnormalities, several features of tumour microenvironment (TME), including cancer-associated mesenchymal stromal cells (MSC), have been recognized to play a key role in OS progression. The pathogenetic function of de novo innervation in TME has been extensively studied in carcinomas but is still an unexplored area of investigation in sarcomas, including OS.

Methods: We evaluated nerve infiltration in tissue samples from a small cohort of human OS (n = 5) and from canine OS (n = 11), a translational model for the human disease, by βIII-tubulin immunostaining. We then analysed nerve-stroma-tumour crosstalk using direct and indirect co-cultures of dorsal root ganglion (DRG) neurons with OS/tumour-associated mesenchymal stromal cells (MSC and cancer-associated fibroblasts, CAF), both under standard and microfluidic conditions. In particular, we investigated the effects of tumour and stromal cells on axonal tropism and outgrowth by measuring neurite recruitment, length, and branches and, vice versa, the impact of neuron-derived secretome on OS cell proliferation and migration. Finally, we assessed the secretion of pro-neurotrophic mediators, including brain-derived neurotrophic factor (BDNF), interleukin-6 (IL-6), and nerve growth factor (NGF), by MSC, CAF, and OS cells. The functional roles of IL-6 and BDNF were also verified by the blocking antibody Tocilizumab (TCZ) and the neutralizing Anti-BDNF antibody.

Results: We provided evidence of OS innervation within and surrounding the tumour in association with mesenchymal stroma that also corresponded to the most proliferative area of the tumour (Ki-67+). In vitro, both MSC and, to a lesser extent, OS cells promoted axonal growth through cytokine (IL-6) and neuromodulator (BDNF) secretion. Extracellular acidosis - a hallmark of OS aggressiveness - amplified IL-6 release by stromal cells, and its pro-neurogenic effect was prevented by IL-6 blockade. In turn, tumour-associated innervation stimulated OS cell proliferation and migration, eventually driving tumour aggressiveness.

Conclusions: We showed, for the first time, that bone-associated nerves, fostered by the OS microenvironment, promote tumour aggressiveness. Interfering with the nerve-tumour axis, particularly with the signalling associated with mesenchymal stroma, offers novel opportunities for OS treatment.

间充质间质驱动骨肉瘤的轴突发生和神经诱导的侵袭性。
背景:骨肉瘤(OS)是最常见的原发性骨恶性肿瘤,是儿童和青少年癌症相关死亡的主要原因。除了基因组异常,肿瘤微环境(TME)的一些特征,包括癌症相关间充质间质细胞(MSC),已被认为在OS进展中起关键作用。TME的新生神经支配的发病功能已经在肿瘤中得到了广泛的研究,但在肉瘤(包括OS)中仍是一个未开发的研究领域。方法:通过β iii -微管蛋白免疫染色,我们评估了一小群人OS (n = 5)和犬OS (n = 11)组织样本中的神经浸润情况,这是人类疾病的转化模型。然后,我们在标准和微流体条件下,使用背根神经节(DRG)神经元与OS/肿瘤相关间充质间质细胞(MSC和癌症相关成纤维细胞,CAF)的直接和间接共培养分析了神经基质-肿瘤串扰。特别是,我们通过测量神经突的招募、长度和分支,研究了肿瘤和基质细胞对轴突向性和生长的影响,反之亦然,研究了神经元源性分泌组对骨肉瘤细胞增殖和迁移的影响。最后,我们评估了MSC、CAF和OS细胞分泌的促神经营养介质,包括脑源性神经营养因子(BDNF)、白细胞介素-6 (IL-6)和神经生长因子(NGF)。阻断抗体Tocilizumab (TCZ)和中和抗体Anti-BDNF也证实了IL-6和BDNF的功能作用。结果:我们提供了肿瘤内部和周围OS神经支配与间充质间质相关的证据,间充质间质间质也与肿瘤最增生的区域(Ki-67+)相对应。在体外,MSC和OS细胞(在较小程度上)都通过细胞因子(IL-6)和神经调节剂(BDNF)的分泌促进轴突生长。细胞外酸中毒——OS侵袭性的一个标志——会放大基质细胞释放的IL-6, IL-6阻断可阻止其前神经发生作用。反过来,肿瘤相关的神经支配刺激OS细胞增殖和迁移,最终驱动肿瘤的侵袭性。结论:我们首次表明,骨相关神经在OS微环境的培养下,促进肿瘤侵袭性。干扰神经肿瘤轴,特别是与间充质间质相关的信号,为骨肉瘤治疗提供了新的机会。
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来源期刊
CiteScore
18.20
自引率
1.80%
发文量
333
审稿时长
1 months
期刊介绍: The Journal of Experimental & Clinical Cancer Research is an esteemed peer-reviewed publication that focuses on cancer research, encompassing everything from fundamental discoveries to practical applications. We welcome submissions that showcase groundbreaking advancements in the field of cancer research, especially those that bridge the gap between laboratory findings and clinical implementation. Our goal is to foster a deeper understanding of cancer, improve prevention and detection strategies, facilitate accurate diagnosis, and enhance treatment options. We are particularly interested in manuscripts that shed light on the mechanisms behind the development and progression of cancer, including metastasis. Additionally, we encourage submissions that explore molecular alterations or biomarkers that can help predict the efficacy of different treatments or identify drug resistance. Translational research related to targeted therapies, personalized medicine, tumor immunotherapy, and innovative approaches applicable to clinical investigations are also of great interest to us. We provide a platform for the dissemination of large-scale molecular characterizations of human tumors and encourage researchers to share their insights, discoveries, and methodologies with the wider scientific community. By publishing high-quality research articles, reviews, and commentaries, the Journal of Experimental & Clinical Cancer Research strives to contribute to the continuous improvement of cancer care and make a meaningful impact on patients' lives.
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