Development of clinically relevant in vivo metastasis models using human bone discs and breast cancer patient-derived xenografts
Development of clinically relevant in vivo metastasis models using human bone discs and breast cancer patient-derived xenografts
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DOI：
10.1186/s13058-019-1220-2
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发表时间：
2019-11-29
影响因子：
7.4
通讯作者：
Ottewell, Penelope
中科院分区：
文献类型：
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作者：
Lefley, Diane;Howard, Faith;Ottewell, Penelope
晚期乳腺癌优先转移到骨;尽管靶向治疗取得了进展，但这种情况仍然无法治愈。研究乳腺癌转移到人骨微环境的临床相关模型的缺乏阻碍了这种疾病的有效治疗方法的发展。为了解决这个问题，我们开发了人源化小鼠模型，其中乳腺癌患者来源的异种移植物（PDX）以低变异性和高频率转移到人骨植入物。方法将人工髋关节置换术后患者的股骨头骨片植入NOD/SCID小鼠皮下，建立人体骨环境模型。对于转移研究，7例患者来源的异种移植肿瘤（PDX：BB3RC 32，ER+ PR+ HER2-; BB2RC 08，ER+ PR+ ER 2-; BB6RC 37，ER-PR- HER2-和BB6RC 39，ER+ PR+ HER2+），MDA-MB-231-luc2，将T47 D-Luc 2或MCF 7-Luc 2细胞注射到第4乳腺导管中，通过荧光素酶成像监测转移并在组织学切片上确认。通过uCT、钙黄绿素摄取和组织形态计量学评估骨完整性、活力和血管形成。采用全基因组芯片、实时荧光定量PCR和免疫组化技术检测肿瘤转移不同阶段的基因/蛋白表达谱。IL-1的重要性在阿那白滞素治疗后得到证实。结果股骨移植为肿瘤细胞转移提供了一个代谢活跃的、人类特异性的部位。4周后，对骨植入物进行血管重建，并显示出活跃的骨重建（如破骨细胞、成骨细胞和钙黄绿素摄取的存在所证明的）。限制骨植入物使用软骨下骨和通过导管内注射引入癌细胞最大限度地转移到人体骨植入物。MDA-MB-231细胞特异性转移至人骨（70%转移），而T47 D、MCF 7、BB 3RC 32、BB 2 RC 08和BB 6 RC 37细胞转移至人骨和小鼠骨。重要的是，人骨是优选的转移部位，尤其是来自ER+ PDX的（100%转移人骨，而20-75%转移至小鼠骨），而ER-ve PDX在20%的人骨和20%的小鼠骨中发生转移。乳腺癌细胞从原发肿瘤发展到骨转移时经历了一系列分子变化，包括IL-1B、IL-1 R1、S100 A4、CTSK、SPP 1和RANK的表达改变。抑制IL-1B信号传导显著减少骨转移。结论我们可靠的和临床相关的人源化小鼠模型为乳腺癌骨转移的建模提供了显着的进步。
Background Late-stage breast cancer preferentially metastasises to bone; despite advances in targeted therapies, this condition remains incurable. The lack of clinically relevant models for studying breast cancer metastasis to a human bone microenvironment has stunted the development of effective treatments for this condition. To address this problem, we have developed humanised mouse models in which breast cancer patient-derived xenografts (PDXs) metastasise to human bone implants with low variability and high frequency. Methods To model the human bone environment, bone discs from femoral heads of patients undergoing hip replacement surgery were implanted subcutaneously into NOD/SCID mice. For metastasis studies, 7 patient-derived xenograft tumours (PDX: BB3RC32, ER+ PR+ HER2-; BB2RC08, ER+ PR+ ER2-; BB6RC37, ER- PR- HER2- and BB6RC39, ER+ PR+ HER2+), MDA-MB-231-luc2, T47D-luc2 or MCF7-Luc2 cells were injected into the 4th mammary ducts and metastases monitored by luciferase imaging and confirmed on histological sections. Bone integrity, viability and vascularisation were assessed by uCT, calcein uptake and histomorphometry. Expression profiling of genes/proteins during different stages of metastasis were assessed by whole genome Affymetrix array, real-time PCR and immunohistochemistry. Importance of IL-1 was confirmed following anakinra treatment. Results Implantation of femoral bone provided a metabolically active, human-specific site for tumour cells to metastasise to. After 4 weeks, bone implants were re-vascularised and demonstrated active bone remodelling (as evidenced by the presence of osteoclasts, osteoblasts and calcein uptake). Restricting bone implants to the use of subchondral bone and introduction of cancer cells via intraductal injection maximised metastasis to human bone implants. MDA-MB-231 cells specifically metastasised to human bone (70% metastases) whereas T47D, MCF7, BB3RC32, BB2RC08, and BB6RC37 cells metastasised to both human bone and mouse bones. Importantly, human bone was the preferred metastatic site especially from ER+ PDX (100% metastasis human bone compared with 20-75% to mouse bone), whereas ER-ve PDX developed metastases in 20% of human and 20% of mouse bone. Breast cancer cells underwent a series of molecular changes as they progressed from primary tumours to bone metastasis including altered expression of IL-1B, IL-1R1, S100A4, CTSK, SPP1 and RANK. Inhibiting IL-1B signalling significantly reduced bone metastasis. Conclusions Our reliable and clinically relevant humanised mouse models provide significant advancements in modelling of breast cancer bone metastasis.