S. A. Lozano Calderón, J. Kim, K. Raskin, J. Schwab, D. Spentzos, Department of Orthopaedic Surgery, Musculoskeletal Oncology Service, Massachusetts General Hospital, Harvard Medical School, Boston, MA Y.-L. Chen, K. Bernstein, T. Delaney, Department of Radiation Oncology, Massachusetts General Hospital, Harvard Medical School, Boston, MA I. Chebib, G. P. Nielsen, V. Deshpande, Department of Pathology, Bone and Soft Tissue Service, Massachusetts General Hospital, Harvard Medical School, Boston, MA R. Rubio, Y. E. Wang, J. Quackenbush, Department of Biostatistics and Computational Biology, Dana Farber Cancer Institute, Boston, MA R. Rubio, Y. E. Wang, J. Quackenbush, Department of Biostatistics, Harvard School of Public Health, Boston, MA C. Garbutt, C. E. Lietz, G. Cote, Department of Medical Oncology, Massachusetts General Hospital, Boston, MA; Department of Medical Oncology, Massachusetts General Hospital, Boston, MA.
Clin Orthop Relat Res. 2019 Sep;477(9):2114-2126. doi: 10.1097/CORR.0000000000000867.
MicroRNAs are small, noncoding RNAs that regulate the expression of posttranslational genes. The presence of some specific microRNAs has been associated with increased risk of both local recurrence and metastasis and worse survival in patients with osteosarcoma. Pathologic fractures in osteosarcoma are considered to be more the manifestation of a neoplasm with a more aggressive biological behavior than the cause itself of worse prognosis. However, this has not been proved at the biological or molecular level. Currently, there has not been a microRNA profiling study of patients who have osteosarcoma with and without pathologic fractures that has described differences in terms of microRNA profiling between these two groups and their correlation with biologic behavior.
QUESTIONS/PURPOSES: (1) In patients with osteosarcoma of the extremities, how do the microRNA profiles of those with and without pathologic fractures compare? (2) What relationship do microRNAs have with local recurrence, risk of metastasis, disease-specific survival, and overall survival in osteosarcoma patients with pathologic fractures?
Between 1994 and 2013, 217 patients were diagnosed and treated at our institution for osteosarcoma of the extremities. Patients were excluded if (1) they underwent oncologic resection of the osteosarcoma at an outside institution (two patients) or (2) they were diagnosed with an extraskeletal osteosarcoma (29 patients) or (3) they had less than 1 year of clinical follow-up and no oncologic outcome (local recurrence, metastasis, or death) (four patients). A total of 182 patients were eligible. Of those, 143 were high-grade osteosarcomas. After evaluation of tumor samples before chemotherapy treatment, a total of 80 consecutive samples were selected for sequencing. Demographic and clinical comparison between the sequenced and non-sequenced patients did not demonstrate any differences, confirming that both groups were comparable. Diagnostic samples from the extremities of 80 patients with high-grade extremity osteosarcomas who had not yet received chemotherapy underwent microRNA sequencing for an ongoing large-scale osteosarcoma genome profiling project at our institution. Six samples were removed after a second look by a musculoskeletal pathologist who verified cellularity and quality of samples to be sequenced, leaving a total of 74 patients. Of these, two samples were removed as they were confirmed to be pelvic tumors in a second check after sequencing. The final study sample was 72 patients (11 patients with pathologic fractures and 61 without). Sequencing data were correlated with fractures and local recurrence, risk of metastasis, disease-specific survival, and overall survival through Kaplan-Meier analyses.
Several microRNAs were expressed differently between the two groups. Among the markers with the highest differential expression (edgeR and DESeq algorithms), Hsa-mIR 656-3p, hsa-miR 493-5p, and hsa-miR 381-3p were upregulated in patients with pathologic fractures, whereas hsa-miR 363, hsa-miR 885-5p, and has-miR 20b-5p were downregulated. The highest differential expression fracture and nonfracture-associated microRNA markers also distinguished groups of patients with different metastasis risk, a well as different disease-specific and overall survival. Furthermore, the profile of pathologic fractures demonstrated a higher differential expression for microRNA markers that were previously associated with a higher risk of metastasis and lower survival rates in patients with osteosarcoma.
In patients who have osteosarcoma, the microRNA profiles of those with pathologic fractures are different than of patients without pathologic fractures. The highest differential expression mircroRNA molecules in patients with pathologic fractures predict also higher risk of metastatic disease as well as worse disease-specific survival and overall survival. Furthermore, we found higher differential expression of microRNAs in the pathologic fracture group previously associated with poor prognosis. The higher risk of metastasis and poorer overall survival in patients with pathologic fractures is inherent to tumor aggressive biologic behavior. It is plausible that the fracture itself is not the direct cause of worse prognosis but another manifestation of tumor biologic aggressiveness. Identification of these molecules through liquid biopsies may help to determine which patients may benefit from surgery before fractures occur. The same technology can be applied to identify patterns of response to conventional chemotherapy, assisting in more specific and accurate systemic therapy.
III, prognostic study.
微小 RNA 是调节翻译后基因表达的小型非编码 RNA。一些特定微小 RNA 的存在与骨肉瘤患者局部复发和转移风险增加以及生存预后较差相关。骨肉瘤病理性骨折被认为更多地是具有更具侵袭性生物学行为的肿瘤的表现，而不是预后更差的原因本身。然而，这在生物学或分子水平上尚未得到证明。目前，还没有对伴有和不伴有病理性骨折的骨肉瘤患者进行 miRNA 谱分析的研究，描述了这两组患者之间 miRNA 谱分析的差异及其与生物学行为的关系。
问题/目的：(1)在肢体骨肉瘤患者中，伴有和不伴有病理性骨折的患者的 microRNA 谱有何不同？(2)在伴有病理性骨折的骨肉瘤患者中，微小 RNA 与局部复发、转移风险、疾病特异性生存率和总生存率有何关系？
1994 年至 2013 年期间，我们机构共诊断和治疗了 217 例肢体骨肉瘤患者。如果患者(1)在外院接受骨肉瘤的肿瘤切除(2 例)或(2)诊断为骨外骨肉瘤(29 例)，或(3)临床随访不足 1 年且无肿瘤转归(局部复发、转移或死亡)(4 例)，则将其排除在外。共有 182 名患者符合条件。其中 143 例为高级别骨肉瘤。在评估化疗前肿瘤样本后，共选择了 80 个连续样本进行测序。测序和非测序患者的人口统计学和临床比较未显示出任何差异，证实两组具有可比性。我们机构正在进行一项大型骨肉瘤基因组分析项目，对 80 例未接受化疗的高级别肢体骨肉瘤患者的肢体诊断样本进行 microRNA 测序。在对肌肉骨骼病理学家进行二次检查以验证要测序的样本的细胞数量和质量后，有 6 个样本被删除，最后总共有 74 名患者。其中，在测序后再次检查时，有 2 个样本被确认为骨盆肿瘤，因此被删除。最终研究样本为 72 例患者(11 例伴有病理性骨折，61 例无)。通过 Kaplan-Meier 分析将测序数据与骨折、局部复发、转移风险、疾病特异性生存率和总生存率相关联。
两组之间有几个 microRNAs 的表达不同。在差异表达最高的标记物(edgeR 和 DESeq 算法)中，hsa-mIR 656-3p、hsa-miR 493-5p 和 hsa-miR 381-3p 在伴有病理性骨折的患者中上调，而 hsa-miR 363、hsa-miR 885-5p 和 has-miR 20b-5p 下调。骨折和非骨折相关的最高差异表达 microRNA 标志物也区分了具有不同转移风险、不同疾病特异性和总体生存率的患者群体。此外，病理性骨折患者的 microRNA 标志物的特征性表达更高，这些标志物与骨肉瘤患者的转移风险和生存率较低相关。
在患有骨肉瘤的患者中，病理性骨折患者的 microRNA 谱与无病理性骨折患者的不同。病理性骨折患者中差异表达最高的 microRNA 分子也预示着更高的转移性疾病风险，以及更差的疾病特异性生存率和总体生存率。此外，我们发现病理性骨折组中与预后不良相关的 microRNAs 表达更高。病理性骨折患者的转移风险和总体生存率较差是肿瘤侵袭性生物学行为的内在表现。骨折本身不太可能是预后较差的直接原因，而是肿瘤生物学侵袭性的另一种表现。通过液体活检识别这些分子可能有助于确定哪些患者可能在骨折发生前受益于手术。同样的技术可以应用于识别对常规化疗的反应模式，从而有助于更具体和准确的系统治疗。
III，预后研究。
