CD8+/FOXP3+-ratio in osteosarcoma microenvironment separates survivors from non-survivors: a multicenter validated retrospective study
CD8+/FOXP3+-ratio in osteosarcoma microenvironment separates survivors from non-survivors: a multicenter validated retrospective study
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DOI：
10.4161/2162402x.2014.990800
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发表时间：
2015-03-01
期刊：
影响因子：
7.2
通讯作者：
Kunz, Pierre
中科院分区：
文献类型：
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作者：
Fritzsching, Benedikt;Fellenberg, Joerg;Kunz, Pierre
骨肉瘤是最常见的原发性骨肿瘤，其特征在于幼年发病、肿瘤异质性和早期肺转移。二十多年来，治疗进展停滞不前。与主要恶性肿瘤不同，诊断时作为预后因素的生物标志物缺失。疾病的罕见性阻碍了研究招募足以超过肿瘤异质性的患者数量。在这里，我们在一个多中心队列中分析了骨肉瘤微环境，以减少肿瘤细胞异质性的影响。我们假设，当通过诊断性活检的全载玻片成像分析时，肿瘤内CD 8（+）T细胞与FOXP 3（+）T细胞的定量比率（CD 8（+）/FOXP 3（+）-比率）提供了强有力的预后信息。我们遵循了肿瘤标志物诊断研究（REMARK）。从纳入的150例病例中，确定了完成治疗的患者，并将其分配到发现队列（2004年之前诊断）或验证队列（2004-2012年诊断）。对CD 8（+）和FOXP 3（+）进行高度标准化的免疫组化，并通过甲基化特异性基因分析进行验证，然后进行全玻片分析和临床结果相关性分析。我们观察到，与CD 8（+）/FOXP 3（+）比值较低的患者相比，CD 8（+）/FOXP 3（+）比值高于中位数（3.08）的患者的估计生存率有所改善（p = 0.000001）。CD 8（+）/FOXP 3（+）比值高于第三四分位数的患者在观察期内无死亡（中位随访时间为69个月）。多变量分析表明，独立于目前的预后因素，包括转移和新辅助化疗的反应。来自独立验证队列的数据证实，CD 8（+）/FOXP 3（+）比值高于3.08的患者生存率提高（p = 0.001）。多变量分析证明，这一观察结果也独立于验证队列中诊断时的预后因素。预处理活检中肿瘤内CD 8（+）/FOXP 3（+）-比值可区分骨肉瘤患者的生存期延长与非生存期。
Osteosarcoma is the most common primary bone tumor characterized by juvenile onset, tumor heterogeneity, and early pulmonary metastasis. Therapeutic improvement stagnates since more than two decades. Unlike major malignancies, biomarkers as prognostic factors at time of diagnosis are missing. Disease rareness hampers study recruitment of patient numbers sufficient to outweigh tumor heterogeneity. Here, we analyzed in a multicenter cohort the osteosarcoma microenvironment to reduce effects of tumor cell heterogeneity. We hypothesized that quantitative ratios of intratumoral CD8(+)T-cells to FOXP3(+)T-cells (CD8(+)/FOXP3(+)-ratios) provide strong prognostic information when analyzed by whole-slide imaging in diagnostic biopsies. We followed recommendations-for-tumor-marker-prognostic-studies (REMARK). From 150 included cases, patients with complete treatment were identified and assigned to the discovery (diagnosis before 2004) or the validation cohort (diagnosis 2004-2012). Highly standardized immunohistochemistry of CD8(+) and FOXP3(+), which was validated by methylation-specific gene analysis, was performed followed by whole-slide analysis and clinical outcome correlations. We observed improved estimated survival in patients with CD8(+)/FOXP3(+)-ratios above the median (3.08) compared to patients with lower CD8(+)/FOXP3(+)ratios (p = 0.000001). No patients with a CD8(+)/FOXP3(+)-ratio above the third quartile died within the observation period (median follow-up 69 mo). Multivariate analysis demonstrated independence from current prognostic factors including metastasis and response to neoadjuvant chemotherapy. Data from an independent validation cohort confirmed improved survival (p = 0.001) in patients with CD8(+)/FOXP3(+)-ratios above 3.08. Multivariate analysis proofed that this observation was also independent from prognostic factors at diagnosis within the validation cohort. Intratumoral CD8(+)/FOXP3(+)-ratio in pretreatment biopsies separates patients with prolonged survival from non-survivors in osteosarcoma.