Gomez-Brouchet Anne, Illac Claire, Gilhodes Julia, Bouvier Corinne, Aubert Sébastien, Guinebretiere Jean-Marc, Marie Béatrice, Larousserie Frédérique, Entz-Werlé Natacha, de Pinieux Gonzague, Filleron Thomas, Minard Véronique, Minville Vincent, Mascard Eric, Gouin François, Jimenez Marta, Ledeley Marie-Cécile, Piperno-Neumann Sophie, Brugieres Laurence, Rédini Françoise
Department of Pathology, IUCT-Oncopole, CHU of Toulouse and University of Toulouse, Pharmacology and Structural Biology Institute, Toulouse, France.
Biostatistics Unit, Claudius Regaud Institut, IUCT-Oncopole, Toulouse, France.
Oncoimmunology. 2017 Aug 24;6(9):e1331193. doi: 10.1080/2162402X.2017.1331193. eCollection 2017.
The French phase 3 trial (OS 2006) testing zoledronic acid, an osteoclast inhibitor, with chemotherapy and surgery did not improve the outcome of patients with osteosarcoma (OS). To understand this unexpected result, the presence of infiltrating immune cells was investigated in 124 pre-therapeutic biopsies of patients enrolled in the trial. The percentage of CD68/CD163 tumor-infiltrating macrophages (TAMs), CD8 lymphocytes, osteoclasts, and the PD1/PDL-1 checkpoint were assessed by immunohistochemistry. M1/M2 macrophage polarization was characterized by pSTAT1/CMAF staining. The expression of these biomarkers was correlated with clinical outcome. No statistical correlations were found with response to chemotherapy. High CD163 levels (>50% of cells per core; 43.8% of patients) were associated with CMAF nuclear expression and significantly correlated with better overall survival ( = 0.0025) and longer metastasis progression-free survival (MPFS, = 0.0315) independently of metastatic status ( = 0.002). Only a trend was observed for patients with high CD68-positive cells ( = 0.0582). CD8 staining was positive in >50% of cases with a median staining of 1%. Lower CD8 levels were associated with metastatic disease at diagnosis and the presence of CD8-positive cells significantly correlated with improved overall survival in zoledronate-treated patients ( = 0.0415). PD1/PDL-1 staining was negative in >80% of cases and was not correlated with outcome. Finally, CD163-positive TAMs and CD8 positive cells are crucial prognostic biomarkers in OS, whereas PD1/PDL-1 checkpoint plays a minor role. For the first time, we described a correlation between CD8 positive cells and survival in zoledronate-treated patients. The immunohistochemical analysis of the microenvironment in biopsies may represent a novel tool for therapeutic stratification.
法国的一项3期试验（OS 2006）对破骨细胞抑制剂唑来膦酸联合化疗及手术进行了测试，结果显示其并未改善骨肉瘤（OS）患者的预后。为了解这一意外结果，研究人员对该试验中124例患者的治疗前活检样本进行分析，调查浸润免疫细胞的存在情况。通过免疫组织化学评估CD68/CD163肿瘤浸润巨噬细胞（TAM）、CD8淋巴细胞、破骨细胞的百分比以及PD1/PDL-1检查点。通过pSTAT1/CMAF染色对M1/M2巨噬细胞极化进行表征。这些生物标志物的表达与临床结果相关。未发现与化疗反应存在统计学相关性。高CD163水平（每个核心中>50%的细胞；43.8%的患者）与CMAF核表达相关，并且与更好的总生存期（P = 0.0025）和更长的无转移进展生存期（MPFS，P = 0.0315）显著相关，且与转移状态无关（P = 0.002）。仅在高CD68阳性细胞的患者中观察到一种趋势（P = 0.0582）。在>50%的病例中CD8染色呈阳性，中位染色为1%。较低的CD8水平与诊断时的转移性疾病相关，并且在唑来膦酸治疗的患者中，CD8阳性细胞的存在与改善的总生存期显著相关（P = 0.0415）。在>80%的病例中PD1/PDL-1染色为阴性，且与结果无关。最后，CD163阳性TAM和CD8阳性细胞是骨肉瘤中关键的预后生物标志物，而PD1/PDL-1检查点作用较小。我们首次描述了唑来膦酸治疗患者中CD8阳性细胞与生存之间的相关性。活检样本中微环境的免疫组织化学分析可能是一种用于治疗分层的新工具。
