肿瘤浸润淋巴细胞对转移性骨肉瘤的安全性及有效性的探索性研究

An exploratory clinical study of the efficacy and safety of tumor-infiltrating lymphocytes in the treatment of metastatic osteosarcoma

目的观察从患者肿瘤组织中提取的肿瘤浸润淋巴细胞(TILs)经体外扩增联合高剂量白细胞介素2(IL-2)回输对转移性骨肉瘤患者的安全性及有效性。方法纳入2019年12月1日至2021年6月20日上海市第一人民医院12例病理诊断为骨肉瘤的受试者。所有患者在接受标准化疗后进展出现转移,并且多线治疗失败。从转移病灶活检取得新鲜肿瘤组织,经体外提取并扩增生产TILs,达到临床治疗量后(109~1011),回输给患者。自体TILs输注后立即给予高剂量IL-2(100000~200000 U/kg)。观察患者不良事件和生存情况。结果1例患者自体TILs制造失败,11例患者采用单次输注自体TILs治疗,男10例,女1例,中位年龄19.9岁(12~33岁)。其中6例采用TILs细胞剂量达1.0×10^(10)以上。11例患者随访1~13个月,耐受良好。报告的最常见的不良事件是发热(10/11)、便秘(3/11)和血谷氨酰转移酶(GGT)增高(3/11)。发热发生率高主要是注射IL-2所致。所有患者经过清除淋巴细胞方案后均经历了淋巴细胞计数的短暂下降和白细胞的减少。4级血液学毒性包括淋巴细胞增多(8/11)、中性粒细胞减少(2/11)和血小板减少(1/11)。未观察到神经毒性。在11例可评价患者中,获得的最佳反应为9例疾病稳定(SD),2例疾病进展(PD)。疾病控制率为9/11,SD中位持续时间>4个月。其中1例SD患者的缓解期超过6个月,肿瘤体积最大缩小超过20%。值得注意的是,受试者9治疗后持续SD状态6个月以上。结论进展期骨肉瘤患者肿瘤组织中可分离出具有体外扩增能力的TILs,通过体外扩增并回输的TILs具有一定的抗骨肉瘤作用。

Objective To explore the safety and efficacy of tumor-infiltrating lymphocytes(TILs)extracted from tumor tissue in patients with pulmonary metastasis of osteosarcoma,the TILs were amplified in vitro to reach clinical dosage and reinfused to the patients combined with high-dose interleukin 2(IL-2).Methods Twelve subjects with pathologically diagnosed osteosarcoma were enrolled from December 2019 to June 20,2021 in Shanghai General Hospital.All subjects progressed with metastasis after standard chemotherapy and failed multiple lines of treatments.Fresh tumor tissue was obtained from the metastatic site and extracted and amplified by Good Manufacturing Practice(GMP)workshop to produce TILs to clinical treatment dosage(109-1011).High-dose IL-2(100000-200000 U/kg)was administered immediately after autogenous TILs infusion to promote the activation,proliferation and antitumor cytolytic activity in vivo.Adverse events(AE)were graded according to Common Terminology Criteria for Adverse Events(CTCAE)standard and tumor response was assessed according to Response Evaluation Criteria in Solid Tumors(RECIST)1.1.Results One patient did not receive treatment due to failure in isolating TILs,total of 11 patients received a single re-infusion of autologous TILs.There were 10 males and 1 female with a median age of 19.9 years(12-33 years).Six of these patients received higher dose levels of 1.0×10^(10) TILs.The 11 patients were followed-up for 1 to 13 months and tolerated well.The most common adverse events reported were fever(10/11),constipation(3/11)and elevated gamma-glutamyl transferase(GGT)(3/11).The high incidence of fever was due to the IL-2 infusion.All patients experienced a transient drop in lymphocyte count and leukopenia leading to non-myeloid ablative lymphocyte clearance.The AE included grade 4 hematologic toxicity,including 8 cases of lymphocytopenia,2 cases of neutropenia and 1 case of thrombocytopenia.No AE of neurotoxicity occurred.Of all the 11 patients,9 patients got stable disease(SD)and 2 patients had progressive disease(PD).The disease control rate was 9/11.The median duration of SD was more than 4 months,and the maximum tumor volume decreased by close to 20%.Patient number 9 had sustained SD status for more than 6 months.Conclusions TILs with in vitro expansion ability could be isolated from tumor tissues of advanced osteosarcoma patients.TILs amplified and reinfused in vitro have anti-osteosarcoma activity.