DataSheet_1_Combination of Bempegaldesleukin and Anti-CTLA-4 Prevents Metastatic Dissemination After Primary Resection or Radiotherapy in a Preclinical Model of Non-Small Cell Lung Cancer.docx

Surgical resection or hypo-fractionated radiation therapy (RT) in early-stage non-small cell lung cancer (NSCLC) achieves local tumor control, but metastatic relapse remains a challenge. We hypothesized that immunotherapy with anti-CTLA-4 and bempegaldesleukin (BEMPEG; NKTR-214), a CD122-preferential IL2 pathway agonist, after primary tumor RT or resection would reduce metastases in a syngeneic murine NSCLC model. Mice bearing Lewis Lung Carcinoma (LLC) tumors were treated with combinations of BEMPEG, anti-CTLA-4, and primary tumor treatment (surgical resection or RT). Primary tumor size, mouse survival, and metastatic disease at the time of death were assessed. Flow cytometry, qRT-PCR, and cytokine analyses were performed on tumor specimens. All mice treated with RT or surgical resection of primary tumor alone succumbed to metastatic disease, and all mice treated with BEMPEG and/or anti-CTLA-4 succumbed to primary tumor local progression. The combination of primary tumor RT or resection and BEMPEG and anti-CTLA-4 reduced spontaneous metastasis and improved survival without any noted toxicity. Flow cytometric immunoprofiling of primary tumors revealed increased CD8 T and NK cells and decreased T-regulatory cells with the combination of BEMPEG, anti-CTLA-4, and RT compared to RT alone. Increased expression of genes associated with tumor cell immune susceptibility, immune cell recruitment, and cytotoxic T lymphocyte activation were observed in tumors of mice treated with BEMPEG, anti-CTLA-4, and RT. The combination of BEMPEG and anti-CTLA-4 with primary tumor RT or resection enabled effective control of local and metastatic disease in a preclinical murine NSCLC model. This therapeutic combination has important translational potential for patients with early-stage NSCLC and other cancers.

手术切除或低分割放疗（hypo-fractionated radiation therapy, RT）可实现早期非小细胞肺癌（non-small cell lung cancer, NSCLC）的局部肿瘤控制，但远处转移复发仍是尚未解决的临床挑战。本研究提出假说：在原发肿瘤接受放疗或手术切除后，联合抗CTLA-4免疫治疗与bempegaldesleukin（BEMPEG; NKTR-214，一种优先结合CD122的IL2通路激动剂），可在同基因小鼠NSCLC模型中减少转移发生。本研究将携带路易斯肺癌（Lewis Lung Carcinoma, LLC）肿瘤的小鼠，通过BEMPEG、抗CTLA-4联合原发肿瘤治疗（手术切除或放疗）的不同组合方案进行干预。实验评估了小鼠的原发肿瘤体积、生存率及死亡时的转移性疾病负荷，并对肿瘤标本开展流式细胞术、定量实时PCR（qRT-PCR）及细胞因子分析。
结果显示，仅接受原发肿瘤放疗或手术切除的小鼠均因转移性疾病死亡，仅接受BEMPEG单药或联合抗CTLA-4治疗的小鼠则因原发肿瘤局部进展死亡。而联合原发肿瘤放疗/切除、BEMPEG与抗CTLA-4的治疗方案，可减少自发性转移并改善小鼠生存率，且未观察到明显毒性反应。对原发肿瘤的流式细胞术免疫分型结果显示，相较于单纯放疗组，联合BEMPEG、抗CTLA-4与放疗的治疗组，肿瘤内CD8阳性T细胞与NK细胞比例升高，调节性T细胞（T-regulatory cells）比例降低。该联合治疗组的肿瘤标本中，与肿瘤细胞免疫易感性、免疫细胞招募及细胞毒性T淋巴细胞活化相关的基因表达水平均显著上调。
在临床前小鼠NSCLC模型中，联合原发肿瘤放疗/切除、BEMPEG与抗CTLA-4的治疗方案可有效控制局部肿瘤与转移性疾病。该治疗组合对早期NSCLC及其他癌症患者具有重要的转化研究价值。