Clubb James H A, Kudling Tatiana V, Heiniö Camilla, Basnet Saru, Pakola Santeri, Cervera Carrascón Víctor, Santos João Manuel, Quixabeira Dafne C A, Havunen Riikka, Sorsa Suvi, Zheng Vincent, Salo Tuula, Bäck Leif, Aro Katri, Tulokas Sanni, Loimu Venla, Hemminki Akseli
Cancer Gene Therapy Group, Faculty of Medicine, University of Helsinki, Helsinki, Finland.
TILT Biotherapeutics Ltd, Helsinki, Finland.
Front Immunol. 2022 Mar 7;13:794251. doi: 10.3389/fimmu.2022.794251. eCollection 2022.
Immune checkpoint inhibitors (ICI) have provided significant improvement in clinical outcomes for some patients with solid tumors. However, for patients with head and neck cancer, the response rate to ICI monotherapy remains low, leading to the exploration of combinatorial treatment strategies. In this preclinical study, we use an oncolytic adenovirus (Ad5/3) encoding hTNFα and hIL-2 and non-replicate adenoviruses (Ad5) encoding mTNFα and mIL-2 with ICI to achieve superior tumor growth control and improved survival outcomes. The effect of Ad5/3-E2F-D24-hTNFa-IRES-hIL-2 was characterized through analyses of virus replication, transgene expression and lytic activity using head and neck cancer patient derived cell lines. Mouse models of ICI naïve and refractory oral cavity squamous cell carcinoma were established to evaluate the local and systemic anti-tumor immune response upon ICI treatment with or without the non-replicative adenovirus encoding mTNFα and mIL-2. We delineated the mechanism of action by measuring the metabolic activity and effector function of CD3 tumor infiltrating lymphocytes (TIL) and transcriptomic profile of the CD45 tumor immune compartment. Ad5/3-E2F-D24-hTNFa-IRES-hIL-2 demonstrated robust replicative capability across all head and neck cell lines screened through potent lytic activity, E1a and transgene expression. , in both ICI naïve and refractory models, we observed improvement to tumor growth control and long-term survival when combining anti-PD-1 or anti-PD-L1 with the non-replicative adenovirus encoding mTNFα and mIL-2 compared to monotherapies. This observation was verified by striking CD3 TIL derived mGranzyme b and interferon gamma production complemented by increased T cell bioenergetics. Notably, interrogation of the tumor immune transcriptome revealed the upregulation of a gene signature distinctive of tertiary lymphoid structure formation upon treatment of murine anti-PD-L1 refractory tumors with non-replicative adenovirus encoding mTNFα and mIL-2. In addition, we detected an increase in anti-tumor antibody production and expansion of the memory T cell compartment in the secondary lymphoid organs. In summary, a non-replicative adenovirus encoding mTNFα and mIL-2 potentiates ICI therapy, demonstrated by improved tumor growth control and survival in head and neck tumor-bearing mice. Moreover, the data reveals a potential approach for inducing tertiary lymphoid structure formation. Altogether our results support the clinical potential of combining this adenovirotherapy with anti-PD-1 or anti-PD-L1.
免疫检查点抑制剂（ICI）已使一些实体瘤患者的临床疗效得到显著改善。然而，对于头颈癌患者，ICI单药治疗的缓解率仍然较低，这促使人们探索联合治疗策略。在这项临床前研究中，我们使用一种编码hTNFα和hIL-2的溶瘤腺病毒（Ad5/3）以及一种编码mTNFα和mIL-2的非复制性腺病毒（Ad5）与ICI联合使用，以实现更好的肿瘤生长控制并改善生存结果。通过使用头颈癌患者来源的细胞系分析病毒复制、转基因表达和裂解活性，对Ad5/3-E2F-D24-hTNFa-IRES-hIL-2的效果进行了表征。建立了未接受过ICI治疗和对ICI耐药的口腔鳞状细胞癌小鼠模型，以评估在使用或不使用编码mTNFα和mIL-2的非复制性腺病毒的情况下，ICI治疗后的局部和全身抗肿瘤免疫反应。我们通过测量CD3肿瘤浸润淋巴细胞（TIL）的代谢活性和效应功能以及CD45肿瘤免疫区室的转录组图谱来阐明其作用机制。Ad5/3-E2F-D24-hTNFa-IRES-hIL-2通过强大的裂解活性、E1a和转基因表达，在所有筛选的头颈细胞系中都表现出强大的复制能力。在未接受过ICI治疗和耐药的模型中，与单药治疗相比，我们观察到将抗PD-1或抗PD-L1与编码mTNFα和mIL-2的非复制性腺病毒联合使用时，肿瘤生长控制得到改善，长期生存率提高。这一观察结果通过显著的CD3 TIL衍生的m颗粒酶b和干扰素γ产生以及T细胞生物能量学增加得到证实。值得注意的是，对肿瘤免疫转录组的研究发现，在用编码mTNFα和mIL-2的非复制性腺病毒治疗小鼠抗PD-L1耐药肿瘤后，一种独特的三级淋巴结构形成基因特征上调。此外，我们在二级淋巴器官中检测到抗肿瘤抗体产生增加和记忆T细胞区室扩大。总之，一种编码mTNFα和mIL-2的非复制性腺病毒增强了ICI治疗效果，这在头颈荷瘤小鼠的肿瘤生长控制和生存改善中得到了证明。此外，数据揭示了一种诱导三级淋巴结构形成的潜在方法。我们的结果共同支持了这种腺病毒疗法与抗PD-1或抗PD-L1联合使用的临床潜力。
