Long noncoding RNA AFAP1-AS1 aggravates immunotherapy resistance in NSCLC by enhancing JAK2 and TYK2 translation

  • J Adv Res. 2026 Apr 24:S2090-1232(26)00358-9. doi: 10.1016/j.jare.2026.04.055.
Yijie Zhang  1 ,  Jiawei Ouyang  1 ,  Qijia Yan  2 ,  Junshang Ge  3 ,  Pan Wu  3 ,  Dan Wang  3 ,  Lvyuan Li  3 ,  Jiarong Zhang  3 ,  Yu Zhong  3 ,  Liting Yang  2 ,  Lei Shi  4 ,  Fang Xiong  5 ,  Ming Zhou  1 ,  Bo Xiang  1 ,  Zhaoyang Zeng  6 ,  Pan Chen  7 ,  Wei Xiong  8 ,  Zhaojian Gong  9
Affiliations
  • 1. NHC Key Laboratory of Carcinogenesis and Hunan Key Laboratory of Cancer Metabolism, Hunan Cancer Hospital and the Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha 410078, Hunan, China; Key Laboratory of Carcinogenesis and Cancer Invasion of the Chinese Ministry of Education, Cancer Research Institute and Xiangya School of Basic Medicine Sciences, Central South University, Changsha 410078, Hunan, China.
  • 2. Key Laboratory of Carcinogenesis and Cancer Invasion of the Chinese Ministry of Education, Cancer Research Institute and Xiangya School of Basic Medicine Sciences, Central South University, Changsha 410078, Hunan, China; Department of Pathology, Xiangya Hospital, Central South University, Changsha 410078, Hunan, China.
  • 3. Key Laboratory of Carcinogenesis and Cancer Invasion of the Chinese Ministry of Education, Cancer Research Institute and Xiangya School of Basic Medicine Sciences, Central South University, Changsha 410078, Hunan, China.
  • 4. Key Laboratory of Carcinogenesis and Cancer Invasion of the Chinese Ministry of Education, Cancer Research Institute and Xiangya School of Basic Medicine Sciences, Central South University, Changsha 410078, Hunan, China; Department of Oral and Maxillofacial Surgery, The Second Xiangya Hospital, Central South University, Changsha 410011, Hunan, China.
  • 5. Department of Pathology, Xiangya Hospital, Central South University, Changsha 410078, Hunan, China.
  • 6. NHC Key Laboratory of Carcinogenesis and Hunan Key Laboratory of Cancer Metabolism, Hunan Cancer Hospital and the Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha 410078, Hunan, China; Key Laboratory of Carcinogenesis and Cancer Invasion of the Chinese Ministry of Education, Cancer Research Institute and Xiangya School of Basic Medicine Sciences, Central South University, Changsha 410078, Hunan, China; FuRong Laboratory, Changsha 410078, Hunan, China.
  • 7. NHC Key Laboratory of Carcinogenesis and Hunan Key Laboratory of Cancer Metabolism, Hunan Cancer Hospital and the Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha 410078, Hunan, China; FuRong Laboratory, Changsha 410078, Hunan, China. Electronic address: [email protected].
  • 8. NHC Key Laboratory of Carcinogenesis and Hunan Key Laboratory of Cancer Metabolism, Hunan Cancer Hospital and the Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha 410078, Hunan, China; Key Laboratory of Carcinogenesis and Cancer Invasion of the Chinese Ministry of Education, Cancer Research Institute and Xiangya School of Basic Medicine Sciences, Central South University, Changsha 410078, Hunan, China; FuRong Laboratory, Changsha 410078, Hunan, China. Electronic address: [email protected].
  • 9. Department of Oral and Maxillofacial Surgery, The Second Xiangya Hospital, Central South University, Changsha 410011, Hunan, China. Electronic address: [email protected].
PMID: 42035912 DOI: 10.1016/j.jare.2026.04.055
Abstract

Background: Lung Cancer is the most common and deadliest malignancy worldwide, with about 85% of cases being non-small cell lung Cancer (NSCLC). Although targeting PD-1/PD-L1 with immune checkpoint inhibitors has revolutionized NSCLC treatment, a substantial proportion of patients still experience intrinsic or acquired resistance. Understanding the mechanisms of immune evasion and resistance and identifying novel therapeutic targets remain critical challenges.

Methods: Using RNA Sequencing, qRT-PCR, and Western blotting, we discovered and validated that the long non-coding RNA (lncRNA) AFAP1-AS1 activates the interferon signaling pathways and upregulates PD-L1 and IDO1 expression in NSCLC cells. Through mass spectrometry analysis combined with Western blotting and sucrose density gradient separation of polyribosomes, it was found that AFAP1-AS1 binds to the translation initiation factor EIF4A1 and promotes the translation of the tyrosine kinases JAK2 and Tyk2. In vitro, CD8+ T cell killing ability after coculture was assessed by flow cytometry. In a mouse subcutaneous tumor model, the immunotherapeutic efficacy of JAK2/Tyk2 inhibitors and a PD-1 monoclonal antibody against AFAP1-AS1-overexpressing tumors was evaluated.

Results: In this study, we demonstrate that elevated AFAP1-AS1 expression correlates with poor response to anti-PD-1 therapy and is inversely associated with CD8+ T-cell infiltration. Mechanistically, AFAP1-AS1 interacts with EIF4A1 to enhance the translation of JAK2 and Tyk2, thereby promoting STAT1 phosphorylation and nuclear translocation, which activate interferon signaling pathways. This cascade upregulates the immunosuppressive checkpoints PD-L1 and IDO1, ultimately suppressing CD8+ T cell cytotoxicity and promoting T cell Apoptosis. Importantly, in preclinical models, pharmacological inhibition of JAK2 and Tyk2 synergistically enhances the efficacy of anti-PD-1 immunotherapy.

Conclusions: Our findings reveal a novel mechanism through which AFAP1-AS1 promotes immunotherapy resistance in NSCLC by binding to the translation initiation factor EIF4A1 to enhance JAK2 and Tyk2 translation. This highlights AFAP1-AS1 and its downstream tyrosine kinases, JAK2/Tyk2, as potential therapeutic targets to improve immunotherapy efficacy in NSCLC.

Keywords
AFAP1-AS1; Immunosuppressive checkpoint; Immunotherapy resistance; Long non-coding RNA(lncRNAs); Non-small cell lung cancer; Tyrosine kinase.
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