Twist1-induced suppression of oncogene-induced senescence in non-small cell lung cancer requires the transactivation domain of Twist1

Audrey Lafargue; Hailun Wang; Sivarajan T Chettiar; Rajendra P Gajula; Amol C Shetty; Yang Song; Brian W Simons; Muhammad Ajmal Khan; Triet Nguyen; Hwai-Wei Tseng; Jinhee Chang; Danielle N Waters; Aaron Chan; Christine Lam; Francesca A Carrieri; Caleb Smack; Nick Connis; Dipanwita Dutta Chowdhury; Katriana Nugent; Ismaeel Siddiqui; Kekoa Taparra; Mohammad Rezaee; Natasha Zachara; Zachary S Morris; Christopher McFarland; Sarki Abba Abdulkadir; Christine L Hann; Phuoc T Tran

doi:10.1016/j.neo.2025.101179

Twist1-induced suppression of oncogene-induced senescence in non-small cell lung cancer requires the transactivation domain of Twist1

Neoplasia. 2025 Aug:66:101179. doi: 10.1016/j.neo.2025.101179. Epub 2025 May 22.

Authors

Audrey Lafargue¹, Hailun Wang², Sivarajan T Chettiar³, Rajendra P Gajula³, Amol C Shetty⁴, Yang Song⁴, Brian W Simons⁵, Muhammad Ajmal Khan⁶, Triet Nguyen⁷, Hwai-Wei Tseng⁶, Jinhee Chang⁷, Danielle N Waters⁶, Aaron Chan⁶, Christine Lam³, Francesca A Carrieri³, Caleb Smack³, Nick Connis⁸, Dipanwita Dutta Chowdhury⁶, Katriana Nugent³, Ismaeel Siddiqui³, Kekoa Taparra³, Mohammad Rezaee³, Natasha Zachara⁹, Zachary S Morris¹⁰, Christopher McFarland¹¹, Sarki Abba Abdulkadir¹², Christine L Hann⁸, Phuoc T Tran¹³

Affiliations

¹ Department of Radiation Oncology and Molecular Radiation Sciences, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, School of Medicine, Baltimore, MD, USA; Department of Radiation Oncology, Division of Translational Radiation Sciences, University of Maryland Baltimore, School of Medicine, Baltimore, MD, USA. Electronic address: alafargue@som.umaryland.edu.
² Department of Radiation Oncology and Molecular Radiation Sciences, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, School of Medicine, Baltimore, MD, USA; GenoImmune Therapeutics, Wuhan, China.
³ Department of Radiation Oncology and Molecular Radiation Sciences, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, School of Medicine, Baltimore, MD, USA.
⁴ Department of Radiation Oncology, Division of Translational Radiation Sciences, University of Maryland Baltimore, School of Medicine, Baltimore, MD, USA; Institute of Genome Sciences, University of Maryland Baltimore, School of Medicine, Baltimore, MD, USA.
⁵ Department of Urology, James Buchanan Urological Institute, Johns Hopkins University, School of Medicine, Baltimore, MD, USA.
⁶ Department of Radiation Oncology, Division of Translational Radiation Sciences, University of Maryland Baltimore, School of Medicine, Baltimore, MD, USA.
⁷ Department of Radiation Oncology and Molecular Radiation Sciences, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, School of Medicine, Baltimore, MD, USA; Department of Radiation Oncology, Division of Translational Radiation Sciences, University of Maryland Baltimore, School of Medicine, Baltimore, MD, USA.
⁸ Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, School of Medicine, Baltimore, MD, USA.
⁹ Department of Biological Chemistry, Johns Hopkins University, School of Medicine, Baltimore, MD, USA.
¹⁰ Department of Human Oncology, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
¹¹ Department of Genetics and Genome Sciences, Case Western Reserve University, School of Medicine, Cleveland, OH, USA.
¹² Department of Urology, The Robert H. Lurie Comprehensive Cancer Center, and Department of Pathology, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.
¹³ Department of Radiation Oncology and Molecular Radiation Sciences, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, School of Medicine, Baltimore, MD, USA; Department of Radiation Oncology, Division of Translational Radiation Sciences, University of Maryland Baltimore, School of Medicine, Baltimore, MD, USA; Department of Urology, James Buchanan Urological Institute, Johns Hopkins University, School of Medicine, Baltimore, MD, USA; Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, School of Medicine, Baltimore, MD, USA. Electronic address: phuoc.tran@umm.edu.

Abstract

Non-small cell lung carcinoma (NSCLC) is a major cause of cancer mortality. High expression of the epithelial-to-mesenchymal transition transcription factor (EMT-TF) Twist1 is strongly associated with metastatic cancers and with treatment resistance. Twist1 can also upregulate O-GlcNAcylation to suppress fail-safe programs such as Kras^G12D oncogene-induced senescence (OIS) that accelerates NSCLC tumorigenesis. We wanted to decipher the critical domains and transcriptional targets required for Twist1 acceleration of lung tumorigenicity. We created a novel genetically-engineered mouse model for autochthonous lung cancer through lung epithelial expression of Kras^G12D oncogene (CR) concomitantly with Twist1^wt (CRT) or a Twist1^F191G transactivation-deficient mutant (CRF191G). Compared to CR and CRF191G, CRT mice had shorter tumor-free survival and more aggressive tumors histologically. CRT lung tumors also showed higher proliferation and lower cell-cycle arrest suggesting that the Twist1 transactivation-domain is important for OIS suppression. Supporting these data, we observed in non-cancer human bronchial epithelial cells (HBECs) that the co-expression of human TWIST1^wt enhanced tumorigenic/invasive programs and could suppress HRas^G12V-induced senescence while co-expressing TWIST1^F187G transactivation-deficient mutant could not. TWIST1^wt co-expression with HRas^G12V in HBECs differentially modulated MYC downstream transcriptional programs. Finally, OIS induction in HBECHRas^G12V-TWIST1^wt was rescued by O-GlcNAcylation inhibition or by treatment with a novel MYC inhibitor MYCi975 or by MYC knockdown. Altogether, these results indicate that the Twist1 transactivation domain is required for Twist1-dependent acceleration of lung tumorigenesis via MYC and nominate MYCi975 as a means to activate latent OIS programs. MYC targeting strategies could limit pro-tumorigenic programs and serve as a therapeutic for TWIST1-overexpressing NSCLCs.

Keywords: GEMM; MYC; Non-Small Cell Lung Cancer; O-GlcNAcylation; Oncogene-Induced Senescence; TWIST1.

MeSH terms

Animals
Carcinoma, Non-Small-Cell Lung* / genetics
Carcinoma, Non-Small-Cell Lung* / metabolism
Carcinoma, Non-Small-Cell Lung* / pathology
Cell Line, Tumor
Cell Proliferation
Cellular Senescence* / genetics
Disease Models, Animal
Epithelial-Mesenchymal Transition / genetics
Gene Expression Regulation, Neoplastic
Humans
Lung Neoplasms* / genetics
Lung Neoplasms* / metabolism
Lung Neoplasms* / pathology
Mice
Nuclear Proteins* / chemistry
Nuclear Proteins* / genetics
Nuclear Proteins* / metabolism
Oncogenes*
Proto-Oncogene Proteins p21(ras) / genetics
Transcriptional Activation*
Twist-Related Protein 1* / chemistry
Twist-Related Protein 1* / genetics
Twist-Related Protein 1* / metabolism

Substances

Twist-Related Protein 1
Nuclear Proteins
Twist1 protein, mouse
TWIST1 protein, human
Proto-Oncogene Proteins p21(ras)