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AF9 targets acetyl-modified STAT6 to diminish purine metabolism and accelerate cell apoptosis during metastasis

Cell Death & Differentiation volume 30, pages 1695–1709 (2023)Cite this article

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Abstract

Cell migration and invasion are two important steps for tumour metastasis, and involved the behaviors including metabolism remodeling and anti-apoptosis. However, it’s still elusive that cancer cells how to antagonize apoptosis during tumour metastasis. In this study, we observed that super elongation complex (SEC) subunit AF9 depletion exacerbated cell migration and invasion but reduced the apoptosis during invasive migration. Mechanically, AF9 targeted acetyl (Ac)-STAT6 at lysine (K) 284 and blocked STAT6 transactivation on the promoter of such genes involved in regulating purine metabolism and metastasis, in turn induced apoptosis of suspended cells. Of note, AcSTAT6-K284 was not induced by IL4 signaling but decreased by limited nutrition which triggered SIRT6 to remove acetyl group at STAT6-K284. The functional experiments proved that AcSTAT6-K284 attenuated cell migration and invasion depending on AF9 expression level. Animal metastatic study further confirmed the AF9/AcSTAT6-K284 axis existed and blocked kidney renal clear cell carcinoma (KIRC) metastasis. In clinical, both AF9 expression and AcSTAT6-K284 were decreased accompanied by the advanced tumour grade and positively correlated with KIRC patients’ survival. Conclusively, we explored an inhibitory axis which not only suppressed tumour metastasis but also could be utilized for drug development to hamper KIRC metastasis.

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Fig. 1: Low expression of AF9 displays a worse prognosis and sustains more aggressive cell migration and invasion.
Fig. 2: AF9 associates with STAT6 and regulates its transactivation during cell migration and invasion.
Fig. 3: Acetyl-modified STAT6 at lysine 284 recruits AF9.
Fig. 4: SIRT6 deacetylates STAT6 acetylation in vitro and in vivo upon limited nutrition stress.
Fig. 5: AF9/AcSTAT6-K284 axis attenuates the expression of genes involved in purine metabolism and metastasis.
Fig. 6: AF9 level and the state of STAT6 acetylation at K284 determine whether STAT6 activates purine biosynthesis to promote anti-apoptosis during invasion.
Fig. 7: AF9/AcSTAT6-K284 axis suppressed metastasis in vivo and prolongs the lifespan of mice and human patients with KIRC.

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Data availability

We provided data accession codes for RNA-sequencing original data in the NCBI BioProject as follows: PRJNA874431. Original western blot was uploaded as Supplementary Material.

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Acknowledgements

The authors thank the CAS Shanghai Institute of Nutrition and Health (SINH) Molecular and metabolism core facility for the metabolite determination and protein synthesis service. This study was supported in part by NSFC grants No. 92053113 and 81570607, National Natural Science Foundation for Young Scholars of China (No. 81902566 and 82103511), and Shanghai Jiaotong University Medical-Engineering Cross Research Fund (No. YG2019QNA53).

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  1. These authors contributed equally: Jialiang Shao, Tiezhu Shi.

Authors and Affiliations

  1. Department of Urology, Shanghai General Hospital, Shanghai Jiaotong University, Shanghai, 200080, China

    Jialiang Shao & Xiang Wang

  2. School of Life Sciences, Guangzhou University, Guangzhou, 510006, China

    Tiezhu Shi, Lianheng Chen, Hua Yu & Xiongjun Wang

  3. Department of Urology, The Affiliated Wuxi No.2 Hospital of Nanjing Medical University, Jiangsu, 214002, China

    Ninghan Feng

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Contributions

SJ, ST, DY and YH designed, performed, and analyzed experiments, prepared the figures. CL provided technical help. WXJ and FN conceived this study, wrote the manuscript and YH rewrote the revised manuscript. WXJ and WX designed and analyzed experiments and wrote the manuscript.

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Correspondence to Hua Yu, Ninghan Feng or Xiongjun Wang.

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All KIRC samples were conducted in accordance with the requirements of Shanghai First People’s Hospital, Shanghai Jiaotong University. The use of clinical samples was approved by the institutional review board of the hospital with the ethical number 2021KSQ367.

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Shao, J., Shi, T., Chen, L. et al. AF9 targets acetyl-modified STAT6 to diminish purine metabolism and accelerate cell apoptosis during metastasis. Cell Death Differ 30, 1695–1709 (2023). https://doi.org/10.1038/s41418-023-01172-7

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