RASSF1A Promotes ATM Signaling and RASSF1A Methylation is a Synthetic Lethal Marker for ATR Inhibitors

Abstract

Aim: The mechanism of RASSF1A in DNA damage repair remains to be further clarified for applying to synthetic lethal strategy. Materials & methods: Eight esophageal cancer cell lines, 181 cases of esophageal dysplasia and 1066 cases of primary esophageal squamous cell carcinoma (ESCC) were employed. Methylation-specific PCR, the CRISPR/Cas9 technique, immunoprecipitation assay and a xenograft mouse model were used. Results:RASSF1A was methylated in 2.21% of esophageal dysplasia and 11.73% of ESCC. RASSF1A was also involved in DNA damage repair through activating Hippo signaling. Loss of RASSF1A expression sensitized esophageal cancer cell lines to ataxia telangiectasia mutated and rad3-related (ATR) inhibitor (VE-822) both in vitro and in vivo. Conclusion:RASSF1A methylation is a synthetic lethal marker for ATR inhibitors.

Tweetable abstract

RASSF1A was methylated in 11.73% of ESCC. RASSF1A is involved in DNA damage repair by activating Hippo signaling. Loss of RASSF1A expression sensitized esophageal cancer cells to the ATR inhibitor, suggesting that RASSF1A methylation is a synthetic lethal marker for ATR inhibitors.

Keywords::

• ATM signaling
• DNA methylation
• RASSF1A
• synthetic lethality
• VE-822

Supplementary data

To view the supplementary data that accompany this paper please visit the journal website at: www.tandfonline.com/doi/suppl/10.2217/epi-2023-0306

Author contributions

A Gao: investigation, methodology, project administration, formal analysis, writing – original draft. P Bai: methodology, validation. M Zhang: methodology, data curation. Y Yao: data curation. JG Herman: data interpretation, editing. Mingzhou Guo: conceptualization, supervision, writing – review and editing, funding acquisition.

Financial disclosure

This work was supported by grants from National Key Research and Development Program of China (2018YFA0208902, 2020YFC2002705), National Natural Science Foundation of China (82272632, 81672138) and Natural Science Foundation of Beijing Municipality (7171008). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

Competing interests disclosure

The authors have no competing interests or relevant affiliations with any organization or entity with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

Writing disclosure

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The study methodologies were in accordance with the principles of the Declaration of Helsinki and the procedure was approved by the institutional review board of the Chinese PLA General Hospital (IRB number: 20090701-015). Informed consents were obtained from all patients. All animal experimental investigations were approved by the Animal Ethics Committee of the Chinese PLA General Hospital (approval number: 2021-X17-07).

Data sharing statement

Data that supports the findings of this study are available from the corresponding author upon reasonable request.

Additional information

Funding

This work was supported by grants from National Key Research and Development Program of China (2018YFA0208902, 2020YFC2002705), National Natural Science Foundation of China (82272632, 81672138) and Natural Science Foundation of Beijing Municipality (7171008). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.