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Genome-Wide CRISPR Screen for Unveiling Radiosensitive and Radioresistant Genes
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Genome-wide CRISPR screening reveals ADCK3 as a key regulator in sensitizing endometrial carcinoma cells to MPA
Zijing Zhang1, Meng Zhang1, Jingyi Zhou2
1State Key Laboratory of Common Mechanism Research for Major Diseases & Department of Medical Genetics, Institute of Basic Medical Sciences & School of Basic Medicine, Chinese Academy of Medical Sciences & Peking Union Medical College, 100005, Beijing, China.
Background:
The effectiveness of conservative treatment of endometrial carcinoma (EC) with oral progesterone therapy, such as medroxyprogesterone acetate (MPA), can be blunted due to primary or acquired resistance, but the underlying mechanisms remain incompletely defined.
Methods:
Genome-wide CRISPR screening was performed to identify potential regulators in response to MPA in Ishikawa cells. Crystal violet staining, RT-qPCR, western blotting, ChIP-qPCR and luciferase assays were employed to elucidate the p53-AarF domain-containing kinase 3 (ADCK3) regulatory axis and its roles in sensitizing EC cells to MPA treatment.
Results:
ADCK3 is identified as a previously unrecognized regulator in response to MPA in EC cells. Loss of ADCK3 in EC cells markedly alleviated MPA-induced cell death. Mechanistically, loss of ADCK3 primarily suppresses MPA-mediated ferroptosis by abrogating arachidonate 15-lipoxygenase (ALOX15) transcriptional activation. Moreover, we validated ADCK3 as a direct downstream target of the tumor suppressor p53 in EC cells. By stimulating the p53-ADCK3 axis, the small-molecule compound Nutlin3A synergized with MPA to efficiently inhibit EC cell growth.
Conclusions:
Our findings reveal ADCK3 as a key regulator of EC cells in response to MPA and shed light on a potential strategy for conservative EC treatment by activating the p53-ADCK3 axis to sensitize MPA-mediated cell death.
Insights
AarF domain-containing kinase 3 (ADCK3) regulates endometrial carcinoma (EC) response to medroxyprogesterone acetate (MPA). Activating the p53-ADCK3 pathway enhances MPA efficacy, offering a new strategy for conservative EC treatment.
Area of Science:
- Gynecologic Oncology
- Molecular Biology
- Cancer Research
Background:
- Conservative treatment of endometrial carcinoma (EC) using oral progesterone like medroxyprogesterone acetate (MPA) faces challenges due to resistance.
- Mechanisms underlying MPA resistance in EC are not fully understood.
Purpose of the Study:
- To identify novel regulators of MPA response in EC cells.
- To investigate the role of these regulators in overcoming MPA resistance.
Main Methods:
- Genome-wide CRISPR screening in Ishikawa EC cells.
- Assays including crystal violet staining, RT-qPCR, western blotting, ChIP-qPCR, and luciferase assays.
- Elucidation of the p53-AarF domain-containing kinase 3 (ADCK3) regulatory axis.
Main Results:
- ADCK3 was identified as a novel regulator of MPA response in EC cells.
- Loss of ADCK3 reduced MPA-induced cell death, primarily by inhibiting ferroptosis via abrogating arachidonate 15-lipoxygenase (ALOX15) activation.
- ADCK3 is a direct downstream target of p53; the compound Nutlin3A combined with MPA inhibited EC cell growth by activating the p53-ADCK3 axis.
Conclusions:
- ADCK3 is a critical regulator of EC cell sensitivity to MPA.
- Activating the p53-ADCK3 axis presents a potential therapeutic strategy to enhance conservative MPA treatment for EC.
