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PPA1 promotes adipogenesis by regulating the stability of C/EBPs

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Abstract

Adipogenesis significantly contributes to healthy adipose tissue expansion in obesity. Increasing adipocyte number or function to alleviate adipose tissue overload could serve as a therapeutic strategy for both lipodystrophy and obesity-related metabolic syndrome. Inorganic pyrophosphatase (PPA1) is an enzyme that catalyzes the hydrolysis of pyrophosphate (PPi) and is involved in many biochemical reactions, but its function in adipose tissue has not been studied previously. In this study, we demonstrated that adipose-specific PPA1 knockout (PPA1AKO) mice showed lipodystrophy and spontaneously developed hepatic steatosis and severe insulin resistance under normal chow diet feeding. PPA1 deficiency suppressed the differentiation of primary adipocyte precursors and 3T3-L1 cells. Notably, PPA1 overexpression can restore inhibited adipogenesis in preadipocytes isolated from db/db mice and type 2 diabetes patients. Mechanistic studies have revealed that PPA1 acts as a positive regulator of early adipocyte differentiation by promoting CCAAT/enhancer-binding proteinβ and δ (C/EBPβ and δ) protein stability. Moreover, the function of PPA1 in adipogenesis is independent of its PPi catalytic activity. Collectively, our in vivo and in vitro findings demonstrated that PPA1 is a novel critical upstream regulator of adipogenesis, controlling adipose tissue development and whole-body metabolic homeostasis.

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Fig. 1: PPA1 ablation results in severe lipodystrophy in vivo.
Fig. 2: Adipose PPA1 ablation in mice resulted in liver steatosis and insulin resistance.
Fig. 3: PPA1AKO mice are resistant to HFD-induced obesity but developed more severe metabolic disorders.
Fig. 4: PPA1 is required for adipocyte differentiation in vitro.
Fig. 5: PPA1 may promote adipocyte differentiation independent of its enzymatic activity.
Fig. 6: Loss of PPA1 inhibited adipogenesis through increasing C/EBPβ and C/EBPδ proteasome-dependent degradation.

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Data availability

The raw RNA-sequencing data reported in this paper have been deposited in the Figshare database (https://doi.org/10.6084/m9.figshare.25679760). All datasets analyzed in the study are available from the corresponding authors on reasonable request. Raw western blots are available in the Supplemental file.

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Acknowledgements

We thank all the lab members for insightful discussion during this study.

Funding

This work was supported by National Key R&D Program of China (2022YFA0806102), National Natural Science Foundation of China (82270874, 82370855, 81970709).

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Authors

Contributions

YY and HYL conceived the project and wrote the manuscript. YYW and YS designed and performed most experiments. YQS performed in vivo studies and analyzed the data. YH and JTW performed in vitro experiments. XH contributed to the discussion and critically reviewed and edited the manuscript. All authors revised and approved the final version of the manuscript. XH is the guarantor of this work and, as such, had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.

Corresponding authors

Correspondence to Haiyan Lin, Ye Yin or Xiao Han.

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The authors declare no competing interests.

Ethics

All animal experiments were approved and complied with the guidelines of the Institutional Animal Care and Use Committee of the Nanjing Medical University, China (Permit Number: IACUC-NJMU 14030178). Human specimens were supplied by the Southeast University Affiliated Zhongda Hospital and written informed consent was obtained from the patients (Permit number: 2020ZDSYLL144-P01 by the Independent Ethics Committee of Southeast University Affiliated Zhongda Hospital).

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Wu, Y., Sun, Y., Song, Y. et al. PPA1 promotes adipogenesis by regulating the stability of C/EBPs. Cell Death Differ (2024). https://doi.org/10.1038/s41418-024-01309-2

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