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DARPP-32 is a novel regulator of M2-macrophage polarization: Sustains STAT6 phosphorylation by directly binding and inhibiting tyrosine phosphatase SHP-1

Oncogene volume 44, pages 4639–4656 (2025)Cite this article

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Abstract

Identifying novel factors driving M2-macrophage polarization may help understand immune modulation in cancers. We identified DARPP-32, a protein phosphatase inhibitor, to be significantly upregulated in FBXW7 knock out THP-1 spontaneously polarized to M2 macrophages. DARPP-32 levels remarkably increased in macrophages treated with different M2-inducers while it copiously decreased upon treatment with M1 inducer Lipopolysaccharide/IFN-γ. Intriguingly, mere DARPP-32 over-expression robustly promoted M2-macrophage polarization in different macrophage cell lines and murine primary bone marrow derived macrophages, even without any external stimulation. Conversely, DARPP-32 depletion hampered the IL-4 induced M2-polarization. Strikingly, DARPP-32 overexpression also dampened the classical M1 activation profile of macrophages in response to LPS, skewing their function towards more anti-inflammatory M2-like phenotype. Mechanistically, Thr34-phosphorylated DARPP-32 positively regulates JAK1-STAT6 signalling by directly binding and inhibiting protein tyrosine phosphatase SHP-1, a key regulator of immune cell signalling and macrophage activation. In line with increased DARPP-32 levels in M2-polarized macrophages, which are often tumor-associated macrophages, we also observed markedly increased DARPP-32 levels in tumor-infiltrating macrophages within primary TNBC tumors. We also observed an increase in DARPP-32 levels in macrophages co-cultured with tumor cells. This is first comprehensive study that demonstrate a novel function of DARPP-32 as a driver/regulator of macrophage polarization towards anti-inflammatory M2-phenotype.

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Fig. 1: Proteomic identification of DARPP-32 from M2 macrophages as a protein induced in differentiated macrophages.
Fig. 2: DARPP-32 is exclusively induced in M2-polarized macrophages.
Fig. 3: DARPP-32 overexpression skews macrophages towards anti-inflammatory M2 phenotype.
Fig. 4: DARPP-32 over-expression drives M2 and inhibits pro-inflammatory functions of M1 Macrophage.
Fig. 5: DARPP-32 sustains JAK1-STAT6 phosphorylation by directly binding and inhibiting protein tyrosine phosphatase SHP-1.
Fig. 6: DARPP-32 by skewing Mϕ towards M2-polarized state promotes immune-suppressive tumor microenvironment and inhibits T-cells activation.
Fig. 7: DARPP-32 expression correlates with higher Tumor Associated Macrophages (TAMs) infiltration.

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Data availability

Data and materials used in this study are available on request to corresponding author.

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Acknowledgements

Senior Research Fellowship (SRF) from the Department of Biotechnology (DBT), India to Sangita & Shivkant is acknowledged. Funding from Department of Biotechnology (DBT)-BioCARe to Swati Srivastava is also acknowledged. Fellowship from University Grants Commission (UGC) to Agniv Shome is acknowledged. We also thank Mr. A. L. Vishwakarma and Ms. Rima Ray Sarkar from FACS facility (Sophisticated and Analytical Instrument Facility, SAIF) and confocal microscopy unit, CSIR-CDRI respectively. CSIR-CDRI Manuscript communication number for this article is 11050.

Funding

This study was supported by funds from the Lady Tata Memorial Trust (LTMT), Mumbai (GAP0239) to AKT.

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Authors and Affiliations

  1. Division of Cancer Biology, CSIR-Central Drug Research Institute, Sector-10, Jankipuram Extension, Sitapur Road, Lucknow, 226031, UP, India

    Sangita Chowdhury, Arppita Sethi, Swati Srivastava, Vishal Upadhyay, Shivkant Mishra, Agniv Shome, Anil Kumar Singh, Shyam Singh & Arun Kumar Trivedi

  2. Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201002, India

    Arppita Sethi, Vishal Upadhyay, Agniv Shome, Anil Kumar Singh & Arun Kumar Trivedi

  3. King George’s Medical University, KGMU, Lucknow, 226003, India

    Madan Lal Brahma Bhatt & Anand Mishra

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Contributions

Experimental design: SC and AKT; Data generation: SC, AS, SS, SKM, AShome, SSingh; Data Analysis: SC, SS, VU, AKS, and AKT; Clinical Sample and data analysis: MLB, AM and AKT; Manuscript preparation: SC, SS and AKT; Fund acquisition and Project monitoring: AKT.

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Correspondence to Arun Kumar Trivedi.

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Chowdhury, S., Sethi, A., Srivastava, S. et al. DARPP-32 is a novel regulator of M2-macrophage polarization: Sustains STAT6 phosphorylation by directly binding and inhibiting tyrosine phosphatase SHP-1. Oncogene 44, 4639–4656 (2025). https://doi.org/10.1038/s41388-025-03610-x

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