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Hiroyuki Tsuchiya¹, Takehiko Hanaki^2,3, Mayu Obora¹

¹Division of Regenerative Medicine and Therapeutics, Department of Genomic Medicine and Regenerative Therapy, Faculty of Medicine, Tottori University, 86 Nishi-cho, Yonago, Tottori, 683-8503, Japan, tottori-u.ac.jp.

Journal of Immunology Research

|April 21, 2026

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This summary is machine-generated.

Researchers identified compounds that reprogram tumor-associated macrophages (TAMs) from a pro-tumor M2 state to an anti-tumor M1 state. This reprogramming enhances anti-PD-1 therapy efficacy by increasing CD8+ T cell infiltration in a mouse model.

Area of Science:

Immunology
Oncology
Pharmacology

Background:

The tumor microenvironment (TME) promotes immune evasion and immunotherapy resistance, partly by recruiting pro-tumor M2-like macrophages.
Tumor-associated macrophages (TAMs) often exhibit an M2-like phenotype within the TME, suppressing anti-tumor immunity.

Purpose of the Study:

To identify compounds (TAM activators) that can reprogram M2-like TAMs to an anti-tumor M1-like phenotype.
To establish a novel screening platform for identifying TAM activators using cancer cell-macrophage heterospheroids.

Main Methods:

Co-culture of liver cancer cells with macrophages to form heterospheroids, mimicking the TME.
Flow cytometry and fluorescence microscopy to assess macrophage phenotype and infiltration.
High-content imaging screen to identify TAM-activating compounds.

Keywords:

heterospheroid high-content imaging analysis macrophage reprogramming screening tumor-associated macrophage

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Main Results:

Heterospheroids suppressed M1 marker induction in macrophages compared to homospheroids.
Two compounds, alprostadil and HX531, were identified as TAM activators.
Alprostadil promoted M1-like TAM polarization, increased CD8+ T cell infiltration, and enhanced anti-PD-1 therapy efficacy in mice.

Conclusions:

Heterospheroid culture effectively recapitulates the immunosuppressive TME and can be used to screen for TAM activators.
Alprostadil shows potential as a novel cancer immunotherapeutic agent by reprogramming TAMs and enhancing T cell responses.
This approach enables the development of new cancer immunotherapies targeting TAMs.