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Published on: November 28, 2019
Context-Dependent Modulation of Macrophage Plasticity by Cordycepin Drives Tumor Regression in Melanoma
Jihae Lim1, Donglan Piao1, Jio Kang1
1College of Pharmacy and Graduate School of Pharmaceutical Sciences, Ewha Womans University, 03760 Seoul, Republic of Korea.
Background And Aim:
Cordycepin (CDC), an adenosine (ADO) analog from Cordyceps mushrooms, exhibits potent anti-tumor and immunomodulatory activities. However, the precise mechanisms governing its effects on macrophage plasticity remain poorly understood. This study aimed to isolate CDC from a high-yielding Cordyceps cultivar, validate its systemic anti-tumor efficacy, and elucidate the mechanobiological cues regulating CDC-driven macrophage functions under varying cell-density states.
Experimental Procedure:
CDC (>98% purity) was isolated and structurally characterized via nuclear magnetic resonance (NMR) and high-resolution electrospray ionization mass spectrometry (HR-ESI-MS). Primary bone marrow-derived macrophages and RAW264.7 cells, cultured under sparse (~30%) or confluent (100%) conditions, were treated with CDC or ADO. We evaluated cell viability, pro-inflammatory cytokine expression, and Nuclear Factor kappa B (NF-κB) p65 signaling, phagocytosis, and migration. The therapeutic potential of CDC-primed macrophages was assessed via in vitro melanoma co-culture and in vivo intratumoral adoptive transfer in B16F10 tumor-bearing mice.
Key Results:
Systemic administration of CDC significantly inhibited melanoma growth in vivo, promoting apoptosis, enhancing macrophage infiltration. We discovered that CDC regulates macrophage functions via a density-dependent "switch": while CDC induced cytotoxicity in sparse cultures, it significantly augmented M1-like cytokine production in confluent states without compromising viability. This density-dependent activation was mediated by the A2A adenosine receptor (A2AR), triggering the Akt-NF-κB p65 signaling axis. Furthermore, CDC upregulated migration- and phagocytosis-associated genes, enhancing tumor cell clearance. Notably, intratumoral injection of CDC-primed macrophages markedly reduced tumor volume and size in vivo.
Conclusions And Implications:
CDC modulates macrophage activation through a unique mechanobiological switch. Under high-density conditions-mimicking the dense tumor microenvironment-CDC enhances A2AR-mediated NF-κB activation, boosting macrophage activation, recruitment, and phagocytosis to facilitate tumor regression. These findings establish CDC as a context-dependent immunomodulator capable of reprogramming macrophages toward a tumoricidal phenotype.
