Symbiotic exclusivity between CLOCK and TFPI2 drives stemness and immunosuppression in glioblastoma models
Fei Zhou, Lizhi Pang, Yang Liu, Fatima Khan, Peiwen Chen
Fei Zhou, Lizhi Pang, Yang Liu, Fatima Khan, Peiwen Chen
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Research Article Immunology Oncology

Symbiotic exclusivity between CLOCK and TFPI2 drives stemness and immunosuppression in glioblastoma models

Abstract

Glioblastoma (GBM) is a highly aggressive brain tumor characterized by extensive crosstalk between glioblastoma stem cells (GSCs) and immunosuppressive microglia, with our previous work identifying CLOCK and TFPI2 as key regulators of this interaction. Here, we uncover a ‘symbiotic exclusivity’ pattern between CLOCK and TFPI2, showing that, despite mutually exclusive amplifications, they sustain symbiotic regulatory interactions in GBM. The CLOCK-BMAL1 complex transcriptionally upregulates TFPI2, while TFPI2-driven hypoxia inducible factor 1 α (HIF-1α) signaling activates nuclear factor k B (NF-kB) P65 to upregulate the CLOCK-BMAL1 complex, creating a positive feedback loop to promote stemness, immunosuppression, and tumor progression. Disrupting the CLOCK-TFPI2 interplay through dual inhibition of their downstream effectors reduces GSC stemness and immunosuppressive microglia, activates antitumor immunity, and synergizes with anti-PD1 therapy to achieve complete tumor regression in 50%–62.5% of tumor-bearing mice. This study uncovers a promising therapeutic strategy for a broader subset of patients with GBM with high expression of either CLOCK or TFPI2, and provides a framework for identifying ‘symbiotic exclusivity’ genes in cancer.

Authors

Fei Zhou, Lizhi Pang, Yang Liu, Fatima Khan, Peiwen Chen

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Figure 6

TFPI2-induced GSC self renewal is regulated by the HIF-1α–NF-κB signaling axis.

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TFPI2-induced GSC self renewal is regulated by the HIF-1α–NF-κB signalin...
(A and B) Immunoblots for CD133 and SOX2 in cell lysates of GSC17 (A) and GSC23 (B) harboring control or TFPI2 overexpression (OE) and treated with or without HIF-1α inhibitor ACF (2 μM). (C and D) Immunoblots for CD133 and SOX2 in cell lysates of control and TFPI2-OE GSC17 (C) and GSC23 (D) treated with or without P65 inhibitor SC75741 (5 μM). (E) In vitro limiting dilution assays in control and TFPI2-OE GSC17 treated with or without ACF (2 μM) or SC75741 (5 μM). (F–H) Representative images (F) and quantification of relative tumorsphere number (G) and size (H) of control and TFPI2-OE GSC17 treated with or without ACF or SC75741. Scale bar: 200 μm. n = 8. (I) In vitro limiting dilution assays in control and TFPI2-OE GSC23 treated with or without ACF (2 μM) or SC75741 (5 μM). (J–L) Representative images (J) and quantification of relative tumorsphere number (K) and size (L) of control and TFPI2-OE GSC23 treated with or without ACF or SC75741. Scale bar: 200 μm. n = 8. (M–P) Representative and quantification of proliferation in GSC17 (M and N) and GSC23 (O and P) harboring control or TFPI2 OE treated with or without ACF (2 μM) or SC75741 (5 μM). n = 3. Data from multiple replicates are presented as mean ± SD. **P < 0.01, ***P < 0.001, 1-way ANOVA test (G, H, K, L, N, and P), 2-way ANOVA test (E and I).