Induced clustering of SHP2-depleted tumor cells in vascular islands restores sensitivity to MEK/ERK inhibition
Yuyi Wang, … , Michael Kruhlak, Giovanna Tosato
Yuyi Wang, … , Michael Kruhlak, Giovanna Tosato
Published March 25, 2025
Citation Information: J Clin Invest. 2025;135(10):e181609. https://doi.org/10.1172/JCI181609.
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Research Article Angiogenesis Oncology

Induced clustering of SHP2-depleted tumor cells in vascular islands restores sensitivity to MEK/ERK inhibition

Abstract

Allosteric inhibitors of the tyrosine phosphatase Src homology 2 domain–containing protein tyrosine phosphatase 2 (SHP2) hold therapeutic promise in cancers with overactive RAS/ERK signaling, but adaptive resistance to SHP2 inhibitors may limit benefits. Here, we utilized tumor cells that proliferate similarly with or without endogenous SHP2 to explore means to overcome this growth independence from SHP2. We found that SHP2 depletion profoundly altered the output of vascular regulators, cytokines, chemokines, and other factors from SHP2 growth-resistant cancer cells. Tumors derived from inoculation of SHP2-depleted, but SHP2 growth–independent, mouse melanoma and colon carcinoma cell lines displayed a typically subverted architecture, in which proliferative tumor cells surrounding a remodeled vessel formed “vascular islands”, each limited by surrounding hypoxic and dead tumor tissue, where inflammatory blood cells were limited. Although vascular islands generally reflect protected sanctuaries for tumor cells, we found that vascular island–resident, highly proliferative, SHP2-depleted tumor cells acquired an increased sensitivity to blockage of MEK/ERK signaling, resulting in reduced tumor growth. Our results show that the response to targeted therapies in resistant tumor cells was controlled by tumor cell–induced vascular changes and tumor architectural reorganization, providing a compelling approach to elicit tumor responses by exploiting tumor- and endothelium-dependent biochemical changes.

Authors

Yuyi Wang, Hidetaka Ohnuki, Andy D. Tran, Dunrui Wang, Taekyu Ha, Jing-Xin Feng, Minji Sim, Raymond Barnhill, Claire Lugassy, Michael R. Sargen, Emanuel Salazar-Cavazos, Michael Kruhlak, Giovanna Tosato

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Figure 5

Kinetics analysis and characterization of tumor island formation.

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Kinetics analysis and characterization of tumor island formation. (A) Re...
(A) Representative images of tumors harvested on days 9, 13, 17, and 21 following inoculation of p-LKO control and SHP2-silenced B16F10 cells. Top panels (scale bars: 200 μm) depict CD31+ vessels and distribution of hypoxia (Hypoxyprobe+); bottom panels (scale bars: 50 μm) show the distribution CD45+ inflammatory cells, Ki67+ proliferating cells, and cell nuclei (DAPI+). (B) Tumor weight of p-LKO and SHP2-silenced B16F10 tumors at the indicated time points; open circles represent individual tumors/mouse. n = 3 (days 9 and 13); n = 4 (days 17 and 21). (C) Quantification of tumor islands. A tumor island is defined as the assembly of live tumor cells around a blood vessel and surrounded by a zone of hypoxia. Quantification was performed by evaluating the entire tumor section through the maximum diameter; open circles represent individual tumors/mouse. n = 3 (days 9 and 13); n = 4 (days 17 and 21). *P < 0.05, by 2-tailed Student’s t test. (D–G) Quantification of CD31+ endothelial cells (D), hypoxic cells (Hypoxyprobe+) (E), CD45+ inflammatory cells (F), and Ki67+ proliferating cells (G) at the indicated time points in control tumors (n = 3, days 9 and 13; n = 4, days 17 and 21) and SHP2-silenced tumors (n = 3, days 9 and 13; n = 4, days 17 and 21). Gray circles: percentage of relative positive area/unit of tumor area; colored dots/circles: relative mean percentage of positive area/tumor. *P < 0.05, **P < 0.01, and ***P < 0.001, by 2-tailed Student’s t test. (H and I) Representative images (scale bar: 50 μm) from immunostaining of p-LKO (n = 3) and SHP2 silenced (n = 3) B16F10 tumor tissues harvested on day 13 for cleaved caspase 3 (cell death), CD31 endothelial cells, and nuclei (DAPI) (H) and quantification of cleaved caspase 3+CD31+ endothelial cells at the indicated time points following tumor cell inoculation (I). Gray circles: relative cleaved caspase 3+CD31+ cells/CD31+ cells analyzed; colored dots: tumor mean (control, n = 3; SHP2-silenced, n = 3). *P < 0.05 and **P < 0.01, by 2-tailed Student’s t test (I) The results in panels B–G and I are presented as means± SEM.