Targeting the αv integrin/TGF-β axis improves natural killer cell function against glioblastoma stem cells
Hila Shaim, … , Amy B. Heimberger, Katayoun Rezvani
Hila Shaim, … , Amy B. Heimberger, Katayoun Rezvani
Published June 17, 2021
Citation Information: J Clin Invest. 2021;131(14):e142116. https://doi.org/10.1172/JCI142116.
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Targeting the αv integrin/TGF-β axis improves natural killer cell function against glioblastoma stem cells

Abstract

Glioblastoma multiforme (GBM), the most aggressive brain cancer, recurs because glioblastoma stem cells (GSCs) are resistant to all standard therapies. We showed that GSCs, but not normal astrocytes, are sensitive to lysis by healthy allogeneic natural killer (NK) cells in vitro. Mass cytometry and single-cell RNA sequencing of primary tumor samples revealed that GBM tumor–infiltrating NK cells acquired an altered phenotype associated with impaired lytic function relative to matched peripheral blood NK cells from patients with GBM or healthy donors. We attributed this immune evasion tactic to direct cell-to-cell contact between GSCs and NK cells via αv integrin–mediated TGF-β activation. Treatment of GSC-engrafted mice with allogeneic NK cells in combination with inhibitors of integrin or TGF-β signaling or with TGFBR2 gene–edited allogeneic NK cells prevented GSC-induced NK cell dysfunction and tumor growth. These findings reveal an important mechanism of NK cell immune evasion by GSCs and suggest the αv integrin/TGF-β axis as a potentially useful therapeutic target in GBM.

Authors

Hila Shaim, Mayra Shanley, Rafet Basar, May Daher, Joy Gumin, Daniel B. Zamler, Nadima Uprety, Fang Wang, Yuefan Huang, Konrad Gabrusiewicz, Qi Miao, Jinzhuang Dou, Abdullah Alsuliman, Lucila N. Kerbauy, Sunil Acharya, Vakul Mohanty, Mayela Mendt, Sufang Li, JunJun Lu, Jun Wei, Natalie W. Fowlkes, Elif Gokdemir, Emily L. Ensley, Mecit Kaplan, Cynthia Kassab, Li Li, Gonca Ozcan, Pinaki P. Banerjee, Yifei Shen, April L. Gilbert, Corry M. Jones, Mustafa Bdiwi, Ana K. Nunez-Cortes, Enli Liu, Jun Yu, Nobuhiko Imahashi, Luis Muniz-Feliciano, Ye Li, Jian Hu, Giulio Draetta, David Marin, Dihua Yu, Stephan Mielke, Matthias Eyrich, Richard E. Champlin, Ken Chen, Frederick F. Lang, Elizabeth J. Shpall, Amy B. Heimberger, Katayoun Rezvani

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Figure 1

GSCs express NK cell receptor ligands and are susceptible to NK cell cytotoxicity.

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GSCs express NK cell receptor ligands and are susceptible to NK cell cyt...
(A) 51Cr release assay showing cytotoxicity of donor-derived NK cells activated overnight with IL-15 (5 ng/mL) against GSCs (blue), K562 cells (black), U87 cell line (green), or healthy human astrocytes (red) (U87: n = 3; K562, GSCs, astrocytes: n = 6). Error bars denote SD. Green asterisks: cytotoxicity against U87 vs. astrocytes. Black asterisks: cytotoxicity against K562 vs. astrocytes. Blue asterisks: cytotoxicity against GSCs vs. astrocytes. (B) Heatmap representing the relative expression of NK cell ligands on GSCs or human astrocytes ranging from blue (low) to red (high). Columns represent the median expression of each receptor (GSC: n = 6; astrocytes: n = 3). (C) Activated HC-NK cells were cocultured with GSCs in the presence or absence of blocking antibodies: anti-NKG2D (blue), anti-DNAM (green), anti-NKp30 (red), or anti–HLA class I (purple). 51Cr release assay against GSCs was assessed (n = 4). Blue asterisks: cytotoxicity against GSCs with or without anti-NKG2D. Red asterisks: cytotoxicity against GSCs with or without anti-NKp30. Green asterisks: cytotoxicity against GSCs with or without anti-DNAM. (D and E) viSNE plots (D) and a comparative mass cytometry heatmap (E) showing the expression of NK cell markers in HC-NK (red), GP-NK (green), and TI-NK cells (blue). Column clustering was identified by FlowSOM. Each row reflects annotation of the expression level for an individual patient. Color scale ranges from blue (lower expression) to red (higher expression) (n = 3). (F) UMAP plot showing clusters for TI-NK versus HC-NK cells by scRNA-seq. (G) Violin plots showing the mRNA expression levels for individual NK cell–related genes in healthy controls (HC-NK cells; blue) and TI-NK cells (red) using scRNA-seq. Markers associated with NK cell activation and cytotoxicity, inhibition, and the TGF-β pathway are presented. Statistical analysis by 2-way ANOVA with Dunnett’s correction for multiple comparisons (A and C) or unpaired t test (G). *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001.