Antiangiogenic immunotherapy suppresses desmoplastic and chemoresistant intestinal tumors in mice
Simone Ragusa, … , Michele De Palma, Tatiana V. Petrova
Simone Ragusa, … , Michele De Palma, Tatiana V. Petrova
Published February 4, 2020
Citation Information: J Clin Invest. 2020;130(3):1199-1216. https://doi.org/10.1172/JCI129558.
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Research Article Angiogenesis Oncology

Antiangiogenic immunotherapy suppresses desmoplastic and chemoresistant intestinal tumors in mice

Abstract

Mutations in APC promote colorectal cancer (CRC) progression through uncontrolled WNT signaling. Patients with desmoplastic CRC have a significantly worse prognosis and do not benefit from chemotherapy, but the mechanisms underlying the differential responses of APC-mutant CRCs to chemotherapy are not well understood. We report that expression of the transcription factor prospero homeobox 1 (PROX1) was reduced in desmoplastic APC-mutant human CRCs. In genetic Apc-mutant mouse models, loss of Prox1 promoted the growth of desmoplastic, angiogenic, and immunologically silent tumors through derepression of Mmp14. Although chemotherapy inhibited Prox1-proficient tumors, it promoted further stromal activation, angiogenesis, and invasion in Prox1-deficient tumors. Blockade of vascular endothelial growth factor A (VEGFA) and angiopoietin-2 (ANGPT2) combined with CD40 agonistic antibodies promoted antiangiogenic and immunostimulatory reprogramming of Prox1-deficient tumors, destroyed tumor fibrosis, and unleashed T cell–mediated killing of cancer cells. These results pinpoint the mechanistic basis of chemotherapy-induced hyperprogression and illustrate a therapeutic strategy for chemoresistant and desmoplastic CRCs.

Authors

Simone Ragusa, Borja Prat-Luri, Alejandra González-Loyola, Sina Nassiri, Mario Leonardo Squadrito, Alan Guichard, Sabrina Cavin, Nikolce Gjorevski, David Barras, Giancarlo Marra, Matthias P. Lutolf, Jean Perentes, Emily Corse, Roberta Bianchi, Laureline Wetterwald, Jaeryung Kim, Guillermo Oliver, Mauro Delorenzi, Michele De Palma, Tatiana V. Petrova

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Figure 2

Inactivation of Prox1 generates desmoplastic, angiogenic, and T cell–excluded tumors.

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Inactivation of Prox1 generates desmoplastic, angiogenic, and T cell–exc...
(A) Apcfl/fl Tp53fl/fl villin-CreERT2 (AP) and Apcfl/fl Tp53fl/fl Prox1flfl villin-CreERT2 (APP) tumor models used in the study. (B) Appearance and weights of tumors and WT cecum and quantitative reverse transcription PCR (qRT-PCR) data for Lgr5 and Krt20. AP (n = 22); APP (n = 15); WT (n = 7). qRT-PCR data were normalized to the AP mean. AP or APP (n = 9); WT (n = 4). Scale bars: 4 mm. (C) Pathways enriched in APP versus AP transcriptomes (n = 6 per genotype). NES, normalized enrichment score. (D) APP tumors were desmoplastic. 3D reconstructions of tumor and WT cecum thick slices. Images show staining for PH3 (green), α-SMA (red), and E-cadherin (white). Scale bars: 50 μm. (E) Quantification of proliferation of stromal and tumor epithelial cells. The α-SMA+ area from D was normalized to the total tumor area and the AP mean. For the α-SMA+ area: AP (n = 10); APP or normal cecum (n = 7). For Ki67+ colocalization: AP or APP (n = 6); normal cecum (n = 4). (F) 3D vascular reconstructions from tumor slices. Images show staining for CD31 (green) and E-cadherin (white). Scale bars: 50 μm. (G) Quantification of vascular parameters and expression of the endothelial marker Cdh5. AP (n = 10); APP (n = 7); normal cecum (n = 7). AP or APP (n = 9); WT (n = 4). Data were normalized to the AP mean. (H) Reduced CD8+ T cell infiltration into APP tumors. Images show staining for CD8+ T cells (green), VE-cadherin (red), and E-cadherin (white). Scale bars: 50 μm. (I) Quantification of CD8+ T cells. AP (n = 10); APP (n = 6); normal cecum (n = 7). (J) APP signature is enriched in PROX1-low human CRCs. Enrichment of the APP versus AP signature in human CRCs (GSE39582; n = 444) was computed using Z scores. Dashed line indicates the linear regression fit. Data represent the mean ± SD. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, and ****P ≤ 0.0001, by 1-way ANOVA with Tukey’s multiple comparisons test (B, E, and G) and grouped analysis by 2-way ANOVA with Tukey’s multiple comparisons test (E).