The Mir181ab1 cluster promotes KRAS-driven oncogenesis and progression in lung and pancreas
Karmele Valencia, … , E. Alejandro Sweet-Cordero, Silvestre Vicent
Karmele Valencia, … , E. Alejandro Sweet-Cordero, Silvestre Vicent
Published December 24, 2019
Citation Information: J Clin Invest. 2020;130(4):1879-1895. https://doi.org/10.1172/JCI129012.
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Research Article Oncology

The Mir181ab1 cluster promotes KRAS-driven oncogenesis and progression in lung and pancreas

Abstract

Few therapies are currently available for patients with KRAS-driven cancers, highlighting the need to identify new molecular targets that modulate central downstream effector pathways. Here we found that the microRNA (miRNA) cluster including miR181ab1 is a key modulator of KRAS-driven oncogenesis. Ablation of Mir181ab1 in genetically engineered mouse models of Kras-driven lung and pancreatic cancer was deleterious to tumor initiation and progression. Expression of both resident miRNAs in the Mir181ab1 cluster, miR181a1 and miR181b1, was necessary to rescue the Mir181ab1-loss phenotype, underscoring their nonredundant role. In human cancer cells, depletion of miR181ab1 impaired proliferation and 3D growth, whereas overexpression provided a proliferative advantage. Lastly, we unveiled miR181ab1-regulated genes responsible for this phenotype. These studies identified what we believe to be a previously unknown role for miR181ab1 as a potential therapeutic target in 2 highly aggressive and difficult to treat KRAS-mutated cancers.

Authors

Karmele Valencia, Oihane Erice, Kaja Kostyrko, Simone Hausmann, Elizabeth Guruceaga, Anuradha Tathireddy, Natasha M. Flores, Leanne C. Sayles, Alex G. Lee, Rita Fragoso, Tian-Qiang Sun, Adrian Vallejo, Marta Roman, Rodrigo Entrialgo-Cadierno, Itziar Migueliz, Nerea Razquin, Puri Fortes, Fernando Lecanda, Jun Lu, Mariano Ponz-Sarvise, Chang-Zheng Chen, Pawel K. Mazur, E. Alejandro Sweet-Cordero, Silvestre Vicent

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Figure 6

Simultaneous reconstitution of Mir181a1 and Mir181b1 rescues the Mir181ab1-knockout phenotype.

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Simultaneous reconstitution of Mir181a1 and Mir181b1 rescues the Mir181a...
(A) Cell proliferation of KLA cells transduced with retroviral vectors containing Mir181a1 and Mir181b1 genomic DNA and treated with adCre or adE for 48 hours assessed by MTS (n = 3). Analysis by ANOVA. wt/wt: wild-type seed sequence of Mir181a1 and Mir181b1. mut/wt: mutated seed sequence of Mir181a1 and wild-type sequence of Mir181b1. wt/mut: wild-type seed sequence of Mir181a1 and mutated sequence of Mir181b1. mut/mut: mutated seed sequence of Mir181a1 and Mir181b1. (B) 3D culture of KLA cells expressing the different Mir181a1 and Mir181b1 constructs. Left: Representative images of KLA miR181 organoids on day 4 after 48 hours of treatment with adE or adCre. Scale bars: 100 μm. Right: KLA miR181 organoid size quantification on day 3 after seeding (n = 14–20) and compared using ANOVA. (C) Average tumor volume of allografts from mouse KLA cells transduced with the different Mir181a1 and Mir181b1 constructs, previously treated with adE or adCre (n = 6 per group), assessed by ANOVA. (D) 3D culture of KPC miR181ko cells transduced with the Mir181a1 and Mir181b1 constructs. Left: KPC miR181ko organoids on day 4 after seeding. Scale bars: 100 μm. Right: Organoid size quantification on day 4 after seeding (n = 8–18) and compared by ANOVA. (E) Average tumor volume of allografts from mouse KPC miR181wt and miR181ko cells (n = 6 per group) and compared using ANOVA. (F and G) Phospho–histone H3 immunofluorescence images and analyses of wt/wt and mut/mut adCre–treated KLA cells (F), and in KPC181ko wt/wt and mut/mut cells (G) 72 hours after seeding (n = 6–8). Results were compared by Kruskal-Wallis test.