HNF4α controls growth, identity, and KRAS inhibitor response in invasive mucinous adenocarcinoma of the lung
Headtlove Essel Dadzie, Yangsook Song Green, Soledad A. Camolotto, Henry U. Arnold, Matthew Gumbleton, Minzhe Guo, Mari Mino-Kenudson, Yutaka Maeda, Benjamin T. Spike, Eric L. Snyder
Headtlove Essel Dadzie, Yangsook Song Green, Soledad A. Camolotto, Henry U. Arnold, Matthew Gumbleton, Minzhe Guo, Mari Mino-Kenudson, Yutaka Maeda, Benjamin T. Spike, Eric L. Snyder
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Research Article Oncology Pulmonology

HNF4α controls growth, identity, and KRAS inhibitor response in invasive mucinous adenocarcinoma of the lung

Abstract

Cellular plasticity is a hallmark of cancer, enabling tumor cells to alter identity and evade therapeutic pressure. In invasive mucinous adenocarcinoma of the lung (IMA), NK2 homeobox 1 (NKX2-1) loss triggers a pulmonary to gastric switch marked by aberrant activation of hepatocyte nuclear factor 4 alpha (HNF4α), a master regulator of gastrointestinal/hepatic differentiation. We show that HNF4α promoted IMA growth and activated a gastric pit cell–like program. Loss of HNF4α enabled forkhead box A1 and A2 (FoxA1/2) transcription factors to bind de novo sites and activate alternative, nongastric identities in IMA. HNF4α also established a mucinous program associated with tolerance to KRAS blockade, and loss of HNF4α enhanced response to KRASG12D inhibition. Mechanistically, HNF4α blocked cell-cycle exit in drug-tolerant persister cells and promoted activity of the antioxidant transcription factor nuclear factor erythroid 2–related factor 2 (NRF2). NRF2 activation partially rescued the effects of Hnf4a deletion on KRASG12D inhibition, whereas NRF2 inhibition enhanced sensitivity to KRASG12D blockade. Thus, HNF4α is a key regulator of growth, identity, and primary response to KRASG12D inhibition in IMA.

Authors

Headtlove Essel Dadzie, Yangsook Song Green, Soledad A. Camolotto, Henry U. Arnold, Matthew Gumbleton, Minzhe Guo, Mari Mino-Kenudson, Yutaka Maeda, Benjamin T. Spike, Eric L. Snyder

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Figure 9

NRF2 mediates HNF4α-dependent resistance to KRAS inhibition in IMA.

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NRF2 mediates HNF4α-dependent resistance to KRAS inhibition in IMA. (A) ...
(A) IPA-predicted upstream transcriptional regulators from DEGs in KNH versus KN GEMM tumors. (B) GSEA showing depletion of the NRF2 gene signature in KNH GEMM tumors. (C) qRT-PCR of Nrf2, Nqo1, and Aldh3a1. ***P < 0.001, by unpaired, 2-tailed Student’s t test. log2Norm, log2-normalized. (D) Spearman correlation between NRF2 activity and HNF4A signature 1 scores in TCGA KRAS-mutant NSCLC; linear regression with 95% CI. Spearman R and P values are shown. (E) HNF4A signature 1 scores in NRF2-low versus NRF2-high tumors. Significance was determined by unpaired, 2-tailed Student’s t test. (F) NRF2 immunoblot of FACS-sorted IMA GEMM tumors. (G) Immunoblot of the indicated proteins in 1311G organoids treated with 1 μM MG132 or vehicle for 2 hours. (H) NRF2 quantification normalized to the loading control following 2 hours of DI-591 treatment. n = 3 independent biological replicates; data indicate the mean ± SD. P = 0.0486, by unpaired, 2-tailed Student’s t test. (I and J) 1311G KNH organoids expressing dox-inducible FLAG-NFE2L2 (NRF2) or empty vector (EV) ± 0.1 μg/mL doxycycline: (I) immunoblot of the indicated proteins (72 hours). (J) qRT-PCR of NFE2L2 and Nqo1 expression (48 hours). Data represent 1 of 3 independent biological replicates and are shown as the mean ± SD. ****P < 0.0001; NS = 0.5102, by 2-way ANOVA with Tukey’s multiple-comparison test. (K) Dose-response curves for 1311G KNH organoids expressing EV or dox-inducible NRF2 treated with BMS-986508 with or without 0.5 μg/mL dox for 72 hours (IC50: EV = 2.5 nM; NRF2 = 33.7 nM). Data represent 1 of 3 biological replicates and are shown as the mean ± SEM. (L) Cell viability of 1311G KN organoids treated with increasing concentrations of ML385 alone or with 10 or 20 nM BMS-986508. Data indicate the mean ± SEM from 1 of 3 biological replicates. **P < 0.01 and ***P < 0.001, by 2-way ANOVA with Tukey’s multiple-comparison test. (M) Bliss synergy score calculated using SynergyFinder. FPKM, fragments per kilobase of transcript per million mapped reads.