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
View: Text | PDF
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

×

Figure 8

HNF4α sustains DTP cells and limits the response to KRASG12D inhibition.

Options: View larger image (or click on image) Download as PowerPoint
HNF4α sustains DTP cells and limits the response to KRASG12D inhibition....
(A) Heatmap showing log2-normalized expression for G/M genes associated with DTP cells in IMA in vivo. (B and C) Dose-response curves for IMA organoids treated with BMS-986508. (B) 1311G (IC50: KN = 47.77 nM; KNH = 5.27 nM) and (C) 429A (IC50: KN = 152.3 nM; KNH = 25.61 nM). IC50 values were calculated by nonlinear regression. Data represent 1 of 3 independent biological replicates and are presented as the mean ± SEM. (D–F) Representative immunoblot analysis of the indicated proteins in 1311G organoids treated with BMS-986508 for 2 hours (n = 2 independent biological replicates) or 72 hours (n = 3), with quantification of pERK normalized to total ERK (tERK). Data are presented as the mean ± SD. (G) Dose-response curves for 1311G organoids treated with RMC-9805 (IC50: KN = 234.3 nM; KNH = 71.17 nM). Data represent 1 of 3 independent biological replicates and are presented as the mean ± SEM. (H) Human-derived organoids (KOR259) transduced with lentiviral dual gRNAs (sgdual) targeting HNF4A P1/P2 isoforms or a nontargeting control (NC), followed by 4 days of selection and treatment with BMS-986508 for 72 hours. IC50 values were 20.5 nM, 13.8 nM, 7.3 nM, and 6.4 nM for the NC and sgdual 1–3, respectively. Data represent 1 of 3 independent biological replicates and are presented as the mean ± SEM. (I) Waterfall plot showing the fold change in tumor volume in 1311G KN and KNH allografts treated with vehicle or BMS-986508. (J) Endpoint tumor volumes from I. ‡P = 0.0126, by unpaired, 2-tailed Student’s t test. (K) Tumor burden in KN and KNH GEMM tumors after 2 weeks of treatment with vehicle or BMS-986508. NS, P = 0.6943; ‡ ‡ ‡P = 0.0002; ****P < 0.0001, by Mann-Whitney U test). (L and M) Representative H&E and HNF4α IHC images. Scale bar: 100 μm. (N–Q) Quantification of pERK (N), MCM2 (O), CCND1 (P), and pRB(S807) (Q) by IHC. ****P < 0.0001 (O); § § §P = 0.0003 (P and Q), by Mann-Whitney U test.