BRD4 promotes endodermal cell fate during mammalian lung development
Hongbo Wen, Derek C. Liberti, Prashant Chandrasekaran, Shahana Parveen, Kwaku K. Quansah, Mijeong Kim, Ana N. Lange, Abigail T. Marquis, Sylvia N. Michki, Annabelle Jin, MinQi Lu, Ayomikun A. Fasan, Sriyaa Suresh, Shawyon P. Shirazi, Lisa R. Young, Jennifer M.S. Sucre, Maria C. Basil, Rajan Jain, David B. Frank
Hongbo Wen, Derek C. Liberti, Prashant Chandrasekaran, Shahana Parveen, Kwaku K. Quansah, Mijeong Kim, Ana N. Lange, Abigail T. Marquis, Sylvia N. Michki, Annabelle Jin, MinQi Lu, Ayomikun A. Fasan, Sriyaa Suresh, Shawyon P. Shirazi, Lisa R. Young, Jennifer M.S. Sucre, Maria C. Basil, Rajan Jain, David B. Frank
View: Text | PDF
Research Article Development Pulmonology

BRD4 promotes endodermal cell fate during mammalian lung development

Abstract

Lung development relies on diverse cell intrinsic and extrinsic mechanisms to ensure proper cellular differentiation and compartmentalization. In addition, it requires precise integration of multiple signaling pathways to temporally regulate morphogenesis and appropriate cell specification. To accomplish this, organogenesis relies on epigenetic and transcriptional regulators to promote cell fate and inhibit alternative cell fates. Using genetic mouse and human embryonic stem cell (hESC) differentiation models, tissue explants, and single-cell transcriptomic analysis, we demonstrated that Bromodomain Containing Protein 4 (BRD4) is required for mammalian lung morphogenesis and cell fate. Endodermal deletion of BRD4 impaired epithelial-mesenchymal crosstalk, leading to disrupted proximal-distal patterning and branching morphogenesis. Moreover, temporal deletion of BRD4 revealed developmental stage–specific defects in airway and alveolar epithelial cell specification with a predominant role in proximal airway cell fate. Similarly, BRD4 promoted lung endodermal cell differentiation into airway lineages in a hESC-derived lung organoid model. Together, these data demonstrate that BRD4 orchestrates early lung morphogenesis and separately regulates cell specification, indicating a multifunctional and evolutionarily conserved role for BRD4 in mammalian lung development.

Authors

Hongbo Wen, Derek C. Liberti, Prashant Chandrasekaran, Shahana Parveen, Kwaku K. Quansah, Mijeong Kim, Ana N. Lange, Abigail T. Marquis, Sylvia N. Michki, Annabelle Jin, MinQi Lu, Ayomikun A. Fasan, Sriyaa Suresh, Shawyon P. Shirazi, Lisa R. Young, Jennifer M.S. Sucre, Maria C. Basil, Rajan Jain, David B. Frank

×

Figure 9

BRD4 bromodomain inhibition sequesters hESC-derived lung endodermal organoids in a SOX2 progenitor state.

Options: View larger image (or click on image) Download as PowerPoint
BRD4 bromodomain inhibition sequesters hESC-derived lung endodermal orga...
(A) IHC for NKX2.1 and PDX1 in DMSO- and JQ1-treated organoids. Scale bars: 100 μm). (B) Quantification of the percentages of NKX2.1+ or PDX1+ organoids between DMSO and JQ1 treatment. Quantification data are represented as mean ± SEM. Two-tailed t tests. n ≥ 6. (C) IHC for SOX2 and SOX9 in DMSO- and JQ1-treated organoids. Scale bar: 100 μm. (D) Quantification of the percentages of organoids containing cells that are SOX2+, SOX9+, and SOX2+SOX9+. Quantification data are represented as mean ± SEM. Two-tailed t test: *P ≤ 0.05, **P ≤ 0.01. n = 4. (E) qPCR analysis of organoids expressing SOX2 or SOX9. Quantification data are represented as mean ± SEM. Two-tailed t test: *P ≤ 0.05. n = 3. (F) IHC for SOX2, KRT5, and SCGB3A2 (left 4 panels) and SOX2, TP63, and TUBB4 (right 4 panels) in DMSO- and JQ1-treated organoids. Boxed areas represent magnified areas shown on the panels to the right of each image. Scale bar: 200 μm, 50 μm (magnified boxes). (G) Quantification of the percentages of undifferentiated organoids. Quantification data represented as mean ± SEM. Two-tailed t test: **P ≤ 0.01. n = 3.