Opposing roles of nuclear receptor HNF4α isoforms in colitis and colitis-associated colon cancer
- University of California, Riverside, United States
- Harvard Medical School, United States
Figures
Figure 1
Differential localization of HNF4α isoforms in mouse colonic crypts.
(A) Schematic of the mouse (and human) Hnf4a gene showing the two promoters (P1 and P2) (top) and the P1- and P2-driven HNF4α isoforms that they express (bottom). The differential N-terminal A/B …
Figure 2
Dysregulation of P1- and P2-HNF4α in mouse models of colitis and colon cancer.
(A) IB of WCE from the distal colon of WT mice treated with 2.5% DSS for 5 days followed by 0 or 3 days recovery, and an analogous region of untreated (Control) mice. Each lane is from a different …
Figure 3 with 1 supplement
Differential susceptibility of HNF4α isoform-specific mice to colitis-associated colon cancer.
(A) Top left, Schematic of Hnf4a exon-swap (i.e., isoform-specific) mice. Top right, IB as in Figure 2A of WCE from the distal colon of the exon-swap mice and their WT controls, probed with the …
Figure 3—figure supplement 1
HNF4α isoform-specific mice subjected to AOM+DSS to induce colitis-associated colon cancer.
(A) Spleen-to-body weight ratio of WTα1 (n=5) and α1HMZ (n=6) mice treated with 10 mg/kg AOM and two cycles of DSS (5 days per cycle) and harvested at ~54 days. (B) Tumor load in WTα7 and α7HMZ male …
Figure 4 with 2 supplements
Differential susceptibility of HNF4α isoform-specific mice to DSS-induced colitis.
(A) Percent mortality of WTα7 (n = 28) and α7HMZ (n = 16) mice treated with 2.5% DSS for 5 days. α7HMZ mice typically died during day 3 to 12 of recovery following DSS treatment. Data pooled from …
Figure 4—figure supplement 1
Increased susceptibility of α7HMZ mice to DSS-induced colitis.
Results from an independent DSS experiment from that shown in Figure 4 A-C. Mice from this experiment were analyzed by IB and IF in Figure 4D. (A) Colon length of WTα7 and α7HMZ mice treated with …
Figure 4—figure supplement 2
Increased inflammation in α7HMZ mice in DSS-induced colitis.
(A) Spleen-to-body weight ratio of α7HMZ (n=7) and α1HMZ (n=7) mice treated for 4 days with 2.5% DSS followed by 18 days of recovery. (B) Crypt length of α7HMZ (n=3) and α1HMZ (n=3) mice treated as …
Figure 5 with 1 supplement
Altered gene expression, interacting proteins, migration and ion transport in HNF4α isoform-specific mice.
(A,B) Comparative Gene Ontology (GO) of genes differentially regulated (≥two-fold) in the distal colon of untreated α1HMZ (A) and α7HMZ (B) mice. (C) Top, Venn diagram of total number of …
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Figure 5—source data 1
Transcriptomic analysis of HNF4α isoform-specific mice.
(A) Top 100 genes DOWN in distal colon of α1HMZ male mice compared to WT controls. (B) Top 100 genes UP in distal colon of α1HMZ male mice compared to WT controls. (C) Top 100 genes DOWN in distal colon of α7HMZ male mice compared to WT controls. (D) Top 100 genes UP in distal colon of α7HMZ male mice compared to WT controls. (E) Up-regulated genes involved in wound healing and immune function enriched in α1HMZ mice. (F) Up-regulated genes involved in cell cycle and DNA repair in α7HMZ mice. (G) Down-regulated genes involved in cell adhesion and ion transport in α7HMZ mice.
- https://doi.org/10.7554/eLife.10903.011
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Figure 5—source data 2
Proteomic analysis of HNF4α isoform-specific mice.
(A) List of proteins that interact with HNF4α in α1HMZ and α7HMZ colons from RIME analysis meeting the criteria described in Figure 5C. (B) Select proteins that interact with HNF4α in α1HMZ and α7HMZ colons from RIME analysis used to prepare the graph in Figure 5C. (C) All peptides that interact with HNF4α in both α7HMZ and α1HMZ colons from RIME analysis in which there are at least 2 positives for each genotype. (D) All peptides that interact with HNF4α preferentially in α7HMZ colons in which there are 2 or more positives for α7HMZ compared to α1HMZ. (E) All peptides that interact with HNF4α preferentially in α1HMZ colons in which there are 2 or more positives for α1HMZ compared to α7HMZ. Figure 5C.
- https://doi.org/10.7554/eLife.10903.012
Figure 5—figure supplement 1
Transcriptomic and BrdU analysis of HNF4α isoform-specific mice.
(A) Total number of genes up- or down-regulated (non log fold change) in α1HMZ and α7HMZ untreated young adult male mice compared to their respective WT controls as determined by Affymetrix Exon …
Figure 6 with 1 supplement
RELMβ knockout decreases susceptibility of α7HMZ mice to colitis.
(A) RELMβ protein level quantified by ELISA in the midcolon homogenate of mice with the indicated genotype treated with 2.5% DSS for 6 days. Genotypes are indicated as Retnlb/Hnf4a. N = 3–5 mice per …
Figure 6—figure supplement 1
Verification of RELMβKO/α7HMZ mice.
(A) Verification of genotype of RELMβ KO mice crossed into α7HMZ mice. Shown is a representative DNA agarose gel of PCR products generated with primers for the Retnlb and Hnf4a loci for 2 mice per …
Figure 7 with 1 supplement
Direct and indirect mechanisms of regulation of RELMβ expression by HNF4α isoforms: impact on DSS sensitivity and recovery.
(A) P2-HNF4α binds the promoter of the RETNLB gene in colonic epithelial cells. Left, schematic of the human RETNLB promoter showing predicted SVM binding sites for HNF4α, as well as sites for NFκB, …
Figure 7—figure supplement 1
Predicted HNF4α binding sites in the human and mouse RELMβ gene and RELMβ reporter assays.
(A–C). Screenshots of UCSC Genome Browser shows putative HNF4α binding sites in human (AB) and mouse (C) RELMβ genes as well as the fragments amplified by the primers for Region 1 (A) and Region 2 (B…
Additional files
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Repoarting Standard
NC3Rs ARRIVE guidelines checklist.
- https://doi.org/10.7554/eLife.10903.018
