Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 May;73(5):1701-1716.
doi: 10.1002/hep.31517. Epub 2021 Mar 16.

17-Beta Hydroxysteroid Dehydrogenase 13 Deficiency Does Not Protect Mice From Obesogenic Diet Injury

Yanling Ma  1   2 , Philip M Brown  1   2 , Dennis D Lin  1   2 , Jing Ma  3 , Dechun Feng  3 , Olga V Belyaeva  4 , Maren C Podszun  1   2 , Jason Roszik  5   6 , Joselyn N Allen  2 , Regina Umarova  2 , David E Kleiner  7 , Natalia Y Kedishvili  4 , Oksana Gavrilova  8 , Bin Gao  3 , Yaron Rotman  1   2
Affiliations

17-Beta Hydroxysteroid Dehydrogenase 13 Deficiency Does Not Protect Mice From Obesogenic Diet Injury

Yanling Ma et al. Hepatology. 2021 May.

Abstract

Background and aims: 17-Beta hydroxysteroid dehydrogenase 13 (HSD17B13) is genetically associated with human nonalcoholic fatty liver disease (NAFLD). Inactivating mutations in HSD17B13 protect humans from NAFLD-associated and alcohol-associated liver injury, fibrosis, cirrhosis, and hepatocellular carcinoma, leading to clinical trials of anti-HSD17B13 therapeutic agents in humans. We aimed to study the in vivo function of HSD17B13 using a mouse model.

Approach and results: Single-cell RNA-sequencing and quantitative RT-PCR data revealed that hepatocytes are the main HSD17B13-expressing cells in mice and humans. We compared Hsd17b13 whole-body knockout (KO) mice and wild-type (WT) littermate controls fed regular chow (RC), a high-fat diet (HFD), a Western diet (WD), or the National Institute on Alcohol Abuse and Alcoholism model of alcohol exposure. HFD and WD induced significant weight gain, hepatic steatosis, and inflammation. However, there was no difference between genotypes with regard to body weight, liver weight, hepatic triglycerides (TG), histological inflammatory scores, expression of inflammation-related and fibrosis-related genes, and hepatic retinoid levels. Compared to WT, KO mice on the HFD had hepatic enrichment of most cholesterol esters, monoglycerides, and certain sphingolipid species. Extended feeding with the WD for 10 months led to extensive liver injury, fibrosis, and hepatocellular carcinoma, with no difference between genotypes. Under alcohol exposure, KO and WT mice showed similar hepatic TG and liver enzyme levels. Interestingly, chow-fed KO mice showed significantly higher body and liver weights compared to WT mice, while KO mice on obesogenic diets had a shift toward larger lipid droplets.

Conclusions: Extensive evaluation of Hsd17b13 deficiency in mice under several fatty liver-inducing dietary conditions did not reproduce the protective role of HSD17B13 loss-of-function mutants in human NAFLD. Moreover, mouse Hsd17b13 deficiency induces weight gain under RC. It is crucial to understand interspecies differences prior to leveraging HSD17B13 therapies.

PubMed Disclaimer

Figures

Figure 1.
Figure 1.. HSD17B13 is expressed in normal hepatocytes.
(A) Gene expression of HSD17B13 in 53 healthy and 33 carcinoma tissues from the GTEx and TCGA databases. Cancer tissues are shown in red and normal tissues are shown in green. Data are sorted by median expression. (B) Tissue distribution of Hsd17b13 in mice. Gene expression of Hsd17b13 was determined in the liver, epididymal fat, inguinal fat, muscle, duodenum, jejunum, ileum, pancrase, kidney, testis, and heart tissues of 20 weeks old C57BL/6 mice by qPCR (n=6, Mean±SEM). (C) HSD17B13 is restrictively expressed within the hepatocyte cluster in human liver determined by single cell sequencing (http://human-liver-cell-atlas.ie-freiburg.mpg.de). (D) Single cell transcriptome data (https://tabula-muris.ds.czbiohub.org) showing the top 12 mouse cell types from 20 organs according to expression level of Hsd17b13, sorted by mean value.
Figure 2.
Figure 2.. Hsd17b13 deficiency induces weight gain in mice fed regular chow diet.
Eight week-old male wild type (WT) and Hsd17b13-knockout (KO) mice were fed regular chow for 12 weeks and sacrified at age 20 weeks, after a 5 hours fast. (A) Body weight of WT and KO mice was measured weekly. (B) Body weight, (C) Lean mass relative to body weight, and (D) Fat mass relative to body weight. (E) Liver weight, (F) Liver-to-body weight ratio, and (G) Liver triglycerides. (H) Hematoxylin & eosin staining of liver sections. Bar indicates 100 μM. (I) Hepatic gene expression, normalized to WT level. Hsd17b13 1–3 and 6–7 denotes two separate regions in the gene. N=6 male mice per group. B-I assessed at sacrifice at age 20 weeks. Mean±SEM; *p<0.05, ****p<0.0001
Figure 3.
Figure 3.. Limited impact of Hsd17b13 deficiency on high fat diet (HFD) liver injury.
Eight week-old male wild type (WT) and Hsd17b13-knockout (KO) mice were fed with HFD for 12 weeks and sacrified at age 20 weeks after a 5 hours fast. (A) Weekly body weight measurements, (B) Liver weights, (C) Liver triglycerides, and (D) Serum alanine aminotransferase (ALT). (E) Hematoxylin & eosin staining of liver sections. Bar indicates 100 μM. (F) Percentage of steatotic hepatocytes. (G) Percentage of macro- and microtsteatotic hepatocytes. (H) Histological inflammation score. (I) Hepatic collagen content by Sirus Red staining (left) or colorimetric assay (right). Arrow indicates representive positive staining. Bar indicates 100 μM. (J) Serum cholesterol, (K) Serum triglycerides, (L) Serum free fatty acids, and (M) Blood glucose. (N) Hepatic gene expression level of fibrosis- and inflammation-related genes. N=6 male mice per group. B-N assessed at sacrifice at age 20 weeks. Mean±SEM; *p<0.05, **p<0.01
Figure 4.
Figure 4.. Hsd17b13 deficiency does not protect mice from Western diet (WD) liver injury.
Eight week-old male wild type (WT) and Hsd17b13-knockout (KO) mice were fed with WD for 16 weeks and sacrified at age 24 weeks after a 5 hours fast. (A) Weekly body weight, (B) Liver weight, (C) Liver triglycerides, and (D) Serum liver enzymes. (E) H&E staining of liver sections. Bar indicates 100 μM. (F) Percentage of steatotic hepatocytes. (G) Percentage of hepatocytes with macro- and micro-steatosis. (H) Histological inflammation score. (I) Hepatic collagen content by Sirius Red staining (left) or colorimetric assay (right). Arrow indicates representive positive staining. Bar indicates 100 μM. (J) Hepatic expression level of fibrosis- and inflammation-related genes. N=7 male mice per group. B-J assessed at sacrifice at age 24 weeks. Mean±SEM
Figure 5.
Figure 5.. Hsd17b13 deficiency does not protect mice from long-term Western diet induced injury.
Eight week-old male wild type (WT) and Hsd17b13-KO (KO) mice were fed Western diet (WD) for 10 months and sacrified after a 5 hour fast. (A) Body weight, (B) liver weight, (C) Liver triglycerides, and (D-E) serum liver enzymes. (F) Hematoxyline & eosin staining of liver sections. Bar indicates 100 μM. (G) Percent of steatotic hepatocytes. (H) Percent of hepatocytes with micro- and macro-steatosis. (I) Histological inflammation score. (J) Hepatic collagen content by Sirius Red staining (left) or colorimetric assay (right). Arrow indicates representive positive staining. Bar indicates 100 μM. (K) Hepatic expression level of fibrosis- and inflammation-related genes. (L) Liver tumor incidence and (M) Liver surface tumor number. N=6 male mice per group. All assessments at sacrifice, after 10 months of WD. Mean±SEM
Figure 6.
Figure 6.. Effect of Hsd17b13 deficiency on alcohol-induced liver disease.
Female wild type (WT, n=8) and Hsd17b13-KO (KO, n=10) mice were given 10 days of Lieber-DeCarli ethanol diet followed by one binge dose of alcohol (ETOH) at age of 18 weeks. (A) Serum ALT level of WT (n=7) and KO (n=10) mice. (B) Liver triglycerides. (C) Liver total cholesterol. (D) Hematoxyline & eosin staining of liver sections. Bar indicates 100 μM. (E) Percentage of steatotic hepatocytes. (F) Histological inflammation score. WT, n=8; KO, n=10 for all experiments unless stated otherwise. Mean±SEM
Figure 7.
Figure 7.. Effect of Hsd17b13 deficiency on the liver lipidome.
Eight week-old male wild type (WT) and Hsd17b-13-KO (KO) were fed high-fat diet for 12 weeks and sacrified at age 20 weeks after a 5 hours fast. Liver samples were analyzed for lipid composition. (A) Total amounts of liver lipids by lipid class, relative to controls. Mean±SEM. P-values from Student’s t-test. (B) Volcano plot of 986 individual liver lipid species. The x-axis denotes the log2 fold change in concentration in KO compared to WT and the y-axis the −log10 of the unadjusted p-value. (C) Heatmap of individual hepatic lipid species by class and fatty acid composition. Color reflects fold-change in concentrations between KO and WT. Red denotes species increased in KO and blue, in WT. (D)Percent of all species within a class that are upregulated in KO (red) or WT (blue). Numbers denote the absolute number of species in each category. Significance calculated by Γ2. TAG, triacylglycerol; DAG, diacylglycerol; MAG, monoacylglycerol; PC, phosphatidylcholine; LPC, lysophosphatidylcholine; PE, phosphatidylethanolamine; LPE, lysophosphatidylethanolamine; PI, phosphatidylinositol; CE, cholesterol ester; SM, sphingomyelin; CER, ceramide; DCER, dihydroceramide; HCER, hexosylceramide; LCER, lactosylceramide. N=6 male mice per group. Animals are the same as depicted in Figure 3.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Younossi Z, Anstee QM, Marietti M, Hardy T, Henry L, Eslam M, George J, et al. Global burden of NAFLD and NASH: trends, predictions, risk factors and prevention. Nat Rev Gastroenterol Hepatol 2018;15:11–20. - PubMed
    1. Loomba R, Schork N, Chen CH, Bettencourt R, Bhatt A, Ang B, Nguyen P, et al. Heritability of Hepatic Fibrosis and Steatosis Based on a Prospective Twin Study. Gastroenterology 2015;149:1784–1793. - PMC - PubMed
    1. Schwimmer JB, Celedon MA, Lavine JE, Salem R, Campbell N, Schork NJ, Shiehmorteza M, et al. Heritability of nonalcoholic fatty liver disease. Gastroenterology 2009;136:1585–1592. - PMC - PubMed
    1. Browning JD, Szczepaniak LS, Dobbins R, Nuremberg P, Horton JD, Cohen JC, Grundy SM, et al. Prevalence of hepatic steatosis in an urban population in the United States: impact of ethnicity. Hepatology 2004;40:1387–1395. - PubMed
    1. Romeo S, Kozlitina J, Xing C, Pertsemlidis A, Cox D, Pennacchio LA, Boerwinkle E, et al. Genetic variation in PNPLA3 confers susceptibility to nonalcoholic fatty liver disease. Nat Genet 2008;40:1461–1465. - PMC - PubMed

Publication types

Substances