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
. 2018 Jul 2;7(7):bio033910.
doi: 10.1242/bio.033910.

Non-invasive assessment of liver disease in rats using multiparametric magnetic resonance imaging: a feasibility study

Anna M Hoy  1 , Natasha McDonald  1 , Ross J Lennen  2   3 , Matteo Milanesi  4 , Amy H Herlihy  4 , Timothy J Kendall  1   5 , William Mungall  6 , Michael Gyngell  4 , Rajarshi Banerjee  4 , Robert L Janiczek  7 , Philip S Murphy  7 , Maurits A Jansen  2   3 , Jonathan A Fallowfield  8
Affiliations

Non-invasive assessment of liver disease in rats using multiparametric magnetic resonance imaging: a feasibility study

Anna M Hoy et al. Biol Open. .

Abstract

Non-invasive quantitation of liver disease using multiparametric magnetic resonance imaging (MRI) could refine clinical care pathways, trial design and preclinical drug development. The aim of this study was to evaluate the use of multiparametric MRI in experimental models of liver disease. Liver injury was induced in rats using 4 or 12 weeks of carbon tetrachloride (CCl4) intoxication and 4 or 8 weeks on a methionine and choline deficient (MCD) diet. Liver MRI was performed using a 7.0 Tesla small animal scanner at baseline and specified timepoints after liver injury. Multiparametric liver MRI parameters [T1 mapping, T2* mapping and proton density fat fraction (PDFF)] were correlated with gold standard histopathological measures. Mean hepatic T1 increased significantly in rats treated with CCl4 for 12 weeks compared to controls [1122±78 ms versus 959±114 ms; d=162.7, 95% CI (11.92, 313.4), P=0.038] and correlated strongly with histological collagen content (rs=0.717, P=0.037). In MCD diet-treated rats, hepatic PDFF correlated strongly with histological fat content (rs=0.819, P<0.0001), steatosis grade (rs=0.850, P<0.0001) and steatohepatitis score (rs=0.818, P<0.0001). Although there was minimal histological iron, progressive fat accumulation in MCD diet-treated livers significantly shortened T2*. In preclinical models, quantitative MRI markers correlated with histopathological assessments, especially for fatty liver disease. Validation in longitudinal studies is required.This article has an associated First Person interview with the first author of the paper.

Keywords: Chronic liver disease; Multiparametric MRI; Rat.

PubMed Disclaimer

Conflict of interest statement

Competing interestsA.H.H., M.G., M.M. and R.B. are employees of Perspectum Diagnostics Ltd, the developer of LiverMultiScan™. A.H.H., M.G. and R.B. hold stock in the company. R.L.J. and P.S.M. are employees of GlaxoSmithKline and hold stock in the company. The remaining authors have no relevant conflicts of interest to declare. This is an academic led and reported study, with industry engagement. The role of Perspectum Diagnostics Ltd was the provision of access to multiparametric MRI methodology and blinded analysis of raw MRI data. Study design and potential conflicts do not affect adherence to policies on sharing data and materials. All study investigations, data analysis, manuscript preparation and decision to submit was undertaken by the academic center.

Figures

Fig. 1.
Fig. 1.
MRI assessment of hepatic fibro-inflammatory injury in rat CCl4 model. (A) Quantification of hepatic collagen content (% area of Picrosirius Red staining). (B) Representative examples of transverse liver MR T1 relaxation maps. (C) T1 relaxation quantification in rats receiving CCl4 or OO for 4 and 12 weeks. (D) Correlation between hepatic collagen content and T1 in rats receiving CCl4 and OO for 12 weeks. Data presented as mean±s.d., analyzed by unpaired t-test (*P<0.05, **P<0.01). Spearman (rs) correlation coefficient was used to examine correlations.
Fig. 2.
Fig. 2.
MRI assessment of hepatic steatosis and steatotic inflammatory injury in rat MCD diet model. (A) Representative examples of transverse liver MR PDFF maps. (B) Quantification of Oil Red O (ORO) staining of hepatic fat. (C) Quantification of hepatic PDFF. (D-F) Correlations between hepatic PDFF and (D) histologically assessed steatosis, (E) steatotic inflammatory injury [N(m)AS] and (F) ORO staining in rats receiving MCD or control diet (N) for 4 and 8 weeks. Data presented as mean±s.d., analyzed by unpaired t-test (**P<0.01, ***P<0.001, ****P<0.0001). Spearman (rs) correlation coefficient was used to examine correlations.
Fig. 3.
Fig. 3.
MRI assessment of hepatic steatosis and steatohepatitis in rat CCl4 model. (A) Representative examples of transverse liver MR PDFF maps. (B) Quantification of Oil Red O (ORO) staining of hepatic fat. (C) Quantification of hepatic proton density fat fraction (PDFF). (D,E) Correlations between hepatic PDFF and (D) histologically assessed ORO staining and (E) steatotic inflammatory injury [N(m)AS] in rats receiving CCl4 or olive oil (OO) for 4 and 12 weeks. Data presented as mean±s.d., analyzed by unpaired t-test (*P<0.05, **P<0.01). Spearman (rs) correlation coefficient was used to examine correlations.
Fig. 4.
Fig. 4.
MRI assessment of hepatic steatosis and steatohepatitis in cilostazol-treated rats on MCD diet. Sprague-Dawley rats received MCD diet for 4 weeks plus cliostazol or vehicle for the last 2 weeks of the model. Animals were scanned at baseline and after 4 weeks. (A) Quantification of hepatic fat by morphometry based on H&E staining. (B) Steatotic inflammatory injury [N(m)AS]. (C) Representative examples of transverse liver MR proton density fat fraction (PDFF). (D) Quantification of hepatic PDFF. (E) Correlation between PDFF and steatotic inflammatory injury [N(m)AS]. Data presented as mean±s.d., analyzed by unpaired t-test. Spearman (rs) correlation coefficient was used to examine correlations.
Fig. 5.
Fig. 5.
MRI assessment of T2*/R2* relaxation in preclinical liver injury models. (A) Representative examples of transverse liver MR T2* maps. (B) Quantification of R2* (1/T2*) in rats receiving CCl4 or olive oil (OO) for 4 and 12 weeks. (C) Quantification of R2* (1/T2*) in rats receiving MCD or control diet (N) for 4 and 8 weeks. (D) Correlation between R2* and PDFF in animals receiving CCl4 or OO injections and MCD or control diets. Data presented as mean±s.d., analyzed by unpaired t-test (****P<0.0001). Spearman (rs) correlation coefficient was used to examine correlations.
Fig. 6.
Fig. 6.
Rat models of chronic liver disease. *1 rat euthanized prematurely due to poor health.
See this image and copyright information in PMC

References

    1. Banerjee R., Pavlides M., Tunnicliffe E. M., Piechnik S. K., Sarania N., Philips R., Collier J. D., Booth J. C., Schneider J. E., Wang L. M. et al. (2014). Multiparametric magnetic resonance for the non-invasive diagnosis of liver disease. J. Hepatol. 60, 69-77. 10.1016/j.jhep.2013.09.002 - DOI - PMC - PubMed
    1. Bergmeyer H. U., Scheibe P. and Wahlefeld A. W. (1978). Optimization of methods for aspartate aminotransferase and alanine aminotransferase. Clin. Chem. 24, 58-73. - PubMed
    1. Bonny J.-M., Zanca M., Boire J.-Y. and Veyre A. (1996). T2 maximum likelihood estimation from multiple spin-echo magnitude images. Magn. Reson. Med. 36, 287-293. 10.1002/mrm.1910360216 - DOI - PubMed
    1. Britton L. J., Subramaniam V. N. and Crawford D. H. G. (2016). Iron and non-alcoholic fatty liver disease. World J. Gastroenterol. 22, 8112-8122. 10.3748/wjg.v22.i36.8112 - DOI - PMC - PubMed
    1. Bydder M., Yokoo T., Hamilton G., Middleton M. S., Chavez A. D., Schwimmer J. B., Lavine J. E. and Sirlin C. B. (2008). Relaxation effects in the quantification of fat using gradient echo imaging. Magn. Reson. Imaging 26, 347-359. 10.1016/j.mri.2007.08.012 - DOI - PMC - PubMed

LinkOut - more resources