Second-Harmonic Generated Quantifiable Fibrosis Parameters Provide Signatures for Disease Progression and Regression in Nonalcoholic Fatty Liver Disease

Nicole Ng; Dean Tai; Yayun Ren; Elaine Chng; Mulugeta Seneshaw; Faridoddin Mirshahi; Michael Idowu; Amon Asgharpour; Arun J Sanyal

doi:10.1177/2632010X231162317

Second-Harmonic Generated Quantifiable Fibrosis Parameters Provide Signatures for Disease Progression and Regression in Nonalcoholic Fatty Liver Disease

Clin Pathol. 2023 Mar 26:16:2632010X231162317. doi: 10.1177/2632010X231162317. eCollection 2023 Jan-Dec.

Authors

Nicole Ng¹, Dean Tai², Yayun Ren², Elaine Chng², Mulugeta Seneshaw³, Faridoddin Mirshahi³, Michael Idowu⁴, Amon Asgharpour³, Arun J Sanyal³

Affiliations

¹ Virginia Commonwealth University School of Medicine, Richmond, VA, USA.
² Histoindex Pte. Ltd, Singapore.
³ Stravitz-Sanyal Institute of Liver Disease and Metabolic Health, Department of Internal Medicine, Virginia Commonwealth University School of Medicine, Richmond, VA, USA.
⁴ Department of Pathology, Virginia Commonwealth University School of Medicine, Richmond, VA, USA.

Abstract

Introduction: The current ordinal fibrosis staging system for nonalcoholic steatohepatitis (NASH) has a limited dynamic range. The goal of this study was to determine if second-harmonic generated (SHG) quantifiable collagen fibrillar properties (qFP) and their derived qFibrosis score capture changes in disease progression and regression in a murine model of NASH, in which disease progression can be induced by a high fat sugar water (HFSW) diet and regression by reversal to chow diet (CD).

Methods: DIAMOND mice were fed a CD or HFSW diet for 40 to 52 weeks. Regression related changes were studied in mice with diet reversal for 4 weeks after 48 to 60 weeks of a HFSW diet.

Results: As expected, mice on HFSW developed steatohepatitis with stage 2 to 3 fibrosis between weeks 40 and 44. Both the collagen proportionate area and the qFibrosis score based on 15 SHG-quantified collagen fibrillar properties in humans were significantly higher in mice on HFSW for 40 to 44 weeks compared to CD fed mice. These changes were greatest in the sinusoids (Zone 2) with further increase in septal and portal fibrosis related scores between weeks 44 and 48. Diet reversal led to decrease in qFibrosis, septal thickness, and cellularity with greatest changes in Zone 2. Specific qFPs associated with progression only, regression only, or both processes were identified and categorized based on direction of fibrosis change.

Conclusion: Complementing recent human studies, these findings support the concept that changes of disease progression and regression can be assessed using SHG-based image quantification of fibrosis related parameters.

Keywords: DIAMOND model; Nonalcoholic fatty liver disease; cirrhosis; collagen; disease activity; fibrosis progression; fibrosis regression; fibrosis stage; histology; mouse models; nonalcoholic steatohepatitis; quantifiable fibrosis parameters; second harmonic generation.