Disclosures: Scott Friedman is a consultant to Galmed

Aramchol Downregulates SCD1 and Induces PPARg

in Hepatic Stellate Cells to Attenuate Cellular Activation and FibrogenesisBrittany Allen1, Jose M. Mato2, Amanda Craig1, David Fernandez-Ramos2, Fernando Lopitz-Otsoa2, Liat Hayardeny3, Augusto Villanueva1 and Scott L Friedman1,

1Division of Liver Diseases, Icahn School of Medicine at Mount Sinai, New York, NY 2CIC Biogune, Bilbao, Spain 3Galmed Pharmaceuticals, Tel Aviv, Israel

1. INTRODUCTION• Activation of hepatic stellate cells (HSCs) drives hepatic fibrosis

through a process that is inhibited by PPARg signaling.

• Aramchol (arachidyl amido cholanoic acid) is a fatty acid-bile acid

conjugate that reduces liver fat content in nonalcoholic fatty liver

disease (NAFLD) and improved nonalcoholic steatohepatitis (NASH)

without worsening fibrosis in a Phase 2b study (Abstract # LB-5).

• Aramchol attenuates fibrosis in two distinct models (MCD diet and

thioacetamide). In mice, Aramchol reduces liver fat by

downregulating the fatty acid synthetic enzyme stearoyl Co-A

desaturase 1 (SCD-1) in hepatocytes.

• The mechanism for Aramchol’s antifibrotic effect is not known;

moreover, although HSCs also store lipids as retinyl esters.

• The role of SCD-1 in HSCs and the impact of Aramchol on SCD-1

activity are unknown. .

3. METHODS• Serum-starved LX2 cells were treated with Aramchol (10 µM) for 24

or 48 hours. Fibrogenic gene expression and SCD-1 protein

expression were measured by qPCR and Western, respectively.

• RNAseq was performed at 24 and 48 h in duplicate. Differential gene

expression was assessed with DSeq2, while gene set enrichment

analysis (GSEA) and gene ontology (GO) analyses assessed

functionality of gene expression changes.

• Primary hepatocytes from C57BL/6 mice were harvested by

perfusion and treated with Aramchol (10 µM) for 48 hours in culture.

PPAR 𝛄 mRNA expression was measured by qPCR.

Figure 1: Aramchol downregulates SCD-1 mRNA and fibrogenic

genes, and upregulates PPAR𝛄 mRNA in HSCs. Based on RT-

PCR (N = 3) *p<0.05, **p<0.01, ***p<0.001.

2. AIM• To investigate the direct anti-fibrotic effect of Aramchol on HSCs

using LX-2, a human hepatic stellate cell line

• To define the mechanism of Aramchol’s effects on HSCs.

4. RESULTS

CONCLUSIONS• Aramchol elevates PPAR𝛄 mRNA and down regulates SCD-1 mRNA and protein and in hepatic stellate cells.

• By RNAseq and pathway analysis, Aramchol downregulates fibrogenic genes that are part of a clinically validated

HSC activation signature, including COL1A1 and ⍺SMA, as well as pathways involved in cholesterol biosynthesis &

homeostasis, and collagen formation.

• Aramchol upregulates PPAR𝛄 mRNA selectively in HSCs and not in hepatocytes.

Figure 5: Aramchol downregulates cholesterol

biosynthesis and collagen formation in HSCs. Gene Set

Enrichment Analysis of LX-2 cells after 24 (A) or 48 (B) hours

of treatment.

Funding support: Galmed Pharmeceuticals

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Figure 2: Aramchol downregulates SCD-1

in HSCs. Western Blot (A) and densitometry

(B) of downregulated SCD-1 protein in HSCs

treated with Aramchol for 24 or 48 hrs. (N=3)

*p<0.05, **p<0.01, ***p<0.001

Figure 3: RNASeq confirms downregulation of fibrogenic genes

and SCD-1 mRNA, and upregulation of PPAR𝛄 mRNA after 48 hrs

of treatment with Aramchol. The cholesterol efflux regulatory protein,

ABCA1, is also upregulated.

Figure 4: Aramchol does not

affect PPAR𝛄 mRNA expression

in primary mouse hepatocytes

after 48 hours of treatment. (N=3)

*p<0.05, **p<0.01, ***p<0.001

V V V 10 10 10 V V V 10 10 10

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