Delineating tesamorelin response pathways in HIV-associated NAFLD using a targeted proteomic and transcriptomic approach

Lindsay T Fourman, Takara L Stanley, James M Billingsley, Shannan J Ho Sui, Meghan N Feldpausch, Autumn Boutin, Isabel Zheng, Colin M McClure, Kathleen E Corey, Martin Torriani, David E Kleiner, Colleen M Hadigan, Raymond T Chung, Steven K Grinspoon, Lindsay T Fourman, Takara L Stanley, James M Billingsley, Shannan J Ho Sui, Meghan N Feldpausch, Autumn Boutin, Isabel Zheng, Colin M McClure, Kathleen E Corey, Martin Torriani, David E Kleiner, Colleen M Hadigan, Raymond T Chung, Steven K Grinspoon

Abstract

NAFLD is a leading comorbidity in HIV with an exaggerated course compared to the general population. Tesamorelin has been demonstrated to reduce liver fat and prevent fibrosis progression in HIV-associated NAFLD. We further showed that tesamorelin downregulated hepatic gene sets involved in inflammation, tissue repair, and cell division. Nonetheless, effects of tesamorelin on individual plasma proteins pertaining to these pathways are not known. Leveraging our prior randomized-controlled trial and transcriptomic approach, we performed a focused assessment of 9 plasma proteins corresponding to top leading edge genes within differentially modulated gene sets. Tesamorelin led to significant reductions in vascular endothelial growth factor A (VEGFA, log2-fold change - 0.20 ± 0.35 vs. 0.05 ± 0.34, P = 0.02), transforming growth factor beta 1 (TGFB1, - 0.35 ± 0.56 vs. - 0.05 ± 0.43, P = 0.05), and macrophage colony stimulating factor 1 (CSF1, - 0.17 ± 0.21 vs. 0.02 ± 0.20, P = 0.004) versus placebo. Among tesamorelin-treated participants, reductions in plasma VEGFA (r = 0.62, P = 0.006) and CSF1 (r = 0.50, P = 0.04) correlated with a decline in NAFLD activity score. Decreases in TGFB1 (r = 0.61, P = 0.009) and CSF1 (r = 0.64, P = 0.006) were associated with reduced gene-level fibrosis score. Tesamorelin suppressed key angiogenic, fibrogenic, and pro-inflammatory mediators. CSF1, a regulator of monocyte recruitment and activation, may serve as an innovative therapeutic target for NAFLD in HIV. Clinical Trials Registry Number: NCT02196831.

Conflict of interest statement

L.T.F. has served as a consultant for Theratechnologies, Inc. T.L.S. has served as a consultant for Theratechnologies, and reports research funding from Novo Nordisk, Inc. J.M.B. and S.J.H.S. report research funding from Boehringer Ingelheim. S.J.H.S. also reports research funding from AstraZeneca. K.E.C. reports research funding from Boehringer Ingelheim and has served as a consultant to Novo Nordisk, Inc, and Bristol Myers Squibb. R.T.C. reports research funding from Abbvie, Gilead, Merck, Bristol Myers Squibb, Boehringer Ingelheim, Janssen, and Roche. S.K.G. has served as a consultant (personal) and currently serves as a consultant through an Institutional Consulting Agreement with Theratechnologies. The Mass General Hospital has a royalty and license agreement with Theratechnologies for Tesamorelin. Dr. Grinspoon is the inventor on a patent for GHRH or Analogues Thereof For Use in Treatment of Hepatic Disease (application 16832128). M.N.F., A.B., I.Z., C.M.M., M.T., D.E.K., and C.M.H. report no conflicts of interest.

Figures

Figure 1
Figure 1
Analysis Schema. A total of 9 plasma proteins were examined, corresponding to top leading edge genes within hepatic gene pathways differentially modulated by tesamorelin. Abbreviations: CASP8, caspase 8; CCL20, C–C motif chemokine ligand 20; CRTAM, cytotoxic and regulatory T-cell molecule; CSF1, macrophage colony stimulating factor 1; CXCL12, C-X-C motif chemokine ligand 12; NCR1, natural cytotoxicity triggering receptor 1; TGFB1, transforming growth factor beta 1; TNFRSF21, tumor necrosis factor receptor superfamily member 21; VEGFA, vascular endothelial growth factor A.
Figure 2
Figure 2
Differential Changes in Plasma VEGFA, TGFB1, and CSF1 by Treatment Status. Tesamorelin led to significant reductions in plasma (A) VEGFA (log2-fold change, mean ± SD, − 0.20 ± 0.35 vs. 0.05 ± 0.34, P = 0.02), (B) TGFB1 (log2-fold change − 0.35 ± 0.56 vs. − 0.05 ± 0.43, P = 0.05), and (C) CSF1 (log2-fold change − 0.17 ± 0.21 vs. 0.02 ± 0.20, P = 0.004) compared to placebo. Bars and error bars indicate mean and standard error of the mean, respectively. Abbreviations: CSF1, macrophage colony stimulating factor 1; TGFB1, transforming growth factor beta 1; VEGFA, vascular endothelial growth factor A.
Figure 3
Figure 3
Relationship of Changes in Plasma VEGFA and CSF1 with Change in NAS Score in Tesamorelin-Treated Participants. Within the tesamorelin-treated arm, reductions in plasma (A) VEGFA (r = 0.62, P = 0.006) and (B) CSF1 (r = 0.50, P = 0.04) were associated with a decrease in NAS score. Linear regression lines with 95% confidence intervals are shown. Abbreviations: CSF1, macrophage colony stimulating factor 1; NAS, NAFLD activity score; VEGFA, vascular endothelial growth factor A.
Figure 4
Figure 4
Relationship of Changes in Plasma TGFB1 and CSF1 with Change in Gene-Level Fibrosis Score in Tesamorelin-Treated Participants. Among tesamorelin-treated participants, declines in plasma (A) TGFB1 (r = 0.61, P = 0.009) and (B) CSF1 (r = 0.64, P = 0.006) were associated with improved gene-level fibrosis score. Linear regression lines with 95% confidence intervals are shown. Abbreviations: CSF1, macrophage colony stimulating factor 1; TGFB1, transforming growth factor beta 1.

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