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Global Translational Medicine GalNAc AGT ASO reduce atherosclerosis

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Figure 1. Antisense oligonucleotides (ASO) targeting hepatic angiotensinogen (AGT) reduced plasma AGT concentrations, systolic blood pressure, and
atherosclerotic lesion area in the thoracic aorta. (A) Experimental protocol. (B) Plasma AGT concentrations on weeks -2, 0, 4, and 11 during ASO
administration. A piecewise mixed-effect model was fitted to examine slopes for distinct time points (-2 to 0 and 0 to 11 weeks). Doses 1, 2.5, and 5 mg/
kg led to profound reductions in plasma AGT concentrations compared to week -2; P < 0.001. (C) Mouse systolic blood pressure (BP) on weeks -2, 3, 7,
and 11. A mixed-effect model was used to compare slopes among the four groups. Doses 1, 2.5, and 5 mg/kg led to significant reductions in systolic BP
compared to week -2; P < 0.001. (D) En face aorta images, scale bar = 1 mm. (E) Atherosclerotic lesion area of the thoracic aorta normalized by total intimal
surface area; N = 6–10 per group.
GalNAc AGT ASO: N-acetylgalactosamine-conjugated antisense oligonucleotides targeting angiotensinogen; PBS: Phosphate-buffered saline;
SQ: Subcutaneous.

2.3. Plasma profiles #: IB59131, IBL-America) after being incubated with

Mouse blood was collected in the presence of exogenous recombinant mouse AGT for 1 h.
ethylenediaminetetraacetic acid (EDTA; final Plasma alanine aminotransferase (ALT) and aspartate
concentration: 1.8 mg/mL) through submandibular aminotransferase (AST) were measured on the Olympus
bleeding during the study. At termination, blood was AU400 clinical analyzer (Olympus, Center Valley, PA)
collected through cardiac bleeding through the right using L-Type ALT.J2 and L-Type AST.J2 reagents and
ventricle. Plasma AGT concentrations were measured calibrators (Fujifilm Healthcare) in Ionis in a blinded
using a mouse AGT enzyme-linked immunosorbent assay manner.
(ELISA) kit (Cat # 245718, Abcam), which detects both
intact AGT and des(AngI)AGT, thereby measuring total 2.4. Quantification of atherosclerosis
AGT. Plasma renin concentrations in mice were measured After euthanasia, mouse aortas were dissected and fixed in
by an enzyme-kinetic assay using an AngI ELISA kit (Cat 10% neutrally buffered formalin overnight. Subsequently,

Volume 2 Issue 1 (2023) 3 https://doi.org/10.36922/gtm.288