The Ischemic Leg Muscle is the Target Organ in PAD
Reduced perfusion causes muscle ischemia and leads to a feed-forward pathway of endothelial injury causing an increase in M1-like macrophage polarization and increased reactive oxygen species, each of which worsens muscle microvascular perfusion and ischemia.
Inflammation Driven by # M1
- Inhibits angiogenesis
- Increase injurious cytokines that access circulation
Greater Endothelial/Vascular Injury in Muscle
- loss microvascular density
- loss of vascular integrity i.e.: greater vascular, permeability
Increased Reactive Oxygen Species (ROS)
A few selected slides will show sites/effects from miR-93 delievry
- ROS directly injures endothelial cells, makes them leaky, and results in them being less able to deliver oxygen and remove toxins from muscle cells.
Patients with PAD have Increased levels of pro-inflammatory cytokines and lower levels of microRNA-93 in affected muscles
Levels of pro-inflammatory M1-macrophage are increased in ischemic muscle biopsies.
Levels of microRNA-93 are decreased in ischemic muscle biopsies.
microRNA-93 promotes formation of functional, non-leaky blood vessels
microRNA-93 significantly reduces levels of ROS under hypoxia/serum starvation (HSS) conditions
H2DCFD = 2′,7′ -dichlorodihydrofluorescein diacetate is a chemically reduced fluorescein and an indicator for reactive oxygen species (ROS) in cells
* Angiogenic properties of miR-93 in EC under HSS published in Hazarika et al. Circulation 2013;127:1818-28.
Above ROS data is included in recently submitted manuscript.
microRNA-93 treatment reduces levels of pro-inflammatory cytokine and M1-macrophage, while increasing M2-marcorphage levels
Pro-inflammatory cytokine levels are decreased in conditioned media from macrophage treated with microRNA-93 (miR93 Mim) or a scrambled oligonucleotide (Scrbd Mim)
microRNA-93 (miR93 Mim) treatment of human monocytes, increases M2-like macrophage while decreasing M1-like macrophage when compared to a scrambled oligonucleotide (Scrbd Mim)
INTELLECTUAL PROPERTY: Broad Protection for Treating PAD
Compositions and methods for treating peripheral arterial disease
- Inventors at the University of Virginia: Brian Annex, Charles Farber, Surovi Hazarika
- US 9,845,465 B2, issued 12/19/17
- European Patent 2885008B, issued 11/7/2018
- Covers the use of both miR-93 and pre-miR-93
- Exclusively licensed from University of Virginia for all fields and all territories.
Methods, Kits, Compositions for Reducing miR106B Activity
- Inventor: Brian Annex
- Assignee: Merand Pharmaceuticals, Inc.
- Provision patent application US 63/148,940 filed on 2/12/2021, with PCT application filed in 2022.