eng Peripheral Artery Disease (PAD) is a chronic vascular condition characterized by the narrowing or obstruction of arteries, primarily in the lower limbs, leading to ischemic complications. Unlike atheromatous PAD, where lipid-rich plaques obstruct blood flow, exists a distinct and more severe subtype, defined as calcific PAD. Calcific PAD is driven by calcification in the medial arterial layer, reducing vascular compliance and causing severe ischemia and a worse clinical profile. Calcific PAD progresses rapidly, resulting in intense pain, reduced mobility, non-healing wounds, and a higher risk of amputation, and mortality than traditional PAD. This condition is particularly prevalent in high-risk populations, including individuals with chronic kidney disease (CKD), due to underlying vascular and metabolic dysfunctions, systemic mineral imbalances, and chronic inflammation. Standard PAD therapies are largely ineffective against MAC due to its non-obstructive nature. As a result, calcific PAD patients face limited treatment options and poorer outcomes. Despite the severe clinical implications, focused research dedicated to understanding and treating this condition remains insufficient and affected patients are frequently excluded from major clinical trials. This issue is further compounded by the absence of preclinical models that accurately replicate the pathophysiology of calcific PAD, making it difficult to explore the underlying mechanisms or test potential therapies. Addressing these gaps in research and treatment is essential to improve outcomes for this vulnerable patient population. This thesis aims to establish an effective preclinical model of PAD driven by cardiovascular calcification (CVC). The primary objective is to replicate key disease features, such as lower limb ischemia status and impaired locomotor function, both induced by ectopic calcification in rats. Additionally, the study evaluates cilostazol, a standard treatment for atherosclerotic PAD, alongside SNF472, an inhibitor of calcification, to compare their therapeutic effects on key disease parameters. Our research involved determining the optimal dosage and induction period of vitamin D3 to achieve a calcification degree capable of simulating the pathological state of the disease. Advanced imaging techniques (histology, micro-computed tomography (µ–CT), and scanning electron microscopy (SEM), were employed for a comprehensive characterization of induced CVC. The thesis encompasses four in vivo trials to establish a robust and reproducible methodology. Ischemic severity was quantified using Laser Speckle Contrast Imaging (LSCI), while treadmill tests were used to evaluate locomotor capacity. Total tissue calcification was assessed through calcium quantification via Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES), supplemented by the analysis of relevant blood biomarkers. Two efficacy studies and one progression study were conducted to compare the therapeutic effects of cilostazol and SNF472, along with an additional study examining the dose-response and administration regimens for SNF472. We have successfully developed a rat model that accurately replicates the pathological condition of the calcific PAD in humans. In this model, we verified the lack of efficacy of cilostazol and demonstrated that SNF472 effectively inhibits CVC, resulting in superior outcomes across all evaluated parameters. These results suggest that SNF472 could be a promising therapeutic candidate for patients suffering from this severe form of PAD.
Marc Blasco Ferrer (Tue,) studied this question.