Update on a brain-penetrant cardiac glycoside that can lower cellular prion protein levels in human and guinea pig paradigms

• Published in: PloS one 2024-09, Vol.19 (9), p.e0308821
• Main Authors: Eid, Shehab, Zhao, Wenda, Williams, Declan, Nasser, Zahra, Griffin, Jennifer, Nagorny, Pavel, Schmitt-Ulms, Gerold
• Format: Article
• Language: English
• Subjects: Adenosine triphosphatase; Analysis; Animal models; Animals; Antibodies; Biology and Life Sciences; Brain; Brain - drug effects; Brain - metabolism; Brain slice preparation; Bupivacaine; Cardenolides - metabolism; Cardenolides - pharmacology; Cardiac glycosides; Cardiac Glycosides - pharmacology; Care and treatment; Cell culture; Cell differentiation; Cell Line; Cell lines; Cell surface; Cells; Cloning; Coronary artery disease; Diagnosis; Dosage and administration; Ethylenediaminetetraacetic acid; Gene therapy; Glioma; Glycosides; Guinea Pigs; Heart diseases; Humans; In vivo methods and tests; Invoices; Lead compounds; Lubrication and lubricants; Lysates; Medicine and Health Sciences; Mice; Neurons; Patient outcomes; Penicillin; Prion diseases; Prion Diseases - drug therapy; Prion Diseases - metabolism; Prion protein; Prions; Proteins; PrPC Proteins - metabolism; Research and Analysis Methods; Scientific equipment and supplies industry; Sodium-Potassium-Exchanging ATPase - metabolism; Swine; Toxicity; Zinc finger proteins
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0308821

Lowering the levels of the cellular prion protein (PrPC) is widely considered a promising strategy for the treatment of prion diseases. Building on work that established immediate spatial proximity of PrPC and Na+, K+-ATPases (NKAs) in the brain, we recently showed that PrPC levels can be reduced by targeting NKAs with their natural cardiac glycoside (CG) inhibitors. We then introduced C4'-dehydro-oleandrin as a CG with improved pharmacological properties for this indication, showing that it reduced PrPC levels by 84% in immortalized human cells that had been differentiated to acquire neural or astrocytic characteristics. Here we report that our lead compound caused cell surface PrPC levels to drop also in other human cell models, even when the analyses of whole cell lysates suggested otherwise. Because mice are refractory to CGs, we explored guinea pigs as an alternative rodent model for the preclinical evaluation of C4'-dehydro-oleandrin. We found that guinea pig cell lines, primary cells, and brain slices were responsive to our lead compound, albeit it at 30-fold higher concentrations than human cells. Of potential significance for other PrPC lowering approaches, we observed that cells attempted to compensate for the loss of cell surface PrPC levels by increasing the expression of the prion gene, requiring daily administration of C4'-dehydro-oleandrin for a sustained PrPC lowering effect. Regrettably, when administered systemically in vivo, the levels of C4'-dehydro-oleandrin that reached the guinea pig brain remained insufficient for the PrPC lowering effect to manifest. A more suitable preclinical model is still needed to determine if C4'-dehydro-oleandrin can offer a cost-effective complementary strategy for pushing PrPC levels below a threshold required for long-term prion disease survival.