<b>Ponatinib and other clinically approved inhibitors of Src and Rho-A kinases</b><b>abrogate dengue virus serotype 2- induced endothelial permeability</b>

<b>Ponatinib and other clinically approved inhibitors of Src and Rho-A kinases</b><b>abrogate dengue virus serotype 2- induced endothelial permeability</b><br>Srishti Rajkumar Mishra ^1#, Ayan Modak¹, Mansi Awasthi^1#, Archana Sobha ², Easwaran Sreekumar^1,3*<br>¹Molecular Virology Laboratory; ²Animal Research Facility, Rajiv Gandhi Centre forBiotechnology (RGCB), Thiruvananthapuram 695014, Kerala, India³Molecular Bioassay Laboratory, Institute of Advanced Virology (IAV), Bio360 LifeSciences Park, Thiruvananthapuram-695317, Kerala, India^#Regional Centre for Biotechnology (RCB), Faridabad, New Delhi<br>*Corresponding author (Easwaran Sreekumar; Molecular Virology Laboratory; RajivGandhi Centre for Biotechnology (RGCB), Thycaud P.O, Thiruvananthapuram695014, Kerala, India; Tel:+91-9447698943; E-mail: [email protected];)<br><br><br>Severe dengue often presents as shock syndrome with enhanced vascular permeability and plasma leakage into tissue spaces. <i>In vitro</i> studies have documented the role of Src family kinases (SFKs) and RhoA-kinases (ROCK) in dengue virus serotype 2 (DENV2)-induced endothelial permeability. Here, we show that the FDA-approved SFK inhibitors Bosutinib, Vandatenib and Ponatinib, as well as the ROCK inhibitors, Netarsudil and Ripasudil significantly inhibit DENV2-induced endothelial permeability. In cultured telomerase immortalized human microvascular endothelial cells (HMEC-1), treatment with these inhibitors reduced the phosphorylation of VE-Cadherin, Src and myosin light chain 2 (MLC2) proteins that were upregulated during DENV2 infection. It also prevented the loss of VE-cadherin from the inter-endothelial cell junctions induced by viral infection. In <i>in-vivo</i> studies using DENV2-infected AG129 IFN receptor-a/b/g deficient mice, ponatinib, when administered 24h post-infection onwards, demonstrated significant benefits in improving body weight, clinical outcomes, and survival rates. While all virus-infected, untreated mice died by day-10 post-infection, 80% of the ponatinib-treated mice survived, and approximately 60% were still alive at the end of the 15-day observation period. The treatment also significantly reduced disease severity factors such as vascular leakage, thrombocytopenia; and restored liver function. Comparable effects were observed even when ponatinib treatment was initiated after symptom onset. The results highlight ponatinib as an effective therapeutic option in severe dengue; and also a similar potential for other FDA- approved SFK and ROCK inhibitors.