The Kline Lab is led by Jeff Kline, MD, a translational researcher in the area of venous thromboembolism. This lab performs clinical and applied laboratory research intended to improve the therapeutic index of all steps of venous thromboembolism diagnosis and treatment.
Located in the Medical Science Building on the IU School of Medicine—Indianapolis campus, the Kline Lab spans 850 square feet of laboratory space and connects via corridor to IU Health University Hospital, where investigators collect samples from patients, which are analyzed in this lab. The three rooms of this lab include a biochemistry/protein production and purification laboratory; a lab equipped to measure hemostasis, coagulation, NOx and cellular metabolism; and a rat surgery area, with dander abatement equipment.
Investigators in this lab work with many different organisms—from an e coli expression vector used to make a mutant plasmin for dissolving blood clots to rats, pigs and humans with pulmonary embolism. The main therapeutic goals are to produce clot lysis while not increasing the bleeding risk and to protect the right ventricle from damage.
Specialized equipment in the Kline Lab includes a GE Healthcare ATKA 25 Pure M1 chromatography system for protein purification; a 96 plate well automated plate reader spectrophotometer (molecular devices) with SoftMax pro data acquisition and a -80C Revco monitored freezer for blood storage; and an STA compact coagulation platform capable of measuring d-dimer, fibrinogen, anti-Xa, protime, PTT, Rivaroxaban estimation, thrombin time, protein C, protein S and any other measurements for thrombophilia (AT, PS, PC), reptilase time, VWF and Lupus Anticoag markers. The lab also has two Haemoscope 5000 thromboelastometry machines with platelet mapping capability; a Sievers nitric oxide analyzer; an Oroboros O2k- high-resolution fluorometry and high fidelity respirometer; a fume hood and a Beckman Coulter Allegra X-14 4L benchtop refrigerated 10,200g max centrifuge; and a fully operational rat surgery area with temperature control, electronic pacing for heart rate control, gas delivery, high-fidelity digital manometer connection for pressure measurements with a DSI archiving system.
This translational research seeks to increase the therapeutic index of current treatment of pulmonary embolism, or rather to develop treatments that work better with less risk than current treatments propose. The broad interest of this project is to understand how PE produces a pathophysiological complex that worsens disease and quality of life. The two main mechanisms by which it does that is by unresolved, or even progressive pulmonary hypertension, which causes right ventricular dysfunction, and the “post PE syndrome” characterized by exercise intolerance and fatigue.
Additionally, PE produces platelet activation, which increases chance of recurrent clots. To study these events, the Kline Lab developed a rat model of PE that shows persistent pulmonary hypertension. Researchers are particularly interested in understanding how soluble guanylate cyclase (sGC) activation, administered in the first weeks after PE, prevents the progression of PE to chronic thromboembolic pulmonary hypertension (CTEPH).
This lab is interested in understanding how PE causes platelet activation followed by reactive oxygen species production, apoptosis, microparticle release and increased platelet turnover. Taken together, these effects contribute to hypercoagulability and possibly recurrent clotting. Researchers harvest platelets from patients without and with PE and study them with multiple methods to test for mechanisms of activation and potential therapeutic routes to reduce their transition to a highly thrombogenic phenotype. Techniques include high fidelity respirometry to study mitochondrial oxygen consumption under different conditions, the use of thromboelastography to measure platelet function in whole blood, and under fluorescent microscopy, using fluroprobes to measure platelet calcium transients in the presence of agonists and antagonists. With this model, investigators have found that increasing sGC activity ameliorates the exaggerated response of platelets from PE.