Inflammation and thrombosis fuel cardiovascular and pulmonary disease: Focus on the interplay of neutrophil inflammasomes with NETs. For many years, we have conducted a successful research program studying the links between thrombosis and inflammation. We plan to continue this line of investigation with the current emphasis on neutrophil extracellular traps (NETs) and the role inflammasomes plays in neutrophils and NETosis. Upon neutrophil activation, peptidylarginine deiminase 4 (PAD4) citrullinates histones and promotes inflammasome assembly needed for effective NETosis and IL-beta production. NETs trap microbes but we have shown a dark side of NETs, i.e., they promote thrombosis, inflammation and age-related heart and lung fibrosis. The central hypotheses of our program are as follows: NETs are involved in the formation of a stable, organized, and vascularized thrombus and breaking up NETs is necessary for thrombolysis. Thrombosis promotes deposition of NETs in both the adjacent vessel wall and in distant organs, leading to post-thrombotic syndrome and an increased systemic pro- coagulant and pro-inflammatory state. Inflammation, activating inflammasome and NET generation as seen in rheumatoid arthritis has systemic consequences such as the development of heart failure with preserved ejection fraction (HFpEF). Inhibiting inflammation by NLRP3 inflammasome or PAD4 inhibitor in mice will reduce the systemic effects and alleviate HFpEF development. We hypothesize further that in many diseases, inhibition of NET formation and reduction of thromboinflammation would be beneficial to the host. We propose to test these hypotheses using mouse and human blood cells, knockout mice and murine disease models we have developed. The work by the “Wagner Lab” is considered innovative and solid; an objective measure is its high citation. Obtaining prolonged funding would free time for more mentoring, innovative thinking, and helpful collegiate activities. In addition, we now study chronic inflammatory and thrombotic diseases and their impact on aging. These experiments take time, and the extended duration of support would assure that we can pursue this exciting research program effectively.