Thrombosis, Hemostasis and Vascular Biology

Our laboratory has two distinct research focuses:

1. Basic science studies of hemostasis and vascular biology with a focus on the physiological function of Tissue Factor Pathway Inhibitor (TFPI)
2. Clinical and translational research studies of iron metabolism in blood donors
   
   

Basic Science Studies:

The blood vessel contains several anticoagulant proteins that prevent the formation of blood clots. The work in our laboratory is focused on one of these proteins called tissue factor pathway inhibitor (TFPI). Humans without TFPI have not been identified and genetically altered mice that do not make TFPI die during embryonic development demonstrating that TFPI has critically important physiological functions.  TFPI is a serine protease inhibitor that exerts anticoagulant activity during the very early stages of blood coagulation. Our laboratory uses a wide variety of biochemical, cellular (tissue culture) and animal models to define and characterize the physiological functions of TFPI. These include studies of the biochemical mechanisms that allow TFPI to inhibit blood coagulation proteases and associate with the surface of blood vessels and platelets, characterization and funtional assessment of different alternatively spliced forms of TFPI and the development of new mouse models to examine the function of TFPI in vivo.  The results of these studies will shed new light on a wide range of human bleeding disorders, such as hemophilia, and diseases that are associated or caused by intravascular blood clots, such as deep venous thrombosis, heart attack, stroke and vascular dementia.

Clinical and Translational Studies:  

Blood donation removes a large amount of iron from the blood donor.  Since blood donation is allowed every 56 days in the United States, repeated donation produces iron deficiency and iron deficiency anemia in many donors.  Thus, blood donors are a unique population of healthy individuals for study of dietary iron absorbtion, hemoglobin production and the adverse consequences of iron deficiency.  We have found that some donors can donate blood repeatedly over many years and not develop iron deficiency anemia suggesting that they are able to absorb dietary iron more efficiently that average.  We perform a variety of studies to define their behavioral, biochemical and genetic characteristics that allow them to donate so frequently and are leading a multi-institutional study to identify the best ways to prevent development of iron deficiency in blood donors.  In addition, we direct the Wisconsin Hub of the National Heart Lung and Blood Institute's Recipient Epidemiology and Donor Evaluation Study (REDS-III).  REDS-III is a multi-institutional group of investigators who perform a number of studies to ensure the safety and availability of blood products in the United States.

Basic Science Grant Support:

• National Heart Lung and Blood Institute R01 Award 08/2002-12/2023 (Prinicpal Investigator)
• Novo Nordisk Research Grant 2007-2020 (Principal Investigator)
• National Institute for Child Health and Development R01 Award 07/2019-06/2023 (Co-Investigator) 

Clinical and Translational Studies Grant Support

• National Heart Lung and Blood Institute Retroviral Epidemiology Donor Study-III (REDS-III) 03/2011-02/2020 (Principal Investigator)
• National Heart Lung and Blood Institute Retroviral Epidemiology Donor Study-IV-Pediatrics (REDS-IV-P) 03/2019-02/2026 (Principal Investigator)
• National Heart Lung and Blood Institute R01 Award 01/2011-12/2015 (Principal Investigator)

Susan Maroney, DVM, PhD
Research Scientist II

Julie Peterson, PhD
Research Scientist II 

Nicholas Martinez 
Research Technologist II

Amy Siebert-McKenzie
Postdoctoral Fellow

Selected basic science publications:

• Warshawsky I, Bu G, Mast A, Saffitz JE, Broze GJ, Jr, Schwartz AL: The carboxy terminus of tissue factor pathway inhibitor is required for interacting with hepatoma cells in vitro and in vivo.  J Clin Invest 1995; 95:1773-1781
• Mast AE, Broze GJ, Jr: Physiological concentrations of tissue factor pathway inhibitor do not inhibit prothrombinase. Blood 1996; 87:1845-1850
• Mast AE, Higuchi DA, Huang Z-F, Warshawsky I, Schwartz AL, Broze GJ, Jr: Glypican-3 is a binding protein on the HepG2 cell surface for tissue factor pathway inhibitor. Biochem J 1997; 327:577-583. PMC1218832
• Lockett JM, Mast AE: The contribution of the region between Gly-160 and the end of the third Kunitz domain to the anticoagulant activity of tissue factor pathway inhibitor. Biochemistry 2002, 41:4989-4997
• Cunningham AC, Hasty KA, Enghild JJ, Mast AE: Structural and functional characterization of tissue factor pathway inhibitor following degradation by matrix metalloproteinase-8. Biochem J 2002, 367:451-458
• Mast AE, Acharya N, Malecha MJ, Hall CL, Dietzen DJ: Characterization of the association of tissue factor pathway inhibitor with human placenta. Arter. Thromb. and Vasc. Biol. 2002, 2099-2104
• Maroney SA, Cunningham AC, Ferrel J, Hu R, Haberichter S, Mansbach CM, Brodsky RA, Dietzen DJ, Mast AE: A GPI-anchored co-receptor for tissue factor pathway inhibitor controls its intracellular trafficking and cell surface expression. J Thromb Hemost 2006, 4:1114-1124
• Donahue BS, Gailani D, Mast AE: Disposition of tissue factor pathway inhibitor during cardiopulmonary bypass. J Thromb Hemost 2006, 4:1011-1016
• Maroney SA, Haberichter SL, Friese P, Collins ML, Ferrel JP, Dale GL, Mast AE: Active tissue factor pathway inhibitor is expressed on the surface of coated platelets. Blood 2007, 109:1931-1937
• Timmer JC, Enoksson M, Wildfang E, Zhu W, Igarashi Y, Denault JB, Ma Y, Dummitt B, Chang YH, Mast AE, Eroshkin A, Smith J, Tao WA, Salvesen GS: Profiling constitutive proteolytic events in vivo. Biochem J 2007, 407:41-48
• Maroney SA, Cooley BC, Sood R, Weiler H, Mast AE: Combined tissue factor pathway inhibitor and thrombomodulin deficiency produces an augmented hypercoagulable state with tissue specific fibrin deposition. J Thromb Hemost, 2008, 6:111-117
• Maroney SA, Ferrel JP, Collins ML, Mast AE: TFPIγ is an active alternatively spliced form of TFPI present in mice but not humans. J Thromb Hemost, 2008, 6:1344-1351
• Maroney SA, Ferrel JP, Pan S, White TA, Simari RD, McVey JH, Mast AE: Temporal expression of alternatively spliced forms of tissue factor pathway inhibitor in mice. J Thromb Hemost, 2009, 7:1106-1113
• White TA, Johnson T, Zarzhevsky N, Tom C, Delacroix S, Holroyd EW, Maroney SA, Singh R, Pan S, Fay WP, van Deursen J, Mast AE, Sandhu GS, Simari RD: Endothelial-derived tissue factor pathway inhibitor regulates arterial thrombosis but is not required for development or hemostasis. Blood, 2010, 116:1787-1794
• Zaman FS, Mast AE, Hall CL: Evaluating surface bound rTFPI through an in vitro model of vessel wall injury. Thrombosis Research, 2011, 127:149-153
• Maroney SA, Mast AE: Tissue factor pathway inhibitor and bacterial infection. J Thromb Haemost, 2011, 9:119-121
• Maroney SA, Cooley BC, Ferrel JP, Bonesho CE, Mast AE: Murine Hematopoietic Cell Tissue Factor Pathway Inhibitor Limits Thrombus Growth. Arterioscler Thromb Vasc Biol, 2011, In Press
• Maroney SA and Mast AE: Tissue factor pathway inhibitor and bacterial infection. Commentary. J. Thromb Hemost, 2011, 9:119-121
• Gilbert GE, Mast AE: Curbing an inhibitor of hemostasis. Commentary. Blood, 2011, 117:5277-5278
• Maroney SA, Ellery PE, Wood JP, Ferrel JP, Bonesho CE, Mast AE: Caveolae optimize tissue factor-factor VIIa activity of cell surface associated tissue factor pathway inhibitor. Biochem J, 2012, 443:259-263
• Maroney SA, Cooley BC, Ferrel JP, Bonesho CE, Nielsen LV, Johansen PB, Hermit MB, Petersen LC, Mast AE: Absence of hematopoietic tissue factor pathway inhibitor mitigates bleeding in mice with hemophilia. Proc Natl Acad Sci USA 2012, 109:3927-3931. PMC3309760.
  * A summary of this paper was published in The Hematologist, July 2012
• Mast AE: Alternatively spliced tissue factor pathway inhibitor: functional implications. Front Biosci, 2011, 3:1457-1462. PMC3549618
• Stephan F, Dienava I, Bulder I, Wouters D, Mast AE, Te Velthuis H, Aarden LA, Zeeleder S: Tissue factor pathway inhibitor is an inhibitor of factor VII activating protease. J Thromb Hemost 2012, 10:1165-1171. PMC3574557
• Maroney SA, Ellery PER, Wood JP, Ferrel JP, Martinez ND, Mast AE: Characterization of human Tissue Factor Pathway Inhibitor-β (TFPIβ) expression and inhibitory activity. J Thromb Hemost 2013, In Press.
• Maroney SA, Mast AE: (2012) Platelet Tissue Factor Pathway Inhibitor Modulates Intravascular Coagulation. Thromb Res, 129 Suppl 2: S21-22. PMC3345333
• Wolberg AS, Mast AE: (2012) Tissue factor and factor VIIa – Hemostasis and beyond. Thromb Res, 129 Suppl 2: S1-4. PMC3336011
• Maroney SA, Ellery PER, Wood JP, Ferrel JP, Martinez ND, Mast AE: Characterization of human Tissue Factor Pathway Inhibitor-beta (TFPI-beta) expression and inhibitory activity. J Thromb Hemost 2013; 11:911-918. PMC3656975
• Wood JP, Bunce MW, Maroney SA, Tracy PB, Camire RM, and Mast AE: Tissue factor pathway inhibitor-alpha inhibits prothrombinase during the initialtion of blood coagulation.  Proc Natl Acad Sci USA 2013; 110:17838-17843. PMC3816478
• Maroney SA, Hansen KG, Mast AE: (2013) Cellular expression and biological activities of alternatively spliced forms of tissue factor pathway inhibitor. Curr Opin Hematol, 20:403-409. PMC3813454
• Sosa I, Ellery PE, Neff A, Mast A, Gailani D: Acquired factor V deficiency in a patient without evidence of a classical inhibitor. Haemophilia 2014; 20:e81-e83. PMC3870044
• Ellery PE, Maroney SA, Martinez ND, Wickens MP, and Mast AE: Translation of human TFPI-beta mRNA is controlled by alternative splicing within the 5' UTR. Arter Thromb and Vasc Biol 2014; 34:187-195. PMC4043743
• Wood JP, Ellery PE, Maroney SA, and Mast AE: Protein S is a cofactor for platelet and endothelial tissue factor pathway inhibitor (TFPI)alpha, but not cell surface associated TFPI. Arter Thromb and Vasc Biol 2014; 34:169-176. PMC4030531
• Elery PE, Maroney SA, Cooley BC, Luyendyk JP, Zogg M, Weiler H, Mast AE: A balance between TFPI and thrombin-mediated platelet activation is required for murine embryonic development.  Blood, 2015; 125:4078-4084. PMC4481595
• Wood JP, Ellery PE, Maroney SA, and Mast AE: Biology of Tissue Factor Pathway Inhibitor.  Blood, 2014; 123:2934-2943. PMC4014837
• Maroney SA, Mast AE: New Insights into the biology of Tissue Factor Pathway Inhibitor. J Thromb Hemost, 2015; Suppl 1:S200-207. PMC4604745
• Wood JP, Baumann-Kreuziger LM, Desai UR, Mast AE: Blocking inhibition of prothrombinse by TFPI: A procoagulant property of heparins. British Journal of Haematology 2016, 175:123-132. PMC5035186
• Mast AE: (2016) Tissue Factor Pathway Inhibitor: Multiple Anticoagulant Activities for a Single Protein. Arter Thromb and Vasc Biol, 36:9-14. PMC4690769
• Peterson JA, Maroney SA, Mast AE: (2016) Targeting TFPI for Hemophilia Treatment. Thromb Res, 141 Suppl 2: S28-30. PMC4950664
  
  

Selected clinical and translational studies publications:

• Mast AE, Blinder MA, Gronowski A, Chumley C, Scott MG: The clinical utility of the serum transferrin receptor level in the evaluation of iron deficiency anemia. Clin Chem 1998; 44:45-51
• Mast AE, Stadanlick JE, Lockett JM, Dietzen DJ: Solvent/detergent treated plasma has decreased antitrypsin activity and absent antiplasmin activity. Blood 1999; 94:3922-3927
• Mast AE, Blinder MA, Lu Q, Flax S, Dietzen DJ: The clinical utility of the reticulocyte hemoglobin content in the diagnosis of iron deficiency. Blood2002, 99:1489-1491
• Mast AE, Blinder MA, Dietzen DJ: Test of the Month: Reticulocyte Hemoglobin Content. Am. J. Hematology 2008, 83:307-310
• Mast AE, Foster T, Pinder HL, Beczkiewicz CA, Bellissimo DB, Murphy AT, Kovacevic S, Wroblewski VJ, Withcher DR: Behavioral, biochemical and genetic analysis of iron metabolism in high intensity blood donors. Transfusion, 2008, 45:2197-2204
• Young S, Fink A, Geiger S, Marabella A, Mast AE, Schellhase KS: Community blood donors knowledge of anemia and design of a literacy appropriate educational intervention. Transfusion, 2010, 50:75-79
• Mast AE, Schlumpf KS, Wright DJ, Custer B, Spencer B, Murphy EL, Simon TL: Demographic correlates of low hemoglobin deferral among prospective whole blood donors. Transfusion, 2010, 50:1794-1802
• Scott EA, Schlumpf KS, Mathew SM, Mast AE, Busch MP, Gottschall JL: Biospecimen repositories: are blood donors willing to participate? Transfusion, 2010, 50:1943-1950
• Cable RG, Glynn SA, Kiss JE, Mast AE, Steele WR, Murphy EL, Wright DJ, Sacher RA, Gottschall JL, Vij V, Simon TL: Iron Deficiency in Blood Donors: Analysis of Enrollment Data from the REDS-II Donor Iron Status Evaluation (RISE) Study. Transfusion 2011, 51:511-522. PMC3050998
• Delaney M, Shellhase KG, Young S, Geiger S, Fink A, Mast AE: Blood center practice and education for blood donors with anemia. Transfusion 2011, 51:929-936. PMC3608117
• Carey PM, High PM, Schlumpf KS, Johnson BR, Mast AE, Rios JA, Simon TL, Wilkinson SL: Donation return time at fixed and mobile donation sites. Transfusion 2012, 52:127-133. PMC3557842
• Mast AE, Lee T-H, Schlumpf KS, Wright DJ, Johnson B, Carrick DM, Cable RG, Kiss JE, Glynn SA, Steele WR, Murphy EL, Sacher R and Busch MP: The impact of HFE mutations on hemoglobin and iron status in individuals experiencing repeated iron loss through blood donation. Br J Haematol 2012, 156:388-401. PMC3508695
• Cable RG, Glynn SA, Kiss JE, Mast AE, Steele WR, Murphy EL, Wright DJ, Sacher RA, Gottschall JL, Tobler LH, Simon TL: Iron Deficiency in Blood Donors: the REDS-II Donor Iron Status Evaluation (RISE) Study. Transfusion 2012, 52:702-711. PMC3618489
• Cable RG, Steele WR, Melmed RS, Johnson B, Mast AE, Carey PM, Kiss JE, Kleinman SH, Wright DJ: The difference between fingerstick and venous hemoglobin and hematocrit varies by sex and iron stores. Transfusion 2012, 52:1031-1040
• Mast AE, Steele WR, Johnson B, Wright DJ, Cable RG, Carey P, Gottschall JL, Kiss JE, Simon TL, Murphy EL: Population-based screening for anemia using first-time blood donors. Am J Hematol 2012, 87:496-502. PMC3525330
• Mast AE, Field JJ: (2012) Iron Chelation Therapy in Patients with Transfusion-Dependent Myelodysplastic Syndrome. Transfusion, 52:2078-2080. PMC3589809
• Brugnara C, Adamson J, Auerbach M, Kane R, McDougall I, Mast A: (2013) Q&A on Iron Deficiency. Clincal Chemistry, In Press
• Spencer BR, Kleinman S, Wright, DJ, Glynn SA, Rye DB, Kiss JE, Mast AE, Cable RG: Restless legs syndrome, pica and iron status in blood donors. Transfusion 2013; 53:1645-1651.
• Mast AE, Schlumpf KS, Wright DJ, Johnson B, Glynn SA, Busch MP, Olbina G, Westermann M, Nemeth E, Ganz T: Hepcidin level predicts hemoglobin concentration in individuals undergoing repeated phlebotomy. Hematologica 2013; 98:1324-1330. PMC3729915
• Kiss JE, Steele WR, Wright DJ, Mast AE, Carey PM, Murphy EL, Gottschall JL, Simon TL, Cable RG: Laboratory parameters for assessing iron deficiency in REDS-II iron status evaluation (RISE) blood donors.  Transfusion 2013; 53:2766-2774. PMC - In Process
• Brugnara C, Adamson J, Auerbach M, Kane R, McDougall I, Mast A: (2013) Q&A on iron deficiency. Clinical Chemistry, 59:740-745
• Mast AE: (2014) Low hemoglobin deferrals in blood donors.  Transfusion Medicine Reviews, 28:18-22. PMC3944402.
• Annen K, Leitch D, Delaney M, Mast AE: The health implications of low hemoglobin deferral in infrequent blood donors.  Transfusion, 2015; 55:86-90. PMC4293201
• Bialowski W, Bryant BJ, Schlumpf KS, Wright DJ, Birch R, Kiss JE, D'Andrea P, Cable RG, Spencer BR, Mast AE: The Strategies to Reduce Iron Deficiency (STRIDE) in blood donors randomized trial: Design, enrollment and early retention.  Vox Sang, 2015; 108:178-185. PMC4300282
• Kiss JE, Brambilla D, Glynn SA, Mast AE, Spencer BR, Stone M, Tobler LH, Kleinman SH, Cable RG: Oral Iron Supplementation After Blood Donation: A Randomized Clinical Trial.  JAMA, 2015; 313:575-583. PMC5094173
• Mast AE Bialkowski W; Bryant BJ, Wright DJ, Birch R, Kiss JE, D’Andrea P, Cable RG, Spencer BR: A randomized, blinded, placebo-controlled trial of education and iron supplementation for mitigation of iron deficiency in regular blood donors. Transfusion 2016, 56:1588-1597. PMC4905782
• Cable RG, Brambilla D, Glynn SA, Kleinman S, Mast AE, Spencer BR, Stone M, Kiss JE: Effect of Iron Supplementation on Iron Stores and Total Body Iron after Whole Blood Donation. Transfusion 2016, 56:2005-2012. PMC4980154
• Chansky MC, King MR, Bialkowski W, Bryant BJ, Kiss JE, D’Andrea P, Cable RG, Spencer BR, Mast AE: Qualitative assessment of pica experienced by frequent blood donors. Transfusion 2017, In Press
• Bloch E, Mast AE, Josephson CD, Klein H, Eder A: (2016) Teenage Blood Donors: Are we asking too little and taking too much? Pediatrics In Press