Francis Owen Blood Research Laboratory
Objectives
The purpose of this N01 contract is to produce and maintain the following dog models of inherited bleeding disorders for independent and collaborative scientific investigations: hemophilia A and hemophilia B with and without inhibitory antibodies to FVIII and FIX, respectively; von Willebrand disease (VWD); FVII-deficiency; and Glanzmann’s Thrombasthenia. Beginning in 1947 with an R01 from NIH/NHLBI, Dr. Kenneth M. Brinkhous and colleagues identified these dogs and demonstrated that (1) they mirror the severe bleeder phenotype present in the respective human disorder and (2) have a strong predictive accuracy for successful translational research.1 In 1999, the reviewers of Dr. Brinkhous’ R01 judged these dogs to be an important national resource that should be made available to the research community and NHLBI support was transitioned to an R24 (T.C. Nichols, PI). Support of this colony has now transitioned to this N01 contract. Since 1999, we have produced bleeder and normal dogs that have been used in collaborations with over 30 teams of investigators, most with NHLBI support, and the work has been reported in over 95 peer-reviewed publications1-96 and Conference Proceedings97-102, several of which were selected for editorial review.103-109 Recently, vectors used in at least 5 human gene therapy trials for hemophilia B and 2 for hemophilia A,4, 5, 110-115 six new recombinant anti-hemophilic proteins,47, 51, 59, 66, 71, 77 and recombinant von Willebrand factor (VWF)116 and IL-1120 for VWD have been tested in these dogs prior to clinical trials. In all cases, the results predicted safety, efficacy, and drug dose responses. The primary benefits of these animals have been to accelerate translatable discovery science in coagulation and hemostasis research and to produce the scientific basis for the safe and efficacious introduction of novel therapeutics into clinical practice.1
Considerable progress has been made over the past 7 decades in understanding the basic science of blood coagulation and hemostasis and in developing new clinical approaches to inherited bleeding disorders. Yet, there are persistent limitations in therapeutic options for people with VWD and hemophilia, especially those with inhibitors. To address these limitations with novel and innovative strategies, our 5 objectives are:
- To maintain a breeding colony of well-characterized dogs with inherited bleeding disorders. We propose to maintain a breeding colony at the Francis Owen Blood Research Laboratory (FOBRL) of the following well-characterized dog models of inherited bleeding disorders: hemophilia A and hemophilia B with and without inhibitory antibodies to FVIII and FIX, respectively; von Willebrand disease (VWD); FVII-deficiency; and Glanzmann’s Thrombasthenia. This N01 contract will ensure that the appropriate number of animals are available to maintain the colony and to produce the requisite number of research animals. This is the only contract that supports production and maintenance of these dogs. To our knowledge, the FOBRL is the only such breeding colony in the USA; the only other is in Canada and has hemophilia A dogs alone.
- To produce affordable purpose-bred research animals with these bleeding disorders in a cost-effective manner. The breeders and normal blood donor animals supported by this N01 will be used to produce and maintain the research animals with inherited bleeding disorders. Animals entered into research studies are supported by separate research grants. The support provided by this contract reduces the cost of research animals by > 70% to investigators with NIH or equivalent nonprofit funding.
- To produce purpose-bred, affordable research animals to serve as blood donors and experimental controls. With support from this N01, we will provide specialized support services for the design and performance of research projects using these dogs including canine blood banking and coagulation analyses. To assure appropriate, optimal, and rigorous experimental design, FOBRL professional staff provide advice, mentoring, and assistance in submitting grants and maintaining inter-institutional congruency.
- To provide specialized support services for the veterinary care of these dogs including canine blood banking and coagulation analyses. The constancy of spontaneous hemorrhages mandates continuous production of canine blood products including recombinant canine FVIII47, fresh and frozen plasma, cryoprecipitate, platelets, red blood cells, and whole blood from normal donor dogs maintained at the FOBRL. Without these products, hemorrhages are usually debilitating or fatal. Also, new hemostatic agents are being developed to support hemophilic dogs with inhibitors including recombinant canine FVIIa62, FIX wild type and gain-of-function Padua76, 117 and FXa-I16L.74, 118 The production, maintenance, and care of these animals require clinical and technical expertise with on site and immediate performance and interpretation of coagulation assays in canine blood and plasma. Specially trained, highly experienced personnel provide this essential daily round the clock care.
- To acquire and characterize new dog models of inherited bleeding disorders. Three new strains are being developed: (1) hemophilia A that develop inhibitory antibodies to canine FVIII in two genetic backgrounds: intron 22 inversion53 or a large multi-exon deletion mutation; (2) hemophilia B with a null mutation that are inhibitor prone (acquired in 2016 with the closure of the colony at UAB)119, 120; and (3) FVII-deficiency for investigations into Tissue Factor-FVIIa biology59, 61, 62, 121, 122 and gene transfer of canine FVII zymogen91 and FVIIa45. These well-characterized dogs provide large animal models in a mixed genetic background to test therapies for rare bleeding disorders for which a sufficient number of patients is unavailable for large clinical trials.123, 124 The molecular defect is known in all strains and will enable novel gene editing research.
References
- Nichols TC, Hough C, Agerso H, Ezban M and Lillicrap D. Canine models of inherited bleeding disorders in the development of coagulation assays, novel protein replacement and gene therapies. J Thromb Haemost. 2016;14:894-905. [PMID:26924758].
- Monahan PE, Samulski RJ, Tazelaar J, Xiao X, Nichols TC, Bellinger DA, Read MS and Walsh CE. Direct intramuscular injection with recombinant AAV vectors results in sustained expression in a dog model of hemophilia. Gene Ther. 1998;5:40-49. [PMID:9536263].
- Zhang Z, Eriksson M, Falk G, Graff C, Presnell SC, Read MS, Nichols TC, Blomback M and Anvret M. Failure to achieve gene conversion with chimeric circular oligonucleotides: potentially misleading PCR artifacts observed. Antisense Nucleic Acid Drug Dev. 1998;8:531-536. [PMID:9918117].
- Herzog RW, Yang EY, Couto LB, Hagstrom JN, Elwell D, Fields PA, Burton M, Bellinger DA, Read MS, Brinkhous KM, Podsakoff GM, Nichols TC, Kurtzman GJ and High KA. Long-term correction of canine hemophilia B by gene transfer of blood coagulation factor IX mediated by adeno-associated viral vector. Nat Med. 1999;5:56-63. [PMID:9883840].
- Snyder RO, Miao C, Meuse L, Tubb J, Donahue BA, Lin HF, Stafford DW, Patel S, Thompson AR, Nichols T, Read MS, Bellinger DA, Brinkhous KM and Kay MA. Correction of hemophilia B in canine and murine models using recombinant adeno-associated viral vectors. Nat Med. 1999;5:64-70. [PMID:9883841].
- Chao H, Samulski R, Bellinger D, Monahan P, Nichols T and Walsh C. Persistent expression of canine factor IX in hemophilia B canines. Gene Ther. 1999;6:1695-1704. [PMID:10516718].
- Wang L, Nichols TC, Read MS, Bellinger DA and Verma IM. Sustained expression of therapeutic level of factor IX in hemohilia B dogs by AAV-mediated gene therapy in liver. Molecular Therapy. 2000;1:154-158. [PMID:10933925].
- Herzog R, Arruda VR, Fischer TH, Read MS, Nichols TC and High KA. Absence of circulating factor IX antigen in hemophilia B dogs of the UNC-Chapel Hill Colony. Thromb Haemost. 2000;84:352-354. [PMID:10959714].
- Fischer TH, Merricks EP, Russell KE, Raymer RA, White GC, Bode AP, Nichols TC and Read MS. Intracellular function in rehydrated lyophilized platelets. Br J Haematol. 2000;111:167-174. [PMID:11091197].
- Arruda VR, Fields PA, Milner R, Wainwright L, De Miguel MP, Donovan PJ, Herzog RW, Nichols TC, Biegel JA, Razavi M, Dake M, Huff D, Flake AW, Couto L, Kay MA and High KA. Lack of germline transmission of vector sequences following systemic administration of recombinant AAV-2 vector in males. Mol Ther. 2001;4:586-592. [PMID:11735343].
- Russell KE, Read MS, Bellinger DA, Leitermann K, Rup BJ, McCarthy KP, Keith JC, Jr., Khor SP, Schaub RG and Nichols TC. Intratracheal administration of recombinant human factor IX (BeneFix) achieves therapeutic levels in hemophilia B dogs. Thromb Haemost. 2001;85:445-449. [PMID:11307812].
- Fischer TH, Merricks E, Bellinger DA, Hayes PM, Smith RS, Raymer RA, Read MS, Nichols TC and Bode AP. Splenic clearance mechanisms of rehydrated, lyophilized platelets. Artif Cells Blood Substit Immobil Biotechnol. 2001;29:439-451. [PMID:11795630].
- Lozier JN, Dutra A, Pak E, Zhou N, Zheng Z, Nichols TC, Bellinger DA, Read M and Morgan RA. The Chapel Hill hemophilia A dog colony exhibits a factor VIII gene inversion. Proc Natl Acad Sci U S A. 2002;99:12991-12996. [PMID:12242334].
- Fischer TH, Merricks EP, Bode AP, Bellinger DA, Russell K, Reddick R, Sanders WE, Nichols TC and Read MS. Thrombus formation with rehydrated, lyophilized platelets. Hematology. 2002;7:359-369. [PMID:12475741].
- Herzog RW, Fields PA, Arruda VR, Brubaker JO, Armstrong E, McClintock D, Bellinger DA, Couto LB, Nichols TC and High KA. Influence of vector dose on factor IX-specific T and B cell responses in muscle-directed gene therapy. Hum Gene Ther. 2002;13:1281-1291. [PMID:12162811].
- Mount JD, Herzog RW, Tillson DM, Goodman SA, Robinson N, McCleland ML, Bellinger D, Nichols TC, Arruda VR, Lothrop CD, Jr. and High KA. Sustained phenotypic correction of hemophilia B dogs with a factor IX null mutation by liver-directed gene therapy. Blood. 2002;99:2670-2676. [PMID:11929752].
- McCarthy K, Stewart P, Sigman J, Read M, Keith JC, Brinkhous KM, Nichols TC and Schaub RG. Pharmacokinetics of recombinant factor IX after intravenous and subcutaneous administration in dogs and cynomologus monkeys. Thromb Haemost. 2002;87:824-830. [PMID:12038784].
- Brinkhous KM, Sandberg H, Widlund L, Read MS, Nichols TC, Sigman J, Oswaldsson U, Schaub RG and Mikaelsson M. Preclinical pharmacology of albumin-free B-domain deleted recombinant factor VIII. Semin Thromb Hemost. 2002;28:269-272. [PMID:12098087].
- Xu L, Gao C, Sands MS, Cai SR, Nichols TC, Bellinger DA, Raymer RA, McCorquodale S and Ponder KP. Neonatal or hepatocyte growth factor-potentiated adult gene therapy with a retroviral vector results in therapeutic levels of canine factor IX for hemophilia B. Blood. 2003;101:3924-3932. [PMID:12531787].
- Olsen EH, McCain AS, Merricks EP, Fischer TH, Dillon IM, Raymer RA, Bellinger DA, Fahs SA, Montgomery RR, Keith JC, Jr., Schaub RG and Nichols TC. Comparative response of plasma VWF in dogs to up-regulation of VWF mRNA by interleukin-11 versus Weibel-Palade body release by desmopressin (DDAVP). Blood. 2003;102:436-441. [PMID:12649145].
- Rathore V, Stapleton MA, Hillery CA, Montgomery RR, Nichols TC, Merricks EP, Newman DK and Newman PJ. PECAM-1 negatively regulates GPIb/V/IX signaling in murine platelets. Blood. 2003;102:3658-3664. [PMID:12893757].
- Ehrhardt A, Xu H, Dillow AM, Bellinger DA, Nichols TC and Kay MA. A gene-deleted adenoviral vector results in phenotypic correction of canine hemophilia B without liver toxicity or thrombocytopenia. Blood. 2003;102:2403-2411. [PMID:12805062].
- Russell KE, Olsen EH, Raymer RA, Merricks EP, Bellinger DA, Read MS, Rup BJ, Keith JC, Jr., McCarthy KP, Schaub RG and Nichols TC. Reduced bleeding events with subcutaneous administration of recombinant human factor IX in immune-tolerant hemophilia B dogs. Blood. 2003;102:4393-4398. [PMID:12933577].
- Arruda VR, Schuettrumpf J, Herzog RW, Nichols TC, Robinson N, Lotfi Y, Mingozzi F, Xiao W, Couto LB and High KA. Safety and efficacy of factor IX gene transfer to skeletal muscle in murine and canine hemophilia B models by adeno-associated viral vector serotype 1. Blood. 2004;103:85-92. [PMID:12969984].
- Harding TC, Koprivnikar KE, Tu GH, Zayek N, Lew S, Subramanian A, Sivakumaran A, Frey D, Ho K, VanRoey MJ, Nichols TC, Bellinger DA, Yendluri S, Waugh J, McArthur J, Veres G and Donahue BA. Intravenous administration of an AAV-2 vector for the expression of factor IX in mice and a dog model of hemophilia B. Gene Ther. 2004;11:204-213. [PMID:14712305].
- Fischer TH, Robbins ME, Bode AP, Nichols TC, Bellinger DE and Schoenfisch MH. Evidence that rehydrated, lyophilized red blood cells are sufficiently deformable for normal microcirculation transit. Microsc Res Tech. 2004;65:62-71. [PMID:15570582].
- Schwaitzberg SD, Chan MW, Cole DJ, Read M, Nichols T, Bellinger D and Connolly RJ. Comparison of poly-N-acetyl glucosamine with commercially available topical hemostats for achieving hemostasis in coagulopathic models of splenic hemorrhage. J Trauma. 2004;57:S29-32. [PMID:15280748].
- Van Cott KE, Monahan PE, Nichols TC and Velander WH. Haemophilic factors produced by transgenic livestock: abundance that can enable alternative therapies worldwide. Haemophilia. 2004;10 Suppl 4:70-76. [PMID:15479375].
- Arruda VR, Stedman HH, Nichols TC, Haskins ME, Nicholson M, Herzog RW, Couto LB and High KA. Regional intravascular delivery of AAV-2-F.IX to skeletal muscle achieves long-term correction of hemophilia B in a large animal model. Blood. 2005;105:3458-3464. [PMID:15479726].
- Haberichter SL, Merricks EP, Fahs SA, Christopherson PA, Nichols TC and Montgomery RR. Re-establishment of VWF-dependent Weibel-Palade bodies in VWD endothelial cells. Blood. 2005;105:145-152. [PMID:15331450].
- Wang L, Calcedo R, Nichols TC, Bellinger DA, Dillow A, Verma IM and Wilson JM. Sustained correction of disease in naive and AAV2-pretreated hemophilia B dogs: AAV2/8-mediated, liver-directed gene therapy. Blood. 2005;105:3079-3086. [PMID:15637142].
- Xu L, Nichols TC, Sarkar R, McCorquodale S, Bellinger DA and Ponder KP. Absence of a desmopressin response after therapeutic expression of factor VIII in hemophilia A dogs with liver-directed neonatal gene therapy. Proc Natl Acad Sci U S A. 2005;102:6080-6085. [PMID:15837921].
- Brunetti-Pierri N, Nichols TC, McCorquodale S, Merricks E, Palmer DJ, Beaudet AL and Ng P. Sustained phenotypic correction of canine hemophilia B after systemic administration of helper-dependent adenoviral vector. Hum Gene Ther. 2005;16:811-820. [PMID:16000063].
- Fischer TH, Thatte HS, Nichols TC, Bender-Neal DE, Bellinger AD and Vournakis JN. Synergistic platelet integrin signaling and factor XII activation in poly-N-acetyl glucosamine fiber-mediated hemostasis. Biomaterials. 2005;26:5433-5443. [PMID:15860200].
- Sarkar R, Mucci M, Addya S, Tetreault R, Bellinger DA, Nichols TC and Kazazian HH, Jr. Long-term efficacy of adeno-associated virus serotypes 8 and 9 in hemophilia A dogs and mice. Hum Gene Ther. 2006;17:427-439. [PMID:16610930].
- McCormack WM, Seiler MP, Bertin TK, Ubhayakar K, Palmer DJ, Ng P, Nichols TC and Lee B. Helper-dependent adenoviral gene therapy mediates long-term correction of the clotting defect in the canine hemophilia A model. J Thromb Haemost. 2006;4:1218-1225. [PMID:16706963].
- Dumont D, Behler RH, Nichols TC, Merricks EP and Gallippi CM. ARFI imaging for noninvasive material characterization of atherosclerosis. Ultrasound Med Biol. 2006;32:1703-1711. [PMID:17112956].
- Xu L, Mei M, Haskins ME, Nichols TC, O’Donnell P, Cullen K, Dillow A, Bellinger D and Ponder KP. Immune response after neonatal transfer of a human factor IX-expressing retroviral vector in dogs, cats, and mice. Thromb Res. 2007;120:269-280. [PMID:17095052].
- Behler RH, Nichols T, Merricks EP and Gallippi CM. A Rigid Wall Approach to Physiological Motion Rejection in Arterial ARFI Imaging: Simulation and In Vivo Demonstration. IEEE Ultrasonics Symposium. 2007; New York, New York:359-364. [PMID:NA].
- Mauldin FW, Jr., Zhu HT, Behler RH, Nichols TC and Gallippi CM. Robust principal component analysis and clustering methods for automated classification of tissue response to ARFI excitation. Ultrasound Med Biol. 2008;34:309-325. [PMID:17913334].
- Ragni MV, Jankowitz RC, Chapman HL, Merricks EP, Kloos MT, Dillow AM and Nichols TC. A phase II prospective open-label escalating dose trial of recombinant interleukin-11 in mild von Willebrand disease. Haemophilia. 2008;14:968-977. [PMID:18680527].
- Fischer TH, Valeri CR, Smith CJ, Scull CM, Merricks EP, Nichols TC, Demcheva M and Vournakis JN. Non-classical processes in surface hemostasis: mechanisms for the poly-N-acetyl glucosamine-induced alteration of red blood cell morphology and surface prothrombogenicity. Biomed Mater. 2008;3:015009. [PMID:18458496].
- Fischer TH, Wolberg AS, Bode AP and Nichols TC. The interaction of factor VIIa with rehydrated, lyophilized platelets. Platelets. 2008;19:182-191. [PMID:18432519].
- Nichols TC, Dillow AM, Franck HW, Merricks EP, Raymer RA, Bellinger DA, Arruda VR and High KA. Protein replacement therapy and gene transfer in canine models of hemophilia A, hemophilia B, von willebrand disease, and factor VII deficiency. ILAR J. 2009;50:144-167. [PMID:19293459].
- Margaritis P, Roy E, Aljamali MN, Downey HD, Giger U, Zhou S, Merricks E, Dillow A, Ezban M, Nichols TC and High KA. Successful treatment of canine hemophilia by continuous expression of canine FVIIa. Blood. 2009;113:3682-3689. [PMID:19109232].
- Milbauer LC, Enenstein JA, Roney M, Solovey A, Bodempudi V, Nichols TC and Hebbel RP. Blood outgrowth endothelial cell migration and trapping in vivo: a window into gene therapy. Transl Res. 2009;153:179-189. [PMID:19304277].
- Sabatino DE, Freguia CF, Toso R, Santos A, Merricks EP, Kazazian HH, Jr., Nichols TC, Camire RM and Arruda VR. Recombinant canine B-domain-deleted FVIII exhibits high specific activity and is safe in the canine hemophilia A model. Blood. 2009;114:4562-4565. [PMID:19770361].
- Behler RH, Scola MR, Nichols TC, Bellinger DA and Gallippi CM. ARFI ultrasound for in vivo hemostasis assessment postcardiac catheterization, part I: preclinical studies. Ultrason Imaging. 2009;31:153-158. [PMID:19771958].
- Behler RH, Scola MR, Nichols TC, Caughey MC, Fisher MW, Zhu H and Gallippi CM. ARFI ultrasound for in vivo hemostasis assessment postcardiac catheterization, part II: pilot clinical results. Ultrason Imaging. 2009;31:159-171. [PMID:19771959].
- Fischer TH, Vournakis JN, Manning JE, McCurdy SL, Rich PB, Nichols TC, Scull CM, McCord MG, Decorta JA, Johnson PC and Smith CJ. The design and testing of a dual fiber textile matrix for accelerating surface hemostasis. J Biomed Mater Res B Appl Biomater. 2009;91:381-389. [PMID:19489008].
- Peters RT, Low SC, Kamphaus GD, Dumont JA, Amari JV, Lu Q, Zarbis-Papastoitsis G, Reidy TJ, Merricks EP, Nichols TC and Bitonti AJ. Prolonged activity of factor IX as a monomeric Fc fusion protein. Blood. 2010;115:2057-2064. [PMID:20056791].
- Obergfell A, Nichols T and Ezban M. Animal models of FVIIa gene expression: their role in the future development of haemophilia treatment. Haemophilia. 2010;16 Suppl 2:24-27. [PMID:20132335].
- Finn JD, Ozelo MC, Sabatino DE, Franck HW, Merricks EP, Crudele JM, Zhou S, Kazazian HH, Lillicrap D, Nichols TC and Arruda VR. Eradication of neutralizing antibodies to factor VIII in canine hemophilia A after liver gene therapy. Blood. 2010;116:5842-5848. [PMID:20876851].
- Arruda VR, Stedman HH, Haurigot V, Buchlis G, Baila S, Favaro P, Chen Y, Franck HG, Zhou S, Wright JF, Couto LB, Jiang H, Pierce GF, Bellinger DA, Mingozzi F, Nichols TC and High KA. Peripheral transvenular delivery of adeno-associated viral vectors to skeletal muscle as a novel therapy for hemophilia B. Blood. 2010;115:4678-4688. [PMID:20335222].
- Haurigot V, Mingozzi F, Buchlis G, Hui DJ, Chen Y, Basner-Tschakarjan E, Arruda VR, Radu A, Franck HG, Wright JF, Zhou S, Stedman HH, Bellinger DA, Nichols TC and High KA. Safety of AAV factor IX peripheral transvenular gene delivery to muscle in hemophilia B dogs. Mol Ther. 2010;18:1318-1329. [PMID:20424599].
- Oldenburg AL, Gallippi CM, Tsui F, Nichols TC, Beicker KN, Chhetri RK, Spivak D, Richardson A and Fischer TH. Magnetic and contrast properties of labeled platelets for magnetomotive optical coherence tomography. Biophys J. 2010;99:2374-2383. [PMID:20923673].
- Hausl MA, Zhang W, Muther N, Rauschhuber C, Franck HG, Merricks EP, Nichols TC, Kay MA and Ehrhardt A. Hyperactive sleeping beauty transposase enables persistent phenotypic correction in mice and a canine model for hemophilia B. Mol Ther. 2010;18:1896-1906. [PMID:20717103].
- Nichols TC, Raymer RA, Franck HW, Merricks EP, Bellinger DA, DeFriess N, Margaritis P, Arruda VR, Kay MA and High KA. Prevention of spontaneous bleeding in dogs with haemophilia A and haemophilia B. Haemophilia. 2010;16 Suppl 3:19-23. [PMID:20586797].
- Petersen LC, Karpf DM, Agerso H, Hermit MB, Pelzer H, Persson E, Nichols TC and Ezban M. Intravascular inhibition of factor VIIa and the analogue NN1731 by antithrombin. Br J Haematol. 2011;152:99-107. [PMID:21083658].
- Knudsen T, Kjalke M, Tranholm M, Nichols TC, Jensen AL and Kristensen AT. Development of a flow cytometric assay for detection of coated platelets in dogs and evaluation of binding of coated platelets to recombinant human coagulation factor VIIa. Am J Vet Res. 2011;72:1007-1014. [PMID:21801056].
- Knudsen T, Kjelgaard-Hansen M, Tranholm M, Wiinberg B, Clausen JT, Hansen JJ, Nichols TC, Kjalke M, Jensen AL and Kristensen AT. Canine specific ELISA for coagulation factor VII. Vet J. 2011;190:352-358. [PMID:21216638].
- Knudsen T, Kristensen AT, Nichols TC, Agerso H, Jensen AL, Kjalke M, Ezban M and Tranholm M. Pharmacokinetics, pharmacodynamics and safety of recombinant canine FVIIa in a study dosing one haemophilia A and one haemostatically normal dog. Haemophilia. 2011;17:962-970. [PMID:21645178].
- Matrai J, Cantore A, Bartholomae CC, Annoni A, Wang W, Acosta-Sanchez A, Samara-Kuko E, De Waele L, Ma L, Genovese P, Damo M, Arens A, Goudy K, Nichols TC, von Kalle C, MK LC, Roncarolo MG, Schmidt M, Vandendriessche T and Naldini L. Hepatocyte-targeted expression by integrase-defective lentiviral vectors induces antigen-specific tolerance in mice with low genotoxic risk. Hepatology. 2011;53:1696-1707. [PMID:21520180].
- Scola MR, Nichols TC, Zhu H, Caughey MC, Merricks EP, Raymer RA, Margaritis P, High KA and Gallippi CM. ARFI ultrasound monitoring of hemorrhage and hemostasis in vivo in canine von Willebrand disease and hemophilia. Ultrasound Med Biol. 2011;37:2126-2132. [PMID:22033127].
- Nichols TC, Bellinger DA, Merricks EP, Raymer RA, Kloos M, DeFriess N, Ragni MV and Griggs TR. Porcine and Canine von Willebrand Factor and von Willebrand Disease: Hemostasis, Thrombosis, and Atherosclerosis Studies. Thrombosis 2010;10.1155/2010/461238:461238. [PMID:22091368].
- Ostergaard H, Bjelke JR, Hansen L, Petersen LC, Pedersen AA, Elm T, Moller F, Hermit MB, Holm PK, Krogh TN, Petersen JM, Ezban M, Sorensen BB, Andersen MD, Agerso H, Ahmadian H, Balling KW, Christiansen ML, Knobe K, Nichols TC, Bjorn SE and Tranholm M. Prolonged half-life and preserved enzymatic properties of factor IX selectively PEGylated on native N-glycans in the activation peptide. Blood. 2011;118:2333-2341. [PMID:21700771].
- Sabatino DE, Lange AM, Altynova ES, Sarkar R, Zhou S, Merricks EP, Franck HG, Nichols TC, Arruda VR and Kazazian HH, Jr. Efficacy and safety of long-term prophylaxis in severe hemophilia A dogs following liver gene therapy using AAV vectors. Mol Ther. 2011;19:442-449. [PMID:21081906].
- Ragni MV, Jankowitz RC, Jaworski K, Merricks EP, Kloos MT and Nichols TC. Phase II prospective open-label trial of recombinant interleukin-11 in women with mild von Willebrand disease and refractory menorrhagia. Thromb Haemost. 2011;106:641-645. [PMID:21833452].
- Karpf DM, Kjalke M, Thim L, Agerso H, Merricks EP, Defriess N, Nichols TC and Ezban M. Pharmacokinetics and ex vivo whole blood clot formation of a new recombinant FVIII (N8) in haemophilia A dogs. Haemophilia. 2011;17:e963-968. [PMID:21682818].
- Fischer TH, Nichols TC, Scull CM, Smith CJ and Demcheva M. Poly-N-acetylglucosamine fibers amplify the effectiveness of recombinant factor VIIA on clot formation in hemophilia B canine blood. J Trauma. 2011;71:S171-175. [PMID:21814114].
- Dumont JA, Liu T, Low SC, Zhang X, Kamphaus G, Sakorafas P, Fraley C, Drager D, Reidy T, McCue J, Franck HW, Merricks EP, Nichols TC, Bitonti AJ, Pierce GF and Jiang H. Prolonged activity of a recombinant factor VIII-Fc fusion protein in hemophilia A mice and dogs. Blood. 2012;119:3024-3030. [PMID:22246033].
- Sabatino DE, Nichols TC, Merricks E, Bellinger DA, Herzog RW and Monahan PE. Animal models of hemophilia. Prog Mol Biol Transl Sci. 2012;105:151-209. [PMID:22137432].
- Scola MR, Baggesen LM, Nichols TC, Key NS and Gallippi CM. A review of current methods for assessing hemostasis in vivo and introduction to a potential alternative approach. Thromb Res. 2012;129 Suppl 2:S57-61. [PMID:22405050].
- Pittman D, Nichols TC, Camire RM, Toso R, Merricks EP, Raymer RA, DeFriess N, Leary B, Parng C, Arkin S and Freubis J. A Factor Xa Variant Restores Hemostasis in Hemophilia A Dog Model. Haemophilia 2012;18:87. [PMID:NA].
- Nichols TC, Franck HW, Franck CT, De Friess N, Raymer RA and Merricks EP. Sensitivity of whole blood clotting time and activated partial thromboplastin time for factor IX: relevance to gene therapy and determination of post-transfusion elimination time of canine factor IX in hemophilia B dogs. J Thromb Haemost. 2012;10:474-476. [PMID: 22482117].
- Finn JD, Nichols TC, Svoronos N, Merricks EP, Bellinger DA, Zhou S, Simioni P, High KA and Arruda VR. The efficacy and the risk of immunogenicity of FIX Padua (R338L) in hemophilia B dogs treated by AAV muscle gene therapy. Blood. 2012;120:4521-4523. [PMID:22919027].
- Nolte MW, Nichols TC, Mueller-Cohrs J, Merricks EP, Pragst I, Zollner S and Dickneite G. Improved kinetics of rIX-FP, a recombinant fusion protein linking factor IX with albumin, in cynomolgus monkeys and hemophilia B dogs. J Thromb Haemost. 2012;10:1591-1599. [PMID:22726310].
- Agerso H, Stennicke HR, Pelzer H, Olsen EN, Merricks EP, Defriess NA, Nichols TC and Ezban M. Pharmacokinetics and pharmacodynamics of turoctocog alfa and N8-GP in haemophilia A dogs. Haemophilia. 2012;18:941-947. [PMID:22812621].
- Ragni MV, Novelli EM, Murshed A, Merricks EP, Kloos MT and Nichols TC. Phase II prospective open-label trial of recombinant interleukin-11 in desmopressin-unresponsive von Willebrand disease and mild or moderate haemophilia A. Thromb Haemost. 2013;109:248-254. [PMID:23238591].
- Pope AG, Wu G, McWhorter FY, Merricks EP, Nichols TC, Czernuszewicz TJ, Gallippi CM and Oldenburg AL. Contrast-enhanced imaging of SPIO-labeled platelets using magnetomotive ultrasound. Phys Med Biol. 2013;58:7277-7290. [PMID:24077004].
- Lozier JN and Nichols TC. Animal models of hemophilia and related bleeding disorders. Semin Hematol. 2013;50:175-184. [PMID:23956467].
- Du LM, Nurden P, Nurden AT, Nichols TC, Bellinger DA, Jensen ES, Haberichter SL, Merricks E, Raymer RA, Fang J, Koukouritaki SB, Jacobi PM, Hawkins TB, Cornetta K, Shi Q and Wilcox DA. Platelet-targeted gene therapy with human factor VIII establishes haemostasis in dogs with haemophilia A. Nat Commun. 2013;4:2773. [PMID:24253479].
- Sherman A, Schlachterman A, Cooper M, Merricks EP, Raymer RA, Bellinger DA, Herzog RW and Nichols TC. Portal vein delivery of viral vectors for gene therapy for hemophilia. Methods Mol Biol. 2014;1114:413-426. [PMID:24557919].
- Nichols TC. Lessons Learned from Animal Models of Inherited Bleeding Disorders. Hematol Educ. 2014;8:39-46. [PMID:26052366].
- Sauna ZE, Lozier JN, Kasper CK, Yanover C, Nichols T and Howard TE. The intron-22-inverted F8 locus permits factor VIII synthesis: explanation for low inhibitor risk and a role for pharmacogenomics. Blood. 2015;125:223-228. [PMID:25406352].
- Nichols TC, Whitford MH, Arruda VR, Stedman HH, Kay MA and High KA. Translational data from adeno-associated virus-mediated gene therapy of hemophilia B in dogs. Hum Gene Ther Clin Dev. 2015;26:5-14. [PMID:25675273].
- Kidd L, Geddings J, Hisada Y, Sueda M, Concannon T, Nichols T, Merricks E and Mackman N. Procoagulant microparticles in dogs with immune-mediated hemolytic anemia. J Vet Intern Med. 2015;29:908-916. [PMID:25871966].
- Cantore A, Ranzani M, Bartholomae CC, Volpin M, Valle PD, Sanvito F, Sergi LS, Gallina P, Benedicenti F, Bellinger D, Raymer R, Merricks E, Bellintani F, Martin S, Doglioni C, D’Angelo A, VandenDriessche T, Chuah MK, Schmidt M, Nichols T, Montini E and Naldini L. Liver-directed lentiviral gene therapy in a dog model of hemophilia B. Sci Transl Med. 2015;7:277ra228. [PMID:25739762].
- Geist RE, DuBois CH, Nichols TC, Caughey MC, Merricks EP, Raymer R and Gallippi CM. Experimental Validation of ARFI Surveillance of Subcutaneous Hemorrhage (ASSH) Using Calibrated Infusions in a Tissue-Mimicking Model and Dogs. Ultrason Imaging. 2016;38:346-358. [PMID:26614530].
- Shetty KA, Merricks EP, Raymer R, Rigsbee N, Nichols TC and Balu-Iyer SV. Soy Phosphatidylinositol-Containing Lipid Nanoparticle Prolongs the Plasma Survival and Hemostatic Efficacy of B-domain-Deleted Recombinant Canine Factor VIII in Hemophilia A Dogs. J Pharm Sci. 2016;105:2459-2464. [PMID:27372547].
- Marcos-Contreras OA, Smith SM, Bellinger DA, Raymer RA, Merricks E, Faella A, Pavani G, Zhou S, Nichols TC, High KA and Margaritis P. Sustained correction of FVII deficiency in dogs using AAV-mediated expression of zymogen FVII. Blood. 2016;127:565-571. [PMID:26702064].
- Lozier JN, Kloos M, Merricks EP, Lemoine N, Whitford M, Raymer RA, Bellinger DA and Nichols TC. Severe Hemophilia A in a Male Old English Sheep Dog with a C->T Transition in Factor VIII Exon 12 Leading to a Premature Stop Codon. Comparative Medicine. 2016;66:1-7. [PMID:27780008 PMCID: PMC5073066 Available on 2017-04-01].
- Siner JI, Samelson-Jones BJ, Crudele JM, French RA, Lee BJ, Zhou S, Merricks E, Raymer R, Nichols TC, Camire RM and Arruda VR. Circumventing furin enhances factor VIII biological activity and ameliorates bleeding phenotypes in hemophilia models. JCI Insight. 2016;1:e89371. [PMID:27734034].
- Herzog RW, Nichols TC, Su J, Zhang B, Sherman A, Merricks EP, Raymer R, Perrin GQ, Hager M, Wiinberg B and Daniell H. Oral Tolerance Induction in Hemophilia B Dogs Fed with Transplastomic Lettuce. Mol Ther. 2017;25:512-522. [PMID:28153098].
- Madsen DE, Nichols TC, Merricks EP, Waters EK and Wiinberg B. Global measurement of coagulation in plasma from normal and haemophilia dogs using a novel modified thrombin generation test – Demonstrated in vitro and ex vivo. PLoS One. 2017;12:e0175030. [PMID:28384182].
- Markusic DM, Nichols TC, Merricks EP, Palaschak B, Zolotukhin I, Marsic D, Zolotukhin S, Srivastava A and Herzog RW. Evaluation of engineered AAV capsids for hepatic factor IX gene transfer in murine and canine models. J Transl Med. 2017;15:94. [PMID:28460646].
- R.E. Geist TCN, E.P. Merricks, M.C. Caughey, C.M. Gallippi, . In Vivo ARFI Surveillance of Subcutaneous Hemorrhage (ASSH) For Monitoring rcFVIII Dose Response In Hemophilia A Dogs. Proceedings of the IEEE International Ultrasonics Symposium; 2012: 2296 – 2299.
- M.R. Scola RHB, T.C. Nichols, M.C. Caughey, E.P. Merricks, R. Raymer, P. Margaritis, K. High, C.M. Gallippi. In vivo Detection of Hemorrhage Rate in Dog Models of Hemophilia and VWD and at Human Femoral Arteriotomy by ARFI Ultrasound. Proceedings of the IEEE International Ultrasonics Symposium; 2011:660-663.
- M.R. Scola TCN, M.C. Caughey, P. Margaritis, K. High, C.M. Gallippi. ARFI Ultrasound for InVivo Monitoring of Soft-Tissue Bleeding and Hemostasis in a Dog Model of Hemophilia A. Proceedings of the IEEE International Ultrasonics Symposium; 2010:1149-1152.
- A.L. Oldenburg, T.H. Fischer, T.C. Nichols, C.M. Gallippi, R. Chhetri and F. Tsui. Contrast to Labeled Rehydrated, Lyophilized Platelets Using Magnetomotive OCT. Biomedical Optics and 3-D Imaging, OSA Technical Digest (CD) (Optical Society of America); 2010:paper BSuF5. ISBN: 978-971-55752-55887-55752.
- R.H. Behler, M.R. Scola, T.C. Nichols, M. Caughey, M.W. Fisher, H. Zhu and Gallippi. CM. In Vivo Hemostasis Detection at Human Femoral Arteriotomy by ARFI Ultrasound. Proceedings of the IEEE International Ultrasonics Symposium; 2009:1879-1882.
- R.H. Behler, T.C. Nichols, E.P. Merricks, and C.M. Gallippi. Reverberation Artifact Rejection in Arterial ARFI Imaging. Proceedings of the IEEE International Ultrasonics Symposium; 2009:2367-2370.
- Linden RM and Woo SL. AAVant-garde gene therapy. Nature Medicine. 1999;5:21-22. [PMID:9883831].
- Ponder K. Gene therapy for hemophilia B in dogs: finally prevention of bleeding, but concerns about inhibitors remain. Blood. 2002;99:2635b-2636. [PMID:NA].
- Lenting PJ and Sixma JJ. DDAVP and interleukin-11: a boosting combination. Blood. 2003;102:415-416. [PMID:NA].
- Arruda VR. Toward gene therapy for hemophilia A with novel adenoviral vectors: successes and limitations in canine models. J Thromb Haemost. 2006;4:1215-1217. [PMID:16706962].
- McVey JH. FVIIa gene delivery: potential treatment for hemophilia? Blood. 2009;113:3649-3650. [PMID:19372262].
- Mischke R. Determination of canine coagulation factor VII. Vet J. 2011;190:305-306. [PMID:22108258].
- Swanson E. Contrast to Labeled Platelets with Magnetomotive OCT for Detecting Vascular Damage. http://www.octnews.org/articles/2474057/feature-of-the-week-112710-contrast-to-labeled-pla/, http://www.octnews.org/articles/2474057/feature-of-the-week-112710-contrast-to-labeled-pla/; 2011.
- Greengard JS, Bodner M, McCormack J, Edwards WR, Sensintaffar JL, Sheridan PL, Phuong T, Mittelstaedt D, Brumm D, Nichols TC, Read M, Brinkhous KM, Jolly DJ and Chang SMW. Sustained expression of human factor VIII from peripheral retroviral gene delivery in rabbits and dogs. Blood (Suppl). 1997;90:240a. [PMID:NA].
- Powell JS, Ragni MV, White GC, 2nd, Lusher JM, Hillman-Wiseman C, Moon TE, Cole V, Ramanathan-Girish S, Roehl H, Sajjadi N, Jolly DJ and Hurst D. Phase 1 trial of FVIII gene transfer for severe hemophilia A using a retroviral construct administered by peripheral intravenous infusion. Blood. 2003;102:2038-2045. [PMID:12763932].
- Manno CS, Pierce GF, Arruda VR, Glader B, Ragni M, Rasko JJ, Ozelo MC, Hoots K, Blatt P, Konkle B, Dake M, Kaye R, Razavi M, Zajko A, Zehnder J, Rustagi PK, Nakai H, Chew A, Leonard D, Wright JF, Lessard RR, Sommer JM, Tigges M, Sabatino D, Luk A, Jiang H, Mingozzi F, Couto L, Ertl HC, High KA and Kay MA. Successful transduction of liver in hemophilia by AAV-Factor IX and limitations imposed by the host immune response. Nat Med. 2006;12:342-347. [PMID:16474400].
- Manno CS, Chew AJ, Hutchison S, Larson PJ, Herzog RW, Arruda VR, Tai SJ, Ragni MV, Thompson A, Ozelo M, Couto LB, Leonard DG, Johnson FA, McClelland A, Scallan C, Skarsgard E, Flake AW, Kay MA, High KA and Glader B. AAV-mediated factor IX gene transfer to skeletal muscle in patients with severe hemophilia B. Blood. 2003;101:2963-2972. [PMID:12515715].
- Nathwani AC, Tuddenham EG, Rangarajan S, Rosales C, McIntosh J, Linch DC, Chowdary P, Riddell A, Pie AJ, Harrington C, O’Beirne J, Smith K, Pasi J, Glader B, Rustagi P, Ng CY, Kay MA, Zhou J, Spence Y, Morton CL, Allay J, Coleman J, Sleep S, Cunningham JM, Srivastava D, Basner-Tschakarjan E, Mingozzi F, High KA, Gray JT, Reiss UM, Nienhuis AW and Davidoff AM. Adenovirus-associated virus vector-mediated gene transfer in hemophilia B. N Engl J Med. 2011;365:2357-2365. [PMID:22149959].
- George LA, Sullivan SK, Giermasz A, Rasko JEJ, Samelson-Jones BJ, Ducore J, Cuker A, Sullivan LM, Majumdar S, Teitel J, McGuinn CE, Ragni MV, Luk AY, Hui D, Wright JF, Chen Y, Liu Y, Wachtel K, Winters A, Tiefenbacher S, Arruda VR, van der Loo JCM, Zelenaia O, Takefman D, Carr ME, Couto LB, Anguela XM and High KA. Hemophilia B Gene Therapy with a High-Specific-Activity Factor IX Variant. N Engl J Med. 2017;377:2215-2227. [PMID:29211678].
- Nichols TC, Merricks E, Muchitsch EM, Baumgartner B, Turecek PL, Ahmad RU, Gritsch H and Schwarz HP. Pharmacokinetics (PK) of rVWF in Dogs and Mice with VWD. XXI ISTH Abstracts. 2007:P-W-197. [PMID:NA].
- French RA, Samelson-Jones BJ, Niemeyer GP, Lothrop CD, Jr., Merricks EP, Nichols TC and Arruda VR. Complete correction of hemophilia B phenotype by FIX-Padua skeletal muscle gene therapy in an inhibitor-prone dog model. Blood Adv. 2018;2:505-508. [PMID:29500218].
- Nichols TC, Merricks EP, Wimsey LA, Pittman D, Miao C, Arruda VA and Camire R. Research_and_Practice_in_Thrombosis_and_Haemostasis p. OC 51.5 | Recombinant Canine and Human FXa- I16LProvide Safe and Efficacious Hemostasis during Major Bleeding Events in Hemophilia A Dogs with High- titer Inhibitory Anti- FVIII Antibodies. Research_and_Practice_in_Thrombosis_and_Haemostasis – Abstracts for 2019 XXVII ISTH Meeting. 2019:109. [PMID:NA].
- Crudele JM, Finn JD, Siner JI, Martin NB, Niemeyer GP, Zhou S, Mingozzi F, Lothrop CD, Jr. and Arruda VR. AAV liver expression of FIX-Padua prevents and eradicates FIX inhibitor without increasing thrombogenicity in hemophilia B dogs and mice. Blood. 2015;125:1553-1561. [PMID:25568350].
- French R, Martin N, Nichols TC, Niemeyer GP, Lothrop CD and Arruda VR. Complete Correction of Severe Canine Hemophilia B By Skeletal Muscle Directed AAV-Based FIX-Padua Gene Therapy in Inhibitor-Prone Dogs. Blood. 2015;126 Abstract 3487. [PMID:NA].
- Knudsen T, Kristensen AT, Sorensen BB, Olsen OH, Stennicke HR and Petersen LC. Characterization of canine coagulation factor VII and its complex formation with tissue factor: canine-human cross-species compatibility. J Thromb Haemost. 2010;8:1763-1772. [PMID:20524980].
- Callan MB, Aljamali MN, Margaritis P, Griot-Wenk ME, Pollak ES, Werner P, Giger U and High KA. A novel missense mutation responsible for factor VII deficiency in research Beagle colonies. J Thromb Haemost. 2006;4:2616-2622. [PMID:16961583].
- Poon MC, Zotz R, Di Minno G, Abrams ZS, Knudsen JB and Laurian Y. Glanzmann’s thrombasthenia treatment: a prospective observational registry on the use of recombinant human activated factor VII and other hemostatic agents. Semin Hematol. 2006;43:S33-36. [PMID:16427383].
- Nurden AT, Pillois X and Wilcox DA. Glanzmann thrombasthenia: state of the art and future directions. Semin Thromb Hemost. 2013;39:642-655. [PMID:23929305].