Original Article
Nonviral Transfer of the Gene Encoding Coagulation Factor VIII in Patients with Severe Hemophilia A
N Engl J Med 2001; 344:1735-1742June 7, 2001
- Abstract
Background
We tested the safety of a nonviral somatic-cell gene-therapy system in patients with severe hemophilia A.
Methods
An open-label, phase 1 trial was conducted in six patients with severe hemophilia A. Dermal fibroblasts obtained from each patient by skin biopsy were grown in culture and transfected with a plasmid containing sequences of the gene that encodes factor VIII. Cells that produced factor VIII were selected, cloned, and propagated in vitro. The cloned cells were then harvested and administered to the patients by laparoscopic injection into the omentum. The patients were followed for 12 months after the implantation of the genetically altered cells. An interim analysis was performed.
Results
There were no serious adverse events related to the use of factor VIII–producing fibroblasts or the implantation procedure. No long-term complications developed, and no inhibitors of factor VIII were detected. In four of the six patients, plasma levels of factor VIII activity rose above the levels observed before the procedure. The increase in factor VIII activity coincided with a decrease in bleeding, a reduction in the use of exogenous factor VIII, or both. In the patient with the highest level of factor VIII activity, the clinical changes lasted approximately 10 months.
Conclusions
Implantation of genetically altered fibroblasts that produce factor VIII is safe and well tolerated. This form of gene therapy is feasible in patients with severe hemophilia A.
Media in This Article
Figure 2
Bleeding Events and Use of Exogenous Factor VIII in Three of the Six Patients.Figure 1
Steps in the Human Factor VIII Gene-Transfer Procedure.Article Activity
- Article
Hemophilia A, an X-linked hemorrhagic disorder due to mutations in the gene that encodes factor VIII, affects 1 in 5000 males.1 Approximately 60 percent of patients with a mutant gene for factor VIII have severe hemophilia (in which the level of factor VIII activity is less than 1.0 percent of normal), whereas the remainder have moderate or mild hemophilia (factor VIII activity, 1.0 to 5.0 percent of normal or more than 5.0 percent of normal, respectively).1 In severe hemophilia, spontaneous bleeding into joints, soft tissues, and vital organs is frequent, whereas in mild hemophilia, bleeding usually occurs only after trauma or surgery.2
Replacement with intravenously administered concentrates of factor VIII controls bleeding in patients with hemophilia A. In patients with severe disease, spontaneous hemorrhage and progression of arthropathy can be prevented by prophylactic administration of exogenous factor VIII to maintain factor VIII activity levels above 1.0 percent of normal.3 Prophylaxis with factor VIII is costly,1 however, and does not guarantee the prevention of hemorrhage.4 Despite considerable improvements in the safety of plasma-derived and recombinant factor VIII concentrates, concern remains about contamination with infectious prions5,6 and small nonenveloped viruses, such as hepatitis A virus7,8 and parvovirus.9-11 Hemophilia A is a suitable candidate disease for gene therapy12 for several reasons: factor VIII production is not regulated in response to bleeding; the broad therapeutic index of factor VIII minimizes the risk of overdoses; delivery of factor VIII into the bloodstream does not require expression of the gene by a specific organ; and even low levels of the protein can be beneficial.
We have developed a nonviral gene-delivery system termed “transkaryotic implantation.” It entails the isolation of somatic cells from a patient, the stable introduction of a therapeutic gene into these cells, the isolation and clonal propagation of a single engineered cell, and the implantation of the clonal cells into the patient.13-15 Mice that received fibroblast implants carrying DNA sequences of the human factor VIII gene produced the human protein; the procedure was safe and resulted in factor VIII activity levels that exceeded 5.0 percent of normal — a level that would be considered therapeutic in patients with severe hemophilia A — for more than one year (unpublished data). These results led us to study the safety of the transkaryotic-implantation system in patients with severe hemophilia A.
Methods
Patients
The study was designed as a single-institution, open-label, phase 1 trial in which autologous fibroblasts that produced human factor VIII were administered to patients with severe hemophilia A. Patients were eligible to be included in the study if they had hemophilia A with a level of factor VIII activity below 2.0 percent of normal; were at least 15 years old; had received at least 50 days of factor VIII therapy before study entry; had six or more bleeding episodes per year; had normal factor VIII clearance; had a hemoglobin level above 12 g per deciliter (7.4 mmol per liter); and had a platelet count above 100,000 per cubic millimeter. Patients with any of the following characteristics were excluded: the presence of a factor VIII inhibitor, as determined by the Bethesda inhibitor assay (which detects antibodies that neutralize factor VIII activity) at the time of enrollment; a history of inducible factor VIII inhibitor; an anticipated requirement for fixed-dose prophylaxis with factor VIII infusions during the study; a history of opportunistic infection or cancer related to the acquired immunodeficiency syndrome; use of investigational therapy for hemophilia within 30 days before enrollment in the study; or clinical evidence of abdominal adhesions or portal hypertension (which would increase the risk associated with the laparoscopic procedure).
The clinical protocol and the informed-consent document were approved by the Center for Biologics Evaluation and Research of the Food and Drug Administration and by the Office of Biotechnology Activities of the National Institutes of Health. Local review and approval were obtained from the Beth Israel Deaconess Medical Center institutional review board, the Harvard University Biosafety Committee, and the Harvard University Human Gene Therapy Advisory Committee. Permission to treat up to 12 patients and to monitor them for up to two years was given. All the patients provided written informed consent.
Figure 1Figure 1
Steps in the Human Factor VIII Gene-Transfer Procedure. shows the steps involved in the study. First, the patients underwent a clinical evaluation in which a complete medical history was obtained and a physical examination, blood and urine tests, electrocardiography, and chest radiography were performed. Patients received diary forms for use at home to record and describe all bleeding episodes and all infusions of factor VIII during the study. The date, time, and site of bleeding were recorded for each episode, as were the number of units of factor VIII administered with each infusion. The reason for factor VIII infusion was assigned to one of the following categories: spontaneous bleeding, injury-related bleeding, prophylaxis, and other reasons (including use for the skin biopsy, the laparoscopy, and other procedures).Skin Biopsy
A full-thickness, elliptical skin-biopsy specimen (1.6 by 0.4 cm) was obtained from the medial surface of the upper arm after the patient received local anesthesia. Dermal fibroblasts were isolated for cell culture and subsequent plasmid transfection. All the patients received an infusion of factor VIII before the biopsy to increase their plasma factor VIII activity level to 100 percent of normal. Factor VIII was administered for an additional three or four days to maintain normal hemostasis.
Laparoscopy
Approximately seven weeks after the skin biopsy, the patients returned to the hospital for another clinical assessment, which was performed the day before cell implantation. Factor VIII was infused just before surgery to increase plasma factor VIII activity levels to approximately 100 percent of normal, and the infusions were continued two or three times daily for one week after surgery. The laparoscopic procedure was performed while the patients were under general anesthesia. A 10-mm trocar was inserted into the peritoneal cavity through an infraumbilical incision with the use of an open technique. After carbon dioxide insufflation, a laparoscope was introduced at this site. Under direct vision, a single, 5-mm operating trocar was inserted into the midabdomen, and through this trocar, a laparoscopic grasper was used to manipulate the greater omentum. A 9-cm (3.5-in.), 20-gauge spinal needle was inserted directly through the abdominal wall under direct vision, and approximately 0.5 ml of the cell suspension was injected through the needle into the greater omentum at each of several sites. After surgery, the patients were observed overnight in the clinical research center. All six patients were discharged home on the first postoperative day.
Clinical assessments at the patients' homes were performed daily by the study nurse on days 2 through 6 after surgery. The patients were scheduled to undergo clinical assessments at the hospital during weeks 1, 2, 3, 4, 6, 8, 12, and 18 and months 6, 9, and 12 after cell implantation, with additional monitoring through year 2.
Preparation of Autologous Fibroblasts Expressing Factor VIII
A plasmid that contained the gene encoding human factor VIII from which the B domain had been deleted and that contained the human fibronectin promoter was introduced by electroporation into dermal fibroblasts that had been isolated from the skin-biopsy specimen. The B-domain coding sequence was not incorporated into the plasmid, because it is not required for the coagulant activity of factor VIII or the interaction of factor VIII with von Willebrand factor16,17; moreover, the presence of the B domain reduces expression of factor VIII in transfected mammalian cells. Fibronectin, an extracellular-matrix protein, is expressed in fibroblasts; its promoter efficiently directs the transcription of B-domain–deleted factor VIII in primary human dermal fibroblasts.
Stably transfected fibroblast clones that had incorporated the plasmid containing B-domain–deleted factor VIII were selected in G418-containing medium, isolated, expanded in nonselective medium, and characterized. Characterization included measurement of factor VIII expression, assessment of cell-growth properties (including soft-agar tumorigenicity assay in vitro), microbial safety, and Southern blotting. Cells from the clone that was designated for implantation were harvested the day before implantation, extensively washed, and introduced into a syringe. Either 100 million or 400 million cloned cells were then administered. During preparation, the fibroblast cultures were aseptically processed in class 100 conditions.18
Factor VIII Assays
Standard one-stage factor VIII clotting-activity assays and Bethesda inhibitor assays based on measurement of the activated partial-thromboplastin time were performed (Automated APTT reagent, Organon Teknika) on an MDA instrument with the use of a standard curve prepared with a normal human pooled plasma calibration standard (Precision Biologic), which was calibrated against a World Health Organization standard. Abnormal controls (Dade and Behring) and normal controls (Precision Biologic), both derived from human plasma, were used to validate the assays. Serum samples from the patients were analyzed for antibodies against factor VIII with two specific enzyme-linked immunosorbent assays. Full-length human factor VIII derived from Chinese-hamster-ovary cells (Recombinate, Hyland–Baxter) or human plasma-derived factor VIII (Hemofil-M, Hyland–Baxter) was used to coat the wells of microtiter plates before the addition of patients' serum. The enzyme-linked immunosorbent assay was developed with a horseradish peroxidase–conjugated goat antihuman antibody. The assay was validated with the use of normal human plasma and abnormal human plasma containing various amounts of factor VIII antibodies (George King Bio-Medical). All specialized blood-coagulation testing was performed at Esoterix Coagulation Laboratories with use of frozen plasma or serum samples that were collected from the patients at all scheduled visits; however, at month 9 only factor VIII activity was tested, and only in Patients 5 and 6.
Results
Characteristics of the Patients and Factor VIII Production by the Implanted Fibroblasts
Table 1Table 1
Characteristics of the Six Patients. shows the characteristics of the six patients. As shown in Table 2Table 2
Total Factor VIII Production by Implanted Autologous Fibroblasts., Patients 1, 2, and 3 each received 100 million cells, and Patients 4, 5, and 6 each received 400 million cells. The total factor VIII production of each of the clones that was implanted is also shown.Safety
The skin biopsy was well tolerated by all six patients, and there were no episodes of bleeding or infectious complications. Laparoscopic implantation of the fibroblasts was also well tolerated. The mean (±SD) duration of surgery was 65±7 minutes. The mean time required for the injection of the fibroblasts was 15±3 minutes. There were no complications related to anesthesia. After the procedure, all six patients had minor abdominal discomfort at the incision site. Other adverse events during the postoperative period were pain referred to the shoulder in five patients, abdominal ecchymoses in two, and low-grade postoperative fever (which did not require treatment) in one. There were no long-term complications associated with surgery.
All the Bethesda inhibitor assays and enzyme-linked immunosorbent assays for antibodies against factor VIII were negative. Preliminary assays for cytotoxic T lymphocytes at selected times through month 12 in Patients 1, 2, and 3 and through month 6 in Patients 4, 5, and 6, in which the patients' lymphocytes were used as the effector cells and transfected autologous fibroblasts expressing B domain–deleted factor VIII were used as the target, were negative; this finding indicates that there was no cellular immune response to the cultured transfected fibroblasts.
One patient with a long-standing history of recurrent paroxysmal atrial tachycardia had two episodes of tachycardia, 7.5 months and 12 months after the implantation procedure. These two serious adverse events were classified as unrelated to the use of the genetically modified fibroblasts or the implantation. There were no other serious adverse events. All six patients had adverse events, most of which consisted of bleeding episodes or pain associated with bleeding. No clinically significant laboratory abnormalities related to the gene therapy were detected.
Clinical Effects
In addition to evaluating the safety of the use of the genetically engineered fibroblasts and of the surgical procedures (skin biopsy and laparoscopy), we appraised several subjective and objective measures of efficacy: bleeding events related to hemophilia, use of exogenous factor VIII for treatment before and during the study, and factor VIII activity levels. Figure 2Figure 2
Bleeding Events and Use of Exogenous Factor VIII in Three of the Six Patients. shows bleeding events and the use of exogenous factor VIII in Patients 1, 3, and 6, all of whom maintained diaries of bleeding and factor VIII use before participation in the study. Patients 2, 4, and 5 did not keep such diaries before entering the study.In Patient 1, the frequency of bleeding and the amount of exogenous factor VIII used did not change after implantation of the transfected fibroblasts. This patient received the implant with the lowest level of factor VIII production in vitro of any of the implants (Table 2), and no clinically important increase in factor VIII activity was detected in his plasma after therapy (Table 3Table 3
Levels of Factor VIII Activity.). Beginning 10 weeks after the implantation of transfected fibroblasts, Patient 3 had no bleeding, and he did not use factor VIII for the treatment of new bleeding for the next 3 months. At week 18, nearly eight weeks after the most recent infusion of factor VIII, his plasma factor VIII activity had increased from less than 0.4 percent of normal to 2.0 percent of normal (Table 3). There were injury-related bleeding episodes, but he reported no spontaneous bleeding. There were several prolonged intervals without bleeding or infusions of factor VIII (Figure 2). Additional measurements of factor VIII activity showed an activity level of 1.0 percent of normal at 6 months and a decline to less than 0.5 percent at 12 months.Patient 6 received the fibroblast clone that produced the largest amount of factor VIII per day in vitro of any of the clones (Table 2). A decreased frequency of bleeding became evident one month after the transfected fibroblasts were implanted (Figure 2). During periods when there was no bleeding or use of exogenous factor VIII, numerous measurements of factor VIII activity showed a persistent increase in activity to a level above that observed at the initial evaluation (Table 3). Like Patient 3, he reported no spontaneous bleeding for approximately 10 months after the implantation (Figure 2). However, there was intermittent injury-related hemorrhage. The frequency of bleeding in this patient increased and spontaneous bleeding recurred during month 12, at which time his level of factor VIII activity had declined to less than 0.5 percent of normal (Table 3).
Table 4Table 4
Monthly Use of Exogenous Factor VIII. shows the average monthly use of factor VIII before and after cell implantation in all six patients. Treatment with infusions of exogenous factor VIII before enrollment in the study was documented by the patients' diaries (Patients 1, 3, and 6) or by pharmacy records (Patients 2, 4, and 5). Use of exogenous factor VIII by Patients 1 and 2 was unchanged after treatment. Use by Patients 3, 4, and 5 was moderately decreased. In Patient 6 there was a considerable reduction in the use of exogenous factor VIII after gene therapy; in the 20 months before therapy, his total use of factor VIII each month ranged from 12,339 to 30,621 U, whereas his use ranged from 2040 to 7098 U per month at months 4, 6, 7, 8, and 10 after gene therapy (Table 4).A standard one-stage assay of factor VIII activity was used to detect changes in factor VIII activity over time (Table 3). Levels of factor VIII activity above pretreatment levels were repeatedly found after treatment in Patients 3, 4, 5, and 6. Patient 3 had a factor VIII activity level of 2.0 percent of normal at week 18 (eight weeks after his most recent factor VIII infusion). It was 1.0 percent of normal at month 6, nine days after a single infusion of factor VIII. These elevated levels were not the result of the use of exogenous factor VIII, because factor VIII activity was undetectable at other points closer in time to a preceding infusion (weeks 2, 6, 8, and 12). Patient 4 had factor VIII activity levels of 0.8 percent and 0.6 percent of normal at weeks 4 and 6, respectively. Although both of these levels were above his initial level before the implantation (less than 0.4 percent of normal), it is unclear whether these small changes are clinically significant. Nevertheless, the appearance of measurable factor VIII coincided with decreased use of exogenous factor VIII. In Patient 5, several measurements of factor VIII activity between week 4 and month 6 after the implantation were above his initial level (which was undetectable). Although the change in the use of factor VIII was small in this patient, there was a general correlation between decreased use of factor VIII through month 10 and the duration of the elevation in factor VIII activity. Patient 6 had the highest measurable level of activity after gene therapy; at week 12, the activity was 4.0 percent of normal. At months 9 and 12, the activity in this patient had decreased to less than 0.5 percent of normal, and spontaneous bleeding recurred.
Discussion
We studied six patients with severe hemophilia A who received autologous fibroblasts that carried the gene that encodes factor VIII. The implantation procedure was safe, and the cells have been well tolerated, a finding that has continued in all patients during a second year of monitoring. In no patient did evidence of factor VIII inhibitor appear at any time during the study. This is important, because inhibitors have the potential to compromise the effectiveness of standard factor VIII–replacement therapy.19
The potential for clinical efficacy of this type of gene therapy is suggested by the increased levels of factor VIII activity in four of the six patients. Before they entered the study, the factor VIII activity levels in all six patients were less than 0.8 percent of normal — a fact that made it possible to detect relatively small increases in factor VIII in plasma after the implantation of the genetically engineered fibroblasts. There was a general correlation between these increases and clinical improvement, such as a decreased frequency of spontaneous-bleeding episodes or decreased use of exogenous factor VIII.
At the relatively low levels of factor VIII activity we observed, several factors may have influenced subjective measures of the clinical efficacy of the gene-therapy procedure we tested. One of these is the presence or absence of active joint disease: treatment of severe joint disease requires persistent, relatively high levels of factor VIII activity. The patients' investment in the therapeutic outcome of the experimental treatment may also have influenced subjective measures of efficacy that suggested therapeutic benefit. However, the correlation between objective and subjective signs of improvement indicates that a prolonged placebo effect was unlikely.
The minimal levels of factor VIII that are required to protect against spontaneous or post-traumatic bleeding are unknown. Protection against bleeding related to trauma or surgery may require high factor VIII levels, in the range of 30 to 100 percent of normal, depending on the nature of the injury.20 In contrast, the clinical features that distinguish between severe and moderate hemophilia suggest that factor VIII activity levels as low as 1.0 to 2.0 percent of normal can protect against spontaneous bleeding. A recent study of prophylactic treatment in patients with severe hemophilia indicated that even levels below 1.0 percent of normal may be sufficient.21 For these reasons, the clinical responses we observed in our patients, who had low plasma levels of factor VIII, are encouraging.
The nonviral ex vivo gene-therapy system we used has several potential advantages over gene therapy based on viral vectors. Patients received a homogeneous clonal population of cells containing a single genetic modification, thereby minimizing the risk of insertional mutagenesis that might occur with viral vectors. In addition, viral vectors can evoke immune responses that may attenuate the effectiveness of subsequent treatments.22 Viral vectors may become infectious by mutation or other mechanisms or may modify the patient's germ line, but these risks are obviated with the use of the nonviral approach. Despite these advantages, our system has noteworthy disadvantages. The implantation procedure is moderately invasive, and the factor VIII–producing autologous fibroblasts must be prepared individually for each patient. Fortunately, there is no technological obstacle to the production of such patient-specific fibroblasts, so the procedure could be feasible if the therapeutic benefit of this approach can be proved in subsequent studies.
Supported by Transkaryotic Therapies.
Drs. Treco and Selden are employees of and own stock in Transkaryotic Therapies.
We are indebted to the patients with hemophilia and their families for their commitment to this investigation and their continuous support and trust; to members of the Departments of Medicine, Surgery, Anesthesia, and Nursing and of the General Clinical Research Center at the Beth Israel Deaconess Medical Center for their contributions; to members of the basic-science and clinical-research teams at Transkaryotic Therapies for their help; and to Dr. Paula Fraenkel of Beth Israel Deaconess Medical Center for her assistance with patient care.
Source Information
From the Center for Hemostasis and Thrombosis Research, Department of Medicine (D.A.R., J.M.O.), and the Department of Surgery (N.E.T.), Beth Israel Deaconess Medical Center and Harvard Medical School, Boston; and Transkaryotic Therapies, Cambridge, Mass. (D.A.T., R.FS.).
Address reprint requests to Dr. Roth at Beth Israel Deaconess Medical Center, Center for Hemostasis and Thrombosis Research, RE-302, 41 Ave. Louis Pasteur, Boston, MA 02115, or at droth@caregroup.harvard.edu.
Appendix
Other members of the Factor VIII Transkaryotic Therapy Study Group were as follows (asterisks indicate a financial interest in Transkaryotic Therapies): J.D. Levine, J. Proper, and B. Furie* (Beth Israel Deaconess Medical Center and Harvard Medical School, Boston) and V.A. Roman,* Z.M. Sabine,* C.W. Phillips,* A.M. Zuliani,* N.A. Savioli,* D. Fisher,* J. Harrington,* M. Borowski,* M.W. Heartlein,* J.C. Lamsa,* E.M. Morrel,* and K.C. Gunter* (Transkaryotic Therapies, Cambridge, Mass.).
- References
References
1
Lozier JN, Kessler CN. Clinical aspects and therapy of hemophilia. In: Hoffman R, Benz E Jr, Shattil S, et al., eds. Hematology: basic principles and practice. New York: Churchill Livingstone, 2000:1883-1904.
2
Richard KA. The diagnosis of hemophilia A and B and von Willebrand's disease. In: Forbes CD, Aledort LM, Madhok R, eds. Hemophilia. London: Chapman & Hall Medical, 1997:53-62.
3
Nilsson IM ,Berntorp E ,Lofqvist T ,Pettersson H . Twenty-five years' experience of prophylactic treatment in severe haemophilia A and B. J Intern Med 1992;232:25-32
CrossRef | Web of Science | Medline4
Manco-Johnson MJ ,Nuss R ,Geraghty S ,Funk S ,Kilcoyne R . Results of secondary prophylaxis in children with severe hemophilia. Am J Hematol 1994;47:113-117
CrossRef | Web of Science | Medline5
Will R ,Ironside J ,Zeidler M , et al. A new variant of Creutzfeldt-Jakob disease in the UK. Lancet 1996;347:921-925
CrossRef | Web of Science | Medline6
Evatt B . Prions and haemophilia: assessment of risk. Haemophilia 1998;4:628-633
CrossRef | Web of Science | Medline7
Vermylen J ,Peerlinck K . Review of the hepatitis A epidemics in hemophiliacs in Europe. Vox Sang 1994;67:Suppl 4:8-11
CrossRef | Web of Science | Medline8
Mariani G ,Di Paolantonio T ,Baklaya R ,Morfini M ,Mannucci PM . Prospective study of the evaluation of hepatitis C virus infectivity in a high-purity, solvent/detergent-treated factor VIII concentrate: parallel evaluation of other markers for lipid-enveloped and non-lipid-enveloped viruses. Transfusion 1993;33:814-818
CrossRef | Web of Science | Medline9
Bartolomei Corsi O ,Azzi A ,Morfini M ,Fanci R ,Rossi Ferrini P . Human parvovirus infection in haemophiliacs first infused with treated clotting factor concentrates. J Med Virol 1988;25:165-170
CrossRef | Web of Science | Medline10
Flores G ,Juarez JC ,Montoro JB , et al. Seroprevalence of parvovirus B19, cytomegalovirus, hepatitis A virus and hepatitis E virus antibodies in haemophiliacs treated exclusively with clotting-factor concentrates considered safe against human immunodeficiency and hepatitis C viruses. Haemophilia 1995;1:115-117
CrossRef | Web of Science11
Yee T ,Cohen BJ ,Pasi KJ ,Lee CA . Transmission of symptomatic parvovirus B19 infection by clotting factor concentrate. Br J Haematol 1996;93:457-459
CrossRef | Web of Science | Medline12
Kaufman RJ . Advances toward gene therapy for hemophilia at the millennium. Hum Gene Ther 1999;10:2091-2107
CrossRef | Web of Science | Medline13
Selden RF ,Skoskiewicz MJ ,Howie KB ,Russell PS ,Goodman HM . Implantation of genetically engineered fibroblasts into mice: implications for gene therapy. Science 1987;236:714-718
CrossRef | Web of Science | Medline14
Selden RF , Skośkiewicz M,Russell PS ,Goodman HM . Regulation of insulin-gene expression: implications for gene therapy. N Engl J Med 1987;317:1067-1076
Full Text | Web of Science | Medline15
Heartlein MW ,Roman VA ,Jiang J-L , et al. Long-term production and delivery of human growth hormone in vivo. Proc Natl Acad Sci U S A 1994;91:10967-10971
CrossRef | Web of Science | Medline16
Toole JJ ,Pittman DD ,Orr EC ,Murtha P ,Wasley L ,Kaufman RJ . A large region (approximately equal to 95 kDa) of human factor VIII is dispensable for in vitro procoagulant activity. Proc Natl Acad Sci U S A 1986;83:5939-5942
CrossRef | Web of Science | Medline17
Eaton DL ,Wood WI ,Eaton D , et al. Construction and characterization of an active factor VIII variant lacking the central one-third of the molecule. Biochemistry 1986;25:8343-8347
CrossRef | Web of Science | Medline18
Federal standard 209E: airborne particulate cleanliness classes in clean rooms and clean zones. Washington, D.C.: General Services Administration, September 11, 1992.
19
Hedner U . Treatment of patients with factor VIII and factor IX inhibitors with special focus on the use of recombinant factor VIIa. Thromb Haemost 1999;82:531-539
Web of Science | Medline20
DiMichele D ,Neufeld JE . Hemophilia: a new approach to an old disease. Hematol Oncol Clin North Am 1998;12:1315-1344
CrossRef | Web of Science | Medline21
Petrini P . What factors should influence the dosage and interval of prophylactic treatment in patients with severe haemophilia A and B? Haemophilia 2001;7:99-102
CrossRef | Web of Science | Medline22
Kay MA ,Manno CS ,Ragni MV , et al. Evidence for gene transfer and expression of factor IX in haemophilia B patients with an AAV vector. Nat Genet 2000;24:257-261
CrossRef | Web of Science | Medline
- Citing Articles (127)
Citing Articles
1
Hideto Matsui. (2012) Endothelial progenitor cell-based therapy for hemophilia A. International Journal of Hematology
CrossRef2
Denise E. Sabatino, Timothy C. Nichols, Elizabeth Merricks, Dwight A. Bellinger, Roland W. Herzog, Paul E. Monahan. 2012. Animal Models of Hemophilia. , 151-209.
CrossRef3
Christopher Luis Chavez, Annahita Keravala, Jacqueline N. Chu, Alfonso P. Farruggio, Vanessa E. Cuellar, Jan Voorberg, Michele P Calos. (2011) Long-term expression of human coagulation factor VIII in a tolerant mouse model using the phiC31 integrase system. Human Gene Therapy111111101256006
CrossRef4
Deepak Raj, Andrew M Davidoff, Amit C Nathwani. (2011) Self-complementary adeno-associated viral vectors for gene therapy of hemophilia B: progress and challenges. Expert Review of Hematology 4:5, 539-549
CrossRef5
Hajime Kurosaki, Masaharu Hiratsuka, Natsuko Imaoka, Yuichi Iida, Narumi Uno, Yasuhiro Kazuki, Chie Ishihara, Yuwna Yakura, Jun Mimuro, Youichi Sakata, Hiroyuki Takeya, Mitsuo Oshimura. (2011) Integration-free and stable expression of FVIII using a human artificial chromosome. Journal of Human Genetics 56:10, 727-733
CrossRef6
Christopher D. Porada, Chad Sanada, Chung-Jung Kuo, Evan Colletti, Walter Mandeville, John Hasenau, Esmail D. Zanjani, Robert Moot, Christopher Doering, H. Trent Spencer, Graça Almeida-Porada. (2011) Phenotypic correction of hemophilia A in sheep by postnatal intraperitoneal transplantation of FVIII-expressing MSC. Experimental Hematology
CrossRef7
A Keravala, C L Chavez, G Hu, L E Woodard, P E Monahan, M P Calos. (2011) Long-term phenotypic correction in factor IX knockout mice by using phiC31 integrase-mediated gene therapy. Gene Therapy 18:8, 842-848
CrossRef8
K. A. HIGH. (2011) Gene therapy for haemophilia: a long and winding road. Journal of Thrombosis and Haemostasis 9, 2-11
CrossRef9
Matthias Titeux, Alain Hovnanian. 2011. Skin Gene and Cell Therapy. , 140.1-140.22.
CrossRef10
Maria-Teresa Alvarez-Román, Monica Martín-Salces, Victor Jiménez-Yuste, Emérito-Carlos Rodríguez-Merchán. 2011. Current and Future Approaches to Gene Therapy in Patients with Hemophilia. , 146-149.
CrossRef11
Barbara Langer, Klaus Cichutek. (2011) Klinische Anwendung der Gentherapie. Bisherige Erfahrungen und Zukunft. Pharmazie in unserer Zeit 40:3, 254-262
CrossRef12
Hideto Matsui, Carol Hegadorn, Margareth Ozelo, Erin Burnett, Angie Tuttle, Andrea Labelle, Paul B McCray, Luigi Naldini, Brian Brown, Christine Hough, David Lillicrap. (2011) A MicroRNA-regulated and GP64-pseudotyped Lentiviral Vector Mediates Stable Expression of FVIII in a Murine Model of Hemophilia A. Molecular Therapy 19:4, 723-730
CrossRef13
Luc JW van der Laan, Yigang Wang, Hugo W Tilanus, Harry LA Janssen, Qiuwei Pan. (2011) AAV-mediated gene therapy for liver diseases: the prime candidate for clinical application?. Expert Opinion on Biological Therapy 11:3, 315-327
CrossRef14
R. R. Montgomery, Q. Shi. (2010) Alternative Strategies for Gene Therapy of Hemophilia. Hematology 2010:1, 197-202
CrossRef15
Inge Petrus, Marinee Chuah, Thierry VandenDriessche. (2010) Gene therapy strategies for hemophilia: benefits versus risks. The Journal of Gene Medicine 12:10, 797-809
CrossRef16
Qizhen Shi, Robert R. Montgomery. (2010) Platelets as delivery systems for disease treatments. Advanced Drug Delivery Reviews 62:12, 1196-1203
CrossRef17
Carol H Miao. (2010) Immunomodulation for inhibitors in hemophilia A: the important role of Treg cells. Expert Review of Hematology 3:4, 469-483
CrossRef18
R. R. MONTGOMERY, P. E. MONAHAN, M. C. OZELO. (2010) Unique strategies for therapeutic gene transfer in haemophilia A and haemophilia B. Haemophilia 16, 29-34
CrossRef19
Zaida Alipio, Dorothy M Adcock, Milton Waner, Teresa M-J O, Louis M Fink, David C Ward, Yupo Ma. (2010) Sustained factor VIII production in hemophiliac mice 1 year after engraftment with induced pluripotent stem cell-derived factor VIII producing endothelial cells. Blood Coagulation & Fibrinolysis 21:5, 502-504
CrossRef20
Steven W. Pipe. 2010. Gene Therapy: Molecular Engineering of Factor VIII and Factor IX. , 239-246.
CrossRef21
Katherine A. High, J. Fraser Wright. 2010. Gene Therapy Trials in Hemophilia A and B. , 231-238.
CrossRef22
Jon P Connelly, Jenny C Barker, Shondra Pruett-Miller, Matthew H Porteus. (2010) Gene Correction by Homologous Recombination With Zinc Finger Nucleases in Primary Cells From a Mouse Model of a Generic Recessive Genetic Disease. Molecular Therapy 18:6, 1103-1110
CrossRef23
Sara Pérez-Luz, Javier Díaz-Nido. (2010) Prospects for the Use of Artificial Chromosomes and Minichromosome-Like Episomes in Gene Therapy. Journal of Biomedicine and Biotechnology 2010, 1-17
CrossRef24
Rui-Jun Su, Angela Epp, Yvette Latchman, Doug Bolgiano, Steven W Pipe, Neil C Josephson. (2010) Suppression of FVIII Inhibitor Formation in Hemophilic Mice by Delivery of Transgene Modified Apoptotic Fibroblasts. Molecular Therapy 18:1, 214-222
CrossRef25
Christopher B Doering, H Trent Spencer. (2009) Advancements in gene transfer-based therapy for hemophilia A. Expert Review of Hematology 2:6, 673-683
CrossRef26
Daniel L. Coutu, Azizeh-Mitra Yousefi, Jacques Galipeau. (2009) Three-dimensional porous scaffolds at the crossroads of tissue engineering and cell-based gene therapy. Journal of Cellular Biochemistry 108:3, 537-546
CrossRef27
Ai Hong Zhang, Jonathan Skupsky, David W. Scott. (2009) Factor VIII Inhibitors: Risk Factors and Methods for Prevention and Immune Modulation. Clinical Reviews in Allergy & Immunology 37:2, 114-124
CrossRef28
Michael Callaghan, Randal J Kaufman. 2009. Haemophilias: Gene Therapy. .
CrossRef29
A. LIRAS, S. OLMEDILLAS. (2009) Gene therapy for haemophilia…yes, but…with non-viral vectors?. Haemophilia 15:3, 811-816
CrossRef30
D. Xu, Z. Alipio, L. M. Fink, D. M. Adcock, J. Yang, D. C. Ward, Y. Ma. (2009) From the Cover: Phenotypic correction of murine hemophilia A using an iPS cell-based therapy. Proceedings of the National Academy of Sciences 106:3, 808-813
CrossRef31
Katherine Bowman, Rita Sarkar, Sanj Raut, Kam W. Leong. (2008) Gene transfer to hemophilia A mice via oral delivery of FVIII–chitosan nanoparticles. Journal of Controlled Release 132:3, 252-259
CrossRef32
B. Peng, P. Ye, B. R. Blazar, G. J. Freeman, D. J. Rawlings, H. D. Ochs, C. H. Miao. (2008) Transient blockade of the inducible costimulator pathway generates long-term tolerance to factor VIII after nonviral gene transfer into hemophilia A mice. Blood 112:5, 1662-1672
CrossRef33
Aiman O. Abbas, Maureen D. Donovan, Aliasger K. Salem. (2008) Formulating poly(lactide-co-glycolide) particles for plasmid DNA delivery. Journal of Pharmaceutical Sciences 97:7, 2448-2461
CrossRef34
P. M. MANNUCCI. (2008) Back to the future: a recent history of haemophilia treatment. Haemophilia 14:s3, 10-18
CrossRef35
PH Tan, SE Janes, AI Handa, PJ Friend. (2008) More trouble ahead; is gene therapy coming of age?. Expert Opinion on Biological Therapy 8:5, 561-567
CrossRef36
H.E. Blum. (2008) Gentherapie von hereditären Lebererkrankungen. Der Gastroenterologe 3:3, 227-232
CrossRef37
Hubert E. Blum. (2008) Molecular therapy and prevention of liver diseases. Virologica Sinica 23:2, 81-92
CrossRef38
Samuel L. Murphy, Katherine A. High. (2008) Gene therapy for haemophilia. British Journal of Haematology 140:5, 479-487
CrossRef39
J. KIMMELMAN. (2007) Staunch protections: the ethics of haemophilia gene transfer research. Haemophilia 0:0, 071115150741001-???
CrossRef40
Jianping Wen, Nong Xu, Anna Li, Jacqueline Bourgeois, Frederick A. Ofosu, Gonzalo Hortelano. (2007) Encapsulated human primary myoblasts deliver functional hFIX in hemophilic mice. The Journal of Gene Medicine 9:11, 1002-1010
CrossRef41
U Griesenbach. (2007) Progress and Prospects: Gene Therapy Clinical Trials (Part 2). Gene Therapy 14:22, 1555-1563
CrossRef42
Hideto Matsui, Masaru Shibata, Brian Brown, Andrea Labelle, Carol Hegadorn, Chandler Andrews, Robert P. Hebbel, Jacques Galipeau, Christine Hough, David Lillicrap. (2007) Ex Vivo Gene Therapy for Hemophilia A That Enhances Safe Delivery and Sustained In Vivo Factor VIII Expression from Lentivirally Engineered Endothelial Progenitors. Stem Cells 25:10, 2660-2669
CrossRef43
Yesim Dargaud, Claude Negrier. (2007) Haemophilia therapies. Expert Opinion on Biological Therapy 7:5, 651-663
CrossRef44
Q. SHI, D. A. WILCOX, S. A. FAHS, J. FANG, B. D. JOHNSON, L. M. DU, D. DESAI, R. R. MONTGOMERY. (2007) Lentivirus-mediated platelet-derived factor VIII gene therapy in murine haemophilia A. Journal of Thrombosis and Haemostasis 5:2, 352-361
CrossRef45
M. Potter, A. Li, P. Cirone, F. Shen, P. Chang. 2007. Artificial cells as a novel approach to gene therapy. , 236-291.
CrossRef46
K. A. High. (2007) Update on Progress and Hurdles in Novel Genetic Therapies for Hemophilia. Hematology 2007:1, 466-472
CrossRef47
Lisa N. Boggio, Craig M. Kessler. 2007. Hemophilia A and B. , 43-59.
CrossRef48
Shu Uin Gan, Oi Lian Kon, Roy Y Calne. (2006) Genetic engineering for haemophilia A. Expert Opinion on Biological Therapy 6:10, 1023-1030
CrossRef49
Edward A. Burton, David J. Fink, Joseph C. Glorioso. 2006. Vectors and Gene Therapy. .
CrossRef50
Katherine P Ponder. (2006) Gene therapy for hemophilia. Current Opinion in Hematology 13:5, 301-307
CrossRef51
D. LILLICRAP, T. VANDENDRIESSCHE, K. HIGH. (2006) Cellular and genetic therapies for haemophilia. Haemophilia 12:s3, 36-41
CrossRef52
S Indraccolo, L Moserle, V Tisato, E Gola, S Minuzzo, V Roni, L Persano, L Chieco-Bianchi, A Amadori. (2006) Gene therapy of ovarian cancer with IFN-α-producing fibroblasts: comparison of constitutive and inducible vectors. Gene Therapy 13:12, 953-965
CrossRef53
Roland W Herzog, Ou Cao, J Nathan Hagstrom, Lixin Wang. (2006) Gene therapy for treatment of inherited haematological disorders. Expert Opinion on Biological Therapy 6:5, 509-522
CrossRef54
Timothy P. O'Connor, Ronald G. Crystal. (2006) Genetic medicines: treatment strategies for hereditary disorders. Nature Reviews Genetics 7:4, 261-276
CrossRef55
Maki Goto, Daisuke Sawamura, Kei Ito, Masataka Abe, Wataru Nishie, Kaori Sakai, Akihiko Shibaki, Masashi Akiyama, Hiroshi Shimizu. (2006) Fibroblasts Show More Potential as Target Cells than Keratinocytes in COL7A1 Gene Therapy of Dystrophic Epidermolysis Bullosa. Journal of Investigative Dermatology 126:4, 766-772
CrossRef56
Jianping Wen, Andrew Gómez Vargas, Frederick A. Ofosu, Gonzalo Hortelano. (2006) Sustained and therapeutic levels of human factor IX in hemophilia B mice implanted with microcapsules: key role of encapsulated cells. The Journal of Gene Medicine 8:3, 362-369
CrossRef57
Mahmoud A. Srour, Thilo Albert, Zaid Aburubaiha, Jan Grupp, Alexandra Schmitt, Rainer Schwaab. (2006) Gene Therapy for Hemophilia*. Transfusion Medicine and Hemotherapy 33:2, 165-168
CrossRef58
Akira Ishiwata, Jun Mimuro, Yuji Kashiwakura, Masanori Niimura, Katsuhiro Takano, Tsukasa Ohmori, Seiji Madoiwa, Hiroaki Mizukami, Takashi Okada, Hiroyuki Naka, Akira Yoshioka, Keiya Ozawa, Yoichi Sakata. (2006) Phenotype correction of hemophilia A mice with adeno-associated virus vectors carrying the B domain-deleted canine factor VIII gene. Thrombosis Research 118:5, 627-635
CrossRef59
Janik Adriaansen, Margriet J. Vervoordeldonk, Sandra Vanderbyl, Gary de Jong, Paul P. Tak. (2006) A novel approach for gene therapy: engraftment of fibroblasts containing the artificial chromosome expression system at the site of inflammation. The Journal of Gene Medicine 8:1, 63-71
CrossRef60
P. H. Tan, C. L. H. Chan, C. Chan, A. J. T. George. (2005) The evolving role of gene-based treatment in surgery. British Journal of Surgery 92:12, 1466-1480
CrossRef61
S. W. PIPE. (2005) The promise and challenges of bioengineered recombinant clotting factors. Journal of Thrombosis and Haemostasis 3:8, 1692-1701
CrossRef62
Jochen Graw, Hans-Hermann Brackmann, Johannes Oldenburg, Reinhard Schneppenheim, Michael Spannagl, Rainer Schwaab. (2005) Haemophilia A: from mutation analysis to new therapies. Nature Reviews Genetics 6:6, 488-501
CrossRef63
Oranuch Thanaketpaisarn, Makiya Nishikawa, Fumiyoshi Yamashita, Mitsuru Hashida. (2005) Tissue-Specific Characteristics of in Vivo Electric Gene: Transfer by Tissue and Intravenous Injection of Plasmid DNA. Pharmaceutical Research 22:6, 883-891
CrossRef64
D. B. BRETTLER. (2005) Gene therapy for hemophilia?. Journal of Thrombosis and Haemostasis 3:6, 1317-1319
CrossRef65
C. HOUGH, D. LILLICRAP. (2005) Gene therapy for hemophilia: an imperative to succeed. Journal of Thrombosis and Haemostasis 3:6, 1195-1205
CrossRef66
Saiho Ko, Ichiro Tanaka, Hiromichi Kanehiro, Hideki Kanokogi, Jun-ichi Ori, Midori Shima, Akira Yoshioka, Alan Giles, Yoshiyuki Nakajima. (2005) Preclinical experiment of auxiliary partial orthotopic liver transplantation as a curative treatment for hemophilia. Liver Transplantation 11:5, 579-584
CrossRef67
L. Xu, T. C. Nichols, R. Sarkar, S. McCorquodale, D. A. Bellinger, K. P. Ponder. (2005) Absence of a desmopressin response after therapeutic expression of factor VIII in hemophilia A dogs with liver-directed neonatal gene therapy. Proceedings of the National Academy of Sciences 102:17, 6080-6085
CrossRef68
N K F Chen, J Sivalingam, S Y Tan, O L Kon. (2005) Plasmid-electroporated primary hepatocytes acquire quasi-physiological secretion of human insulin and restore euglycemia in diabetic mice. Gene Therapy 12:8, 655-667
CrossRef69
Yun Chau Long, S. Jaichandran, Liam Pock Ho, Sim Leng Tien, Soo Yong Tan, Oi Lian Kon. (2005) FVIII gene delivery by muscle electroporation corrects murine hemophilia A. The Journal of Gene Medicine 7:4, 494-505
CrossRef70
C. M. Kessler, J. C. Gill, G. C. White, A. Shapiro, S. Arkin, D. A. Roth, X. Meng, J. M. Lusher. (2005) B-domain deleted recombinant factor VIII preparations are bioequivalent to a monoclonal antibody purified plasma-derived factor VIII concentrate: a randomized, three-way crossover study. Haemophilia 11:2, 84-91
CrossRef71
Guy Young, Louis Aledort. (2005) Therapy for haemophilia: recent advances and goals for the future. Expert Opinion on Emerging Drugs 10:1, 173-184
CrossRef72
Weila Wang, Michael Merchlinsky, John Inman, Basil Golding. (2005) Identification of a novel immunodominant cytotoxic T lymphocyte epitope derived from human factor VIII in a murine model of hemophilia A. Thrombosis Research 116:4, 335-344
CrossRef73
Amy L. Dunn, Thomas C. Abshire. (2004) Recent advances in the management of the child who has hemophilia. Hematology/Oncology Clinics of North America 18:6, 1249-1276
CrossRef74
Marie-Helene Rodriguez, Jean-Luc Plantier, Nathalie Enjolras, Muriel Rea, Marylene Leboeuf, Georges Uzan, Claude Negrier. (2004) Biosynthesis of FVIII in megakaryocytic cells: improved production and biochemical characterization. British Journal of Haematology 127:5, 568-575
CrossRef75
M. K. L. Chuah, D. Collen, T. Vandendriessche. (2004) Preclinical and clinical gene therapy for haemophilia. Haemophilia 10:s4, 119-125
CrossRef76
Jay Lozier. (2004) Gene therapy of the hemophilias. Seminars in Hematology 41:4, 287-296
CrossRef77
C. Negrier. (2004) Gene therapy for hemophilia? Yes. Journal of Thrombosis and Haemostasis 2:8, 1234-1235
CrossRef78
Kelly M. Axsom, Catherine S. Manno. (2004) Treatment of Hemophilia With Genetic Engineering. Transfusion Alternatives in Transfusion Medicine 6:2, 46-54
CrossRef79
A. L. Nicolle, K. L. Talks, J. Hanley. (2004) The Genetics of bleeding disorders: a report on the UK Haemophilia Centre Doctors' Organisation annual scientific symposium, 10th October 2003. Haemophilia 10:4, 390-396
CrossRef80
A. C. Nathwani, A. M. Davidoff, E. G. D. Tuddenham. (2004) Prospects for gene therapy of haemophilia. Haemophilia 10:4, 309-318
CrossRef81
J. M. Teitel, D. Barnard, S. Israels, D. Lillicrap, M.-C. Poon, J. Sek. (2004) Home management of haemophilia. Haemophilia 10:2, 118-133
CrossRef82
Paul L F Giangrande. (2004) Blood Products for Hemophilia. BioDrugs 18:4, 225-234
CrossRef83
HONGJUN WANG, CLEMENS A. VAN BLITTERSWIJK, MARION BERTRAND-DE HAAS, ARNOLD H. SCHUURMAN, EVERT N. LAMME. (2004) IMPROVED ENZYMATIC ISOLATION OF FIBROBLASTS FOR THE CREATION OF AUTOLOGOUS SKIN SUBSTITUTES. In Vitro Cellular & Developmental Biology - Animal 40:8, 268
CrossRef84
R. J. Kaufman. (2003) Good things come in small packages for hemophilia. Journal of Thrombosis and Haemostasis 1:12, 2472-2473
CrossRef85
D. A. Wilcox, Q. Shi, P. Nurden, S. L. Haberichter, J. B. Rosenberg, B. D. Johnson, A. T. Nurden, G. C. White Ii, R. R. Montgomery. (2003) Induction of megakaryocytes to synthesize and store a releasable pool of human factor VIII. Journal of Thrombosis and Haemostasis 1:12, 2477-2489
CrossRef86
G. M. Bhopale, R. K. Nanda. (2003) Blood coagulation factor VIII: An overview. Journal of Biosciences 28:6, 783-789
CrossRef87
Frank Park. (2003) Correction of Bleeding Diathesis Without Liver Toxicity Using Arenaviral-Pseudotyped HIV-1–Based Vectors in Hemophilia A Mice. Human Gene Therapy 14:15, 1489-1494
CrossRef88
Carol H. Miao, Xin Ye, Arthur R. Thompson. (2003) High-Level Factor VIII Gene Expression In Vivo Achieved by Nonviral Liver-Specific Gene Therapy Vectors. Human Gene Therapy 14:14, 1297-1305
CrossRef89
Pontus Blomberg, CI Edvard Smith. (2003) Gene therapy of monogenic and cardiovascular disorders. Expert Opinion on Biological Therapy 3:6, 941-949
CrossRef90
Dieter C. Gruenert, Emanuela Bruscia, Giuseppe Novelli, Alessia Colosimo, Bruno Dallapiccola, Federica Sangiuolo, Kaarin K. Goncz. (2003) Sequence-specific modification of genomic DNA by small DNA fragments. Journal of Clinical Investigation 112:5, 637-641
CrossRef91
Roland W Herzog, Valder R Arruda. (2003) Update on gene therapy for hereditary hematological disorders. Expert Review of Cardiovascular Therapy 1:2, 215-232
CrossRef92
K Smith. (2003) Gene therapy: theoretical and bioethical concepts. Archives of Medical Research 34:4, 247-268
CrossRef93
P. M. Mannucci. (2003) Hemophilia: treatment options in the twenty-first century. Journal of Thrombosis and Haemostasis 1:7, 1349-1355
CrossRef94
T. VandenDriessche, D. Collen, M. K. L. Chuah. (2003) Gene therapy for the hemophilias. Journal of Thrombosis and Haemostasis 1:7, 1550-1558
CrossRef95
Catherine S. Manno. (2003) The promise of third-generation recombinant therapy and gene therapy. Seminars in Hematology 40, 23-28
CrossRef96
Q Shi, D.A Wilcox, S.A Fahs, P.A Kroner, R.R Montgomery. (2003) Expression of human factor VIII under control of the platelet-specific αIIb promoter in megakaryocytic cell line as well as storage together with VWF. Molecular Genetics and Metabolism 79:1, 25-33
CrossRef97
Paula HB Bolton-Maggs, K John Pasi. (2003) Haemophilias A and B. The Lancet 361:9371, 1801-1809
CrossRef98
J-L. Plantier, N. Enjolras, M-H. E. Rodriguez, J-M. Masse, E. M. Cramer, C. Negrier. (2003) The P-selectin cytoplasmic domain directs the cellular storage of a recombinant chimeric factor IX. Journal of Thrombosis and Haemostasis 1:2, 292-299
CrossRef99
Susana Ortiz-Urda, Qun Lin, Cheryl L. Green, Douglas R. Keene, M. Peter Marinkovich, Paul A. Khavari. (2003) Injection of genetically engineered fibroblasts corrects regenerated human epidermolysis bullosa skin tissue. Journal of Clinical Investigation 111:2, 251-255
CrossRef100
X. Ye, K. R. Loeb, D. W. Stafford, A. R. Thompson, C. H. Miao. (2003) Complete and sustained phenotypic correction of hemophilia B in mice following hepatic gene transfer of a high-expressing human factor IX plasmid. Journal of Thrombosis and Haemostasis 1:1, 103-111
CrossRef101
A. Van Damme, M. K. L. Chuah, F. Dell'accio, C. De Bari, F. Luyten, D. Collen, T. VandenDriessche. (2003) Bone marrow mesenchymal cells for haemophilia A gene therapy using retroviral vectors with modified long-terminal repeats. Haemophilia 9:1, 94-103
CrossRef102
G. C. White. (2003) Antenatal cell gene transfer: an immunological Trojan horse?. Journal of Thrombosis and Haemostasis 1:1, 14-15
CrossRef103
HOWARD A. PEARSON. (2002) History of Pediatric Hematology Oncology. Pediatric Research 52:6, 979-992
CrossRef104
Catherine S. Manno. (2002) Gene therapy for bleeding disorders. Current Opinion in Hematology 9:6, 511-515
CrossRef105
Michel Alcalay, Adeline Deplas. (2002) Rheumatological management of patients with hemophilia. Part I: joint manifestations. Joint Bone Spine 69:5, 442-449
CrossRef106
Colleen Notley, Annie Killoran, Cherie Cameron, Kimberly Wynd, Christine Hough, David Lillicrap. (2002) The Canine Factor VIII 3′-Untranslated Region and a Concatemeric Hepatocyte Nuclear Factor 1 Regulatory Element Enhance Factor VIII Transgene Expression In Vivo. Human Gene Therapy 13:13, 1583-1593
CrossRef107
Paul E. Monahan, Gilbert C. White. (2002) Hemophilia gene therapy: Update. Current Opinion in Hematology 9:5, 430-436
CrossRef108
R Ardaillou. (2002) La thérapie génique : sa place actuelle et son avenir. La Revue de Médecine Interne 23:8, 679-682
CrossRef109
T. Tonn, C. Herder, S. Becker, E. Seifried, M. Grez. (2002) Generation and Characterization of Human Hematopoietic Cell Lines Expressing Factor VIII. Journal of Hematotherapy <html_ent glyph="@amp;" ascii="&"/> Stem Cell Research 11:4, 695-704
CrossRef110
Rikke Christensen, Flemming Güttler, Thomas G Jensen. (2002) Comparison of epidermal keratinocytes and dermal fibroblasts as potential target cells for somatic gene therapy of phenylketonuria. Molecular Genetics and Metabolism 76:4, 313-318
CrossRef111
C. S. Lengsfeld, T. J. Anchordoquy. (2002) Shear-induced degradation of plasmid DNA. Journal of Pharmaceutical Sciences 91:7, 1581-1589
CrossRef112
KATHERINE HIGH. (2002) Gene-Based Approaches to the Treatment of Hemophilia. Annals of the New York Academy of Sciences 961:1, 63-64
CrossRef113
CHRISTOPHER L. KARP, RICHARD WENSTRUP, DAVID L. BUTLER. (2002) Immunological Barriers (Opportunities?) to the Use of Bioengineered Tissue. Annals of the New York Academy of Sciences 961:1, 337-340
CrossRef114
K. Kelley, I. Verma, G. F. Pierce. (2002) Gene therapy: reality or myth for the global bleeding disorders community?. Haemophilia 8:3, 261-267
CrossRef115
Kazuhiro Oka, Lawrence Chan. (2002) Recent advances in liver-directed gene therapy for dyslipidemia. Current Atherosclerosis Reports 4:3, 199-207
CrossRef116
Y. Israel, F. T. Crews, R. G. Thurman, G.- C. Tu, E. Garver, B. Ponnappa, E. Karahanian, R. Rubin, B. Hoplight, M. Sethna, R. Hanes, M. B. Wilkie, M. D. Wheeler. (2002) Gene and Antisense Delivery in Alcoholism Research. Alcoholism: Clinical and Experimental Research 26:4, 582-585
CrossRef117
Carmen Garc
a-Mart
CrossRef118
Sunitha Govindaswamy, Jim Chandler, Ray Latimer, Alain Vuylsteke. (2002) Management of the patient with coagulation disorders. Current Opinion in Anaesthesiology 15:1, 19-25
CrossRef119
J.M. Kaminski, K. Nguyen, M. Buyyounouski, A. Pollack. (2002) Prostate cancer gene therapy and the role of radiation. Cancer Treatment Reviews 28:1, 49-64
CrossRef120
Christopher E. Walsh. (2002) Gene therapy for the hemophilias. Current Opinion in Pediatrics 14:1, 12-16
CrossRef121
Giuseppe Novelli. (2002) Genome Medicine: Gene Therapy for the Millennium. Pharmacogenomics 3:1, 15-18
CrossRef122
Makiya Nishikawa, Mitsuru Hashida. (2002) Nonviral Approaches Satisfying Various Requirements for Effective in Vivo Gene Therapy. Biological & Pharmaceutical Bulletin 25:3, 275-283
CrossRef123
K. John Pasi. (2001) Gene therapy for haemophilia. British Journal of Haematology 115:4, 744-757
CrossRef124
Miller, Daniel G., Stamatoyannopoulos, George, . (2001) Gene Therapy for Hemophilia. New England Journal of Medicine 344:23, 1782-1784
Full Text125
Mannucci, Pier M., Tuddenham, Edward G.D., . (2001) The Hemophilias — From Royal Genes to Gene Therapy. New England Journal of Medicine 344:23, 1773-1779
Full Text126
&NA;. (2001) Factor VIII gene therapy: promising phase I results in haemophilia A. Inpharma Weekly &NA;:1292, 9
CrossRef127
Sven Bj??rkman, Erik Berntorp. (2001) Pharmacokinetics of Coagulation Factors. Clinical Pharmacokinetics 40:11, 815-832
CrossRef
