Recombinant factor IX secreted by transduced human keratinocytes is biologically active

Gerrard, Ann J.; Austen, D. E. G.; Brownlee, G. G.

doi:10.1046/j.1365-2141.1996.d01-1923.x

Cited by 12 publications

(4 citation statements)

References 8 publications

(13 reference statements)

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“…In parallel, many other cell types have been shown to express functional FIX following ex vivo transduction with FIX retroviral vectors. These cell types include myoblasts25, 26, 143, endothelial cells23, hematopoietic cells29, 144 hepatocytes145, keratinocytes18, 19, 146, 147 and gut epithelial cells148. Transplantation of these engineered cells in vivo often resulted in detectable but transient levels of FIX, reminiscent of the fibroblast transplantation experiments.…”

Section: Gene Therapy For Hemophilia Bmentioning

confidence: 99%

Gene therapy for hemophilia

2001

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Hemophilia A and B are X-chromosome linked recessive bleeding disorders that result from a deficiency in factor VIII (FVIII) and factor IX (FIX) respectively. Though factor substitution therapy has greatly improved the lives of hemophiliac patients, there are still limitations to the current treatment that have triggered interest in alternative treatments by gene therapy. Significant progress has recently been made in the development of gene therapy for the treatment of hemophilia A and B. These advances parallel the technical improvements of existing vector systems including MoMLV-based retroviral, adenoviral and AAV vectors, and the development of new delivery methods such as lentiviral vectors, helper-dependent adenoviral vectors and improved non-viral gene delivery methods. Therapeutic and physiologic levels of FVIII and FIX could be achieved in FVIII- and FIX-deficient mice and hemophilia dogs by different gene therapy approaches. Long-term correction of the bleeding disorders and in some cases a permanent cure has been realized in these preclinical studies. However, the induction of neutralizing antibodies often precludes stable phenotypic correction. Another complication is that certain promoters are prone to transcriptional inactivation in vivo, precluding long-term FVIII or FIX expression. Several gene therapy phase I clinical trials are currently ongoing in patients suffering from severe hemophilia A or B. No significant adverse side-effects were reported, and semen samples were negative for vector sequences by sensitive PCR assays. Most importantly, some subjects report fewer bleeding episodes and occasionally have very low levels of clotting factor activity detected. The results from the extensive preclinical studies in normal and hemophilic animal models and encouraging preliminary clinical data indicate that the simultaneous development of different strategies is likely to bring a permanent cure for hemophilia one step closer to reality.

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Section: Gene Therapy For Hemophilia Bmentioning

confidence: 99%

Gene therapy for hemophilia

2001

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“…The second approach implicates a direct injection of the vector containing the transgene in target tissues or in the blood stream. Several potential target cells were shown to be able to produce an active FIX such as hepatocytes (9), fibroblasts (10), myoblasts (11,12), keratinocytes (13) and stromal cells (14).…”

Section: Introductionmentioning

confidence: 99%

Expression of Coagulation Factor IX in a Haematopoietic Cell Line

Rodriguez

Enjolras²,

Plantier³

et al. 2002

Thromb Haemost

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SummaryWe have developed a gene therapy project for haemophilia B which aims to express factor IX (FIX) in haematopoietic lineage. Haematopoietic stem cells and subsequent megakaryocyte-derived cells represent the target cells of this approach. Our speculation is that platelets can deliver the coagulation factor at the site of injury, and subsequently correct the haemostasis defect. In order to direct FIX expression in cells from the megakaryocytic lineage, we designed a FIX cassette where the FIX cDNA was placed under the control of the tissue-specific glycoprotein IIb (GPIIb) promoter. In stably transfected HEL cells, FIX production was higher when driven by the GPIIb promoter compared to the CMV promoter. Using a cassette containing both the GPIIb promoter and a truncated FIX intron 1, FIX synthesis was dramatically increased in HEL cells. Northern blot analysis demonstrated an increase in FIX mRNA amounts, which paralleled with an increase of FIX antigen in the culture supernatants. Using a one-stage clotting assay and an activation by FXIa and FVIIa/TF, the HEL-derived recombinant FIX was shown to be a biologically active protein. This recombinant protein exhibited a 60-kDa molecular mass and was more heterogeneous than plasma immunopurified FIX (Mononine®). The molecular mass difference could be partly explained by a different glycosylation pattern. The GPIIb promoter appears therefore to be a very attractive sequence to specifically direct FIX production in the megakaryocytic compartment of hematopoietic cells. These data also demonstrate that hematopoietic cells may represent potential target cells in an approach to gene therapy of haemophilia B.

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“…Multiple different cell lines have been transduced with factor IX including primary fibroblasts (St Louis & Verma, 1988; Palmer et al , 1989; Axelrod et al , 1990), myoblasts (Dai et al , 1992; Yao & Kurachi, 1992; Yao et al , 1994), hepatocytes (Armentano et al , 1990), haemopoietic stem cells (Hao et al , 1995), endothelial cells (Yao et al , 1991), gut epithelial cells (Lozier et al , 1997) and keratinocytes (Gerrard et al , 1993, 1996; Page & Brownlee, 1997, White et al , 1998) ex vivo using factor IX‐containing retroviruses. Functional factor IX is produced by many cell types despite the need for complex post‐translational gamma carboxylation required for full function of factor IX.…”

Section: Gene Therapy Studies In Haemophilia Bmentioning

confidence: 99%

Gene therapy for haemophilia

Pasi

2001

Br J Haematol

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Recombinant factor IX secreted by transduced human keratinocytes is biologically active

Cited by 12 publications

References 8 publications

Gene therapy for hemophilia

Gene therapy for hemophilia

Expression of Coagulation Factor IX in a Haematopoietic Cell Line

Gene therapy for haemophilia

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