Catabolism of thymidine in human blood platelets purification and properties of thymidine phosphorylase

Des̀granges, Claude; Razaka, G; Rabaud, M.; Bricaud, H

doi:10.1016/0005-2787(81)90174-x

Cited by 98 publications

(58 citation statements)

References 33 publications

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“…For transduction of murine hematopoietic progenitors, 5Â10 5 Lin À cells per well, obtained as detailed below, were seeded in 24-well plates at 5Â10 5 cells per ml in Iscove's modified Dulbecco's medium (Invitrogen) supplemented with 10% heat-inactivated fetal bovine serum, 2 mM L-glutamine, 1 U l À1 penicillin, 1 U l À1 streptomycin and recombinant murine growth factors (rmSCF 10 ng ml À1 , rmTPO 10ngml À1 , rmIL3 10ngml À1 and rmFlt3 10 ng ml À1 ) (ImmunoTools, Friesoythe, Germany). Lentiviruses were added at 100 MOI and cells were centrifuged at 300 g, 321C for 90 min.…”

Section: Vector Constructionmentioning

confidence: 99%

“…4 TP catalyzes the phosphorolysis of thymidine (dThd) or deoxyuridine (dUrd) nucleosides to the corresponding bases, thymine or uracil, and deoxyribose-1-phosphate. 5 In MNGIE patients, mutations in TYMP markedly reduce TP activity, resulting in a systemic accumulation of dThd and dUrd. Plasma and tissue dThd and dUrd, which are undetectable (o0.05 mM) in normal individuals, reach micromolar concentrations in MNGIE patients.…”

Section: Introductionmentioning

confidence: 99%

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Hematopoietic gene therapy restores thymidine phosphorylase activity in a cell culture and a murine model of MNGIE

et al. 2011

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Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is an autosomal recessive disorder caused by mutations in the TYMP gene, which encodes thymidine phosphorylase (TP). TP dysfunction results in systemic thymidine (dThd) and deoxyuridine (dUrd) overload, which selectively impair mitochondrial DNA replication. Allogeneic hematopoietic transplantation has been used to treat MNGIE patients; however, this approach has serious adverse effects, including the toxicity of myeloablative conditioning, graft rejection and graft-versus-host disease. With the aim of testing the feasibility of gene therapy for MNGIE, we transduced TP-deficient B-lymphoblastoid cells from two MNGIE patients, with lentiviral vectors carrying a functional copy of the human TYMP DNA coding sequence. This restored TP activity in the cells, which reduced the excretion of dThd and dUrd and their concentrations when added in excess. Additionally, lentiviral-mediated hematopoietic gene therapy was used in partially myeloablated double Tymp/Upp1 knockout mice. In spite of the relatively low levels of molecular chimerism achieved, high levels of TP activity were observed in the peripheral blood of the transplanted mice, with a concomitant reduction of nucleoside concentrations. Our results suggest that hematopoietic gene therapy could be an alternative treatment for this devastating disorder in the future.

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Section: Vector Constructionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Hematopoietic gene therapy restores thymidine phosphorylase activity in a cell culture and a murine model of MNGIE

et al. 2011

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“…TP is one of two pyrimidine phosphorylases in the base and nucleoside salvage pathway. Human TP and PD-ECGF share the same tissue distribution: platelets, placenta and some tumours (Kubilus et al, 1978;Desgranges et al, 1981;Shaw et al, 1988a, b;Yoshimura et al, 1990;Usuki et al, 1989;Ishikawa et al, 1989). The vast majority of human blood TP is localised in the platelet cytosol where one of its normal physiological roles is to reg-ulate thymidine homeostasis (Shaw et al, 1988 a, b).…”

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confidence: 99%

Thymidine phosphorylase activity of platelet‐derived endothelial cell growth factor is responsible for endothelial cell mitogenicity

Finnis

Dodsworth

Pollitt

et al. 1993

European Journal of Biochemistry

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Recombinant human platelet-derived endothelial cell growth factor, expressed in the yeast Saccharomyces cerevisiae was purified to greater than 98 % purity by anion-exchange and hydroxyapatite chromatography. It was shown to possess thymidine phosphorolytic activity in vitro (pH optimum, pH 5.3; K,, 0.11 mM; V, , , 12.5 mmol min-' mg-'; turnover number, 9.4 s -I ) . Covalent modification simultaneously inhibited the enzymatic and mitogenic properties of the protein, while interaction with a cell-surface receptor was not required to stimulate mitogenesis. Purified Escherichia coli thymidine phosphorylase was also mitogenic toward endothelial cells. It is proposed that platelet-derived endothelial cell growth factor is human thymidine phosphorylase which promotes endothelial cell proliferation by reducing thymidine levels that would otherwise be inhibitory to endothelial cell growth.

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“…D-dRib also displays angiogenic activity. Addition of thymidine to platelets that have high levels of TP gives rise to thymine and D-dRib in the extracellular medium (18,19). Consequently, D-dRib is considered an effector of TP-related angiogenesis in endothelial cells.…”

Section: Discussionmentioning

confidence: 99%

Angiogenic Factor Thymidine Phosphorylase Increases Cancer Cell Invasion Activity in Patients with Gastric Adenocarcinoma

Lee

Rha

et al. 2008

Molecular Cancer Research

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We investigated the biological role of thymidine phosphorylase (TP), an angiogenic factor, in gastric cancer cell migration and invasion and explored a therapeutic approach for high TP-expressing tumors using TP enzymatic inhibitor (TPI) and rapamycin. We established TP cDNA overexpressing gastric cancer cell lines (MKN-45/TP and YCC-3/TP) and did invasion and adhesion assays with Matrigel-coated transwell membranes. The related signal pathway using recombinant human TP (rhTP), deoxy-D-ribose (D-dRib), and signal pathway inhibitors (wortmannin, LY294002, and rapamycin) was investigated. First, AGS and MKN-1 gastric cancer cell lines showed dose-dependent up-regulation of invasiveness through Matrigel following treatment with rhTP or D-dRib. TP-overexpressing cancer cell lines displayed increased migration and invasion activity, which doubled with rhTP and D-dRib treatment. This activity depended on the enzymatic activity of TP, and TP stimulated the adhesion of cancer cells onto Matrigel and induced actin filament remodeling. Finally, we showed that this activity is related to increased phosphatidylinositol 3-kinase activity in TP-overexpressing cells and that combination treatment with rapamycin and TP enzymatic inhibitor produces an additive effect to abrogate TP-induced invasion. Taken together, TP increases the migration and invasion of gastric cancer cells, especially in TP-expressing cells. Therapies targeting TP might diminish the propensity for invasion and metastasis in gastric cancer. (Mol Cancer Res 2008;6(10):1554 -66)

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Catabolism of thymidine in human blood platelets purification and properties of thymidine phosphorylase

Cited by 98 publications

References 33 publications

Hematopoietic gene therapy restores thymidine phosphorylase activity in a cell culture and a murine model of MNGIE

Hematopoietic gene therapy restores thymidine phosphorylase activity in a cell culture and a murine model of MNGIE

Thymidine phosphorylase activity of platelet‐derived endothelial cell growth factor is responsible for endothelial cell mitogenicity

Angiogenic Factor Thymidine Phosphorylase Increases Cancer Cell Invasion Activity in Patients with Gastric Adenocarcinoma

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