Preclinical Toxicity Evaluation of Erythrocyte-Encapsulated Thymidine Phosphorylase in BALB/c Mice and Beagle Dogs: An Enzyme-Replacement Therapy for Mitochondrial Neurogastrointestinal Encephalomyopathy

Levene, Michelle; Coleman, David; Kilpatrick, Hugh; Fairbanks, Lynette D.; Gangadharan, Babunilayam; Gasson, Charlotte; Bax, Bridget E.

doi:10.1093/toxsci/kfs278

Cited by 35 publications

(24 citation statements)

References 20 publications

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“…This hypothesis is supported by one study reported a decade ago in which increased expression of TYMP in human hepatocellular carcinomas was correlated with high incidence of portal vein thrombosis 41 . This hypothesis is also supported by a recent study, in which perfusion of erythrocyte-encapsulated TYMP to mice resulted in thrombi in the lungs 42 . Interestingly, Aspirin, an anti-platelet drug, inhibits TYMP production in a human monocyte cell line, THP1 cells 43 , suggesting that Aspirin may also inhibit TYMP production in platelet and thus results in the anti-thrombotic effect.…”

Section: Discussionsupporting

confidence: 67%

Thymidine Phosphorylase Participates in Platelet Signaling and Promotes Thrombosis

Gigante

Pérez‐Pérez

et al. 2014

Circulation Research

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Rationale Platelets contain abundant thymidine phosphorylase (TYMP), which is highly expressed in diseases with high risk of thrombosis, such as atherosclerosis and type II diabetes. Objective Test the hypothesis that TYMP participates in platelet signaling and promotes thrombosis. Methods and Results By using a ferric chloride (FeCl3) induced carotid artery injury thrombosis model, we found time to blood flow cessation was significantly prolonged in Tymp−/− and Tymp+/− mice compared to wild type (WT) mice. Bone marrow transplantation and platelet transfusion studies demonstrated that platelet TYMP was responsible for the antithrombotic phenomenon in the TYMP deficient mice. Collagen-, collagen-related peptide (CRP)-, adenosine diphosphate-and/or thrombin-induced platelet aggregation were significantly attenuated in Tymp+/− and Tymp−/− platelets, and in WT or human platelets pretreated with TYMP inhibitor KIN59. Tymp deficiency also significantly decreased agonist-induced P-select in expression. TYMP contains an N-terminal SH3 domain binding proline-rich motif and forms a complex with the tyrosine kinases Lyn, Fyn and Yes in platelets. TYMP-associated Lyn was inactive in resting platelets, and TYMP trapped and diminished active Lyn after collagen stimulation. Tymp/Lyn double haploinsufficiency diminished the antithrombotic phenotype of Tymp+/− mice. TYMP deletion or inhibition of TYMP with KIN59 dramatically increased PECAM-1 tyrosine phosphorylation and diminished CRP or collagen induced AKT phosphorylation. In vivo administration of KIN59 significantly inhibited FeCl3 induced carotid artery thrombosis without affecting hemostasis. Conclusion TYMP participates in multiple platelet signaling pathways and regulates platelet activation and thrombosis. Targeting TYMP might be a novel anti-platelet and anti-thrombosis therapy.

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Section: Discussionsupporting

confidence: 67%

Thymidine Phosphorylase Participates in Platelet Signaling and Promotes Thrombosis

Gigante

Pérez‐Pérez

et al. 2014

Circulation Research

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“…However, enzyme replacement has been attempted for MNGIE using purified thymidine phosphorylase enzyme encapsulated in the patient's own red blood cells (Moran et al ., ), so far without long‐term clinical success. Very recently, a preclinical toxicity evaluation of this system has been performed in mice and Beagle dogs, and was complicated by infusion‐related immune responses in both species (Levene et al ., ). Finally, one study described the construction of active thymidine phosphorylase encapsulating nanoreactors as novel enzyme delivery vehicles (De Vocht et al ., ) but clinical utility of this method has not yet been reported in either cell or animal models of MNGIE.…”

Section: Therapies Still In Developmentmentioning

confidence: 97%

Development of pharmacological strategies for mitochondrial disorders

Kanabus¹,

Heales

Rahman

2014

British J Pharmacology

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Mitochondrial diseases are an unusually genetically and phenotypically heterogeneous group of disorders, which are extremely challenging to treat. Currently, apart from supportive therapy, there are no effective treatments for the vast majority of mitochondrial diseases. Huge scientific effort, however, is being put into understanding the mechanisms underlying mitochondrial disease pathology and developing potential treatments. To date, a variety of treatments have been evaluated by randomized clinical trials, but unfortunately, none of these has delivered breakthrough results. Increased understanding of mitochondrial pathways and the development of many animal models, some of which are accurate phenocopies of human diseases, are facilitating the discovery and evaluation of novel prospective treatments. Targeting reactive oxygen species has been a treatment of interest for many years; however, only in recent years has it been possible to direct antioxidant delivery specifically into the mitochondria. Increasing mitochondrial biogenesis, whether by pharmacological approaches, dietary manipulation or exercise therapy, is also currently an active area of research. Modulating mitochondrial dynamics and mitophagy and the mitochondrial membrane lipid milieu have also emerged as possible treatment strategies. Recent technological advances in gene therapy, including allotopic and transkingdom gene expression and mitochondrially targeted transcription activator-like nucleases, have led to promising results in cell and animal models of mitochondrial diseases, but most of these techniques are still far from clinical application.

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“…β-Galactosidase - [21] β-Glucocerebrosidase (β-glucosidase) Gaucher disease [21,37,38,[44][45][46]250] β-Glucuronidase Syndrome Slaya [251] l-Phenylalanine ammonia lyase Phenylketonuria [48,252,253] Phenylalanine hydroxylase [50,254] Uricase (uratoxidase) Uric acid removal [255,256] Urease, urease + alanine dehydrogenase Urea utilization [257][258][259] Adenosine deaminase Severe combined immunodeficiency caused by deaminase deficiency [27,[55][56][57][58]260] Thymidine phosphorylase Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) [56,246,261,262] Glutamate dehydrogenase Hyperammonemia [59,60,72] Glutamine synthetase [73,74] Glutamate dehydrogenase + alanine aminotransferase [11,63] Arginase Hyperammonemia due to arginase deficiency [263] Alcohol dehydrogenase…”

Section: Active Substance Application Referencesmentioning

confidence: 99%

Erythrocytes as Carriers: From Drug Delivery to Biosensors

et al. 2020

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Drug delivery using natural biological carriers, especially erythrocytes, is a rapidly developing field. Such erythrocytes can act as carriers that prolong the drug's action due to its gradual release from the carrier; as bioreactors with encapsulated enzymes performing the necessary reactions, while remaining inaccessible to the immune system and plasma proteases; or as a tool for targeted drug delivery to target organs, primarily to cells of the reticuloendothelial system, liver and spleen. To date, erythrocytes have been studied as carriers for a wide range of drugs, such as enzymes, antibiotics, anti-inflammatory, antiviral drugs, etc., and for diagnostic purposes (e.g., magnetic resonance imaging). The review focuses only on drugs loaded inside erythrocytes, defines the main lines of research for erythrocytes with bioactive substances, as well as the advantages and limitations of their application. Particular attention is paid to in vivo studies, opening-up the potential for the clinical use of drugs encapsulated into erythrocytes.Pharmaceutics 2020, 12, 276 2 of 44 property of RBCs allows to load them with biologically active substances of different molecular weights. For these reasons, erythrocytes are promising biocompatible cells for drug delivery.The methods for incorporating various substances into red blood cells differ in the way that substances penetrate the cells. The cause of permeability may be the pores' formation in the cell membrane due to a physical exposure (high voltage electric pulse [10,11] or ultrasound [12]). Drug molecules can also enter the RBCs by endocytosis in the presence of certain chemical compounds (for example, primaquine [13], vinblastine, chlorpromazine, hydrocortisone or tetracaine [14,15]), or using the cell-penetrating peptides bounded to the compound that should be encapsulated [16]. However, the most popular are different variants of osmotic methods.In some cases, RBCs are first exposed to a hyperosmotic pulse of a low molecular weight substance that penetrates very well through the cell membrane (for example, dimethyl sulfoxide (DMSO) [17,18] or glucose [19,20]). After washing the cells, which decreases the external concentration of these compounds and creates a gradient of their concentration between both sides of the RBC membrane, the target drug is introduced into the external volume. Water with this drug begins to enter into the cells to decrease the osmotic pressure there. The process ends when the gradient of DMSO or glucose disappears. The pores close and part of the drug remains into RBCs. Other, the most popular of the osmotic methods are hypoosmotic. These methods are based on creating a hypotonic environment around RBCs, which causes swelling of the cells and opening pores in the cellular membrane, through which therapeutic compounds can penetrate RBCs. Then, a hypertonic solution is introduced into the cell suspension. The pores close, the cells restore their original size, trapping the drug molecules inside the cell. Osmotic methods are divided into severa...

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Preclinical Toxicity Evaluation of Erythrocyte-Encapsulated Thymidine Phosphorylase in BALB/c Mice and Beagle Dogs: An Enzyme-Replacement Therapy for Mitochondrial Neurogastrointestinal Encephalomyopathy

Cited by 35 publications

References 20 publications

Thymidine Phosphorylase Participates in Platelet Signaling and Promotes Thrombosis

Thymidine Phosphorylase Participates in Platelet Signaling and Promotes Thrombosis

Development of pharmacological strategies for mitochondrial disorders

Erythrocytes as Carriers: From Drug Delivery to Biosensors

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