Lentiviral RNAi-induced downregulation of adenosine kinase in human mesenchymal stem cell grafts: A novel perspective for seizure control

Cell therapies based on focal delivery of the inhibitory neuromodulator adenosine were previously shown to provide potent seizure suppression in animal models of epilepsy. However, hitherto used therapeutic cells were derived from rodents and thus not suitable for clinical applications. Autologous p...

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Veröffentlicht in:Experimental neurology 2007-11, Vol.208 (1), p.26-37
Hauptverfasser: Ren, Gaoying, Li, Tianfu, Lan, Jiang Quan, Wilz, Andrew, Simon, Roger P., Boison, Detlev
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container_end_page 37
container_issue 1
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container_title Experimental neurology
container_volume 208
creator Ren, Gaoying
Li, Tianfu
Lan, Jiang Quan
Wilz, Andrew
Simon, Roger P.
Boison, Detlev
description Cell therapies based on focal delivery of the inhibitory neuromodulator adenosine were previously shown to provide potent seizure suppression in animal models of epilepsy. However, hitherto used therapeutic cells were derived from rodents and thus not suitable for clinical applications. Autologous patient-derived adenosine-releasing cell implants would constitute a major therapeutic advance to avoid both xenotransplantation and immunosuppression. Here we describe a novel approach based on lentiviral RNAi mediated downregulation of adenosine kinase (ADK), the major adenosine-removing enzyme, in human mesenchymal stem cells (hMSCs), which would be compatible with autologous cell grafting in patients. Following lentiviral transduction of hMSCs with anti-ADK miRNA expression cassettes we demonstrate up to 80% downregulation of ADK and a concentration of 8.5 ng adenosine per ml of medium after incubating 10 5 cells for 8 h. hMSCs with a knockdown of ADK or cells expressing a scrambled control sequence were transplanted into hippocampi of mice 1 week prior to the intraamygdaloid injection of kainic acid (KA). While mice with control implants expressing a scrambled miRNA sequence or sham treated control animals were characterized by KA-induced status epilepticus and subsequent CA3 neuronal cell loss, animals with therapeutic ADK knockdown implants displayed a 35% reduction in seizure duration and 65% reduction in CA3 neuronal cell loss, when analyzed 24 h after KA-injection. We conclude that lentiviral expression of anti-ADK miRNA constitutes a versatile tool to generate therapeutically effective adenosine releasing hMSCs, thus representing a model system to generate patient identical autologous adult stem cell grafts.
doi_str_mv 10.1016/j.expneurol.2007.07.016
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While mice with control implants expressing a scrambled miRNA sequence or sham treated control animals were characterized by KA-induced status epilepticus and subsequent CA3 neuronal cell loss, animals with therapeutic ADK knockdown implants displayed a 35% reduction in seizure duration and 65% reduction in CA3 neuronal cell loss, when analyzed 24 h after KA-injection. We conclude that lentiviral expression of anti-ADK miRNA constitutes a versatile tool to generate therapeutically effective adenosine releasing hMSCs, thus representing a model system to generate patient identical autologous adult stem cell grafts.</abstract><cop>Amsterdam</cop><pub>Elsevier Inc</pub><pmid>17716659</pmid><doi>10.1016/j.expneurol.2007.07.016</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record>
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subjects Adenosine
Adenosine - metabolism
Adenosine kinase
Adenosine Kinase - genetics
Adenosine Kinase - metabolism
Animals
Biological and medical sciences
Cell therapy
Down-Regulation
Epilepsy
Excitatory Amino Acid Agonists
Genetic Vectors
Headache. Facial pains. Syncopes. Epilepsia. Intracranial hypertension. Brain oedema. Cerebral palsy
Hippocampus - pathology
Hippocampus - physiopathology
Human mesenchymal stem cells
Humans
Kainic Acid
Lentivirus
Lentivirus - genetics
Male
Medical sciences
Mesenchymal Stem Cell Transplantation
Mesenchymal Stromal Cells - enzymology
Mesenchymal Stromal Cells - metabolism
Mice
Mice, Inbred C57BL
Nervous system (semeiology, syndromes)
Neurology
Neuroprotection
RNA Interference
RNAi
Status epilepticus
Status Epilepticus - chemically induced
Status Epilepticus - pathology
Status Epilepticus - physiopathology
Status Epilepticus - surgery
Transduction, Genetic
title Lentiviral RNAi-induced downregulation of adenosine kinase in human mesenchymal stem cell grafts: A novel perspective for seizure control
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