X-Linked inhibitor of apoptosis protein gene-based neuroprotection for the peripheral nervous system.
OBJECTIVE
The recently discovered X-linked inhibitor of apoptosis protein (XIAP) is among the most potent inhibitors of programmed cell death. In the current experiment, we examine the potential of adenoviral XIAP gene delivery to protect neurons of the peripheral nervous system using in vitro models of amyotrophic lateral sclerosis (ALS) and diabetic neuropathy.


METHODS
XIAP complementary deoxyribonucleic acid was fused in frame with the green fluorescent protein sequence and cloned into a first generation adenoviral vector. The impact of XIAP gene expression on glutamate-induced apoptosis was measured in the neuronal SH-SY5Y cell line with immunohistochemistry for active caspase-3 and with cell density assays. Next, the effect of XIAP expressing neurons on the survival of uninfected neighboring neurons was measured. Finally, the impact of XIAP gene expression on glutamate-induced apoptosis was assessed in embryonic motor neuron and dorsal root ganglion cultures.


RESULTS
XIAP gene expression reduced the percentage of active caspase-3 positive SH-SY5Y neurons and preserved cell density after glutamate exposure. In heterogeneously infected cultures, cells infected with XIAP were protected, but uninfected neighboring cells were not. In primary E15 models, inhibition of proapoptotic effects was demonstrated after glutamate insult in motor neurons and glucose insult in dorsal root ganglion cells.


CONCLUSIONS
XIAP gene delivery through the neurosurgical delivery of viral vectors may provide a means for neuroprotection in ALS and diabetic neuropathy.
Garrity-Moses
Mary E
ME
Cleveland Clinic Foundation, Lerner Research Institute, Department of Neuroscience and Center for Neurological Restoration, Cleveland, Ohio 44195, USA.
Teng
Qingshan
Q
Krudy
Christina
C
Yang
Jun
J
Federici
Thais
T
Boulis
Nicholas M
NM
D000690:Amyotrophic Lateral Sclerosis / Q000378:metabolism / Q000473:pathology*; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002452:Cell Count; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003929:Diabetic Neuropathies / Q000378:metabolism / Q000473:pathology*; D005240:Feasibility Studies; D005727:Ganglia, Spinal / Q000187:drug effects; D015870:Gene Expression; D018014:Gene Transfer Techniques; D018698:Glutamic Acid / Q000506:poisoning; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D009046:Motor Neurons / Q000187:drug effects; D018696:Neuroprotective Agents / Q000494:pharmacology; D017933:Peripheral Nervous System / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D051636:X-Linked Inhibitor of Apoptosis Protein / Q000235:genetics* / Q000494:pharmacology
D018696:Neuroprotective Agents; D011993:Recombinant Fusion Proteins; D051636:X-Linked Inhibitor of Apoptosis Protein; D049452:Green Fluorescent Proteins; D018698:Glutamic Acid; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases