Expression of peptide NAP in rat retinal Müller cells prevents hypoxia-induced retinal injuries and promotes retinal neurons growth.
NAP (NAPVSIPQ) is a short peptide derived from activity-dependent neuroprotective protein (ADNP) sequence, whose potent and direct neuroprotective capabilities have been widely accepted. However, due to the high risk and inconvenience of intraocular injections, NAP is difficult to be clinically administered as therapeutic agent in treating retinal diseases. Currently, stable transfection of this octapeptide into cells has not been reported, partly because of its small size and lacking of 5' signal sequence. Here, we have developed a novel NT4-NAP fusion gene by attaching the 5' nonfunctional preproregion of neurotrophin 4 (NT4) to NAP cDNA. Recombinant adeno-associated virus was established to introduce NT4-NAP construct into cultured rat retinal Müller cells (RMC), resulting in sustained high level NAP production from stable transfection. Functional analyses of RMC cells transfected with NAP revealed the remarkably reduced cytotoxicity and apoptosis of the cells under hypoxia. Furthermore, coculturing of transfected RMC-NAP cells with primary rat retinal neural cells offer marked protection to the latter against hypoxia induced cellular damages. Together our data indicate that stable transfection of NAP into retinal Müller cells with constant NAP production is possible. NAP produced from cellular transfection maintained its biological neuroprotective activities. This targeted gene expression may provide an effective treatment for retinal diseases in the near future.
Zheng
Yuping
Y
Department of Ophthalmology, 2nd Affiliated Hospital, School of Medicine, Xi'an JiaoTong University, No. 36 Xiwy Road, Xi'an, Shaanxi 710004, China.
Zeng
Hao
H
She
Huaning
H
Liu
Hui
H
Sun
Naixue
N
D000818:Animals; D015687:Cell Hypoxia; D049109:Cell Proliferation; D002478:Cells, Cultured; D000229:Dependovirus / Q000235:genetics; D050939:Gene Fusion; D015316:Genetic Therapy; D009414:Nerve Growth Factors / Q000235:genetics; D018696:Neuroprotective Agents; D009842:Oligopeptides / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012160:Retina / Q000166:cytology / Q000378:metabolism*; D012164:Retinal Diseases / Q000628:therapy*; D055351:Retinal Neurons / Q000502:physiology*; D014162:Transfection
D009414:Nerve Growth Factors; D018696:Neuroprotective Agents; D009842:Oligopeptides; C425904:davunetide; C073922:neurotrophin 4