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High Transgene Expression by Lentiviral Vectors Causes Maldevelopment of Purkinje Cells In Vivo

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Abstract

Lentiviral vectors are promising as gene-transfer vehicles for gene therapy targeted to intractable brain diseases. Although lentiviral vectors are thought to exert little toxicity on infected cells, the adverse influence of viral infection on vulnerable developing neurons has not been well studied. Here, we examined whether lentiviral vector infection and subsequent transgene expression affected the morphological and functional maturation of vigorously developing cerebellar Purkinje cells in vivo. Lentiviral vectors expressing GFP under the control of the murine stem cell virus (MSCV) promoter were injected into the cerebellar cortex of neonatal rat pups. Three weeks after treatment, GFP-expressing Purkinje cells were compared with control Purkinje cells from phosphate-buffered saline-injected rats. Analysis of the dendritic tree showed that total dendrite length in GFP-expressing Purkinje cells was almost 80% that in control Purkinje cells. Electrophysiological examination showed that short-term synaptic plasticity at parallel fiber–Purkinje cell synapses and climbing fiber–Purkinje cell synapses was significantly altered in GFP-expressing Purkinje cells. In contrast, maldevelopment of infected Purkinje cells was substantially attenuated when lentiviral vectors with much weaker promoter activity were used. These results suggest that the maldevelopment of Purkinje cells was mainly caused by subsequent expression of a high amount of GFP driven by the strong MSCV promoter. Thus, the use of lentiviral vectors carrying a strong promoter may require particular precautions when applying them to neurological disorders of infants.

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Acknowledgments

The lentiviral vector and MSCV promoter were kindly provided by St. Jude Children’s Research Hospital and the American National Red Cross, respectively. This work was supported in part by a Grant-in-Aid from the Japan Society for the Promotion of Science (JSPS), MEXT; grants from Research on Measures for Intractable Diseases (Ataxic Diseases and Neurodegenerative Diseases) from the Ministry of Health, Labour, the Sumitomo Foundation, Takeda Science Foundation and Uehara Memorial Foundation (to H. Hirai). We thank Dr. Hosoi for critical reading of the manuscript. The authors declare that they have no competing financial interests.

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Authors and Affiliations

  1. Department of Neurophysiology, Gunma University Graduate School of Medicine, Maebashi, Gunma, 371-8511, Japan

    Yusuke Sawada, Go Kajiwara, Akira Iizuka, Kiyohiko Takayama, Anton N. Shuvaev, Chiho Koyama & Hirokazu Hirai

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  1. Yusuke Sawada
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  2. Go Kajiwara
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  3. Akira Iizuka
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  4. Kiyohiko Takayama
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  5. Anton N. Shuvaev
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  6. Chiho Koyama
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  7. Hirokazu Hirai
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Correspondence to Hirokazu Hirai.

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Yusuke Sawada and Go Kajiwara contributed equally to this work.

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Sawada, Y., Kajiwara, G., Iizuka, A. et al. High Transgene Expression by Lentiviral Vectors Causes Maldevelopment of Purkinje Cells In Vivo. Cerebellum 9, 291–302 (2010). https://doi.org/10.1007/s12311-010-0161-1

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