This presentation explains how we intend to generate a Gene Therapy tool to cure the neurodegenerative disease Huntington’s disease (HD). Our work is aimed at the creation of Herpes Simplex Virus type1 (HSV) -based vectors as potential therapy for HD. The results of our experiments will indicate 1) which step of the synthesis pathway of the neuroprotective hormone melatonin is impaired in HD cells, and 2) if restoring, or even increasing, levels of melatonin endogenously produced in HD cell can have a neuroprotective effect.

1. Department of Neurological Surgery
“Toward a Gene
Therapy for
Huntington Disease”
Daniela Leronni, PhD
Department of Neurological Surgery
Dr. Robert Friedlander lab

2. Department of Neurological Surgery
Gene Therapy
• Gene therapy is a treatment modality intended to change the
expression of a gene (or genes) involved in a pathway altered by
the disease.
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From Collins M and Thrasher A, Proc. R. Soc. B 2015

3. Department of Neurological Surgery
Herpes Simplex Virus Biology
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From Burton EA, Fink DJ and Glorioso JC,
DNA and Cell Biology, 2002

4. Department of Neurological Surgery
Herpes Simplex Virus replication
Regulation of gene expression during lytic infection
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From Burton EA, Fink DJ and Glorioso JC,
DNA and Cell Biology, 2002

5. Department of Neurological Surgery
Herpes Simplex Virus
-gene therapy-
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Advantages as a gene therapy vector:
•broad host cell range
•highly infectious
•transduction of non-dividing cells
•latent behavior of the virus may be exploited for the stable long-term expression
of therapeutic transgenes in neurons
•multiple or large therapeutic transgenes can be accommodated, by replacing
dispensable viral genes

6. Department of Neurological Surgery
• HD is an autosomal dominant neurodegenerative disease due to an extended CAG
repeat in the gene encoding for the protein huntingtin (htt).
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Huntigton Disease
-Huntington’s Chorea-
Figure created by Serafi Cambray
for eurostemcell.org

7. Department of Neurological Surgery
Melatonin
-the darkness hormone-
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• Melatonin has been shown to be neuroprotective in cellular models of HD and to
decrease mortality in a mouse model of HD.
• Melatonin has been proposed as a neuroprotective agent on the basis of its ability to
function as a free radical scavenger
• Melatonin can exert a wide range of antiapoptotic effects, mainly targeting
mitochondria, but it can also enhance cell survival pathways leading to cell rescue.
Modified from Schwartz JR and Roth T. Drugs. 2006

8. Department of Neurological Surgery
Melatonin Synthesis Pathway
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From “Konturek SJ, Konturek PC and Brzozowski T, J. Physiol. Pharmacol. 2006”
(ADC)

9. Department of Neurological Surgery
Aims
The results of our experiments will indicate
•1) which step of the melatonin synthesis
pathway is impaired in HD, and
•2) if restoring, or even increasing, levels of
melatonin produced in HD cells can have a
neuroprotective effect.
05/31/18

10. Department of Neurological Surgery
HSV-vectors building
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NATNAT
ADCADC
NATNAT T2AT2A ADCADC
NATNATT2AT2AADCADC

11. Department of Neurological Surgery
Preliminary Results in vitro
-Q111 cell line survival assay-
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12. Department of Neurological Surgery
Conclusions
• HSV infection has a protective effect on
Q111 under stress.
• The protective effect is due to the
overexpression of the two enzymes.
• The two enzymes have to be both
overexpressed.
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13. Department of Neurological Surgery
JDNI7
20wpi
Future prospective –in vivo studies-

14. Department of Neurological Surgery
Aknowledgement
• Dr. Friedlander lab
– Wang, Jingjing
– Oxana Baranova
• Dr. Glorioso lab
– Dr. Justus Cohen
– Dr. Yoshitaka Miyagawa (University of Tokyo)
– Dr. Gianluca Verlengia (University of Ferrara)
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