Gene therapy and analysis of pathology in neural cell cultures

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Gene therapy and analysis of pathology in neural cell cultures:
Variant Infantile Batten Disease
Stephanie M. Hughes, Nicole J. Neverman, Hollie E. Peacock, Katie M.Hope, Department of Biochemistry, Brain Health Research Centre, University of Otago, Dunedin.
David N. Palmer, Lucy A Barry, Nadia L. Mitchell, Janet Xu, Faculty of Agriculture and Life Sciences, Lincoln University, Canterbury, New Zealand.
Contact: stephanie.hughes@otago.ac.nz David.Palmer@lincoln.ac.nz
INTRODUCTION
 Mutations in CLN5 and CLN6 have
been identified in three breeds of
sheep.
 Our laboratories have developed
neural cultures from these sheep to
investigate early pathology and test
gene therapy vectors.
BDSRA 2012
WHAT THIS MEANS
FOR THERAPY
KEY PROJECTS
CLN5 CELLS HAVE
CHARACTERISTIC
BATTEN DISEASE
PATHOLOGY
CLN5
CLN6
CLN6 CELLS SHOW EARLY CHANGES IN
CELLULAR STRUCTURES
There is a change in the structure of the part
of the cell where CLN6 functions, shown
here by reduced green labeling in the CLN6
affected cells.
The degradation of material in lysosomes
depends on the lysosome having a acidic
pH (indicated by red fluorescence in these
images) is reduced in CLN6 affected cells.
Gene therapy vectors are now being
used to test therapeutic potental in
the sheep models
Neural cells from CLN5-affected sheep (left)
show characteristic accumulation of storage
material (black staining top), and green
bottom, not seen in normal control cells
Cells isolated from fetal sheep brain can
be grown in a dish and used to analyze
disease processes and test gene therapy
vectors
Neural cell cultures can be used to
analyse pathology and used to
screen candidate drugs or gene
therapies.
CLN5
Expression
CLN5 (green) expressed by a gene therapy
vector.
VIRUSES CAN BE MODIFIED
TO CARRY GENES such as
CLN5 OR CLN6 INTO
NEURONS IN CELL CULTURE
AND INTO THE BRAIN
CLN6
Acknowledgements:
We thank the Neurological
Foundation of New Zealand,
Otago Medical Research
Foundation and the Otago
School of Medical Sciences for
funding.
CLN6 (green) expressed by a gene therapy
vector.
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