Gene therapy and analysis of pathology in neural cell cultures: Variant Infantile Batten DiseaseStephanie 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: firstname.lastname@example.orgDavid.Palmer@lincoln.ac.nz CLN5 CLN6 BDSRA 2012 Key Projects What this means For Therapy 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. CLN5 cells have characteristic Batten disease pathology 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 cell cultures can be used to analyse pathology and used to screen candidate drugs or gene therapies. 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 Expression CLN5 CLN6 VIRUSES CAN BE MODIFIED TO CARRY GENES such as CLN5 OR CLN6 INTO NEURONS IN CELL CULTURE AND INTO THE BRAIN Acknowledgements: We thank the Neurological Foundation of New Zealand, Otago Medical Research Foundation and the Otago School of Medical Sciences for funding. CLN5 (green) expressed by a gene therapy vector. CLN6 (green) expressed by a gene therapy vector.