Neuroinflammation in Juvenile Batten Disease and the Role of an Anti-inflammatory Treatment Tammy Kielian, Venkata R Kakulavarapu, Megan Bosch, Amy Aldrich, Maria Burkovetskaya, and Nikolay Karpuk

Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska, 68198, tkielian@unmc.edu

KEY PROJECTS WHAT THIS MEANS FOR THERAPY

ACKNOWLEDGEMENTS - Forest Laboratories, Inc./Actavis/AstraZeneca - Rachel Fallet and Jessica Odvody for excellent technical

assistance - Dr. Jonathan Cooper and Nanet Willumsen- King’s College

London - NINDS 1R21NS084392-01A1, Bee For Battens- The Saoirse

Foundation

INTRODUCTION

• Juvenile Batten Disease mainly affects brain function in

children severely impairing their behavior, intellectual

ability, and day-to-day activities. Brain inflammation has

been suggested as being one potential factor

contributing to neuron loss in Juvenile Batten Disease.

• Several brain cells including microglia, astrocytes, and

neurons appear dysfunctional in Juvenile Batten Disease.

• Our laboratory’s main focus is to understand how brain

inflammation, which is harmful to brain function, occurs

in Juvenile Batten Disease.

• We use the CLN3∆ex7/8 mouse model of Juvenile Batten

Disease in our research and are currently testing the

effectiveness of roflumilast, an anti-inflammatory drug,

to determine if reducing neuroinflamation can result in

slowing disease progression and symptoms.

Microglia Astrocytes Neurons

• Mouse models of Juvenile Batten Disease display early signs of microglia & astrocyte activation

• In mice, microglia and astrocyte activation is apparent within 1-3 months of age; however, neuronal death is significantly delayed in comparison (~ 12 months)

• These findings suggest that chronic glial activation and subsequent inflammation may influence neuron survival

Morales et al., Front. Cell. Neurosci., 2014

• Microglia protect against foreign insults and clear dead cells/debris in the brain

• In response to insults,

microglia become activated and produce inflammatory cytokines, such as TNF-a and IL-1b

• These inflammatory

mediators can have neurotoxic effects, and may contribute to JNCL progression.

A

B

• Figure A shows abnormal cytokine production in CLN3∆ex7/8 microglia that kills neurons

• Figure B shows that TNF-a production can be reduced in these microglia when treated with an anti-inflammatory compound

• Roflumilast (Daliresp©) is a FDA approved phosphodiesterase-4 (PDE4) inhibitor to reduce the risk of exacerbations in patients with chronic obstructive pulmonary disease (COPD)

• PDE4 inhibitors have been found

in other neurological studies to reduce brain inflammation by inhibiting microglial and astrocyte activation and attenuating microglial proinflammatory cytokine production

• We administered Roflumilast to CLN3∆ex7/8 and normal mice orally, once a day, for 6 months starting at 1 month of age

• After 3 months of drug treatment, motor function in CLN3∆ex7/8 mice was significantly improved, as indicated by an increased amount of time mice were able to stay on the spinning rotarod

• Additionally, astrocyte and microglia activation has been significantly lowered following drug treatment (not shown)

• Astrocytes have many functions in the brain and are constantly nurturing neurons to help them function properly

• In Juvenile Batten Disease,

astrocytes become hyperactive, which can be seen by increased Glial Fibrillary Acidic Protein expression (in green above)

• As a result, they do not properly support neurons resulting in neuronal death

• Neurons are brain cells that are critically involved in all of our physical and mental activities

• Neurons send electrical signals through synapses via specialized structures called axons and dendrites

• In Juvenile Batten Disease,

some of these neurons die early, which disrupts communication in the brain and eventually leads to cognitive and motor decline

Kielian Laboratory

• This image shows two different types of synapses, inhibitory and excitatory

• In CLN3∆ex7/8 mice , there is an increase in excitatory and decrease in inhibitory synapses

• This disrupted ratio can cause neuronal death and seizure activity

Inhibitory Excitatory

• GFAP, as indicated by the intensity of red staining, is increased in the brains of CLN3∆ex7/8 mice

• The Visual Cortex (VC) is responsible for processing information from the eyes

• The thalamus (TH) is involved in sleep regulation and is also responsible for relaying sensory and motor signals to other brain regions

Overview

Rotarod Motor activity