Characterization of the Immune Environment in the Post-Mortem Spinal Cord Tissue of ALS Patients and Controls Restricted; Files Only

File, Brittany (2017)

Permanent URL: https://etd.library.emory.edu/concern/etds/xk81jm07g?locale=en
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Abstract

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease that causes progressive muscle weakness and ultimately death due to respiratory failure. There is no cure for ALS, and its causes are unknown. Neuroinflammation is a notable pathological finding present at the site of motor neuron death in spinal cords of patients with ALS, and it is suggested that inflammation may have both protective and deleterious effects on neuronal survival. Microglia are innate immune cells that function as actors in immune surveillance of the central nervous system (CNS) and can become active contributors to neuroinflammation. With activation, microglia undergo morphological changes that are recognized during neuropathological evaluation. While it is hypothesized that the immune system first responds to aid and repair damaged motor neurons, it has also been suggested that as ALS progresses the once neuroprotective response shifts to a self-sustaining cycle of neurotoxicity and cell death. This project extends such research and investigates the presence of inflammatory markers, specifically microglia, in spinal cords collected at autopsy from patients dying with ALS. In addition, we investigated the possible effect on microglial reactivity in patients that participated in a clinical trial of intraspinal transplantation of stem cells for ALS. The aims of this study are two-fold: 1) to compare microglial activation in the spinal cords of ALS patients versus age-matched disease and normal (no neurological disease) controls; and 2) to determine whether the injection of neural stem cells alters the local immune environment in post-mortem tissue of the ALS patients.

Table of Contents

1. Abstract………………………………………………………………....................... 1

2. Introduction…………………………………………………………………………. 2

3. Materials and Methods…………………………………………………………….... 9

4. Results………………………………………………………………………………. 13

5. Discussion………………………………………………...………………………… 22

6. Limitations and Future Directions………………………………………………….. 27

7. Figures & Tables……………………………………….…………………………… 29

Table 1. Disease characteristics for the four subject groups............................... 29

Figure 1. Differences in density of microglial populations noted between medial and lateral anterior horn………………………………………………………..….. 30

Figure 2. Annotating the region of interest (anterior horn) in Aperio ImageScope…………………………………………………………………… 31

Figure 3. Quantitative analysis of microglia using the object counts………… 32

Figure 4. Quantitative analysis of microglia using the object area…………… 32

Figure 5. Quantitative analysis of microglia using positive pixel count……… 33

Figure 6. Qualitative analysis of microglial populations in control spinal cords……………………………………………………………….……….…. 34

Figure 7. Qualitative analysis of microglial populations in ALS spinal cords cord……………………..…………………………………………………….. 34

Table 2. A summary table of an initial analysis of non-disease control

vs ALS spinal cords………………………………………………………..… 35

Figure 8. Microglial population analyses within varying region in

non-disease control spinal cord…………………………………………….... 36

Figure 9. Microglial analyses of the rostral spinal cord in non-disease

control, disease control, and ALS patients…………………………………. 37

Figure 10. Analysis of microglia area in the rostral spinal cord in

non-disease control and ALS patients…………………………………….… 38

Figure 11. Microglial population analyses of the caudal spinal cord

in non-disease control, disease control, and ALS patients…………………. 39

Figure 12. Microglial population analyses of the caudal spinal cord

in non-disease control and ALS patients……....…………………………… 40

Figure 13. A comparison of injected and non-injection spinal cord

regions in case 5…………………………… ……………………………… 41

Figure 14. Stem cell treated sections of spinal cord exhibit regions

of clearing………………………………………………………………….. 42

Table 3. A summary table of an initial analysis of stem cell

positive vs negative regions……………………………………………….. 43

Figure 15. Microglial population analyses of stem cell positive vs negative regions……………………………………………………………………... 44

Figure 16. Correlation analyses within stem cell positive regions………… 45

Table 4. Quantitative analysis results for individual non-disease

control, control, and ALS cases……………………………………………. 46

Table 5. Quantitative analysis results for individual stem cell cases……… 47

8. References ……………………………………………………………………… 48

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