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Genetic Modified Cell Therapy for Amyotrophic Lateral Sclerosis (ALS)
Tianyi Cai
PBIO-4500
12/02/2014
Overview and Futures of ALS
• A fatal neurodegenerative disorder causes death of motor neurons
• Patients develop paralysis rapidly and die due to respiratory insufficiency
in 3-5 years after diagnosis
• 10% ALS are familial (fALS)
• 20% of fALS cases are linked to mutation for SOD1 (over-expressed)
allele containing Gly93.
• Traditional pharmacological therapies have largely failed so far
Design and Mechanism of the Experiment
• The researchers found that glucagon-like peptide 1 (GLP-1) exhibits anti-
oxidation and is neuroprotective against excitotoxicity.
• To avoid systemic administration, the researchers decided to deliver GLP-1
directly into the central neurvous system.
• Researchers investigative tested an encapsulated cell line which can produce
GLP-1 to replace an osmotic minipump which may cause more complications.
• Scientists modified a mesenchymal stromal cell (MSC) line to produce GLP-1.
• An encapsulated GM MSC is intracerebroventricular injected in experimental
animals to test the results.
Materials and Methods
• Experimental animals
– G93A transgenetic familial ALS mice (Figure 1.)
• Cell Line and Alginate microcapsules
– Cell lines: hMSC cell line was immortalized by transduction with
the human Telomarase Reverse Transcriptase
(hTERT) gene. Then, cells are transfected with a
plasmid vector encoding GLP-1 fusion gene.
– Microcapsules: Cells are embedded in a 160 μm diameter spherical
alginate matrix. One matrix contains about 94
cells. (Figure 2.)
Figure 1. Analysis of post-mortem brain tissue at day 110 of SOD1 (G93A) mice.
Knippenberg S, Thau N, Dengler R, Brinker T, et al. (2012) Intracerebroventricular Injection of Encapsulated Human Mesenchymal Cells Producing Glucagon-Like Peptide 1 Prolongs Survival in a Mouse Model of ALS. PLoS ONE 7(6): e36857. doi:10.1371/journal.pone.0036857http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036857
Figure 2. Alginate encapsulated GLP-1 producing mesenchymal cells.
Knippenberg S, Thau N, Dengler R, Brinker T, et al. (2012) Intracerebroventricular Injection of Encapsulated Human Mesenchymal Cells Producing Glucagon-Like Peptide 1 Prolongs Survival in a Mouse Model of ALS. PLoS ONE 7(6): e36857. doi:10.1371/journal.pone.0036857http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036857
Materials and Methods
• Method
– Drill a 1.4 mm hole on the head of each mouse.
– Inject microcapsules which were treated by the cell solution into brain through
the hole.
• Treatment group and vehicle control group
– One group of mice was treated by microcapsules contains MSC which can
produce GLP-1.
– The other group mice was treated by empty microcapsules.
Result
• Survival Study (Figure 3, Figure 4)
– General conditional
– Survival percentage
– Weight
– Rotarod performance
– Step length analysis
– Runtime analysis
• Histological analysis
Figure 3. Effects of GLP-1 treatment on survival times, general condition and weight measurements.
Knippenberg S, Thau N, Dengler R, Brinker T, et al. (2012) Intracerebroventricular Injection of Encapsulated Human Mesenchymal Cells Producing Glucagon-Like Peptide 1 Prolongs Survival in a Mouse Model of ALS. PLoS ONE 7(6): e36857. doi:10.1371/journal.pone.0036857http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036857
Figure 4. Effects of GLP-1 treatment on rotarod performance and footprint analyses.
Knippenberg S, Thau N, Dengler R, Brinker T, et al. (2012) Intracerebroventricular Injection of Encapsulated Human Mesenchymal Cells Producing Glucagon-Like Peptide 1 Prolongs Survival in a Mouse Model of ALS. PLoS ONE 7(6): e36857. doi:10.1371/journal.pone.0036857http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036857
Figure 5. Immunhistological analysis of MAP2 in spinal cord tissue of SOD1 (G93A) mice.
Staining against microtubule associated protein 2 was stronger in animals treated with GLP-1 MSC (right) compared to control group (Left).
Conclusion
According to the data, we can conclude that intracerebroventricular injection
of encapsulated hMSC-GLP1 exhibits neuroprotective potential in SOD1-
G93A mice.
Discussion
• Advantages
– Compare to the only medicine, Riluzole, hMSC-GLP1 delays the
disease onset and weight loss.
– This cell therapy has less side effects compare to the other therapies.
• Limitation
– Has to be treated before the disease onset.
– Surgery may cause other systemic inflammation.
Reference
Knippenberg S, Thau N, Dengler R, Brinker T, Petris; Intracerebroventricular injection of encapsulated human mesenchymal cells producing glucagon-like peptide 1 prolong survival in a mouse model of ALS. PLoS One, 2012.
Lewis C, Suzuki M; Therapeutic applications of mesenchymal stem cells for amyotrophic lateral sclerosis. Stem Cell Research & Therapy, 2014, 5.
Keifer O, O’Connor D, Boulis N; Gene and protein therapies utilizing VEGF for ALS. Pharmacology & Therapeutics, 2014, 141.
Gao A, Peng Y, Deng Y, Qing H; Potential therapeutic applications of differentiated inducedpluripotent stem cells in the treatment of neurodegenerative diseases. Neuroscience, 2013, 228.
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