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.

Thank you!