Gene Therapy for

Rheumatoid Arthritis
By Ethan Cadaoas, Mateo Eusebio, and Tyler Leighton
What is RA?
Rheumatoid Arthritis is a chronic inflammatory disorder that affects mainly joints. It
is also an autoimmune disorder, meaning that the immune system mistakenly
attacks tissue. Unlike wear and tear damage from osteoarthritis, RA affects the
lining of joints, causing swelling, joint deformation, and bone erosion.
Why should you care about RA?
About 0.3% to 1% of the world population is affected by RA. It tends to occur
between the ages 20 and 40 and mostly affects women. It disabling disease which
causes great pain and deformations. Within 10 years of getting RA, around 50% of
patients in developed countries are unable sustain a full-time job. Additionally, RA
increases the chances of a stroke and heart attack. RA is a common disabling and
life destroying disorder that can become possible life threatening.
Why RA hasn't been cured yet.
This is because the cause of RA is unknown for the most part. Scientists are having
trouble finding whether it is of bacterial etiology or viral etiology that causes the
immune system to attack the body’s tissues. Due to scientists not knowing the
cause a cure can not be developed.
Problems with current treatments
There are also problems with the anti-rheumatic treatments, or DMARDs. These
drugs help treat the disease by suppressing the immune system to stop the
inflammation in the joints that RA is causing. This suppression of the immune
system can cause other issues with the body, such as fevers and in rare cases, liver
and blood problems. With these blood problems come higher risk of infection due
to the decreasing white blood cells.
What is gene therapy?
When a gene is defective, it loses the ability to function properly. During gene
therapy, scientists insert a copy of the healthy gene in place of the faulty one in
order to express healthy functions. In order to insert the copy, they require the
assistance of a vector, which are plasmids that are artificially modified to move and
insert the gene into a cell. The healthy gene uses the vector to transport itself into
the cell, where the healthy gene can be inserted into the chromosome.
How gene therapy treats RA
Our idea is to use the IL 10 gene. It produces the IL 10 cytokine, a small protein, that
has anti inflammatory properties. It will be used to treat the inflammation and
swelling of the joints. This gene would be inserted into a DNA plasmid, which would
then be inserted into a CD4+ T-Cell or helper T cell that has been extracted from
the patient. The T-Cell will then be injected into the joints affected by RA.
Why it's better than other treatments
It is currently impossible to cure RA due to the cause of it being unknown. Our
solution will not cure RA, but suppresses the immune system. Helper T cells are the
ideal for the delivery of immunoregulatory molecules due to these cells having
specific homing properties. Which will cause them to migrate to the site of
inflammation and express the IL-10 cytokines, which will suppress inflammation and
lesions. Helper T cells are a non viral vector, which means they are not as effective
as viral. However, they are saver compared to viral, since they can cause infections
Non viral vectors are also cheaper to produce than viral vectors. Compared to
DMARDs which the suppress the whole immune system, the IL-10 cytokine doesn't
fully suppress the immune system and allows it to still function properly and it
suppressed inflammation.
Our Action Plan
Some funding would be a good start to be able to operate. Next we’d need to
produce the genes that have to do with a non viral solution. After that, we would
conduct preliminary clinical trials on mice affected by RA to see if the non viral gene
is safe and effective. Once we pitch the idea and get support from someone, we
can distribute the product to people suffering from RA. For each individual’s
treatment to work properly and avoid any further complications with the immune
system, we would need to use that person's own T cells for the delivery of the non
viral vector.
Procedure Pt. 1
Testing the T Cell Migration with mice
1. Get a sample size of 20 mice with RA
2. Take the blood sample of mice with rheumatoid arthritis
3. Separate the CD4+ T-Cells from the blood samples
4. Inject DNA Plasmid with pKiwi gene, to make them glow
5. Clone T Cells
6. Inject mice with T Cells
7. Observe cells in a dark environment to see if they move to the inflamed
joints
Procedure Pt. 2
Testing IL 10 effectiveness on RA with mice
1. Get a sample size of 20 mice with RA
2. Take the blood sample of mice with rheumatoid arthritis
3. Separate the CD4+ T-Cells from the blood samples
4. Inject T Cells with IL 10
5. Clone T Cells
6. Inject DNA Plasmid with bioluminescence gene into point of inflammation,
and through gene expression let it spread
7. Once the area is covered inject T Cells with IL 10
8. Observed what happens with the bioluminescence inflammation areas
and if it shrinks or not
9. If it shrinks we know the IL 10 reduces inflammation
Procedure Pt. 3
Testing T Cell with IL 10 on humans with RA
1. Get a sample size of 100 people with RA
2. Take the blood sample of each person
3. Separate the CD4+ T-Cells from the blood samples
4. Inject DNA Plasmid with IL 10
5. Cultivate and clone T Cells
6. Inject people with new modified T-Cells
7. Based on how long it takes for T Cells to travel to the inflamed area, we
would wait and observe the effects of the genes for the same amount of
time as the previous test
Works Cited
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