1. qPCR ANALYSIS OF GENE EXPRESSION IN SALIVARY GLANDS OF NOD MICE TREATED BY
BONE MAROW CELLS
Baraka Gathere*1, Jacqueline Garcia*1, RAMIN RABII 2, SHEN HU 2
UCLA SCHOOL OF DENTISTRY
Our study suggests that bone marrow cell treatment may
successfully repair damaged salivary glands in NOD
mice. We found that the treatment is effective due to
inhibiting inflammation while also improving the
regeneration and differentiation of salivary gland cells.
Abstract
The goal and purpose of our lab is to determine what
expressive genes in HSCT are responsible for treating
Sjögren's symptoms.
Our lab would find a way to treat the 1.4 million people in
the U.S. alone that have Sjögren's Syndrome.
Introduction: Sjögren’s syndrome is an inflammatory autoimmune disease that causes the
immune system to attack the exocrine glands that produce saliva and tears. Objective: We are
going to test the treatment of Sjögren’s syndrome to determine if the treatment results in coercing
moisture-producing glands to produce more amylase and reduce the genetic expression of
inflammatory genes. One of the more successful treatments for this syndrome is hematopoietic
stem cell transplantation (HSCT). HSCT is the transplantation of multiple hematopoietic stem
cells or blood, usually derived from bone marrow, peripheral blood stem cells, or umbilical cord
blood. Methods: We transcribed 5 samples of mRNA from a NOD mouse into cDNA using a
Thermal Cycler 480 machine. Three of the samples coded for primary Sjögren’s with the gene,
Sjögren’s syndrome antigen B no poly a. After that, we took the samples and placed the wells
into a 7500 fast real-time PCR system and ran a program that would track the changes of the
cDNA. Results: To be determined after acquiring our data. Discussion: Sample 2 was a
reference gene, and the other samples were what we read to attain our results. Sample 836 and
847 matched our hypothesis; but sample 4 showed a decrease in amylase and sample 841
showed an increase in the sss antigen. Conclusion: On previous studies conducted, mice who
had oral cancer with similar symptoms to that of Sjögren's were treated with bone marrow cells,
and they had a successful recovery from the disease and treatment. This can lead to finding a
cure or treatment for other syndromes and/or diseases, like cancer.
Figure 1. qPCR analysis of vimentin (VIM), parotid secretary protein (PSP), amylase (AMY),
aquaporin 1 (AQP1), lacrimal androgen-binding protein (LABP), caspase 8 associated
protein 2 (CASP8AP2), Sjogren syndrome antigen (SSA) and calcium binding protein (CAL) in
salivary gland cells of the NOD mice after bone marrow cell extract treatment.
.
Fig. 2: Bone marrow cell extract (soup)
treatment helps improve the saliva flow rate
in NOD mice
We transcribed 5
samples of mRNA
from a salivary cell
from a NOD mouse
into cDNA and then
used Real-Time RT-
PCR to make
numerous copies.
♦Three of the
samples were coded
as the genetics of
primary Sjögren's
syndrome (Sjögren's
syndrome antigen B
no poly a), while the
other two served as
standards for the
base line.
Bone marrow cell extract treatment improves the salivary
flow rates in NOD mice.
Salivary amylase and parotid secretary protein genes
were found to be up-regulated whereas the Sjogren
syndrome antigen gene was down-regulated in the NOD
mice treated by bone marrow cells.
Inflammatory cytokines were down-regulated whereas
several growth factors were up-regulated in the NOD
mice when treated by bone marrow cells.
.
Thank you for your
attention.
We would like to thank
our parents and family for
their constant support.
We would like to thank
Dr. Hu, Ramin Rabii,
Kaori Misuno and the
rest of the team in Dr.
Hu's lab for teaching us
and helping us with our
experiments
Thank you Ed Meyer for
answering our questions
pertaining to the project.
Fig.3: qPCR analysis of growth
factors/cytokines in salivary gland cells of the
NOD mice after bone marrow cell extract
(BMCE) treatment (n=4). * p<0.05
.
Sjögren’s syndrome causes the immune system to attack and
inflame the salivary and lacrimal glands.
The syndrome is extremely painful due to significant
reduction of saliva and tear production.
•There is no cure for Sjögren’s syndrome, but there is
treatment for it
Background Information
Materials and Methods
Purpose
Results Discussion
Conclusion
Acknowledgments