1. RESEARCH POSTER PRESENTATION DESIGN © 2015
www.PosterPresentations.com
Materials:
Cell Lines:
 HFF-1: Homo sapiens Fibroblast Foreskin Cells
 Bg-1: Homo sapiens Ovarian Cancer Cells
Promoters assembled with GFP Reporter
Methods:
Materials and Methods Results Conclusion
The data in this round of experiments indicates that there is no
difference in promoter activity between glucose-deprived and non-
glucose-deprived cells. Neither is there a difference between cancer
and non-cancer cells. However, before definite conclusions can be
made about promoter activity in glucose-deprived environments,
further experiments will be conducted.
According to the Glucose Reduction Cell Viability Studies, there
appears to be a response to glucose deprivation. This is illustrated by
the cells’ rebound in viability after the glucose drops below a critical
concentration. Accurately identifying the concentration where the
cells respond is necessary for the promoter activity chronic glucose
shock study. Several optimum concentrations were present, and
0.03125 mg/ml was chosen for experimentation.
Therefore, in the Promoter Activity Chronic Glucose Shock
experiments, the lack of increase in promoter activity may be due to
the use of a sub-optimum glucose concentration. According to the
data other promising glucose concentrations to test are 0.06 and
0.125 mg/ml of glucose. Additionally, in the HFF-1 glucose
reduction cell viability study, the third trial due to experimental
reasons yielded different values than the first two which skewed the
composite graph. This trial can be re-run to more definitively
determine a glucose concentration to use for further chronic shock
experimentation.
References
1. Zhang X, Turner C, and Godbey WT. Comparison of Caspase Genes
for the Induction of Apoptosis following Gene Delivery. Mol
Biotechnol. (2009) 41: 236-246.
2. Zhang X and Godbey WT. Preclinical Evaluation of a Gene therapy
Treatment for Transitional Cell Carcinoma. Cancer Gene Ther. (2011)
18: 34-41.
3. Xuguang Chen. Advanced transitional Cell Carcinoma
Treamtments Via Expression-Targeted Gene Delivery and Minicell
Technology. Diss. Tulane University, 2014. Print. 135-168.
4. Baluchamy, S., et al. Differential oxidative stress gene expression
profile in mouse brain after proton exposure. In Vitro Cellular &
Developmental Biology- Animal, 46, 718-725
Acknowledgments
Funding for this project was supported by the Center for
Engaged Learning & Teaching and the Louisiana Alliance for
Minority Participation, Tulane University, New Orleans, LA.
Department of Chemical and Biomolecular Engineering, Tulane University, New Orleans , USA
Christopher Chedid, Zoe Poncher, and W. T. Godbey
Reduced Glucose Effect on Promoter Activity
Glucose Reduction Cell Viability Studies
Plated 100,000 cells/well
    
     
HFF-1
Bg-1
Replaced Medium with
DMEM D-glucose Dilutions
24 hrs
24 hrs
Trypsinize
  
  
 Count with
Hemocytometer
Promoter Activity Chronic Glucose Shock Studies
Plated 100,000 cells/well
    
     
HFF-1
bg-1
16 hrs
Transfect Cells with
PEI/DNA Solution
pcmv pgrp ppolh
pznf
Pro-
moter-
less
2 hrs
HFF-1 Bg-1
Shock
Replaced Medium with
Reduced Glucose Conc.
Control (Non-shock)
Replaced Medium with
Normal Glucose Conc.
Future Work
Trypsinize at various
time points and build
activity profiles
through FACS
measurements
 Additional Promoter Activity Chronic Glucose
Shock Studies at Higher Glucose Concentration
 Oxidative Stress Shock Study on the effect of
pcmv, pgrp94, ppolh, Pznf394, and ptxnll promoter
activity
 Heat Shock Study and UV shock study on the
effect of previously mentioned promoter activity
 Additional studies exploring other promoters
reactions to glucose, oxidative and heat stressed
environments
 pcmv: Cytomegalovirus  ppolh: Polymerase (DNA directed)
eta
 pgrp94: Endoplasmin  Pznf394: Zinc finger protein
Glucose Reduction Cell Viability Studies
Promoter Activity Chronic Glucose Shock Studies
0
10
20
30
40
50
60
%Expression Promoters
Promoter Expression at 24 Hours
Bg-1 Shock
Bg-1 No Shock
HFF-1 Shock
HFF-1 No Shock
Normal medium:
4.5 g/L
Glucose Conc.
Shock medium:
.03125 mg/mL
Glucose Conc
0
5
10
15
20
25
30
35
Cells/ml(x104)
Glucose Concentration (g/L)
HFF-1 Glucose Reduction Viability Study
0
10
20
30
40
50
0 0.125 0.25 0.5 1 2
Cells/ml(x104)
Glucose Concentration (g/L)
Bg-1 Glucose Reduction Viability Study
 The data reveals a trend of consistent or increased cellular
growth in medium with lower glucose concentrations
 Proves both normal and cancerous cells can survive prolonged
immersion in a glucose poor environments, such as that found
near cancer cell colonies
Special thanks to our research advisor, Dr. W.T. Godbey for all
his assistance and guidance for this research project. Our
discussions consisting of lab techniques, previous research
experiences, and group work at unbearably early hours, made
this project both enjoyable and educational.
Expression Targeted Gene Therapy treats illnesses by altering
gene expression and uses expression-targeting to specifically affect
diseased cells rather than healthy cells in the body. Expression-
targeting is achieved by introducing a regulatory element, such as a
promoter, to the gene being delivered. Promoters are used to
increase expression of a gene. Specific promoters shown to be
overly active in certain stressed environments will cause the cells
undergoing such stress to express the therapeutic gene while
unstressed cells will remain unaffected.
In order to target specific cells, the chosen promoters must only
be active in distinct conditions. Glucose deprivation, heat stress,
hypoxia, and oxidative stress are several specific stresses in which
promoters can be activated. Furthermore, these stresses are caused
by inflammation which is associated with many ailments including
cancer. So, finding promoters that are specific to the before-
mentioned stresses opens many doors to improved treatment for
numerous illnesses including cancer. This improves upon the
current treatment of chemotherapy, a drug therapy infamous for its
side effects due to unspecificity.
The three promoters studied in the following round of
experiments are hypothesized to be active in specified stresses due
to their original functions in the body. Endoplasmin (grp94),
Polymerase (DNA directed) eta (polh), and zinc finger protein (znf
394) are respectively hypothesized to activate in glucose stress, UV
stress, and oxidative stress. Promoter activity in low glucose stress
was studied in the following experiments. To emphasize the
applicability of this treatment to cancer, experiments were
performed using both cancer and non-cancer cells to compare
promoter activity and determine if a significant difference exists.
Introduction
Inflammation and
cancer cause
stressed
environments in
tissues
Promoter-controlled genes
introduced into the body,
activate in stressed areas and
provide therapeutic effects to
treat inflammation or cancer
 Determine optimum glucose concentration to use in
experiments
 Determine effect of glucose deprivation stress on
promoter activity in cancer and non-cancer cells
Objectives
Promoter
Low
Glucose
Environment
Promoter
Low glucose conditions will increase
specific promoters’ activity most
prominently in cancer cell lines
Hypothesis