Potential of Captopril and Olmesartan to

Decrease Radiation Induced Pulmonary Fibrosis

Madisen Klink
Dr. Susanna Driscoll, MD
Viterbo University
Background

● Radiation treatment may be used to treat lung cancer

○ Fibrosis is a possible side effect
● Up to 43% of patients develop
pulmonary fibrosis
● Median survival after diagnosis is
2.5–3.5 years
● Fibrosis is particularly harmful in
lungs
● Increased survival rate of cancer patients
Indicators of Fibrosis
Renin-Angiotensin System

● Importance of ACE
ACE Inhibitors and ARBs

● ACE inhibitors vs ARBs

○ Currently used to treat
hypertension and
heart failure
● AT1 receptor properties
● AT2 receptor properties
Specific Aim

Specific Aim: Determine the effect of captopril or olmesartan to decrease the amount of
fibrosis markers present in irradiated lung tissue cells
● Hypothesis: Irradiated lung tissue cells treated with captopril or olmesartan will
produce fewer ROS
Methodology

● Grew rat lung fibroblast cells

○ Passaged into four flasks
● Plated and treated in six-well plates 24 hours before irradiation
○ Irradiated rat lung fibroblast cells vs control cells
■ No treatment, treated with captopril, treated with olmesartan
● Irradiated cells
○ Trials 1 and 2 were irradiated at 5 gray
○ Trial 3 was irradiated at 10 gray
● Flow cytometry to detect the presence of ROS three days after irradiation
Data Analysis

● Comparisons between trials and treatment groups

ROS Results

A: irradiated vs. non-

irradiated negative
control cells

B: irradiated vs. non-

irradiated positive
control cells
 ROS Results
A: irradiated captopril cells vs irradiated
negative control cells (left) or irradiated
positive control cells (right)

B: irradiated olmesartan cells vs irradiated

negative control cells (left) or irradiated
positive control cells (right)

C: non-irradiated captopril cells vs non-

irradiated negative control cells (left) or non-
irradiated positive control cells (right)

D: non-irradiated olmesartan cells vs non-

irradiated negative control cells (left) or non-
irradiated positive control cells (right)
ROS Breakdown

● Superoxide
dismutase (SOD)
converts ROS into
hydrogen peroxide
● Catalase (CAT)
converts hydrogen
peroxide into water
Summary

● No visible difference in ROS levels between irradiated and non-irradiated cells

● No visible difference in ROS levels between different treatment groups
● Assay able to detect ROS presence
● Adjust protocol to account for window of elevated
ROS levels
Future Work

● Repeat the current assay as it is with single dose of 5 or 10 gray with or without
ACE-I/ARB, but measure ROS within 10 hours of irradiation and at 14 and 21 days
later
● Fractionated dose of 10 gray with 0.5 gray daily 5 days per week for 20 week days
with or without ACE-I/ARB, with ROS measured within 10 hours of irradiation and
at 14 and 21 days later
● qPCR for collagen type I gene expression
Acknowledgements

● Dr. Susanna Driscoll, MD

● Ben Fleuchaus
● Dr. Scott Cooper, PHD
● Dr. Luke Bussiere, PHD
● Easton Halverson
● Viterbo University
● University of Wisconsin La Crosse
Questions?
References
● Shrimpton, A. J., Walker, S. L. M., & Ackland, G. L. (2020). Angiotensin converting enzyme inhibitors and angiotensin receptor blockers. BJA Education, 20(11), 362-367.
https://https://doi.org/10.1016/j.bjae.2020.07.004
● Ding, N., Li, J. J., & Sun, L. (2013). Molecular Mechanisms and Treatment of Radiation-Induced Lung Fibrosis. Current Drug Targets, 14(11), 1347-1356.
https://10.2174/13894501113149990198
● Pulmonary Fibrosis - Symptoms and Causes. (2018). Mayo Clinic. Retrieved 10/25/2022, from
https://www.mayoclinic.org/diseases-conditions/pulmonary-fibrosis/symptoms-causes/syc-20353690
● Straub, J. M., New, J., Hamilton, C. D., Lominska, C., Shnayder, Y., & Thomas, S. M. (2015). Radiation-induced fibrosis: mechanisms and implications for therapy. Journal of
Cancer Research & Clinical Oncology, 141(11), 1985-1994. https://10.1007/s00432-015-1974-6
● Chopra, M., Beswick, H., Clapperton, M., Dargie, H. J., Smith, W. E., & McMurray, J. (1992). Antioxidant Effects of Angiotensin-Converting Enzyme (ACE) Inhibitors: Free
Radical and Oxidant Scavenging are Sulfhydryl Dependent, but Lipid Peroxidation is Inhibited by Both Sulfhydryl- and Nonsulfhydryl-Containing ACE Inhibitors. Journal of
Cardiovascular Pharmacology, 19(3), 330-340. https://10.1097/00005344-199203000-00005
● Richter, K., & Kietzmann, T. (2016). Reactive oxygen species and fibrosis: further evidence of a significant liaison. Cell and Tissue Research, 365(3), 591-605.
https://10.1007/s00441-016-2445-3
● Harikrishnan, V., Titus, A. S., Cowling, R. T., & Kailasam, S. (2019). Collagen receptor cross-talk determines α-smooth muscle actin-dependent collagen gene expression in
angiotensin II–stimulated cardiac fibroblasts[MMK1] . Journal of Biological Chemistry, 294(51), 19723-19739. https://https://doi.org/10.1074/jbc.RA119.009744
● Ghosh, S. N., M.D, Zhang, R., M.D, Fish, B. L., B.A, Semenenko, V. A., Ph.D, Li, X. A., Ph.D, Moulder, J. E., Ph.D, Jacobs, E. R., M.D, & Medhora, M., Ph.D. (2009). Renin-
Angiotensin System Suppression Mitigates Experimental Radiation Pneumonitis. International Journal of Radiation Oncology, Biology, Physics, 75(5), 1528-1536.
https://10.1016/j.ijrobp.2009.07.1743
● Medhora, M., Gao, F., Fish, B. L., Jacobs, E. R., Moulder, J. E., & Szabo, A. (2012). Dose-modifying factor for captopril for mitigation of radiation injury to normal lung.
Journal of Radiation Research, 53(4), 633-640. https://10.1093/jrr/rrs004
● Molteni, J. E. Moulder, E. F. Cohen, W. F. Ward, B. L. Fish, J. M. Taylor, L. F. Wolfe, L. Brizio-Molteni, P. Veno, A. (2000). Control of radiation-induced pneumopathy and lung
fibrosis by angiotensin-converting enzyme inhibitors and an angiotensin II type 1 receptor blocker. International Journal of Radiation Biology, 76(4), 523-532.
https://10.1080/095530000138538
● Delanian, S., Baillet, F., Huart, J., Lefaix, J. -., Maulard, C., & Housset, M. (1994). Successful treatment of radiation-induced fibrosis using liposomal CuZn superoxide
dismutase: clinical trial. Radiotherapy and Oncology, 32(1), 12-20. https://10.1016/0167-8140(94)90444-8
● Shimura, T., Ando, T., Narao, M., Sasatani, M., Kamiya, K., & Ushiyama, A. (2020). Mechanism of turnover or persistence of radiation-induced myofibroblast in vitro. Cell
Cycle, 19(23), 3375-3385. https://10.1080/15384101.2020.1848063
● Wang, Y., Branicky, R., Noë, A., & Hekimi, S. (2018). Superoxide dismutases: Dual roles in controlling ROS damage and regulating ROS signaling. The Journal of Cell