Progress in ovarian cancer using the
laying hen model.
I was contacted recently from the Department of Defense ovarian cancer research program for an update on our progress. We were first funded by the DOD in 2006 so this marks the 10th anniversary of working in the chicken model of ovarian cancer-- the only natural model of spontaneous ovarian cancer that replicates the human disease. Our current efforts are focusing on development of biomarkers for early detection by mining the large archive we have of chicken tissues, collected from long-term longitudinal studies that enable us to look before, during and after ovarian cancer starts. We are looking at the estrogen metabolite index (EMI), serum prostaglandin E2 (PGE2), soluble E-cadherin and micro RNA 200. This is the report I generated for the DOD-- 20 papers and over 3 million dollars in extramural funding support. I think the DOD got their money's worth, and receiving that first ovarian cancer grant from the DOD (a.k.a. Congressional Directed Medical Research Program) is what hatched the whole research endeavor in chicken ovarian cancer.
For the first
15 years of my independent career the focus of my research was on the role of
inflammation and oxidative stress in the regulation of male reproductive
function. A colleague of mine at UIC
recruited me to help with his ovarian cancer project to help understand the
role of oxidative stress and inflammation in the etiology of ovarian
cancer. He made the comment that he’d
heard the only really good animal model for ovarian cancer was the laying hen. I had a colleague at UIUC who was the world expert
in chicken reproductive physiology so I approached Dr. Janice Bahr and she
agreed to help provide access and her expertise so that we could look at
ovarian cancer in the chicken. What made
all of the success we’ve had in this model possible, was the pilot grant I
received from Department of Defense Ovarian Cancer Research Program
(OCRP). The funding enable me to
collaborate with Dr. Barua at Rush University, who had expertise with chickens,
knew about ovarian cancer, but had no funding.
With our collective expertise we have helped to establish the hen model
as the best natural model of spontaneous ovarian cancer. Notably, we published a high impact review
article in Oncogene describing the hen model, to help the model gain wider
acceptance.
Research
findings based directly on CDMRP funded project.
1)
CYP1B1,
the “P450 of cancer” is highly expressed in the post-ovulatory follicle of the
normal hen ovary likely induced by the oxidative stress, inflammation and
estrogenic microenvironment in the ovary after ovulation when the surface of
the ovary is undergoing wound healing.
We hypothesize that the enzyme, a known estrogen hydroxylase, is
positioned to oxidatively activate estrogens at the site where primary ovarian
cancer originates. {Zhuge, Y., Ansenberger, K., Mahon
2)
CYP1B1,
CYP1A1 and CYP3A4, which are P450 enzymes that hydroxylate estrogen in a
region-specific way, are modulated by diets supplemented with flaxseed or the
phytoestrogen lignan component of flaxseed, secoisolariciresinol. CYP1B1 and CYP3A4 whose actions potentiate
estrogen’s oncogenic effects, are down regulated by the flax diets, while
CYP1A1 which renders estrogens harmless, is induced by these diets. This work has led to the discovery that
2-methoxy-estradiol is a potent pro-apoptotic and anti-proliferative agent,
that likely contributes directly to the anti-cancer effects of flaxseed. { Dikshit, A, Adriao-Gomes Filho, M, Eilati,
E, McGee, S, Small C, Gao, C, Klug, T, Hales DB, “Flaxseed reduces the
pro-carcinogenic microenvironment in the ovaries of normal hens by altering the
prostaglandin and estrogen pathways in a dose dependent manner” British Journal of Nutrition 113:1384-1395
(2015); Dikshit, A, Gao C, Small, C,
Hales, KH, Hales, DB “Flaxseed and its components cause differential
effects on estrogen receptor expression and signaling pathways in the ovary of
pre-cancerous laying hens.” J Steroid
Biochemistry and Molecular Biology (in press 2016) DOI: 10.1016/j.jsbmb.2016.02.028.}
Based on
our observations from the DOD funded project, we hypothesized that the
inflammation and oxidative stress that results from ovulation, and induces
CYP1B1, may be a druggable target. We
reasoned that if we could feed the hens a diet rich in natural antioxidants
that target ovary, then the diet may reduce ovulation-associated inflammation
and ameliorate ovarian cancer. Omega-3
polyunsaturated fats were attractive because they were known to reduce oxidative
stress and inflammation and at the same time be cardioprotective. We found that the most effective way to
deliver omega-3 to the hen was to feed them flaxseed. Because hens fed flaxseed accumulate omega-3
in their yolk, we knew then that the ovary must be exposed to the omega-3. We found that flaxseed supplemented diets do
indeed reduce the severity and incidence of ovarian cancer in the hens.
These
findings have spawned several new research projects and helped me to garner
several extramural awards. Notably, we
have been able to parlay work with the hen into two clinical studies. We observed that the flax affects estrogen
metabolism and determined that we could measure the different estrogen
metabolites in the hens. To test the
clinical utility of this finding, we collaborate with the OB/GYN department at
SIU School of Medicine, with Dr. Laurent Brard.
(Coincidently I first met Dr. Brard when we were both on the OCRP grant
review panel in Reston, VA). When women
report to the clinic with an undiagnosed adnexal mass, Dr. Brard collects urine
and sends it to my laboratory. We
analyze the estrogen metabolites and have determined the a shift in the ratio
of 2 to 16 hydroxylated estrogens, with an increase in 16 relative to 2, is
associated with gynecological pathology.
Based on these findings we are actively applying for funding to
determine estrogen metabolite index can be used as biomarker for detection of
ovarian and endometrial cancers.
We determined
that flaxseed slows the progression of ovarian cancer by targeting inflammatory
prostaglandins {Ansenberger, K., Richards, C., Barua, A.,
Bahr, J.M., Luborsky, J.L., Hales, D.B. “Decreased severity of ovarian cancer and
increased survival in hens fed a flaxseed enriched diet for one year” Gynecological Oncology 117:341-347 (2010);
Eilati, E, Pan, L, Bahr, JM, Hales, DB
“Age dependent increase in Prostaglandin pathway coincides with onset of
ovarian cancer in laying hens” Prostaglandins, Leukotrienes
& Essential Fatty Acids 87:177-184 (2012); Eilati,
E, Bahr, JM, Hales, DB “Long Term
Consumption of Flaxseed-Enriched Diet Decreases Ovarian Cancer Incidence and
Prostaglandin E2 in Hens.” Gynecologic Oncology 130:620-628 (2013); Eilati, E, Zhuge, Y, Hales, KH,
Ansenberger Fricano, K, Rui, Y, van Breemen, RB, Hales, DB “Flaxseed enriched diet-mediated reduction in ovarian
cancer severity is correlated to the reduction of prostaglandin E2 in laying
hen ovaries.” Prostaglandins, Leukotrienes & Essential Fatty Acids
89:179-87 (2013)}
Based on these
studies, we hypothesized that flaxseed supplementation may help to prevent
recurrence of ovarian cancer after women have undergone cytoreductive surgery
and chemo-therapy. These first line
therapies are highly effective—initially, and after women recover from the
surgery and chemo-therapy, they are essentially disease and symptom free for an
average of 2 years. Unfortunately the
majority of these women get recurrent disease and many ultimately succumb
because the recurrent cancer has become chemo-resistant. Our clinical trial is designed to provide
women in remission 20 grams of flaxseed per day in hope that this will delay or
ideally prevent the recurrence of the disease.
{https://clinicaltrials.gov/ct2/show/NCT02324439
}. We obtained funding from
the Simmons Cancer Institute at SIU School of Medicine for the initial study
and are applying for NCI funding for this trial.
Original
peer-reviewed journal articles stemming from initial CDMRP funding for research
using the chicken model of ovarian cancer.
1)
Stammer,
K., Edassery, S.L., Barua, A., Bitterman, P., Bahr, J.M., Hales, D.B., Luborsky, J. “Selenium-Binding Protein 1 expression in
ovaries and ovarian tumors of the laying hen, a spontaneous model of human
ovarian cancer.” Gynecologic Oncology 109(1):115-21
(2008)
2)
Hales, D.B, Zhuge, Y., Lagman, JA,
Ansenberger, K, Mahon, C, Barua, A., Luborsky, J., Bahr, JM. “Cyclooxygenase
Expression and Distribution in the Normal Ovary and Their Role in Ovarian
Cancer in Gallus Domesticus” Endocrine
33:235-24(4 2008)
3)
Barua,
A., Edassary, S.L., Bitterman, P., Abramowicz, J.S., Dirks, A., Bahr, J.M., Hales, D.B., Bradaric, M.J.,
Luborsky, J.L. “Prevalence of
anti-tumor antibodies in laying hen model of human ovarian cancer” Int J Gyn Cancer 19(4):500-507 (2009)
4)
Barua A,
Bitterman P, Abramowicz J, Bradaric MJ, Edassery SL, Dirks A, Hales DB, Bahr JM and Luborsky JL,
(2008) Histopathology of ovarian tumors in laying hens, a preclinical model of
human ovarian cancer. International
Journal of Gynecological Cancer: 19(4):531-539 (2009)
5)
Zhuge, Y., Ansenberger, K., Mahon
6)
Barua
A, Abramowicz JS, Bitterman P, Bahr JM, Hales
DB, Luborsky JL. OP17.08: Transvaginal ultrasound predicts ovarian tumor
associated neo-angiogenesis. Ultrasound Obstet Gynecol; 32:370 (2008)
7)
Ansenberger,
K., Zhuge, Y., Lagman, J.A., Richards, C., Barua, A., Bahr, J.M., Hales, D.B., “E-cadherin Expression in
Ovarian Cancer in the Laying Hen, Gallus
Domesticus, compared to Human Ovarian Cancer” Gynecologic Oncology 113:
362-369(2009)
8) Barua, A., Bitterman,P., Bahr, J.M.,
Bradaric, M., Hales, D.B. Luborsky,
J., Abramowicz, J. "Detection of tumor associated neo-angiogenesis by
Doppler ultrasound during early stage ovarian cancer in laying hens: A
preclinical model of human spontaneous ovarian cancer" American Journal of
Ultrasound in Medicine (JUM) 29:173–182 (2010)
9)
Ansenberger,
K., Richards, C., Barua, A., Bahr, J.M., Luborsky, J.L., Hales, D.B. “Decreased
severity of ovarian cancer and increased survival in hens fed a flaxseed
enriched diet for one year” Gynecological
Oncology 117:341-347 (2010)
10) Barua A, Bitterman P, Bahr JM, Basu S,
Sheiner E, Bradaric MJ, Hales DB,
Luborsky JL, Abramowicz JS. Contrast-enhanced sonography depicts spontaneous
ovarian cancer at early stages in a preclinical animal model. Journal of
Ultrasound in Medicine.30:333-45 (2011)
11) Eilati, E, Pan, L, Bahr, JM, Hales, DB “Age dependent increase in
Prostaglandin pathway coincides with onset of ovarian cancer in laying hens” Prostaglandins, Leukotrienes
& Essential Fatty Acids
87:177-184 (2012)
12)
Machado,
SA, Bahr, JM, Hales, DB, Braundmeier, AD, Quade, BJ, Nowak, RA
“Validation of the Aging Hen (Gallus gallus domesticus) as an Animal Model for
Uterine Leiomyomas” Biology of
Reproduction 87 (4):1-11 (2012)
13) Eilati, E, Bahr, JM, Hales, DB “Long Term Consumption of
Flaxseed-Enriched Diet Decreases Ovarian Cancer Incidence and Prostaglandin E2
in Hens.” Gynecologic Oncology 130:620-628
(2013)
14) Eilati, E, Zhuge, Y, Hales, KH,
Ansenberger Fricano, K, Rui, Y, van Breemen, RB, Hales, DB “Flaxseed enriched diet-mediated reduction in ovarian
cancer severity is correlated to the reduction of prostaglandin E2 in laying
hen ovaries.” Prostaglandins, Leukotrienes & Essential Fatty Acids 89:179-87
(2013)
15) Eilati, E, Small, CF, McGee, SR, Kurrey,
NK, Hales, DB “Anti-inflammatory effects of fish oil in
ovaries of laying hens target prostaglandin pathways.” Lipids in Health and Disease 12 (1):152-163
(2013).
16) Lengyel, E, Burdette, JE, Kenny, HA,
Matei, D, Pilrose, J, Haluska, P, Nephew, KP, Hales, DB, Stack, MS “Review: Epithelial ovarian cancer
experimental models” Oncogene 33:3619-3633
(2014)
17)
Hales, K.H, Speckman S.,
Kurrey, N.K., Hales, D.B. “Uncovering Molecular Events Associated with
Chemosuppressant Effects of Flaxseed: A Microarray Analysis in Laying Hen Model
of Ovarian Cancer BMC Genomics 15 (1):709-713 (2014)
18)
Dikshit, A, Adriao-Gomes Filho, M, Eilati, E, McGee,
S, Small C, Gao, C, Klug, T, Hales DB, “Flaxseed reduces the
pro-carcinogenic microenvironment in the ovaries of normal hens by altering the
prostaglandin and estrogen pathways in a dose dependent manner” British Journal of Nutrition 113:1384-1395
(2015)
19)
Dikshit, A, Gao C, Small, C, Hales, KH, Hales, DB
“Flaxseed and its components cause differential effects on estrogen receptor
expression and signaling pathways in the ovary of pre-cancerous laying hens.” J Steroid Biochemistry and Molecular Biology
(in press 2016) DOI:
10.1016/j.jsbmb.2016.02.028
Davis,
JE, Cain, J, Small, C, Hales, DB “Therapeutic effect of flax-based diets
on fatty liver in aged
