Source: CLEMSON UNIVERSITY submitted to
SPONTANEOUS OVARIAN ADENOCARCINOMA IN AGING AND AGED COMMERCIAL LAYING HENS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
0220849
Grant No.
(N/A)
Project No.
SC-1700373
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Aug 15, 2009
Project End Date
Sep 30, 2014
Grant Year
(N/A)
Project Director
Birrenkott, G.
Recipient Organization
CLEMSON UNIVERSITY
(N/A)
CLEMSON,SC 29634
Performing Department
Animal & Veterinary Sciences
Non Technical Summary
Ovarian adenocarcinoma is the second most prevalent tumor found in laying hens (Goss, 1940). More recently (Alfonso et al., 2005) reported that approximately 45% of all aged commercial laying hens had tumors of ovarian or oviductal origin. Ovarian adenocarcinoma is the fifth leading cause of death in women (Jemal et al., 2007). In spite of the high incidence and severity of the problem ovarian adenocarcinomas are extremely difficult to detect and treat early. This is one of the reasons that there is such a high mortality rate associated with these tumors. Much additional basic and applied scientific discovery is needed to find ways to detect, treat and prevent this condition. The result of additional knowledge in this area could help to avoid pain and suffering in hens and women. The chicken hen is the idea model for this particular spontaneous pathology because of the large number of ovulations that she undergoes during her lifetime. No other animal model is as applicable to the study of this problem in poultry and women.
Animal Health Component
100%
Research Effort Categories
Basic
(N/A)
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3013210102050%
3013210116010%
3113210102025%
3113210116015%
Knowledge Area
311 - Animal Diseases; 301 - Reproductive Performance of Animals;

Subject Of Investigation
3210 - Egg-type chicken, live animal;

Field Of Science
1020 - Physiology; 1160 - Pathology;
Goals / Objectives
The overall objectives of this study are to: 1) Characterize age-related changes in ovarian follicular dynamics and blood flow during initial and subsequent laying cycles in hens (Gallus domesticus). 2) Define changes in the cellular relationships in the wall of the post-ovulatory follicle (POF) of aging hen using histological assessments. 3) Culture avian ovarian surface epithelial cells from hens of different ages, co-culture with granulosa cells and evaluate changes in growth rates, molecular and cellular markers for carcinomas. Outcomes or Projected Impacts - Finding a non-invasive way to measure and monitor hen ovarian cancer would allow for selection of breeding birds, perhaps, less likely to pass this trait on to their offspring. This would result in better hen health and egg productivity. This would positively affect the entire commercial egg laying and result in less mortality and greater egg production at a lower cost. Objectives two and three will result in a better understanding of the interaction of ovarian cells in the development and progression of ovarian cancer. The impact of this knowledge would be the development of new treatments prevention or treatment of chicken or for human ovarian cancer. This would have a significant economic impact on commercial egg production and human health care costs. It would also alleviate pain, suffering and mortality associated with this cancer - having profound social and animal welfare implications.
Project Methods
Animals - Single-comb white Leghorn (SCWL) pullets and hens will be grown and housed at the Morgan Poultry Center at Clemson University with photoperiods and diets controlled for optimum egg production. In addition, commercial laying hens may be donated at the end of their laying cycles for one-time collection of tissues. All procedures and protocols will be under the auspices of the Clemson University Animal Care and Use Committee (IACUC) and conform to the Association for Assessment and Accreditation of Laboratory Animal Care (AALAAC) standards. Objective 1. SCWL pullets at the beginning of their first laying cycle will be monitored weekly by color Doppler ultrasonography. Ovarian dynamics (follicular sizes) and blood flow will be acquired digitally using a trans-abdominal (non-invasive) approach. Oviductal scans for the presence of tumors and blood flow will also be performed and recorded at this time. Hens will be examined throughout the first laying cycle, force-molted and then continue to be examined through the second reproductive year. Special attention will be paid to any build up ascites within the peritoneal cavity - a sequelea of ovarian adenocarcinoma. Objective 2. SCWL hens at various time periods of their first and second reproductive years will be sacrificed and ovarian tissue, specifically the post-ovulatory follicle (POF) will be collected and fixed for subsequent histological examination. Sections of POF will stained and examined for age-related changes (in the hen and in the time since ovulation) in the configuration (involution) of the ovarian surface epithelium (OSE) in relation to the lumen of the empty follicular shell (granulosa cells, basal lamina, theca cells and stroma). If differences or anomalies, such as temporal affects on spatial changes in the configuration of the microenvironment of the OSE, basal lamina and follicular cells, are found during the aging of the hen or of the POF - additional laser capture dissecting microscopy will be done. Objective 3. Ovarian surface epithelial (OSE) cells from dominant (F1) follicles of young and aged (> 1 year old) hens will be harvested by a combination of mechanical and enzymatic digestion. The technology for the isolation and culture of OSE cells in the hen has been published (Giles et al., 2006). Recent work on the isolation of immature (white follicle) oocytes suggests that this single-cell layer can be enzymatic ally removed (see progress report 2008/01 -2008/12 from CSREES ALA015-030, Dr. W. D. Berry). There is already a published abstract which suggests a regional change in the OSE of the largest ovarian follicles of the hen (Giles and Johnson, 2002). Such regional differences will be further explored in the co-culture experiments with granulosa cells. Statistical Analysis - The Experimental Statistics faculty within the department of Applied Economics & Statistics will be consulted on all matters pertaining to experimental design, pre- and post-hoc analysis of data.

Progress 08/15/09 to 09/30/14

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? No further accomplishments since 2013 progress report due to reassignment of principal investigator. How have the results been disseminated to communities of interest? No further accomplishments since 2013 progress report due to reassignment of principal investigator. What do you plan to do during the next reporting period to accomplish the goals? No further accomplishments since 2013 progress report due to reassignment of principal investigator.

Impacts
What was accomplished under these goals? No further accomplishments since 2013 progress report due to reassignment of principal investigator.

Publications


    Progress 01/01/13 to 09/30/13

    Outputs
    Target Audience: This research and other work associated with this avian/human reproductive adenocarcinoma is of interest to other scientists and the public. This is especially true because it is one of the leading causes of death in women and chickens are the only spontaneous model for this condition. This material has been presented in three classes (300+ students), high school science students visiting Clemson University and through poster presentations at the IPPE (International Processing and Production Expo) in Atlanta, GA (attended by 25,000+ USA and international people interested in poultry and science). Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? The project has provided research training for many undergraduate students. Approximately half of the students on this project are now in veterinary schools or working in the poultry industry. How have the results been disseminated to communities of interest? The results have been disseminated at a national scientific meeting, a published abstract, in classroom discussions and at a national poultry industry meeting (IPPE, Atlanta, GA) with 25,000+ attendees. What do you plan to do during the next reporting period to accomplish the goals? I plan to initiate a new undergraduate research project to work on this problem in more detail (following up on our preliminary work) and to submit a publication based on the published abstract.

    Impacts
    What was accomplished under these goals? Preliminary data has been obtained and compared against our previously published work. The preliminary results have been presented at a national meeting and an international exposition.

    Publications

    • Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Creative inquiryUndergraduate research on the hen as a model for ovarian adenocarcinoma. G. Birrenkott* and J. Fain, Clemson University, Clemson, SC. Poult. Sci. 91 (Supplement 1): 102


    Progress 01/01/12 to 12/31/12

    Outputs
    OUTPUTS: A comprehensive review of our previous published and unpublished work on ovarian development in chickens and guinea fowl (Research Note: Ovarian Follicular Growth and Maturation in the Domestic Pigeon and Guinea Fowl (Numida meleagris), G. P. BIRRENKOTT, M. A. SHOOP, K. COOPER, and M. WIGGINS, Poult Sci 1988 67:1783-1786; doi:10.3382/ps.0671783) was undertaken to reestablish techniques and procedures to more accurately monitor ovarian follicular development as a model for human ovarian adenocarcinoma. This included attendance at the 2012 Poultry Science Association annual meeting (Athens, GA) to discuss our results with staff from the University of Illinois who have a very active research program in hen ovarian cancer. PARTICIPANTS: Dr. Glenn Birrenkott was the principal investigator of this project. Dr. Jillian Fain, Senior Lecturer in Animal & Veterinary Sciences helped with the ultrasonography. A total of eight Animal & Veterinary Sciences undergraduates participated in the Creative Inquiry project. TARGET AUDIENCES: The target audiences include not only scientists and laypersons interested in the biology of ovarian development and adenocarcinoma but also faculty interested in finding ways to engage students in scientific research. The development of research topics, acquisition and analysis of data with output to targeted audiences involves the skill sets that are deemed important in our undergraduate students (critical thinking, ethical judgement and communication). PROJECT MODIFICATIONS: The primary change will be developing commitments from students to participate in Creative Inquiry projects that span multiple semesters. This will provide for more complex experiments on a biological phenomenon that develops over time.

    Impacts
    Using undergraduates in a Creative Inquiry research project on development of ovarian adenocarcinoma in hens enhanced their knowledge of avian biology and the scientific method. As noted in our presentation (below) this limits experiments to a single semester and is less applicable to a topic like avian adenocarcinoma that may take years to develop. The ultrasonography data clearly showed differences in opacity between the various yolk layer during follicular development similar to those observed with lipid soluble dyes in our 1988 work.

    Publications

    • Birrenkott, G. P., and J. Fain, 2012. Creative inquiry Undergraduate research on the hen as a model for ovarian adenocarcinoma. Poultry Science 91:103 (Suppl. 1).


    Progress 01/01/11 to 12/31/11

    Outputs
    OUTPUTS: Undergraduate students in a Clemson University Creative Inquiry project participated in this project where we collected data on the ultrasonography of necropsied SCWL hen ovaries. Normal ovaries with discernible metastasizing adenocarcinomas were examined along with soft-shell, hard-shell and broken-out eggs. A library of ovarian images of normal and adenocarcinomas was compiled. Ultrasonography of laid eggs could sometimes distinguish fine structures of the yolk such as the blastodisc, latebra and neck of the latebra. PARTICIPANTS: Not relevant to this project. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.

    Impacts
    The data from this year's necropsy and ultrasonography project has refined our ability to detect ovarian and extraovarian structures in the laying hen. The library of post-mortem images will now be used to guide our in situ ultrasonography and make inferences about the status of the ovaries of older laying hens in various stages of production and those undergoing the early stages of ovarian adenocarcinoma.

    Publications

    • No publications reported this period


    Progress 01/01/10 to 12/31/10

    Outputs
    OUTPUTS: Several experiments were conducted and analyzed to get preliminary data on ovarian follicular parameters of chicken hens and cattle. This work was facilitated by undergraduate Creative Inquiry projects. Weekly training sessions in gross anatomy, ultrasonography and data collection were held during the semester. All of the students used this research project as part of their undergraduate degree program. The concept and preliminary data were shared with poultry industry managers in South Carolina and Georgia. These industry professionals made suggestions and indicated a possibility of future donations and collaborative work. PARTICIPANTS: In addition to the PI (Dr. Glenn Birrenkott), six undergraduate Animal & Veterinary Sciences students were involved in a Creative Inquiry project each regular semester to study the literature associated with ovarian dysfunctions and helped to collect the data in this summary. TARGET AUDIENCES: Biologists interested in ovarian function and dysfunction, especially in birds and women might be interested in the results. Poultry producers, specifically table-egg producers may be interested in the results relative to their aged or force-molted hens. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

    Impacts
    Preliminary experiments were performed on SCWL laying hens to determine the feasibility of using color Doppler ultrasonography to determine ovarian status in cadavers and in vivo. Distinct differences in the opacity of the ovarian white and yellow yolk material were observed. This difference was also found in soft-shell eggs and yolks from broken-out eggs. Based on these observations and necropsies of normal hens and those with ovarian adenocarcinoma; an increase in intra-abdominal opacity (area and intensity) should be noted. Preliminary data were also collected on glucose concentrations during ovarian follicular development. Normal hens had blood glucose concentrations of about 300 mg%. It was difficult to accurately measure glucose in these yolky follicles, but concentrations were lower than would be expected by simple diffusion. Future experiments will focus on quantitative and qualitative ultrasonography opacity in the body cavity and ovaries of hens with ovarian adenocarcinoma. Color Doppler ultrasonography will also be used to determine blood flow characteristics to the normal and cancerous ovarian tissue and associated metastasis. Specific glucose receptors in layers of the hen's ovarian follicle, especially the ovarian germinal (surface) epithelium and how these change during folliculogenesis and development of ovarian adenocarcinoma should reveal why glucose is not in higher concentrations in the yolk. This may also relate to our unpublished observation that bovine cystic follicles have higher glucose concentrations than normal-sized follicles and historical data that diabetic women may have a higher incidence of polycystic ovarian syndrome - another ovarian disorder in women and domesticated animals.

    Publications

    • No publications reported this period