As the field of oncofertility continues to advance, the Oncofertility Consortium seeks to support various high-risk, high-gain initiatives in order to facilitate the launch of early-stage ideas into future grant-funded projects. Applications for pilot projects are evaluated each year, with only a few awarded with financial support and access to Oncofertility Consortium resources. A total of 8 pilot projects have been backed by the Oncofertility Consortium, with additional projects planned each year.
Funded Projects, Current and Past:
- Improving Tamoxifen Adherence by Addressing Fertility Concerns
- A Novel Approach to Assess Direct and Estradiol-Mediated Effects of Ovulation Induction Regimens and Breast Cancer Development
- The Male Perspective: Survey for Preservation of Adolescent Reproduction
- Oncofertility Communication Initiative: Assessing the Short Documentary Format for Oncofertility Communications
- Development of Blood Spot Assays for Long-term Longitudinal Studies of Reproductive Status of Cancer Survivors
- Deriving Oocytes from Embryonic Stem Cells
- The Effects of GDF9 Levels on TZP Reorganization and Oocyte Competence in Growing Follicles Cultured in Alginate
- Identification of Developmental Markers and Culture Environments
- Role of GDF-9 in Human Folliculogenesis
- Does Gleevac Block Programmed Cell Death in Primary and Primordial Mouse Follicles Via the p63 Pathway?
- Gene Expression in Small Antral Follicles Derived in Vivo and From Encapsulated 3-Dimensional Culture in Macaques
- Comparing Infertility and Cancer Research, Development and Social Understandings
- The Road to Adulthood: How Do Young Adults Make Health and Fertility-Related Decisions
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1. Improving Tamoxifen Adherance by Addressing Fertility Concerns
Site: Northwestern University
Principal Investigator: Jacqueline S. Jeruss, MD, PhD
Study Description
The study objective is to evaluate physician- and patient-level factors, including fertility, concerns that affect adherence to tamoxifen for young women diagnosed with breast cancer. An estimated 23,000 women under age 45 are diagnosed with breast cancer annually, of whom roughly 70% have estrogen-receptor positive (ER+) breast cancer. For these women, a 5-year course of daily tamoxifen results in a 47% reduction in annual recurrence risk and a 26% reduction in annual mortality. Despite such substantial benefits, adherence and persistence are low: 20% of patients fail to reach the optimal adherence threshold during the first year of treatment, and at 5 years, up to 50% of patients have discontinued treatment. Young women consistently have been shown to have lower rates of adherence to hormonal therapy when compared to women between the ages of 50-70. It is necessary to address both clinician and patient barriers to tamoxifen adherence. We hypothesize that, for young breast cancer patients, a unique set of patient- and provider-level factors, including fertility concerns, contributes to the poor tamoxifen adherence observed in this patient population.
2. A Novel Approach to Assess Direct and Estradiol-Mediated Effects of Ovulation Induction Regimens and Breast Cancer Development
Site: Northwestern University
Principal Investigator: Jacqueline S. Jeruss, MD, PhD
Study Description
The major goal of this project is to determine if ovulation induction through medical intervention causes breast epithelial cells to progress to neoplasia, either directly by inducing proliferation or indirectly by increasing stromal angiogenesis in a 3-dimensional culture system. In addition, investigators are determining if prior infertility treatment increases the risk of breast cancer in a cohort of women with BRCA 1/2 mutations or a family history of breast cancer.
Progress and Results
Clomiphene, commonly used as a treatment for infertility, acts by blocking the estrogen receptor (ER), though it has been hypothesized that this drug also effects cell proliferation in vitro by estrogen independent mechanisms. To analyze the effects of clomiphene treatment on breast cancer development we used 3 well-established breast cancer cell lines. MCF-10A cells, which approximate normal immortalized mammary epithelium and are ER negative, showed a slight increase in colony area in response to estrogen and a slight increase in colony number in response to clomiphene during 10 days of treatment. MCF-7 cells, which are estrogen receptor (ER) positive and well-differentiated breast cancer cells, showed a significant increase in colony area in response to estrogen treatment and a slight increase in colony number in response to clomiphene. HCC 1937 cells, which are ER negative and harbor a BRCA1 mutation, showed a slight increase in colony number n response to estrogen. Proliferation assays, 3D culture, and cell cycle analysis studies are in progress to determine the response of these cells to clomiphene, FSH, estradiol, and hCG. We will also use a mouse model with mammary epithelial hyperplasia to assess how infertility treatment affects the progression of hyperplasia to malignancy. Lastly, we continue to accrue BRCA 1/2 carriers or patients with a strong family history of breast cancer to determine the risk of developing breast cancer following infertility treatment. Publications from this and related research from Dr. Jeruss are:
Jeruss J S. Discussing Fertility Preservation with Breast Cancer Patients. Cancer Treatment and Research. 2010; 156: 461-6. PMID: 20811855.
Jeruss JS, Mittendorf EA, Tucker SL, Gonzalez-Angulo AM, Buchholz TA, Sahin AA, Cormier JN, Buzdar AU, Hortobagyi GN, Hunt KK. Staging of breast cancer in the neoadjuvant setting. Cancer Res. 2008 Aug 15;68(16):6477-81. PMID: 18701468.
Weiss MS, Peñalver Bernabé B, Bellis AD, Broadbelt LJ, Jeruss JS, Shea LD. Dynamic, large-scale profiling of transcription factor activity from live cells in 3D culture. PLoS One. 2010 Nov 17;5(11):e14026. PMID: 21103341.
3. The Male Perspective: Survey for Preservation of Adolescent Reproduction
Site: Northwestern University
Principal Investigator: Robert Brannigan, MD
Study Description
Understanding and overcoming the barriers to fertility preservation is essential to optimize the reproductive health of young cancer patients. The overall aim of this project is to develop and execute a national survey (Survey for Preservation of Adolescent Reproduction, SPARE) of pediatric oncologists to assess perspectives on fertility preservation in both pre-pubertal (age 1-12) and pubertal (age 13-18) cancer patients, including willingness to discuss fertility, knowledge of current fertility preservation methods, and awareness of the American Society of Clinical Oncology published fertility preservation recommendations (ASCOR) regarding sperm cryopreservation strategies and referral to a fertility specialist. Through SPARE, we sought to obtain post-ASCOR data from pediatric oncology specialists nationwide.
Progress and Results
A total of 209 respondents, primarily consisting of pediatric oncologists, responded to the SPARE survey, providing data on both male and female pediatric oncology fertility preservation knowledge, attitudes, and practices. Our work on SPARE revealed that fertility preservation in both male and female adolescents is hampered by a lack of knowledge among treating physicians of the American Society for Clinical Oncology recommendations on this topic, as well as numerous barriers to the delivery of fertility preservation case. This project also provided insight into the gender disparity that exists in the approach that clinicians utilize in treating male and female adolescents. We conclude that pediatric oncologists are strongly motivated to preserve fertility in male pediatric cancer patients but barriers to semen preservation and referral to fertility specialists exist. Additional promotion of the ASCO fertility preservation recommendations is essential to optimize reproductive health of young cancer patients. This pilot project sucessfully resulted in a publication, as follows:
Köhler TS, Kondapalli LA, Shah A, Chan S, Woodruff TK. Results from the survey for preservation of adolescent reproduction (SPARE) study: gender disparity in delivery of fertility preservation message to adolescents with cancer. Brannigan RE. J Assist Reprod Genet. 2010 Nov 26.
4. Oncofertility Communication Initiative: Assessing the Short Documentary Format for Oncofertility Communications
Site: Northwestern University
Principal Investigator: Barbara O’Keefe, PhD
Co-Investigator: Sean Zehnder, PhD
Study Description
Video portrayals of cancer patients offer special opportunities to shape the way audiences think about issues related to a diagnosis of cancer. We hypothesize that video portrayals of patients that dramatize their maturation into complex, multidimensional human beings will lead audiences of all types to cognize other cancer patients in more complex, multidimensional, and historicized ways, and that this broader framing will in turn change decision-making. The purpose of this project is to pilot both (a) the use of short documentary films for increasing the salience of fertility issues in decision-making about cancer treatment (oncofertility) and (b) methods for designing, disseminating, and evaluating messages about oncofertility. A long-term goal is to use the knowledge gained via this pilot to build a sustained research and development program on oncofertility communications.
Progress and Results
The first part of our pilot project was to produce three short documentaries about cancer survivors and their issues they have faced related to having biological children. Two Chicago-based professional videographers were hired and helped to produce a high definition video that portrays individual patient experiences with cancer. These videos tells to story of three unique cancer survivors: Gayle, a survivor of adult cancer who opted to participate in a new research protocol with the hope of one day being able to conceive another child; Wesley, a marathoner and business man who was diagnosed with Hodgkin’s Lymphoma as a young man; and Connie, a survivor of childhood cancer who is now struggling with infertility caused by her cancer treatment. These videos will help to develop evidence-based guidelines for web communication with patients and others and assist in the promotion of information about oncofertility. View the videos.
5. Development of Blood Spot Assays for Long-term Longitudinal Studies of Reproductive Status of Cancer Survivors
Site: Northwestern University
Principal Investigator: Teresa K. Woodruff, PhD
Co- Investigators: Clarisa Gracia, MD, Laxmi Kondapalli, MD, and Thomas McDade, PhD
Study Description
The blood spot assay is a method for collecting samples for hormone measurements. Dried blood spot (DBS) samples, which are collected by a minimally invasive finger prick, require little collection time, no post-collection processing, have low biohazard risk, and facilitate sample storage and transport. In this pilot project, DBS assays will be developed to measure markers of reproductive function (inhibin B, AMH, and FSH). These hormone profiles can then be used to assess reproductive potential in cancer patients. The aims of this project are to:
- Validate measurements of inhibin B, AMH and FSH in media and dried blood spots
- Assess inhibin B, AMH and FSH in pools of serum samples collected from early follicular phase, mid follicular phase and luteal phase
- Assess inhibin B, AMH and FSH in blood spots taken during one complete menstrual cycle and compare the results to a serum-based assay
Progress and Results
We have developed an enzyme immunoassay protocol for AMH using DBS derived from finger stick whole blood samples, and are currently working to improve the assay sensitivity. We have also begun collecting a set of 100 matched plasma/dried blood spot samples from menstruating reproductive aged women at the University of Pennsylvania. These samples will allow us to compare results obtained from DBS with gold-standard clinical methods using serum/plasma from venipuncture. Using Bland-Altman plots and regression analyses, we will inspect for agreement and bias across the assay range and generate a conversion formula for calculating plasma equivalents based on DBS results.
6. Deriving Oocytes from Embryonic Stem Cells
Site: Oregon National Primate Research Center
Principal Investigator: Shoukhrat M. Matalipov, PhD
Co-Investigator: Mary B. Zelinski, PhD
Postdoctoral Fellow: Araceli Valle Prieto
Project Description
Recent research suggests that embryonic stem cells (ESCs) could serve as a source of viable gametes, since ESCs can proliferate indefinitely in an undifferentiated pluripotent state and, when allowed, can differentiate into any cell type of an adult body. The goal of this proposal is to evaluate the potential of monkey ESCs to contribute to the female germ line lineage and to explore the feasibility of deriving functional oocytes suitable for use in assisted reproductive technologies (ARTs). We hypothesize that primate ESCs derived by somatic cell nuclear transfer can form female germ cells and gametes upon spontaneous or directed differentiation. In this pilot project, we propose:
- To develop optimal culture conditions for directed differentiation of monkey ESCs to the female germ cell lineage
- To evaluate three-dimensional (3-D) culture for directed differentiation of monkey ESC-derived follicles to functional oocytes.
Progress and Results
We have developed genetically tagged monkey ESCs under the control of germ cell-specific promoters that will allow us to detect and select early germ cells upon induced differentiation.
7. The Effects of GDF9 Levels on TZP Reorganization and Oocyte Competence in Growing Follicles Cultured in Alginate
Site: Northwestern University
Principal Investigator: Teresa K. Woodruff, PhD
Co-Investigators: Susan L. Barrett, MS, PhD
Project Description
Small secondary ovarian follicles are capable of growing in a 3D alginate matrix to generate fertilizable eggs that can give rise to live offspring in mice. Though this system has been proven successful, the percentage of competent oocytes generated from each pool of cultured follicles remains low. It is known that somatic cell-oocyte interactions, namely via transzonal projections (TZPs), are necessary for proper follicle growth and oocyte development, and that these interactions can be compromised during prolonged follicle culture. Studies have also indicated that GDF9, an oocyte maternal-effect gene, plays a role in maintaining these physical connections between somatic cells and the oocyte. The goals of this pilot study are 1) to evaluate the organization and maintenance of TZPs in growing follicles cultured in our 3D alginate system in mice, primates, and humans and 2) to determine if higher levels of secreted GDF9 correlate with well-organized somatic cell-oocyte interactions, successful follicle growth, and oocyte competency. These studies will elucidate if all follicles have the potential to become dominant follicles producing competent oocytes.
8. Identification of Developmental Markers and Culture Environments
Site: Northwestern University
Principal Investigators: Lonnie Shea, PhD, and Linda Broadbelt, PhD
Project Description
In developing cultures systems for growing follicles, there are at least two significant challenges: i) determining if follicles are developing appropriately within the culture environment, and ii) providing key factors at the appropriate time to promote follicle growth and maturation. In this pilot project, we will analyze microarray data obtained from mouse ovarian follicles at different stages to identify factors that vary in expression levels between stages and can, therefore, be used as markers of growth progression. We will also use bioinformatics tools to identify key biological pathways that are active during the transition between the multiple follicle stages. This analysis will assist us in identifying factors that could be added to the culture medium to promote follicle growth at specific stages. Results from these studies will be translated to the culture of nonhuman primate and human follicles. The strategy developed in this proposal will provide a foundation for identifying environments that maximize the growth and development of immature follicles and will identify mechanisms through which the environment influences follicle maturation.
9. Role of GDF-9 in Human Folliculogenesis
Site: University of California, San Diego
Principal Investigator: Shunichi Shimasaki, PhD
Co-Investigator: Heidi Cook-Andersen, PhD
Project Description:
Genetic studies in female mice and ewes have demonstrated that the oocyte-specific growth and differentiation factor-9 (GDF-9) regulates female fertility. In humans, a high incidence of mutations in the GDF-9 gene has been found in women with premature ovarian failure (POF) and mothers of dizygotic (DZ) twins. In addition, GDF-9 mRNA levels are reduced in growing human oocytes in polycystic ovary syndrome (PCOS) and PCO ovaries from primordial follicle recruitment through the small Graafian follicle stage. Despite these important clinical observations, almost nothing is known about the mechanism by which GDF-9 controls ovarian function in women. In this pilot project, we will use our purified recombinant human GDF-9 (rhGDF-9) to determine the biological function of the rhGDF-9 in follicle growth and development in human ovaries. We will also determine the biological function of the mutant GDF-9 proteins described in POF women and mothers of DZ twins. The results of these studies will provide the first insight into understanding the molecular and cellular mechanisms of how ovarian function in women is governed physiologically by GDF-9 during reproductive years.
10. Does Gleevac Block Programmed Cell Death in Primary and Primordial Mouse Follicles Via the p63 Pathway?
Site: Northwestern University, Feinberg School of Medicine
Principal Investigator: Teresa K. Woodruff, PhD
Co-Investigator: Takeshi Kurita, PhD, Lonnie D. Shea, PhD, So-Youn Kim, PhD
11. Gene Expression in Small Antral Follicles Derived in Vivo and From Encapsulated 3-Dimensional Culture in Macaques
Site: Oregon National Primate Research Center
Principal Investigator: Mary B. Zelinski, PhD
Co-Investigators: Richard L. Stouffer, PhD, and Jing Xu, PhD
12. Comparing Infertility and Cancer Research, Development and Social Understandings
Site: Northwestern University, Center for Bioethics
Co-Investigator: Lisa Campo-Engelstein, PhD, and Sarah Rodriguez, PhD
13. The Road to Adulthood: How Do Young Adults Make Health and Fertility-Related Decisions
Site: Northwestern University, Feinberg School of Medicine
Principal Investigator: Karrie Snyder, PhD
This research was supported by the Oncofertility Consortium®, funded by the National Institutes of Health through the NIH Roadmap for Medical Research, Grant UL1DE19587 and UL1DE019587.