Team:MIT/Integrated Practices

• BACKGROUND
• EXPERIMENTS
  Synthetic Promoters microRNA Recombinase Proof of Concept Collaborations
• FUTURE WORK
  Circuit Design Clinical Applications
• HUMAN PRACTICES
  Parts Toolbox Accessible Protocols Integrated Human Practices Public Outreach Interlab Study
• NOTEBOOK
  Lab Notebook Parts Mammalian Parts Collection Attributions Safety
• AWARDS
  HP Silver HP Gold Integrated HP Basic Part
• TEAM

Integrated Human Practices

Our circuit design was informed by several meetings with experts in the fields of endometriosis and synthetic biology.

June 13 - Asgi Fazleabas Visit

Asgi Fazleabas is a researcher and professor of Obsterics, Gynecology and Reproductive Biology at Michigan State University. He gave a talk to our team in early June, enlightening us about an important characteristics of endometriosis: estrogen dominance, progesterone resistance, and differing miRNA profiles. We learned progesterone resistance is, simplified, due to a lack of progesterone receptors in the cells. Here, “progesterone receptor” refers to the PGR-B isoform, the one that activates due to progesterone and binds to DNA transcription factors. This isoform is downregulated in ectopic endometrial cells. Since progesterone was a biomarker we initially designed our circuit to sense, we searched for another way for our circuit to sense a high level of progesterone present in the cell’s environment. Professor Fazleabas, doing research on a baboon model, found that in endometriosis, estrogen responsive genes are upregulated constantly, while progesterone responsive genes are always downregulated. The reason estrogen is present in abnormally high levels in ectopic cells is due to the way it’s metabolized. E2(estradiol), what we are referring to when we say “estrogen” is metabolized in the cells by a chemical reaction that needs a certain form of progesterone present to occur (research this reaction a little more so you can be more scientific). When progesterone is scarce, a buildup of unmetabolized estrogen occurs in the cell. Additionally, Asgi educated us on the fact that ectopic endometrial cells make a lot of aromatase, the enzyme that catalyzes the chemical reaction that produces E2. We thought: if we cannot sense progesterone levels in diseased cells, maybe instead we can sense for unusually high amount of estrogen?

July 11 - Meeting with Ron Weiss and Linda Griffith

Linda Griffith is a professor of Biomedical and Mechanical Engineering at MIT and the director of the MIT Center for Gynepathology Research. She advised our team throughout the summer and fall as we made crucial decisions about our circuit and experimental design. Professor Griffith gave us insight to the clinical situation most endometriosis patients are in. Most likely, a symptomatic women who has seen a doctor is already taking some type of hormone regulator, whether it be an IUD or pill. Therefore, the patient is not cyclic like we assumed in our original circuit design and would be in a constant high-progestin state (except for endometriotic lesions, since they are progesterone resistant). To further complicate the challenge of hormone sensing universally across diseased and healthy cells in the reproductive system, endometriotic lesions are consistently producing their own estrogen due to upregulated factors that cause the chemical reaction to occur. Instead, we abandoned the original circuit entirely that assumes all cells go through a normal estrogen and progesterone high phase, and instead decided to sense for: 1) abnormally high levels of estrogen and 2) progesterone resistance during a relative estrogen low phase.

July 21 - Kevin Osteen

Talk about meeting

July 27 - Harriet Fitzgerald

Talk about meeting