Revision as of 01:20, 19 October 2016

• 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

Clinical Applications

Developing a diagnosis

Endometriosis is a complex disease – any diagnostic needs to account for a variety of factors. We opted for a three-pronged approach, sensing progesterone resistance and miRNA dysfunction while maintaining temporal specificity. Each of these components represents a key hallmark of the disease, and when put together can create a sensitive diagnostic tool.

Implementing a Diagnosis

We initially contemplated creating an in-vivo diagnostic, but changed our minds after speaking with Professor Linda Griffith (MIT). Realistically, inserting a genetic circuit into a human (likely through the use of a viral vector) is many years away from being a possibility due to major safety concerns.

Implementing an in-vitro diagnostic that can be used on an endometrial biopsy is more feasible. An endometrial biopsy is a 5-15 minute procedure that can be done in a doctor’s office without anesthesia and is commonly used when testing for endometrial cancer. While not pain-free, endometrial biopsies are considerably less invasive than laparoscopic surgery, the current diagnostic method for endometriosis.

Due to the nature of the procedure, the cells collected are eutopic endometrial cells, not the ectopic cells associated with endometriosis. However, the literature suggests that the eutopic endometrium of women with endometriosis displays different molecular markers than that of healthy women. Notably, the eutopic endometrium of patients with endometriosis displays progesterone-resistance – a key identifier we used in designing our circuit. Additionally, there are differences in miRNA profiles between the eutopic endometrium of healthy women and that of women with endometriosis.

Of course, an in-vitro sample does not experience the natural hormone cycle. However, the tissue from the biopsy can be dosed with estrogen and progesterone to simulate the menstrual cycle, taking advantage of our proposed circuit architecture. We have characterized our promoters by dosing cells with progesterone and estrogen and have had success.

An in-vitro approach is considerably safer than using a viral vector for in-vivo diagnosis. The literature supports the notion that the molecular markings of eutopic endometrial cells can be used to identify patients with and without endometriosis. Dosing tissue from an endometrial biopsy with hormones allows us to test these molecular differences and use our diagnostic in an in-vitro setting. With a less invasive diagnostic method, we hope that the diagnostic process can be expedited.

Synthetic Biology & Medicine - How SynBio Approaches Can Transform Diagnostics

Our approach of using a genetic circuit to sense diseased cells presents possibilities beyond endometriosis. Similar strategies can be used to diagnose other diseases... KEEP GOING WITH THIS https://2016.igem.org/Team:MIT/Clinical_Applications