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Team:Slovenia/Idea
From 2012.igem.org
Idea
For this year's Team Slovenia iGEM project we wanted to exploit the advantages of synthetic biology for an advanced biological medical therapy which could overcome some of the difficulties connected to current application of biopharmaceuticals.
Each disease and its corresponding therapy is a story of its own, but we sought to design a synthetic biological platform for drug application that could be used to treat several different diseases and include safety components that could be useful in different therapeutic applications.
Our solution was to engineer mammalian cells, so that they would produce and deliver biological drugs inside the organism, as if the drugs were produced by the host cells themselves. We planned to incorporate genetic switches into the engineered cells allowing controlled production of one or several desired therapeutics, depending on the stage of the disease. The engineered cells would be incorporated in microcapsules that may be implanted into the affected tissue to ensure an immune-privileged environment for the therapeutic cells. The semi-permeable capsule allows the flux of nutrients and effector molecules through the capsule’s pores but prevents cell escape and dissemination throughout the body. For the unlikely case of cells escaping capsules, we designed a safety tag ensuring destruction of cells outside microcapsules by the patient's immune system. This form of therapy could be terminated at any given time by an outside signal inducing microcapsule degradation and therapeutic cell apoptosis in which case no traces of the exogenous material should remain in the tissue.
The main features of our proposed therapeutic device:
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| Figure 1. Microencapsulated engineered cells for the production and delivery of biological therapeutics. |
An important advantage of the microencapsulated cell approach is that the therapeutic protein can reach therapeutic concentrations locally in the affected tissue, while maintaining a low systemic concentration, thus avoiding systemic side effects, such as systemic immunosupression. Production of protein therapeutics by cells also eliminates the need for the repeated invasive administration of therapeutics, which are typically applied by injections. Repeated application of therapeutics into the liver, brain or other complex tissues, where local delivery could be beneficial, is quite difficult. We aimed to develop an implantible cellular device for the local, regulated and safe delivery of therapeutic proteins. The usual obvious solution would be gene therapy, which has, despite its early promises, not delivered many successful clinical applications because of the problems of viral delivery and variable effectiveness caused by the random site of integration. Long term survival of encapsulated cells beyond months may be problematic with current technology and their efficiency may decrease, therefore we should focus on the therapy of acute conditions.
The desired properties of synthetic cellular devices for the next generation of cell-based therapies
- Control over the selection of different states of therapeutic cells for production of different drug combinations during different stages of therapy should be possible from the outside by a physician
- The ability to inactivate and destroy cells of the device at the end of the therapy, leaving as little remains as possible and elimination of any cells that might have escaped from microcapsules
The synthetic biological components that needed to be developed
Universal orthogonal bistable toggle switch for mammalian cells
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| Figure 2. Regulation of the several different states of therapeutic cells based on several orthogonal toggle switches. |
Safety mechanisms for therapy with microencapsulated cells
For the safe therapeutic implementation of microencapsulated engineered cells in the organism we envisioned that we should be able to cease the therapy without having to physically remove the capsules.
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| Figure 3. Safety mechanisms that we planned to introduce for the application of microencapsulated engineered mammalian cells in therapy. |
Microcapsule degradation - Most of the microcapsules used in literature and therapy are composed of polymerized alginate that is biocompatible but cannot be degraded in mammalian tissues. Alginate lyase harvested from a marine microorganism effectively degrades alginate, so secretion of this enzyme from cells should degrade the capsules and allow their resorption by phagocytic cells.
Termination - We should be able to inactivate the implanted cells at the end of the therapy or at any other stage if the therapy has to be terminated, regardless of the cause. We introduced a safety mechanism that would enable the controled induction of apoptosis of the implanted cells that is leak-free until the selected moment. Therefore we could inactivate therapeutic cells and enable their resorption without causing inflammation.
Escape tag - ensures that any cell that may have escaped from a microcapsule is eliminated by the immune system of the host organism. The escaped cells would most likely be recognized as foreign in any case but in order to provide the maximal safety measures we decided to introduce an additional tag that would alert the natural killer cells of the host immune response.
Next: The Switch >>
