Blood clotting cascade is like a relay race where the runners are clotting factors. In Haemophilia A, factor 9, after receiving a baton from factor 11, is racing to form a clot that will stop a bleed in a patient. However, the next member of the race, factor 8, is missing!
The Radboud iGEM team finds a solution to save the day, our improved Haemophilia treatment, InfinityF∞.
Factor 8 is a large glycoprotein which participates in the intrinsic pathway of the coagulation cascade.
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In the world, more than 400,000 male patients are living with HemA. HemA also accounts for 80% of hemophilia cases. A chance for a newborn male to acquire Haemophilia A is 1 in 5,000–10,000 males with a high risk of patient mortality when undergoing a surgery. Without treatment, even small bleeds can become dangerous and in serious cases, patients have a high risk of spontaneous bleeding [2, 3].
And, even though this approach has its benefits - it can be done by the patient themselves and there are possibilities for an implantable port for easy administration - there are still some limitations.
Injections are invasive and uncomfortable, which should always be important to consider when developing a treatment. On top of that, a Haemophilia patient has to make frequent injections - every 48 hours - to keep their FVIII levels on a normal level. The frequent injections are in part the result of the high clearance of factor VIII from the circulation with a half-life of 10-12 hours. This causes the factor VIII concentration in the blood to fluctuate a lot, spiking up after injection, then quickly going down until the next injection.
Currently, there are multiple engineered recombinant factor VIII products available, but these are way more expensive than the cheapest variety of FVIII, which is purified from blood products and already has an annual cost of $300,000!
InfinityF∞ - an intranasal mRNA delivery system for Factor VIII using ionizable lipid nanoparticles.
We are hoping that this new system will give a more controlled release of factor VIII. The coolest thing about it is that the nanoparticles are small enough to be delivered via an intranasal route.
This means patients do not need to get injections anymore, and they can just use a nasal spray!
InfinityF delivers the mRNA sequence for factor VIII encapsulated by a lipid nanoparticle that serves as a carrier for the medicine. When the nanoparticle reaches its target in the body, the cells will take it up into a lysosome and break the LNP preserving the genetic sequence inside. After that, the cell produces the protein factor VIII from the mRNA and releases it into the blood.
Ionizable lipids are crucial for LNP preparation, and they provide effective mRNA encapsulation, as well as safe LNP delivery in the body. Their role is crucial, since they change their charge due to protonation under acidic pH (become positively charged), or deprotonation under the neutral pH (become not charged). This feature makes it possible to encapsulate the mRNA more efficiently, as well as their safe delivery in a blood neutral pH environment.
The LNPs are delivered via the nasal spray delivery system by the use of hydrogel, composed of poloxamer 407, poloxamer 188, NA-CMC, and almotriptan. This hydrogel provides a biphasic release pattern: the quick release for 30 minutes (this provides an acute release of a medicine upon administration) and a sustained release for a 5 hour period (this maintains a loading dose of the medicine).
LNPs are administered into the bloodstream to travel to the liver and target LSECs with the help of surface proteins. Once inside the cells, the LNPs will release the mRNAs, leading to the production of coagulation factor VIII. This approach enables patients to produce their own coagulation factor VIII, eliminating the need for frequent injections of plasma-derived or recombinant proteins.
The factor VIII mRNA is produced with in vitro transcription using factor VIII pcDNA as a template. Our ionizable lipid nanoparticles contain synthetic lipids as well as an ionizable lipid, which is essential for the release of the mRNA in the cells.
The factor VIII mRNA is produced with in vitro transcription using factor VIII pcDNA as a template. Our ionizable lipid nanoparticles contain synthetic lipids as well as an ionizable lipid, which is essential for the release of the mRNA in the cells.
While the current Hemophilia medicine has a high clearance with a half-life of 10-12 hours, thanks to the biphasic release pattern of hydrogel, applied in InfinitiF∞, patient can rely on the temporarily release of the medicine, prolonging the time of its circulation in the bloodstream, and providing patient with a longer duration of effect of the medicine.
Production of protein from mRNA occurs gradually and not in all cells at the same time, spreading out the high concentration spike that occurs after injection of FVIII.
Costs are very important when speaking about medical treatment. Since the current Haemophilia medicine utilizes purified protein from human plasma, the price skyrockets to big numbers!
Current price of Haemophilia medicine (in the USA): 8000 euro/week [4].
Estimated price of LNP dosage for InfinityF∞ medicine (for an average patient of 60 kg): 160 euro/week. Estimated price of mRNA production for InfinityF∞ medicine (for an average patient of 60 kg): 790 euro per batch. Primary estimation for the Total price of InfinityF∞ medicine with mRNA production costs included: 950 euro/week.
The Radboud iGEM team was started by a group of enthusiastic students that were looking to broaden their horizon and start something new. We hope to introduce the iGEM competition to Radboud university while also putting Radboud university on the map within the iGEM competition.
As a new team, we spent a lot of time navigating the organisational aspects within the university as well as familiarizing ourselves with the systems of the competition. With our efforts, we hope to pave the way for future Radboud iGEM teams.
We are beyond excited to showcase InfinityF∞ as the first team to represent Radboud University within the iGEM competition, and it is with passion and determination that we say: