Our project represents an attempt for therapeutically managing the condition of obesity. Although we believe that obese people should not be socially isolated and blamed for their weight and appearance, it is crucial to remember at what cost obese people are living with this condition. The most serious consequences of obesity on health are hypertension, diabetes, myocardial infarction and major cardiovascular events. In particular, diabetes, a consequence of caloric excess, shows a direct association with other comorbidities, such as hypertension which is positively correlated due to vessel damage . Excessive weight or obesity (BMI ≥25 kg/m2) was found in more than 60% of the adults surveyed in Russia, approaching prevalence levels in the USA, and higher than the prevalence reported in any recent survey in the European Union .
From survey: “I have been struggling with a high amount of Ghrelin and Leptin for a long time, so the idea of a probiotic that can regulate those hormones would be very useful”
We think that finding the treatment for obesity can eventually decrease the number of deaths from the health conditions (listed earlier) in our country and in the whole world as well. It is our main motivation to develop this project. Obese people do struggle every day, sensations of appetite and satiety are blunt at some level, there is a trick that their brain plays out before them. However, there are several therapy types available. Dietary therapy with increased exercise activity, surgical interventions, weight reducing drugs. Most of them expect a great strength of will from patients or lead towards life-long health limitations . Pharmacological options currently available are fairly limited and can be used only for short periods of time . That’s why we believe in the idea of creating such an easy to use type of therapeutics which is actually a native protein of a human organism and has a potential of giving a life-long lasting effect.
Obesity is clearly an issue here in Russia, so we decided to not only practically develop a therapeutic solution to such a condition, but also to raise awareness about the significance of healthy nutrition among young people. While managing our social media (Instagram and VKontakte) we saw a lot of people interested in our project idea, so we decided to carry out a survey which included several questions regarding people’s health, eating habits, and thoughts on GMO and living therapeutics. This way we have both learned about our potential audience demand and analyzed the current state of the audience’s health.
“Rate the influence of eating habits (e.g., not overeating at night, having a good breakfast, not taking long breaks between meals) in relation to health from 1 to 5”
But this is not the only improvement to our project thanks to this survey. Our audience was generally interested and motivated to use such bacteria. These people are open to the idea, but based on several audience opinions we re-enter the discussion about the safety of LEAP2BRAIN:
“Concerned about safety and use of it as a ‘diet pill’ to encourage unhealthy weights”
“It could help many people control their weight, a possible problem I see is if the flora could accept this organism or what is the interaction with the different types of bacterial flora or where could it spread.”
“If it's safe and has a kill switch, then why not.”
The last one has led us to integrating a kill switch in the genetic construct which expresses chimeric protein. Indeed, the ability to instantly stop the treatment is crucial. We believe that through next successful experimental steps the level of approval by the audience will be increased.
Scientific aspect of the project
Living therapeutics is a small developing field, intranasal probiotics are even rarer, including the iGEM competition. Stepping out of our comfort zone we plan to advance the development of this field. Sharing our story on social media we engaged a lot of people into the problems of synthetic biology. We hope that even our small audience, though luckily international, can have a positive impact in terms of educating people from different backgrounds in their daily life. We’re happy to see synthetic biology developing and its flourishing.
Adding to the scientific aspect of our work we have started the research on how cell-penetrating peptides affect the membrane permeability for a protein “from the inside”. During our project we have planned to make a control cell line secreting LEAP2 + Myc-tag without LMWP. Even though we claimed before only about the practical solution to obesity, we’ve also set the goal to get into research and bring new knowledge to the scientific world as well.
Throughout the project development we had a system of expert sessions where we discussed
difficulties of the idea development with experts from different biological fields. We’d like to thank them:
Primary PI. Sergei Sedykh, PhD in Biochemistry, researcher at laboratory of repair enzymes, Institute of chemical biology and fundamental medicine, ICBFM SB RAS.
Secondary PI. Maria Galyamova, the head of the Infrastructure center “HealthNet”, head of the project support group of the national technology initiative.
Dr. Nina Tikunova, doctor of biological sciences, lead scientist, the head of the laboratory of molecular microbiology,
ICBFM SB RAS.
Dr. Dmitry Zharkov, corresponding member of RAS, doctor of biological sciences, associate professor, the head of the laboratory of genomic and protein engineering, ICBFM SB RAS.
Dr. Maxim Filipenko, PhD, chief researcher, the head of the pharmacogenomics laboratory, ICBFM SB RAS.
Dr. Grigoriy Stepanov, PhD, the head of laboratory of genome editing, ICBFM SB RAS.
Special thanks to our web-developer.
Although there’s a lot of things we have discussed throughout our sessions, the main project switch happened
regarding in vitro experiments. As our project is a proof-of-concept, combining the idea of living therapeutics and
nose-to-brain delivery of macromolecules, ideally we needed to model and test both of these aspects. Originally,
provided we demonstrated secretion of the LEAP2 fusion protein from bacteria, we were planning to move straight onto
in vivo murine models to measure the success of N2B delivery
using immunohistochemical techniques. Needless to say, at the time we were also concerned with the ethical aspect.
For a more comprehensive analysis, we would have first tested the delivery of the purified fusion
protein, using a protocol such as the one used by Hansen et. al., 2013 . Only under the condition that this
step worked, would we have considered moving on to actually planting the modified bacteria into the nose
(ideally not the E.coli we used to demonstrate the secretion of our protein, but a native bacterium found in
the microbiota of the nose (i.e. Neisseria spp). Obviously, the second step was unrealistic for the timeframe
of the competition, however, should the concept of combining living therapeutics and N2B delivery continue its development in the real world of science, unfortunately, animal testing would be necessary at some point to test for the safety of our treatment. In fact, our survey showed that the safety of our
innovative therapy was the main area of interest for both scientists and average citizens.
The article, on which we based our idea of making a fusion protein with the cell-penetrating peptide, LWMP,  states that the mechanism by which their fusion protein travels to the hypothalamus via the olfactory and trigeminal nerves is unclear. It is speculated that endocytosis and macropinocytosis play an important role, however, the intra-tissue penetration mechanisms are “largely unknown” and most likely occur via an extracellular mechanism.
De Lorenzo, Antonino et al. “Why primary obesity is a disease?.” Journal of translational medicine vol. 17,1 169. 22 May. 2019, doi:10.1186/s12967-019-1919-y
Kontsevaya A, Boytsov S et al. “Overweight and Obesity in the Russian Population: Prevalence in Adults and Association with Socioeconomic Parameters and Cardiovascular Risk Factors”. Obes Facts. 2019;12(1):103-114. doi: 10.1159/000493885.
Wirth, Alfred et al. “The prevention and treatment of obesity.” Deutsches Arzteblatt international vol. 111,42 (2014): 705-13. doi:10.3238/arztebl.2014.0705
Ruban, Aruchuna et al. “Current treatments for obesity.” Clinical medicine (London, England) vol. 19,3 (2019): 205-212. doi:10.7861/clinmedicine.19-3-205
Hanson, L. R., Fine, J. M., Svitak, A. L., & Faltesek, K. A. (2013). Intranasal administration of CNS therapeutics to awake mice. Journal of visualized experiments : JoVE, (74), 4440. https://doi.org/10.3791/4440
Lin, Tingting et al. “Nose-to-brain delivery of macromolecules mediated by cell-penetrating peptides.” Acta pharmaceutica Sinica. B vol. 6,4 (2016): 352-8. doi:10.1016/j.apsb.2016.04.001