Team:Sydney Australia/Medals

Medal Criteria

Select each category to view more information on how we fulfilled medal criteria.

Bronze Medal Criteria

How did we achieve this?

 

We registered and will be attending the Giant Jamboree in November.

 

We have met all the deliverables requirements:


Our project attributions page can be found on our wiki


We contributed by participating in the interlab study

Silver Medal Criteria

How did we achieve this?
 

PART VALIDATION

 

We validated our parts by showing that they work. We performed an ELISA Assay testing for properly folded insulin, and detected expression of Cytoplasmic Proinsulin, Cytoplasmic Winsulin, Ecotin Proinsulin and YncM Winsulin in the cell fractions we expected them to be.

 

We also performed a glucose uptake assay to identify if the insulin and Winsulin we produced was functional.

 

Multiple insulin-sensitive cell lines were tested for glycogen synthesis and glucose oxidation upon treatment with human insulin and with our insulin analogues. It was found that Ecotin-Proinsulin, Cytoplasmic-Proinsulin and YncM-Winsulin all induced an increase in glycogen synthesis and glucose oxidation, therefore are stimulating the insulin-receptor and functioned as we expected!

See our Results page for more information

 

COLLABORATION

 

We collaborated with many other iGEM teams in order to get where we are today. We helped the Sao Paulo State University team with their work, as they were also focusing on producing insulin. We helped them with data in Australia, and they in turn told us more about the Brazilian situation, as well as sharing knowledge on our Bacillus work and using the modelling program MatLab.

 

We also communicated with Bristol University, and upon learning they had developed a tool for editing the iGEM wiki, we gave it a go and sent them our feedback. They returned the favour by providing us with data about insulin prices in England, which we were finding hard to come by!

 

We worked with Cologne-Dusseldorf to contribute to their post-card distribution idea, by both producing a post-card and by distributing their postcards once they arrived. This collaboration worked both – we all helped one another by distribution each others’ project ideas to otherwise unreasonable parts of the world, a fantastic experience.

 

We talked with Auckland University over skype multiple times, and were able to help each other with working on our wikis. We were also excited to host them at our University when a couple of their team members flew over to visit the SBA conference.

 

We also worked with Chennai by exchanging protocols and tips for the lab.

 

Finally, we worked with Uppsala by participating in their webinar series about the ethics of synthetic biology and intellectual property, which was a fantastic learning experience – both by us contribution our experiences and hearing what other teams from around the world had to say about our work.  This work contributed significantly to how we approached our work; particularly with how intellectual property affects the ability of iGEM teams to contribute to the open source registry.  

 

See our Collaborations page for more information

 

HUMAN PRACTICES

 

As we were developing a therapeutic agent, there were various things that we needed to address to ensure that we delivered a safe, ethical, useful product for the world. On our Human Practices page we have discussed these issues and how they have impacted our project in its development. Furthermore, we went out to the general public to discover how close to home the issues we were learning about really are. This survey, though we only had a limited response, gave us great insight into the reality of diabetes treatment in the world. These results then made us reconsider our understanding of the Social Injustices caused by high priced medications.

 

Gold Medal Criteria

How did we achieve this?

 

INTEGRATED HUMAN PRACTICES

In our project it was very important to talk to multiple groups to gain a complete understanding of the issue of insulin accessibility: Type I diabetics dependent on insulin for health and survival, physicians prescribing the insulin, concerned citizens and members of organizations interested in helping those in need.

 

Our integrated human practices can be broken into three categories: project design, applied design and entrepreneurship.

 

Project Design:

  • Anthony Di Franco from the USA, founder of the OpenInsulin project alerted us to the issue and jumpstarted the project as did Meow-Ludo Meow-Meow was another connection linking us to the OpenInsulin project, via his group at BioFoundry

  • Neil Donelan from Insulin For Life suggested that we did not need to focus all of our efforts on a thermostable insulin, as human proinsulin is generally stable without refrigeration – so we considered the option of generating proinsulin in a cheap manner

  • Jeni, a pharmacist, warned us of the dangers of modifying our insulin excessively, as endocrinologists prefer not to diagnose biosimilars that may induce an allergic reaction in patients

  • Len Mancini from Maxwell’s Patent and Trademark Attorneys helped us in the design of our Winsulin sequence, to ensure we weren’t infringing on any existing pateitns and ensure we can safely submit our sequence to be Open Source

  • Edwina Wang – a Type I diabetic alerted us of the importance of modelling our insulin in order to predict whether it will be short or long-acting. We found that our modelling predicted it would be short acting.

 

Applied design:

  • Dr David Beran is an expert on the insulin market working at the Geneva University Hospitals who alerted us to the fact that it will be difficult to enter the insulin market, due to expensive qualifications and licenses required

  • So we went to George XX from OpenInsulin who alerted us to the possibilities of crowd-funding, to aid a jump-start of this Open Source project

 

Entrepreneurship:

  • Mike Nichols from INCUBATE hub at the University of Sydney educated us on the possibilities of creating a start-up company that could fund the initial production of Winsulin and/or a more cheaply produced human proinsulin
    • He also pointed out the value in us targeting a country with an especially high need for insulin, in order to break into the market

  • Dr Narcyz Ghinea from The University of Sydney, an expert in access to high cost medicines, assisted us with data on where the money for clinical trials is sourced from, giving us the idea of a potential use of government grants for this open-sourced project

  • Neil Donelan from Insulin For Life pointed out the value in producing insulin in the countries that need it most, reducing the costs and problems associated with transport

 

See our Integrated Human Practices page for more information

 

PART IMPROVEMENT

We improved previous parts and submitted them to the registry and have listed them in our parts page

 

Improved Parts

BBa_K2417000 Ecotin-Proinsulin

BBa_K2417001 Cytoplasmic-Proinsulin

 

which were improving on

BBa_M39904 Human Insulin cDNA

BBa_M1877 human insulin

 

Previous parts in the registry encoding ‘Human Inuslin’ were either incomplete or did not match human insulin when entered into a BLAST database. It seems as though there were few teams attempting to create human proinsulin (A, B and C-peptide chains) on a single vector, and so our addition of the complete verified human proinsulin coding sequence is an improvement on these previous parts. Furthermore, we have added an N-terminal His tag to our proinsulin constructs, allowing ease of purification using affinity chromatography. Our addition of an arginine residue between the His tag and the proinsulin protein allows simple one-step cleavage and removal of both the C-peptide and the His tag, as all have the same trypsin cleavage site (arginine residue).

 

The addition of the Ecotin tag assists in the expression of proinsulin by inducing its transport to the periplasm, an oxidative environment with reduced proteases, ideal for folding proteins with disulphide bonds such as proinsulin. Furthermore, periplasmic expression allows for a simplified purification procedure, as only the periplasmic fraction is required.

 

We have also used an extended ribosome binding site with high efficiency of recruitment, which is an improvement on parts entered that were only protein coding sequences.

 

 

MODELLING

We created multiple models and used them to direct the path of our project

 

We created models of three types: experimental, physiological and economic. In the experimental model we wanted to examine the complexities of optimizing recombinant insulin production from microbes, so we examined the practicality of multiple expression systems.

 

A model was produced that took into consideration the multiple energy costs within a cell, and examined the predicted success of multiple expression systems: cytoplasmic expression, periplasmic expression and secretion into the surrounding media. From this model we predicted periplasmic expression to produce the highest yield.

 

This first model was based on the experimental work in the lab. We next produced another model that would predict the behavior of our recombinant insulin under human physiological conditions. From this model we predict that our insulin analogue Winsulin will be relatively fast acting compared to human insulin.

 

Finally, we developed economic models examining the pricing of insulin across different markets.

 

 

DEMONSTRATE YOUR WORK

We performed an assay mimicking physiological conditions whereby cells were tested with the insulins we produced (that were previously confirmed via ELISA assay).

 

We experimented on multiple cell lines, all of which were receptive to insulin, and measured their levels of glycogen synthesis and glucose oxidation after treatment. It was observed that when tested with Ecotin-Proinsulin, Cytoplasmic-Proinsulin and YncM-Winsulin all the cells showed more glycogen synthase and glucose oxidation activity than at basal levels with no insulin added.

 

Since this mimics the role that insulin plays in the human body, we have demonstrated the functionality of our parts and the success of our project.

 

Special Medal Criteria

Our team also is eligible for a number of special awards: for Integrated Human Practices, Education and Public Engagement, Model, Entrepreneurship and Applied Design.

 

INTEGRATED HUMAN PRACTICES

We utilized input that we received from online communities (insulin users) and professional advisors, to direct our project towards developing a regular Human Insulin due to its ubiquity on the global market, homology to the endogenously produced protein and freedom from intellectual property constraints.

 

Key Communications:

  • We listened to insulin users (via our survey, and communication with charity organisations involved in the distribution of insulin) who told us that ease of use and accessibility was important. From this we tried to include in our design of a novel analogue thermostability, an increased pI to result in maintenance of baseline blood glucose.
    • It also motivated us to try and simplify the purification and production of our Winsulin construct in order to reduce the expense associated with insulin, and make it more accessible to those in need

  • We also used weekly meetings with the OpenInsulin team to educate ourselves on the realities of open-source production, and develop our potential plan for moving forward.

  • When ReaGent (from Ghent, Belgium) integrated their views into the team we benefited from sharing global perspectives.

  • We spoke to patent attorneys at Maxwell's Patent and Trademark Attorneys, whose feedback we fed into the design of our Winsulin construct so as not to encroach on any existing patents

  • Our communication with Dr Narcyz Ghinea from The University of Sydney, an expert in access to high cost medicines, assisted us with data on where the money for clinical trials is sourced from, giving us the idea of a potential use of government grants for this open-sourced project

 

EDUCATION AND PUBLIC ENGAGEMENT

 

Engaging with the wider community was a huge focus of our project – both by alerting them to the issue of insulin accessibility and to the massive potential found in synthetic biological solutions

 

Key Activities:

  • We participated in the Synthetic Biology Australasia 2017 conference by presenting a talk on our project, opening up a discussion with the Australasian synthetic biologist community

  • We partook in the National Science Week at the Australian Museum by running a stall for the Joint Academic Microbiology Seminars (JAMS) group. This event was organized to educate school-aged children on the possibilities of science, especially the biological sciences

  • We participated in the Northern Sydney Science Hub, an event held to educate the public on the variety of microorganisms, and to break the stigma of perceived uncleanliness surrounding them

  • We invited students from the Mount St Benedicts School into our labs to teach them some basic biochemical and microbiological techniques, to inspire them to consider undertaking a STEM degree

  • We also joined a weekly meeting with Counter Culture Labs to gain feedback on our project, as their lab aims towards a similar goal

  • Our social media presence has been maintained throughout the year, through Facebook, Snapchat and Twitter, allowing us to access the broader community and the community of insulin users alike with our work, and to receive useful feedback at the same time

 

 

MODEL

Our modelling approach was multi-faceted to address the complex issue of insulin accessibility, which we broke into three main divisions: production, physiological activity and economic status

 

  1. To characterise the experimental/production of insulin, we analysed three expression systems: cytoplasmic, periplasmic and secreted
    • We interrogated the yield of folded protein predicted by each model, which allowed us to gain insights into the best way to optimize protein production
  2. For our physiological model, we identified a link between the thermodynamic and computationally predicted properties of insulin analogues and their action profiles
    • We used this to predict characteristics of our novel insulin analogue, Winsulin
  3. For our economic modelling we attempted to elucidate the pressures causing insulin price inflation
    • We did this via a 50 country analysis on insulin expense and a 10 country breakdown on affordability relative to income bracket
    • We used in-depth case studies to attempt to explain anomalies within the data and provide context to outliers

 

 

ENTREPRENEURSHIP

We communicated with online Diabetes support communities to identify issues with insulin pricing

  • We identified open-source manufacturing of insulin and the pump associated with it as an interesting, novel solution to tackling the current issues of insulin pricing
  • We worked on developing a novel single-chain insulin analogue with simplified purification properties to help reduce the costs of production, and so reduce costs to the consumer

 

We designed four tiers of functional business models to pursue in the future:

  1. Open-source solutions
  2. Small-medium business
  3. Large corporations
  4. State/Public investment

We also communicated with various experts in the field of startup business and the pharmaceutical industry in order to inform our production of these business models

 

APPLIED DESIGN

We designed a novel Single-Chain Analogue based on our research surrounding the issue of insulin affordability. Through:

  • Case studies
  • Economic analysis
  • Business model design
  • Community outreach
  • Intellectual Property Rights

 

We spoke to multiple specialists to inform these views, including the authors of the ACCISS report and members of the OpenInsulin project