Communication is Key

Our approach to designing AtheroSHuffle included vast prior research and expertise before we finalized our goal. This stems from our team’s belief that a solution cannot be made first before the problem is found, but rather the solution works within an existing problem’s conditions. Because we wanted to tackle the problem of rising cardiovascular diseases, we needed to make sure our solution would be implemented from the root cause of this problem, and not simply mitigate the effects. This is why our team has identified certain core values to integrate into all parts of our project and a mission statement to communicate these goals.

brain
Index
Creating a Mission Statement
Our Values and Goals
Integrity
Novel
Teamwork
Environment
Global Engagement
Responsiveness
Accessibility
Transparency
Ethics

Creating a Mission Statement

"AtheroSHuffle represents a collaboration between science and society…"

We talked to experts in the field, major stakeholders, the general public, and other iGEM teams to design a device that meets the needs of our target audience of everyday consumers by prioritizing accuracy as well as convenience. In addition to our device, we recognize that in order to create a lasting change in cardiovascular health, we need to raise awareness about the seriousness of this issue in the world today. Hence, we conducted efforts geared towards education about atherosclerosis, cardiovascular health, and synthetic biology to the general public of all ages.

…to proactively combat the complications of cardiovascular health"

It is not enough to detect cardiovascular disease once it has done irreversible damage. Rather it is important that we detect it while there is still time to reverse or slow the condition. This is the purpose of our device, AtheroSHuffle. Cardiovascular health encompasses a complex biological system reliant on many factors. There are a wide variety of symptoms and the disease impacts many parts of the body. Proactive measures allow users the opportunity to prevent further complications and ensure the best quality of life.

Our Values and Goals

I

Integrity

We’ve worked hard throughout the design process of AtheroSHuffle to keep the needs of our users in mind and to ensure users are given honest and accurate results regarding their cardiovascular health. We designed an app to analyze the results of an individual’s blood sample test and provide honest feedback and health recommendations. We also worked with medical professionals as well as a clinical dietitian for heart and vascular patients to make sure we are basing our design off of honest and up-to-date information.

Novel

Our team spent the summer developing an innovative atherosclerosis diagnosis test focusing specifically on an individual’s oxLDL blood levels. Through conversations with healthcare experts and medical experts, we learned of the value of detecting biomarkers associated with atherosclerosis besides biomarkers, like C-reactive protein (CRP), that are already being tested in order to confirm the accuracy of diagnoses. The novelty of our research arises from the antibodies we’ve designed specific to the oxLDL biomarker on our lateral flow assay test strip.

N

T

Teamwork

Teamwork comes when each member feels they are supported and their voice matters. Our team strives to maintain a challenging, yet cooperative atmosphere. Each team member has a role and when everyone does their part, we are able to create a system of checks and balances to create a trusting device that encourages general consumers to contemplate their cardiovascular health. We believe work can be play, which allows us to invest as much as we can give. No matter a short or long work day, we always had smiles on our face because we recognized the importance of the impact our project would have on society. This was only possible through the supportive environment we created as a team.

Environment

With the rising use in single-use plastics and medical waste, our team wanted to lessen our impact on the environment. Although our current test strip has been prototyped in plastic, we have researched and planned to implement biodegradable materials for the cassette of our test strip. In addition, our LFA optimization works to include the minimal amount of materials and antibodies needed to still provide reliable results. Future projects would include creating a device similar to a diabetes digital reader that allows this test to be reusable. This project, however, can only be done once the research done in our current research is analyzed.

E

G

Global Engagement

Synthetic biology is used in a vast number of fields such as agriculture, manufacturing, diagnostics, therapeutics, etc. Many similar techniques, however, are used despite the different areas of interest. To learn about the different applications of these techniques, we met with different iGEM teams from around the world (IISER Pune 2, Lithuania, URochester, Bio-Brussels etc.), and having these domestic and non-domestic collaborations allowed us to learn from different cultural standpoints. We learned about technologies that other teams used that we both found would be useful to the direction of our project. For example, we met with the lead of the Vilnius 2020 iGEM team and she led us to the Biodot Airjet machine that would help us immobilize antibodies to our test strip.

Our global engagement continued with our partnership with the IISER Pune 2 team to create a blog that would reach high school students around the world. We laid the groundwork for the blog posts and website and it will soon be available on a public domain. We aim to encourage students to harbor an interest in STEM through our blog from a global perspective.

Responsiveness

We maintained an open dialogue with experts in the field and affected stakeholders throughout our AtheroSHuffle development process. Through conversation, we learned what individuals in the cardiovascular field felt needed to be emphasized in our research efforts. For example, after meeting with a representative from Tury Health, we understood the importance of not only knowing atherosclerosis and information specific to our project, but also the importance of expanding our efforts and understanding cardiovascular health as a whole. This feedback shaped the conversations we had during team meetings, and specifically integrated human practices and outreach meetings, as we adjusted our research efforts to cover a greater scope of heart health.

R

A

Accessibility

With accessibility at the forefront of our design, we decided to create a test strip due to its relative ease of use and affordability. With the COVID pandemic, people became accustomed to using test strips to monitor their health. It also drastically increased the market for lateral flow assays and test strips, which advantageously refined the methods of production. While we recognize that our device will not be in mass production soon, we have designed a device that is affordable and accessible to all.

In addition, we have incorporated small changes such as changing the typical test strip color from red to black, to create a sharper contrast and including read-aloud features on our app for those who may have visual difficulties.

Our HP efforts also led us to the decision to create educational tools to raise awareness of the disease. We recognize that many people are unaware of atherosclerosis and therefore, as many experts pointed out, accessibility is first achieved through education of the topic. In an effort to expand the scope of our audience, we also translated our blog to many different languages besides English such as Spanish, French, Hindi, Arabic, etc.

Transparency

From the start of our project to the end, we wanted to keep the commitment to remain transparent and be an open source platform. Our wiki details our weekly progress in wet lab and all of our findings are well documented. We are eager to share our findings as well as information regarding future projects that could strengthen our design with iGEM and the research community as a whole in hopes that our work could aid in the efforts toward improving atherosclerosis diagnosis. We’ve also kept an open dialogue with medical and biosafety professionals to ensure our design provides a safe and effective product for users to benefit from.
In terms of our transparency with users, we recognize the limitations of our design and stress that our device is a screening device for use in conjunction with traditional diagnostic methods. When a positive result is received, we encourage users to speak with medical experts to share their concerns.

T

E

Ethics

As a team, we all agreed that ethics was a priority in the design of the project and therefore, we started off with the question: Is synthetic biology a viable answer to the issue at hand? Many people are skeptical of synthetic biology as a solution. We met with many experts to answer this question and ultimately, the response was the same: The increasing issue of socioeconomic disparity, accessibility, and high risk of cardiovascular disease requires the use of synthetic biology to detect atherosclerosis in a proactive manner. With this reassurance, we began to design our device. This led to the decision of using E. Coli SHuffle as a chassis to create our antibodies. There are many ethical implications with traditional methods of antibody production that utilize animals. Therefore, we employed these engineered E. Coli to create our antibodies in an ethical manner.

Standing on the Shoulders of Giants

As a student-led research team, knowledge of the area in which we are researching is crucial. Many of us come from intellectual backgrounds rooted in STEM, so while these concepts of genetics, engineering, diagnostics, mathematical modeling, and biology are not new to us, much help is needed for us to properly execute and better inform our project design. As students at a university renowned for research, we have endless resources and experts in applicable fields very close by. We have taken advantage of the connections we have with our PI, Dr. Keith Kozminski, and other UVA faculty members, as well as the ability to grow connections with other experts, and intend to continue this practice throughout the aftermath of AtheroSHuffle. It is through these connections with those who have dedicated their career to being just as curious as the community of iGEM, that we are able to go into this project not only with ambition to detect early signs of arterial plaque build-up, but with the tools to do so.

scientist
Index
Survey
Dr. Bindu Kalesan
Dr. Jay Brown
Ieva Lingyte
Dr. Norbert Leitinger
Ms. Jennifer Kershner

Survey

Although we had consulted numerous experts on the design of our product, the general public was an important stakeholder to determine how we can best tackle the rise of cardiovascular disease as a whole. Researchers cannot release a public product without understanding their primary consumers. In addition to marketability purposes, it is helpful to understand the awareness levels of atherosclerosis, its risk factors, and differing levels of health consciousness of different demographics.

To consult the public, we created an online survey to assess health attitudes to provide us with information on the usability of our device. In general, respondents were asked about their exercise and nutrition lifestyle choices, as well as non-private information on their family health. The second half of the survey asked about their potential use of our product.

Questions ranged from multiple choice to sliding range estimates. The survey was distributed to mutual adult contacts of advisors and students, in addition to in-person data collection at local shopping centers.

To minimize bias and create a purposeful survey, our questions were reviewed by multiple people, including Sarah Robinson from UVA’s Centery for Survey Research, and Ashley Williams, an IRB coordinator. Data from the survey was analyzed for statistical differences with T-tests.

Survey Analysis

Dr. Bindu Kalesan

We approached Dr. Bindu Kalesan, CEO of Tury Health, to gain the perspective of a biotechnologist from a company with many of the same goals and values as ours. She praised our use of synthetic biology to solve this issue of atherosclerosis detection and saw huge potential for the device as a proactive measure.

Initial Status

Our team solely focused on the details of atherosclerosis, such as disease progression, important biomarkers, and current methods of detection.

Interaction

A major takeaway from the meeting was her emphasis on our knowledge of the subject. While she recognized the importance of understanding the details of atherosclerosis and synthetic biology, she stressed the importance of fully understanding the details of cardiovascular disease and health as a whole. Once we understand the larger problem, we can fully understand the extent and implications of the specific problem on society.

Post-Interaction Status

We backtracked our research to fully learn about cardiovascular health, which proved to be helpful in understanding the mechanisms of atherosclerosis and the development of the app.

Additional Notes

She encouraged us to think about the next steps once a person is diagnosed with atherosclerosis. She classified our device as a primary intervention step, but encouraged us to think about the secondary and tertiary intervention steps. This is important to acknowledge as it helps us introduce our device seamlessly in the existing sequence of steps.

Dr. Jay Brown

At the very beginning of our AtheroSHuffle journey, we spoke with Dr. Jay Brown, a professor of microbiology and immunology at the University of Virginia. We spoke with him around the same time that we spoke with Dr. Kalesan.

Initial Status

We were focused on finding specific wet lab protocols and lost sight of the bigger picture. We concentrated on the details of detection of atherosclerosis.

Interaction

Dr. Brown explained the different types of heart disease, treatment methods, antibody structure, and antibody production methods via interesting anecdotes. He especially emphasized the importance of being familiar with the problem we are tackling and the current state-of-the-art. This would help us explain the issue and device clearly and succinctly to the public.

Post-Interaction Status

We started to hold weekly journal clubs, where we would read background articles and present the papers we read to the team.

Ieva Lingyte

The Vilnius-Lithuania 2020 iGEM team constructed a lateral flow assay (LFA) to detect bacteria infection in recirculating aquaculture systems. Therefore, we reached out to their team lead, Ieva Lingyte to learn more about how they constructed their LFA.

Our meeting with Ms. Lingyte. (From left to right: Ieva Lingyte, Alyssa Dioguardi, Yilun Zhou, Isha Patel.)

Initial Status

We were solely focused on the detection of atherosclerosis and the development of an accessible and easy to use screening test for consumers to use proactively.

Interaction

Ieva explained how one of their contributions encouraged them to look at prevention and treatment as well as detection because detection does not solve the ultimate issue of bacteria infections but rather only one aspect of it. Therefore, her team added two more prongs to their solution and she encouraged us to also look at the next steps after diagnosis.

Post-Interaction Status

Given that our device detects the effects of a medical condition, we realized that we would not be able to propose a viable treatment plan (especially since there already are treatment plans if detected). Therefore, we decided to create our app, AtheroShuffle, to promote a heart healthy lifestyle in both a preventive and primary treatment intervention manner to reduce the risk of further development into serious cardiovascular complications. The app would record test results, assess risk through a predictive model (inspired by a model developed in Dr. Coleen McNamara’s lab from UVA), and heart healthy lifestyle tips.

Dr. Norbert Leitinger

Dr. Norbert Leitinger is a professor in the Department of Pharmacology at the University of Virginia whose research interests are in the role of lipid oxidation products in inflammation and vascular immunology in various diseases, such as fatty liver disease and atherosclerosis.

First Meeting

Initial Status

We did not fully understand the structure of our antigen, oxLDL. This made it difficult to model where on the antigen we should target our antibody to bind to, and therefore, we were not able to identify the test line antibody.

Interaction

Dr. Leitinger provided us with essential current information about the field of atherosclerosis as a whole. He, first, gave us a background of some of the influential scientists in the field, specifically Dr. Joseph Witzum, who discovered the E06 antibody that most labs researching oxidized phospholipids use today. We originally were targeting the ApoB protein on the surface of the oxLDL, but Dr. Leitinger pointed out that most researchers in atherosclerosis today are targeting the oxidized phospholipids with the E06 antibody and that we should look into antibodies that also bind to these oxidized phospholipids. He offered his structures of oxLDL to us and then offered to show us his lab at a later time and introduce us to his grad student, Clint Upchurch.

Members of our team meeting with Dr. Leitinger. (From left to right: Alex Heise, Justin Orchard-Hayes, Isha Patel, Ivory Tang, Yilun Zhou, Peneeta Wojcik, Yasir Mahboba, Dr. Leitinger, Clint Upchurch.

Post-Interaction Status

We learned about the general structure of oxLDL and a general idea of antibodies to look into.

Second Meeting

After a month, we reconvened with Dr. Leitinger to discuss the recent progress we have made and put forward some queries we had at the current moment.

Initial Status

Dr. Leitinger reaffirmed that oxLDL serves as a promising biomarker for atherosclerosis (AS) as it has a predictive relationship with AS. We still had not narrowed down what antibodies to use because we were unsure of the epitope we should target on oxLDL. We had two major contenders, Anti-Phosphocholine McPC603 and anti-oxLDL IK17 (from papers Dr. Leitinger recommended) but we were not sure which one to use.

Interaction

Upon explaining our dilemma, Dr. Leitinger explained the advantage of using both antibodies together. One antibody targeted MDA-modified apoB, while the other targeted oxidized phospholipids (OxPLs). He pointed out a potential caveat to be aware of: oxPL structure is altered when over-oxidized and certain antibodies that recognize oxPL will no longer be able to recognize over-oxidized oxPL. Therefore, he recommended we use both to assure that the antigen will be detected.

Post-Interaction Status

We decided to use both Anti-Phosphocholine McPC603 and anti-oxLDL IK17 as our test line antibodies to ensure accuracy and visibility.

Ms. Jennifer Kershner

Biosafety is a crucial keystone consideration in any synthetic biology research. Hence, we spoke with Ms. Jennifer Kershner, a biosafety officer at the University of Virginia, to discuss the safety of our device and specifically, the employment of a kill switch.

Initial Status

We were considering implementing a kill switch for our bacteria to prevent unwanted release of bacteria.

Interaction

She assured us that as long as no bacteria leave the lab, a kill switch is not needed. She also explained that because our concentration of antibodies is low, we will not need to take any biosafety precautions for this.

Post-Interaction Status

We decided to leave out the kill switch per Ms. Kershner’s recommendation.

Additional Notes

To read about the full interaction, read the Biosafety section on the Project Design page

Collin Marino

Collin Marino was a member of the Virginia iGEM 2020 and 2021 teams and has a lot of expertise with synthetic biology

Initial Status

We ran into some difficulties with our ligation protocol and seeked out help from a previous iGEM member, Collin Marino.

Interaction

Collin gave advice on BioBricking, restriction digest, and ligation protocols.

Post-Interaction Status

After meeting with Collin, we were able to troubleshoot our ligation reactions with a refined BioBrick design. This helped us move forward in our wet lab work.

Additional HP Contacts