Project Management

Design Goals:

Design Constraints

Ethics, Health, and Safety

Engineering Standards

• Standard of care (SOC) treatment for advanced HNSCC includes lymphatic resection to prevent metastasis. Novel treatments must be compared to SOC to evaluate efficacy.

• ISO 12052:2017 - DICOM standards (relevant to IMARIS imaging analysis software)

• ASTM F3354-19 and ISO 22442-1 - tissue decellularization standards.

Ethical, Environmental, and Societal Concerns:

• We avoided doing in vivo work because our experiment lacked the safety data to ensure its success in mice. We also minimized the number of mice tested with as is recommended by the IACUC.

• Because of COVID-19, we had to delegate the two sub-projects requiring lab work to two members who had access, while the other two members worked remotely on the modeling software IMARIS. This required leadership within subprojects and active teamwork across different subprojects to ensure all members were active participants. As such, this encouraged us to be self-initiating with sub-projects, but also to openly communicate data and knowledge with each member to allow for collaboration.

Innovation and Entrepreneurship

• The general research community for lymphatics and immunology lacks a foundational model of murine lymphatic networks, which our team had the potential to fulfill with the innovative idea to use IMARIS to map lymphatic networks. The collaboration between translational scientists and engineers also allowed for the creation of an engineered organ with functionality and patient translation in mind at all stages of the design process.

Risk Assessment and Mitigation

Annotations Key

P = prevent; risk needs concerted team action to prevent from happening;

R = reduce; risk needs attention so that likelihood of occurrence and/or severity of effect are reduced

T = transfer; others will assume the risk, i.e. by buying insurance

A = accept; there are minimal consequences

Technical Risks

The technical skills required for implantation of decellularized lymph nodes has not yet been discussed, however is a current challenge general decellularized organs face (P). In addition, it is very challenging to standardize a decellularization protocol due to varied sizes of lymph nodes (all of which are on the millimeter scale) (P). Team members must learn how to operate new software to analyze mice CT scans to map the lymph node positions (P). Lastly, the team must rely on confirming histological analysis with a professional (A).

Resource-Related Risks

Unfortunately, there are current delays to in vivo testing due to difficulties in optimizing the decell protocol (P). We will need a constant supply of mouse lymph nodes for decellularization, characterization, and in vivo tests which depend on the availability of our mentor (R). In addition, certain equipment or reagents that are needed for our experiments are in high demand due to the COVID-19 pandemic, and if we are unable to attain more of these, this could limit our project. For example, PPE and pipette tips have been backordered at several companies. (P)

Safety-Related Risks

If the decellularization protocol does not completely remove genetic material, the implant may cause an immune response in the mice (R). The device must be kept sterile and implanted properly to avoid adverse effects to the mouse (R). Lastly, team members must follow proper lab safety procedures (including wearing PPE, taking caution with chemicals, and maintaining social distancing in the lab due to COVID-19) to avoid risks associated with in-person lab work.

Environmental Risks

The project consumes a very large amount of mouse lymph nodes, due to the low yield of successfully decellularized samples (ie. fully intact, undamaged) (A)

Social Risks

Because this current project will take place in a mouse model, there is little social risk at the present moment. In the future, the end goal of this project would be to implant donated and decellularized lymph nodes (from either human donors or animals) into patients. However, this depends on patients’ willingness to accept donated tissues/cellular therapies. This may also be a concern in terms of finding a source of donor tissue, as that has not yet been established (T)

Catastrophic Events

A member of the team becomes infected with COVID-19 (P)

Mitigation Plan for Most Significant Risks

1. A team member may become infected with COVID-19 (P). To avoid this catastrophe, all of us will be taking steps to protect ourselves individually by doing all group work virtually, staying home as much as possible, and quarantining when appropriate. The Gutkind lab and Christman lab are operating at 50% capacity, requiring each member to undergo daily symptom screening before entering the lab. Every lab member is masked at all times, and lab air is regularly filtered.

2. An improperly crafted device will bring safety risks to mouse model (P). Every effort must be undertaken to ensure in vivo experiments meet ethical guidelines. To ensure safety, the device will be properly characterized through benchtop and in vitro experiments before implantation in vivo. Upon any serious adverse reactions in vivo, the experiment will be halted and either the device removed or the animal euthanized, depending on severity.

3. Implantation of decellularized lymph nodes brings technical risks that must be addressed (P). We will discuss surgical protocols with our mentor to understand more about the physical requirements regarding the implantation of a replacement lymph node (in our case, a decellularized lymph node) We will also look into the literature on surgical implantation of different types of decellularized tissues to see how the general procedure would work. Iterative attempts to optimize the correct protocol regarding implantation of lymph nodes into the mouse model will be conducted.

4. Standardization of decellularization protocol to ensure efficacy at all varied sizes of lymph nodes (R).By establishing a general protocol and testing it on several lymph nodes of varying sizes, we will characterize each one to determine the size range within which we can confidently say the decellularization has been successful. Additionally, while this risk may increase our consumption of mouse lymph nodes, we will attempt to mitigate this by reusing “unsuccessful” (i.e. damaged) decellularized lymph nodes for characterization and successful ones for implantation.

Project Timeline (Gantt Chart)