On March 2013, Charles Konsitzke, Associate Director of the UW Biotechnology Center (UWBC) and the Center for Genomic Science Innovation (CGSI), received an unusual request: to help identify the remains of a missing-in-action (MIA) soldier – PFC. Lawrence S. Gordon – tracked down by a civilian researcher through historical investigation. Konsitzke, who comes from a large military family, was intrigued and engaged the sequencing team within the UWBC. Using DNA analysis, the sequencing team successfully confirmed PFC. Lawrence S. Gordon’s identity the following year. This fortuitous collaboration led to the conception and creation of the UW Missing in Action Recovery and Identification Project (UW MIA RIP).
Since then, the project has expanded its analysis of environmental DNA (eDNA) to connect with historical investigations so as to identify and recover MIA soldiers. While this technology is traditionally applied in ecological contexts to measure biodiversity or detect the presence of a particular species, the UW MIA RIP is utilizing eDNA analysis to locate and identify the missing remains. This process analyzes environmental samples (such as soil or ocean floor sediment) at potential resting sites for human DNA, to pinpoint the location and identity of the remains.
The project has recovered the remains of two other soldiers – U.S. Army Air Force pilots 1st Lt. Frank Fazekas and 2nd Lt. Walter B. “Buster” Stone – and is currently working on the recovery of a WWII American service member in Northern Europe.
The UW MIA RIP is a cross-disciplinary project that involves several steps that precede DNA analysis, from investigating historical records to searching for eyewitness accounts at suspected remain sites. These investigations can take years.
“At that time, we will be meeting with local residents and possible eyewitnesses, looking at the site to see if we can find any disturbance or material, such as aircraft wreckage, and once we concluded with the scouting mission,” explains Konsitzke, now the team lead for the project. “Once we conclude with the scouting mission, we then determine to proceed or not proceed with the recovery mission, which means we have our team of faculty, staff and students to go into the field during the summer period, where we excavate the site and recover remains.”
Running the tests
Konsitzke explains that this process can be a challenge. He remembers going to a site in northern France were a plane crashed into a farm field, and the farmers filled the resulting crater with everything that they could find to relevel the ground. “It was WWII so there was a lot of dead animals, a lot of garbage, so we were going through a lot of skeletons – pig skeletons, rabbit skeletons, horse skeletons – to get to where our plane was,” shares Konsitzke. The team has a forensic anthropologist on-site to help identify the remains.
After locating and collecting soil samples, isolated eDNA is sequenced and compared to human DNA, confirming that human remains are near the sampling site. Once collected, DNA from the human remains is sequenced and compared to samples provided by relatives, sometimes with the help of genealogy tools such as Ancestry.com, which matches the remains to the MIA soldier.
Challenges and Solutions
Remains can be extremely degraded depending on the environment in which they are located. For example, soil with a high pH level can make the remains very fragile, almost like mush. Additionally, even if the remains are still preserved, the DNA may be degraded, which adds another obstacle to be overcome. “You modify the protocol, investigate multiple pathways; in a way it is its own research project” explains Konsitzke. “You attempt different pathways, have successes, fallbacks, and just continue to work through that process.”
The Biotechnology Center plays a central role in these cases, as very degraded samples require sensitive methods and lab environments clean of other DNA contaminants. Currently, the eDNA soil samples are taken to a 3-phase clean room meant for handling small amounts of degraded material. However, the team is currently testing new methods that may facilitate the recovery process.
The project hopes to expand their current eDNA protocols to capture human DNA from samples before sequencing. “When people typically do eDNA sampling they look at everything in that sample. We are taking everything else out and focusing specifically on that human output”, explains Konsitzke.
That would allow the team to identify sample ancestry and ethnicity in the field. Konsitzke considers this an important step, because some countries object to grave disinterment for individuals native to the home country. Determining ancestry of the remains in the field would allow the recovery to proceed.
While so far the project focuses on recovering the remains of soldiers that fell over half a century ago during World War II, Konsitzke says that the impact of the recoveries helps families heal from emotional trauma that has persisted for generations. “All that pain and sorrow [of an MIA soldier] has been passed to their kids, and their kids… They took on this weight of recovery, and when you have that closure it is a spiritual moment.”
The project not only impacted families, but also changed how missing-in-action programs operate. The UW MIA RIP was the first academic institution to partner with the Defense POW/MIA Accounting Agency (DPAA), an organization in the Department of Defense agency whose mission is to recover American military personnel listed as prisoners of war or missing in action. Currently, there are over 82,000 MIA U.S. service members. Now, the DPAA uses the UW MIA RIP as a model, and partners with over fifty other institutions, academic or non-profit, for MIA identifications.
Part of the success of the project is due to the diversity of volunteers that make up the team. “Our team has MBA students, economists, mechanical engineers, historians, law school students, anthropologists, archeologists, etc. Every student has a passion and having that melting pot of knowledge gives us those ‘aha!’ moments, where they’ll say, ‘hey have you ever tried that?’ and it’s just outstanding,” shares Konsitzke. He explained that anyone can volunteer to join their program. Students or staff with a passion for World Wars, history, or any of the conflicts are encouraged to apply through their website.