Maleth II

Maleth II

The Maleth Program, also known as Project Maleth is the first Maltese space bioscience program carried out by SpaceOMIX, that sent skin tissue samples from diabetic foot ulcer (DFU) patients to be studied on the International Space Station (ISS) in 2021. Maleth II is now on its way to following up on these findings on its second space research mission. The mission, enabled by the ICE Cubes service, a commercial service provider in partnership with ESA, is designed to replicate and validate the observations of the Maleth I mission, as well as conduct additional research on other cell types.

Professor Joseph Borg, a molecular geneticist and biomedical laboratory scientist at the University of Malta, is the lead of Project Maleth. The project focuses on research to improve precision-based medicine in the treatment of Diabetic Foot Ulcers (DFUs). The study will look at human skin microbiome samples from Type 2 diabetic patients who have DFUs that are resistant to treatment. The term “microbiome” refers to all microorganisms that live in or on the human body, including bacteria and fungi. The researchers wanted to send these samples to space, specifically the International Space Station (ISS) because microgravity and the harsh conditions of space provide a unique opportunity to better study the resistance of such microorganisms. A combined total of eleven skin tissue samples were sent to investigate the effects of spaceflight on DFU skin microbiomes and their resistance to harsh environments through Maleth I and II.

Why space research is a game changer?

Access to the space environment, microgravity, and high radiation present itself as a lifetime opportunity to investigate various biology experiments and fundamental biomedical science questions, which can have implications for real-world clinical problems back on Earth.

Microgravity affects all levels of biological organization, including cells, tissues, organs, and organisms, in novel and useful ways, sometimes in ways that allow medical, biotech, and other problems on Earth to be better addressed. The advantages of microgravity enable us to produce medicines, materials, and living tissue that would be impossible to produce in the nominal Earth’s gravity environment. Therefore, following the successful return and analysis of samples from Maleth I, the results of which are expected to be published by the end of the year, Project Maleth successful sent Maleth II, a second follow-up mission to the ISS. Maleth II as aforementioned is designed to replicate and validate the findings of Maleth I by sending microbiome samples from DFU patients again. In addition to the human skin tissue samples, yeast cell samples were also positioned inside the bio-cube payload on the ISS for further study.

How will the Maleth II mission be carried out?

The investigation from Maleth I, which sent skin microbiome samples from a subset of Type 2 diabetic patients with diabetic foot ulcers (DFUs) that are resistant to treatment, to be studied aboard the International Space Station, is continued in Maleth II, also known as the Follow-up Study of Human Skin Tissue Microbiome Studies and Yeast Cells in Space (ISS). Maleth II’s goal is to assess the degree of repeatability seen in comparison to the earlier Maleth I mission.

Patients with DFU who were resistant to treatment had skin samples submitted to the ISS. To learn more about the samples’ environmental adaptation and transformation, ground-based (analogue) and space-based cultures are used. The complete microbial profile of DFUs in Malta is still the research’s main objective. As a result, both ground-based and space-based samples will be analyzed utilizing next-generation metagenomic sequencing (where the genes of the host and bacteria are sequenced) and metabolomic analysis (where the components of bacterial metabolism are identified).

The outcomes are then contrasted with those of a typical bacteria culture in order to identify the various bacterial species that are present in infected ulcers and their environmental and antibiotic resistance.

Yeast (Saccharomyces cerevisiae) cell samples will also be flown to the ISS for a different investigation in addition to the skin samples. The fact that yeast cells are “eukaryotes,” or the same type of cells that humans are comprised of with DNA packaged as chromosomes and inside a real nucleus, sets them apart from bacteria and makes them an interesting new frontier in research.

This ground-breaking study’s objective is to see how the cells respond to the environment in space. Genetic sequencing will be performed on every single cell, and the outcomes will be compared between Day 1 space samples, Day 60 space samples, and samples that were cultured on Earth. Understanding changes at the single cell level is a really novel way to do science and comprehend the genetic basis of life. Single-cell sequencing offers substantial resolution, unlike any other technology to date.

How will the Maleth II experiment be carried out in the ISS?

The Bio-Cube, developed by the ICE Cubes service provided by Space Applications Services, contains the experiment. The ICE Cubes facility, which is part of the Columbus module of the International Space Station and run by the European Space Agency, is where the cube is kept (ESA). The Bio-Cube will be powered once it is set up in the ICE Cubes facility, enabling real-time interactivity and the direct following of onboard imagery as one of the distinctive characteristics offered by the ICE Cubes service.

Maleth II’s cube includes an aluminium outside framework rather than a plastic one like Maleth I had, providing a more durable house for the experiment. With the exception of an improved onboard computer and control software, the electronic internals remained essentially the same as those in Maleth I and the main two reasons for choosing the Ice Cubes platform are:

  • The first is the ICE Cube’s real-time interaction and monitoring capabilities. Professor Borg and his team can always see what’s going on inside the Cube. This is where Arkafort Ltd. steps in. Data connectivity and sample monitoring of the Cube are made possible by data links provided by the ICE Cubes service and relayed to the Mission Control Centre managed by Arkafort Ltd. in Malta. The cube’s onboard computer is a Raspberry Pi running a Linux operating system and some proprietary control software that allows it to be operated remotely.
  • The second feature of the ICE Cubes service is the fast track. It took less than 11 months for Maleth I to make contact and return samples from space to Earth for analysis.

Inside the cube contains five independent human skin tissue samples, each with its own unique microbiome, as well as one lyophilized Bakers’ yeast sample. The human skin tissue samples are suspended in a Phosphate Buffered Saline (PBS) solution, which is known (and has been tested) to keep both the human skin and, more importantly, the microbiome alive and well in suspension. The investigation is monitored 24/7 during mission duration by onboard cameras that look over the cuvettes/samples inside the Cube at Arkafort’s Mission Control Center in Malta. All planned molecular work is only done before and after spaceflight (on sample return, approximately after 45 to 50 days on the space station).

As an outcome, what will Maleth II’s goal be?

Maleth II Project aims to identify biomarkers (such as mRNA, a small single-stranded piece of DNA) that could potentially be targeted for better patient management and treatment, improving laboratory and healthcare services, and directly affecting the quality of life of Maltese diabetic patients and their families.

The goal is to learn more about how these treatment-resistant ulcers behave and how to treat them more effectively. Previous research has shown that the space environment causes bacterial resistance to antibiotics. Much remains unknown about how bacteria acquire resistance and studying these bacteria in space in conjunction with genetic sequencing may help us better understand the mechanisms underlying resistance development. Furthermore, other bacteria may be present, preventing the proper healing of these ulcers. By identifying all the bacteria present, it is possible to determine whether there are microorganisms that can be used to predict prognosis. Finally, the microbiome of healed ulcers can be compared to those that did not heal, revealing which bacteria may be involved with chronicity.

All data is deposited in NASA’s Gene Lab database, an open-access analysis platform for bioscience research that maximizes the global use of its data.

How can Maleth II be beneficial for Space and/or Earth applications?

Important to mention are the enhancements of Maleth II, which comprises biological duplicates (i.e., replicas from different time points, as it is the second time that this same study is being done in space) in addition to technical replicates (i.e., copies of the same kind of material).

Biological replicates are used to show whether the outcomes of the initial experiment can be accurately replicated in subsequent studies. Researchers will thus be much more confident in the outcomes if this is the case. Additionally, Maleth II will have more metagenomics sequencing.

Regarding Earth applications, has a lot of potential for individuals with Type 2 Diabetes Mellitus who suffer from difficult-to-treat diabetic foot ulcers. If several biomarkers are discovered using this method, it could revolutionize molecular therapy and precision-based medicine for the treatment of these ulcers and guarantee a higher standard of living for patients and their families.

Additionally, being Malta’s first biomedical science experiment in orbit, the project works with Esplora, the Malta Council for Science and Technology, as well as elementary and secondary schools in Malta, to increase local awareness of vital STEAM-related issues.

On the other hand, for Space applications, it relates to any future astronauts who embark on longer missions to the moon and/or Mars may benefit from improved space flight by identifying biomarkers that are present in the human skin microbiota. To ensure the viability and success of future space missions to low Earth orbit and beyond, it is necessary to better understand how the mouth, gut, and skin microbiomes behave and interact in space, and to be used in potential therapy in space missions.

SpaceOMIX dedication to humanity

To further our purpose and work on some of humanity’s most significant concerns and aspirations to date, SpaceOMIX will continue to proudly collaborate with top universities and organizations. The team will move on to the upcoming missions under a new, as-yet-unannounced program after the third and final mission under the Maleth program, and after completing SpaceOMIX’s second voyage to the International Space Station in 2022.

In this emerging field of genome biology and space science, SpaceOMIX stands out because our core group is working to establish a mission control center to monitor and follow remote communications with the payloads, as well as our own genetic engineering laboratory for processing samples from the payload and our own mechanical and electrical space engineers who will design the payload’s internal mechanics.

The Maleth Project is funded by the Ministries for Foreign and European Affairs, as well as the Parliamentary Secretary for Youth, Research, and Innovation under the Ministry for Education of Malta. The mission is being facilitated by the University of Malta Research Innovation, Development Trust (RIDT) who will be supporting a PhD candidate, Ms Christina Gatt, and other students at the University of Malta as well as all the science being conducted in this second mission. The commercial sector, which comprises MeDirect, Malta’s first digital bank, and Singleron Biotechnologies, Cologne, Germany, are supporting the Maleth II launch.

The mission control center is managed by Arkafort Ltd., and it will communicate with the ICE Cubes platform, which is run by Space Applications Services in the ISS’s Columbus Module, directly.

The mission is enabled by ESA through the ISS commercial partnership initiative, which facilitates access to exploration infrastructure to private companies such as Space Application Services.


Other Missions

Maleth I

Project Maleth I was the first ever Maltese mission in space. It focuses on research to improve precision-based medicine in treating diabetic foot ulcers more efficiently.