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NASA NTRSDate not supplied

Lunar Explorer Instrument for space biology Applications (LEIA): An overview of planned science concept of operations

Radiation and reduced gravity pose biological risks to crewed deep space exploration. To better understand deep space radiation biology, the LEIA mission will measure charged particles and fast neutrons as well as yeast growth, metabolic rate, and bioengineered production of carotenoids at the lunar surface. LEIA will be delivered to the south polar region of the Moon by the Commercial Lunar Payload Services (CLPS) program. The LEIA science concept of operations was developed to isolate radiation and partial gravity from other environmental conditions experienced by the payload. Due to CLPS integration requirements, there will be at least eight months from loading yeast into the payload until activation on the lunar surface. Replicate yeast strains will be loaded in a randomized complete block design to minimize batch differences in desiccation tolerance and positional effects in the microfluidics culture system. Temperature and relative humidity will be controlled throughout integration and flight to maintain yeast viability and to enable measurement of yeast growth parameters within the lunar surface operations timeframe, with the ground control matching environmental conditions where possible. LEIA is co-manifested with the European Space Agency’s PROSPECT mission, which will be operating a drill during yeast growth. Vibration translated through the lunar lander will be mitigated and quantified to account for potential impacts on LEIA optical measurements and yeast growth rate. Total space radiation dose, including the transit exposure from Earth to the Moon, will be measured to obtain more accurate charged particle and fast neutron dose rates on the lunar surface. These radiation data will also yield ground truth radiation dose experienced by the yeast during the mission. Quantifying these environmental factors impacting the LEIA payload will allow more accurate ground control experiments to better isolate the biological responses to radiation and reduced gravity at the lunar surface.

Lunar Surface Mission↗
NASA NTRSDate not supplied

Yeast strain development to test in-space bioproduction in the Lunar Explorer Instrument for space biology Applications (LEIA) mission

The Lunar Explorer Instrument for space biology Applications (LEIA) is investigating the effects of lunar radiation and gravity on yeast viability, growth, and metabolism. LEIA is part of the CP-22 Commercial Lunar Payload Services (CLPS) surface mission to the south polar region of the Moon. The biological payload will test genetic factors that are likely to influence the tolerance of yeast for deep space and lunar surface radiation. LEIA is also investigating in-space production of β-carotene in bioengineered yeast. This carotenoid is both an antioxidant and pro-vitamin A- an essential human micronutrient. We report progress on engineering carotenoid-expressing strains to test the impacts of strain background and specific genetic variants on growth and production of β-carotene. To test for enhanced sensitivity to the LEIA mission environment, we generated gene knockouts for the RAD51 DNA damage repair locus as well as the SOD1, SOD2, and TSA1 reactive oxygen species (ROS) defense enzymes. We are also generating strains expected to increase tolerance to abiotic stressors and ROS. To be included in the biology payload, each strain needs to satisfy a series of requirements to be compatible with the mission concept of operations. The LEIA mission will conduct experiments using an autonomous light emitting diode optical detection system and microfluidics incubator to quantify growth, metabolism, and carotenoid production. Strains must produce sufficient carotenoids for bioproduction to be detectable with this optical system. Cells will be loaded into microfluidics cards, desiccated, and stored for 8-12 months prior to the initiation of lunar surface operations. The CLPS lander will operate for one lunar day, and strains will need to grow to stationary phase within 96 hours to ensure that telemetry of LEIA data to Earth can be completed. Genetic variants also need to display expected phenotypes within these optical detection, storage, and growth cycle constraints.

Yeast Engineering↗
NASA NTRSDate not supplied

The BioSensor Instrument Beyond LEIA: a Versatile Platform for Lunar Biology

Introduction: The BioSensor is a deep-space-compatible automated microfluidic culturing instrument. While originally designed to measure the effects of deep space radiation on yeast growth for the BioSentinel mission, it has the potential to host a diverse range of life science experiments with single- and/or multi-celled organisms and can be adapted to interface with a diverse range of platforms in both crewed and uncrewed settings. It is therefore a leading candidate for hosting life sciences experimentation associated with a lunar surface habitat. BioSensor design: The function of the BioSensor is to monitor the growth and metabolic activity of samples in batch fluidic culture, without the need for crew involvement. The current configuration houses organisms in 16 wells within 16 microfluidic cards, accommodating a total of 256 samples, replicates, and controls. Each well has an optical system including three LEDs and a photodiode detector to measure absorbance at three wavelengths, enabling measurement of optical density, color change in dyes such as alamarBlue, and bioproduction of pigmented compounds. Organisms are loaded into fluidic wells and air-dried for storage during integration, launch, and transit, then activated by the introduction of culture medium from storage bags via manifolds that fill one card at a time. Temperature is controlled by individual card heaters, and timing of all activities (fluidics fills, optical measurements, temperatures) is directed by an experiment script. The self-contained BioSensor payload is roughly 4U in volume; with electrical/mechanical/thermal interface, e.g. for operation on ISS or a lunar lander, as well as a linear energy-transfer (LET) charged-particle radiation spectrometer, the entire system is closer to 6U. BioSentinel and LEIA: Flown on the ISS and in a deep-space free flyer for BioSentinel, the BioSensor has been modified for use in the LEIA mission, including improvements to reduce the sensitivity to lengthy launch delays. LEIA will monitor yeast growth in the radiation and reduced-gravity environment of the lunar surface no earlier than 2026, on a CLPS lander [4]. Changes include accommodating additional culture media and an additional LED color for a new biological assay (bioproduction of carotenoids-- dietary antioxidants), as well as modifications to the housing to allow late-load biology changeout and improved isolation between fluidics and electronics. Future prospects: Future work with the BioSensor, beyond LEIA, will include expanding the range and diversity of organisms and assays accommodated. Preliminary work has demonstrated the growth of Arabidopsis seedlings in BioSensor fluidic cards, including optical measurements of growth rate over time. Minor modifications could allow measurement of phenotypes related to photosynthetic capacity in both plants and cyanobacteria. The experimental capabilities of the BioSensor could be dramatically increased by introducing the capability for fluorescence measurements, and/or the design of novel biological assays using luminescence. The BioSensor can also be adapted for new platforms and experiment settings; in addition to free-flyer, ISS, and CLPS lander, a preliminary design concept has been developed for crewed deployment directly to the lunar surface. The instrument could be accommodated inside a lunar habitat, where its automated operation would make it an excellent candidate for experiments from fundamental investigations into the response of organisms to lunar surface conditions to applied-science purposes such as screening engineered strains of various organisms for bioproduction capability.

J A Lee↗
NASA NTRSDate not supplied

Metabolic Vessel for Impedance Spectroscopy and Electrochemistry (MVISE): The Ground Mapping Unit for the Lunar Explorer Instrument for Space Biology Applications (LEIA)​

The BioSensor payload on the upcoming LEIA platform aboard a CLPS lander will carry yeast to the moon to study response to radiation and lunar gravity. The LEIA BioSensor is designed to monitor metabolic activity using absorbance in conjunction with alamarBlue for measuring colorimetric changes as proxy measurement for redox potential. The science data returned from small spacecraft mission modules like LEIA is limited as it relies solely on optical measurements, necessitating a corresponding ground mapping unit that is equipped with multiple electrochemical sensors for accurate mapping of the optical data and operates fully automatically. This technology development work discusses the extensive design and optimization efforts put into the ground mapping unit, MVISE. MVISE is a custom designed, 3D-printed vessel with an agitation system, with six different electrochemical sensor probes, and ports for sample collection and a pressure release valve. The sensors provide real-time data, with dry absorbance measurements aligning with LEIA flight hardware and wet measurements demonstrating invasive sensor design enabling a comparison between the two setups. The 3D printer resin was tested for mechanical robustness, biocompatibility, and resistance to autoclave sterilization. The inner walls were coated with food-grade epoxy, ensuring a smooth finish to prevent microbial lodging and dye staining. The MVISE has successfully passed a week-long leak test and is now undergoing active biology tests. The MVISE prototype will be prepared for radiation tests, with three identical units being tested for varying radiation levels and culture compositions at the NASA Space Radiation Laboratory in November 2024. The integrated sensor approach proposed in this work will enable the accurate mapping of the BioSentinel/LEIA optical flight data to six sensor parameters on the ground unit for better science data return and will enable the first effort to evaluate classical biochemical sensor measurements by comparing and contrasting their responses.

Chinmayee Govinda Raj↗
NASA NTRSDate not supplied

LEIA: An Investigation of Radiation Risks to Biology at the Lunar South Pole

Radiation and reduced gravity pose biological risks to crewed deep space exploration. At the cellular level, radiation damage can be amplified by reduced gravity. Empirical evidence on cellular responses to beyond low Earth orbit (BLEO) environments is imperative to develop effective countermeasures for crew health and in-space biomanufacturing. The Lunar Explorer Instrument for Space Biology Applications (LEIA) project is developing an instrument suite to be delivered to the south polar region of the Moon by the Commercial Lunar Payload Services (CLPS) program. This presentation will provide an overview of the LEIA hardware, experiments, and mission timeline. The LEIA instruments include the BioSensor, the ARES charged particle detector, and the Mini-FND. The BioSensor is an autonomous light emitting diode (LED)-based spectrophotometer and microfluidic incubator. The BioSensor activates yeast cultures and can measure cell growth, metabolic activity, and carotenoid production. The ARES is a Timepix-based charged particle radiation detector that measures dose, dose rate, and linear energy transfer spectra. The Mini-FND is a fast neutron detector that measures albedo neutron flux and energy spectra. Combined, these instruments will be used for yeast genetics experiments to quantify growth, metabolism, and synthetic biology-enabled production of human nutrients, while taking real time measurements of biologically relevant radiation exposure on the lunar surface. These data will be used to test the importance of selected DNA damage repair and reactive oxygen species defense pathways in mitigating cellular damage from lunar surface radiation.

Yeast↗
NASA NTRSDate not supplied

LEIA: An Investigation of Radiation Risks to Biology at the Lunar South Pole

Radiation and reduced gravity pose biological risks to crewed deep space exploration. At the cellular level, radiation damage can be amplified by reduced gravity. Empirical evidence on cellular responses to beyond low Earth orbit (BLEO) environments is imperative to develop effective countermeasures for crew health and in-space biomanufacturing. The Lunar Explorer Instrument for Space Biology Applications (LEIA) project is developing an instrument suite to be delivered to the south polar region of the Moon by the Commercial Lunar Payload Services (CLPS) program. This presentation will provide an overview of the LEIA hardware, experiments, and mission timeline. The LEIA instruments include the BioSensor, the ARES charged particle detector, and the Mini-FND. The BioSensor is an autonomous light emitting diode (LED)-based spectrophotometer and microfluidic incubator. The BioSensor activates yeast cultures and can measure cell growth, metabolic activity, and carotenoid production. The ARES is a Timepix-based charged particle radiation detector that measures dose, dose rate, and linear energy transfer spectra. The Mini-FND is a fast neutron detector that measures albedo neutron flux and energy spectra. Combined, these instruments will be used for yeast genetics experiments to quantify growth, metabolism, and synthetic biology-enabled production of human nutrients, while taking real time measurements of biologically relevant radiation exposure on the lunar surface. These data will be used to test the importance of selected DNA damage repair and reactive oxygen species defense pathways in mitigating cellular damage from lunar surface radiation.

Yeast↗
NASA NTRSDate not supplied

Microbial Optical Data Processing: A Key Step in the Metabolic Assessment of Lunar Explorer Instrument for Space Biology Applications (LEIA) and Biosentinel’s Payload Data

The BioSensor payload platform on BioSentinel and LEIA autonomously collects optical data from microbial model organisms in liquid culture. The BioSensor is designed to monitor metabolic activity using absorbance measurements of cell density and alamarBlue, a readily available colorimetric redox indicator dye. BioSentinel, a pioneering NASA CubeSat, uses yeast to study deep space radiation. LEIA investigates radiation and lunar gravity response. The experimental setup includes 16 wells equipped with three LEDs (570, 630, and 850 nm) and their corresponding photodetectors. One well is a calibration control without biology while the rest have desiccated cultures. Autonomous rehydration initiates the experiment. Data from the BioSensor are received from the flight and ground units, enabling comparison to uncover location-based metabolic rate variations. This study presents a Python Jupyter notebook developed for efficient data processing of multiple CSV files containing date and time columns, temperature, and well illumination data. It offers a user-friendly interface while maintaining computational power, automatically recognizing and iteratively processing data files in a user-input path. A Hampel filter with a short window eliminates outlier artifacts from sensor dropout. Because absorbance is a relative measurement, conversion from raw illumination requires defining a “blank” value, so the first data points are averaged to provide the necessary denominator. A cube-root function correction mitigates undesired drift caused by air pockets during the fluidic card filling phase, maintaining optical path length consistency. Beer-Lambert's law is applied to further convert absorbance values to cell and dye form concentrations, the desired science parameters. The processed data are saved and visualized as SVG plots. Future plans include extracting specific science parameters from the processed data like growth rate and metabolic rate, and identification of features corresponding to metabolic and phenotypic shifts such as starvation, shifts from aerobic to anaerobic growth, and osmotic stresses.

Space biology↗
NASA NTRSDate not supplied

The Lunar Explorer Instrument for Space Biology Applications (LEIA) Pre-Flight Tests: Mechanical Considerations​

The BioSensor payload on the upcoming LEIA platform aboard the CLPS (CP-22) lander will carry yeast to the Moon's south pole to study the effects of lunar radiation and gravity. From Earth, BioSensor will transport desiccated yeast within 16 fluidic cards, each containing 16 fluidic wells of 100 microliters volume. At the start of the experiment, these microbes will be autonomously rehydrated on the Moon, and optical absorbance measurements at three wavelengths will be performed across all 256 wells. Absorbance measurements in fluidic wells are susceptible to noise and artifacts due to bubble formation during well rehydration and microbial growth. BioSensor, as a biofluidics system, faces significant data anomalies caused by payload vibrations, particularly from the operation of the co-located PROSPECT’s ProSEED drill, to assess lunar resources by penetrating the subsurface. To evaluate the impact of drill vibrations, fully assembled and filled fluidic cards were subjected to sine sweep signals in the x, y, z planes, ranging from 5 - 2560 Hz at g-loads of 0.25 and 0.5, with on-board accelerometer for localized vibration measurement. Wells with various fill conditions, including partial fills with different bubble sizes and completely filled wells, were tested. Results indicated that small bubbles adhered to well walls at high frequencies but were dislodged at low frequencies, causing data anomalies. Medium and large bubbles exhibited higher interference with optical data due to lower surface tension and variable bubble surface flexion. Overall, it was found that irrespective of the fill type, low-frequency vibrations were most detrimental to optical readings. Understanding the drill schedule and correlating it with BioSensor timestamps will help identify periods when BioSensor data is unreliable, a crucial step for accurately interpreting the microbial response data and ensuring the validity of the BioSensor's scientific measurements on the lunar surface.

Chinmayee Govinda Raj↗
NASA NTRSDate not supplied

Yeast Strain Development and Hardware Testing in Preparation of a Lunar BioSensor

With Artemis missions underway, it is clear we are going back to the Moon to stay. Before sending Astronauts for long-duration missions, it is crucial to understand the technological and biomedical countermeasures needed to protect them before they get there. We can use knowledge gained from biological CubeSats to guide the next generation of experiments to support human habitation on the Moon. Lunar Explorer Instrument for space biology Applications (LEIA) is NASA’s latest BioSensor, adapted BioSentinel, the only CubeSat to travel Beyond Low Earth Orbit. BioSentinel launched on Artemis I and is currently >50 million kilometers from Earth (as of July 2024). LEIA aims to identify biological responses to the Lunar environment, which unprotected against would pose a threat to astronauts (cancer, cardiovascular disease, neurological impairment). The suite of instruments within LEIA detects Lunar radiation using two on-board radiation sensors (ARES charged particle detector, Mini-Fast Neutron Detector), then monitors real-time biological responses to the Lunar environment via an autonomous microfluidic system, fit with 3-LED emitter and detector boards and the alamarBlue metabolic indicator dye. LEIA will use a genetic approach in addition to synthetic biology to test counter-measure production in space, with the goal to inform and protect astronauts for future Moon missions. We have conducted preliminary tests in preparation for launch to the anticipated South Pole of the Moon, optimizing the biology (strain down-selection, desiccation tolerance, radiation sensitivity) and improving the hardware (including a blue LED to detect the beta-carotene countermeasure product). Our team will discuss these findings in several parts – an overview of the LEIA mission (Mark Settles), adapting flexible CubeSat platforms for deep-space applications (Sergio Santa Maria, Kira Rienecker), developing new technologies to support LEIA ground studies (Chinmayee Govinda Raj), and yeast strain development and hardware testing in preparation for LEIA (presented here).

synthetic biology↗
NASA NTRSDate not supplied

Developing Flexible Instruments for Biological Missions Beyond Low Earth Orbit

As the future of spaceflight focuses on human exploration beyond low Earth orbit (BLEO), space biology experiments using model organisms are becoming increasingly important. NASA’s Artemis missions seek to build technologies that enable extended crewed flights into deep space, a region not travelled by humans for over 50 years. From the Apollo missions and ground experiments, it is known that BLEO galactic cosmic radiation can cause damage to DNA and proteins, as well as an increased risk of cancer to astronauts. NASA’s latest biosensor technology, the Lunar Explorer Instrument for space biology applications (LEIA), builds upon the viable and cost-effective platform of CubeSats to take biology experiments back to the Moon. Stationed on the lunar South Pole, LEIA will collect valuable in-situ radiation data, and use yeast to study the response to combined partial gravity and radiation stressors, as well as provide a proof-of-concept of on-demand bio nutrient production as a countermeasure for future crewed missions. Directly enhancing the abilities of the BioSentinel CubeSat mission, the main components of LEIA are the two radiation sensors and the 4U BioSensor, composed of 16-microfluidic cards housing dried yeast cells. The payload is fully contained and autonomous, directly sending data back to Earth without the need for sample return. In addition, the thermal environment of the BioSensor is optimized to keep cells alive for the pre-launch period and duration of the Artemis III mission, while also running on limited power supply. This talk highlights how the development of flexible instruments, like LEIA, enables human exploration into deep space. The technology developed with LEIA can be used as a stepping-stone for establishing a sustained lunar surface habitation and beyond, as humans continue to venture into space.

Payne Elizabeth Turney↗
NASA NTRSDate not supplied

Use of Tardigrade Proteins for Enhancing Cellular Stress Tolerance on the Moon

With sights set on returning a human presence to the Moon and plans to explore even farther beyond, it is imperative we understand the impact of deep space radiation, partial gravity, and the lunar surface on biology. The need for countermeasures to protect future astronauts from the unique conditions of space is ever-pressing and an assessment of bioproduction capabilities in space is a crucial step for sustaining long-term missions. The Lunar Explorer Instrument for space biology Applications (LEIA) aims to investigate the cell’s response to the lunar environment and test strategies for enhancing cellular tolerance on the Moon, utilizing the budding yeast Saccharomyces cerevisiae as a biosensor. S. cerevisiae serves as an excellent model organism due to its widespread use as an analog for human cells, well-characterized set of genetic tools, and ability to survive desiccation. However, these cells must remain desiccated for up to one year prior to launch and the radiation-sensitive strains designed for LEIA will likely be more vulnerable to desiccation effects. In this work, we introduce tardigrade proteins known for their protective mechanisms into potential LEIA strains to test genetic strategies for improving desiccation and radiation tolerance. Tardigrades are extremotolerant animals well-known for their ability to survive in harsh environments. Notably, they can survive extreme desiccation and radiation due to the upregulation of intrinsically disordered proteins (IDPs). It has been shown that integration of these tardigrade IDPs, such as cytosolic abundant heat-soluble (CAHS) proteins, into yeast can confer increased desiccation tolerance in synergy with trehalose. Our ongoing testing and assessment of bioengineered radiation and desiccation tolerance will help us gain insight into optimizing strain design and selection for protecting LEIA’s biological payload as well as genetic engineering strategies for enhancing bioproduction potential for future missions beyond low Earth orbit.

Genetics↗
NASA NTRSDate not supplied

Engineered Yeast to Test Risks for Human Exploration of the Lunar Surface

Jessica W. Chau, Natalie N. Ball, Aditya Hindupur, Sandra T. Vu, Jennifer Gil Acevedo, Lauren C. Liddell, Chinmayee Govinda Raj, Gentry, Sergio R. Santa Maria, A. Mark Settles Crewed exploration of the Moon carries risks of long duration exposure to reduced gravity and to deep space radiation. The Lunar Explorer Instrument for space biology Applications (LEIA) investigates the effects of increased radiation and reduced gravity on yeast viability and growth in a Commercial Lunar Payload Services (CLPS) surface mission to the south polar region. LEIA conducts yeast genetics experiments to quantify growth, metabolism, and synthetic biology-enabled production of human nutrients, while taking real time measurements of biologically relevant radiation exposure on the lunar surface. We have engineered beta-carotene producing yeast strains to test the importance of selected DNA damage repair and reactive oxygen species (ROS) defense pathways in mitigating cellular damage from lunar surface radiation. Carotenoids are important dietary antioxidants, and beta-carotene is pro-vitamin A, which is needed for vision and immune function. Carotenoids are sensitive to ROS produced by ionizing radiation and NASA is testing on-demand production of carotenoids from yeast in the BioNutrients space flight experiments. In LEIA, we test the effects of deep space on carotenoid yield in engineered yeast strains. The LEIA team uses CRISPR-Cas9 to engineer yeast to express carotenoids as well as to generate loss-of-function mutations. We are generating mutations in the RAD51 DNA damage repair locus and three genes that function to reduce oxidative damage to the cell: SOD1, SOD2, and TSA1. These strains are tested for carotenoid production using microfluidics and LED spectroscopy to allow remote sensing of cellular growth and carotenoid levels. Keywords: synthetic biology, oxidative stress tolerance, biosensors, space radiation, beyond low Earth orbit, lunar surface, CRISPR/Cas9, gene editing, desiccation, carotenoids.

synthetic biology↗
NASA NTRSDate not supplied

Use of Tardigrade Proteins for Enhancing Cellular Stress Tolerance on the Moon

With sights set on returning a human presence to the Moon and plans to explore even farther beyond, it is imperative we understand the impact of deep space radiation, partial gravity, and the lunar surface on biology. The need for countermeasures to protect future astronauts from the unique conditions of space is ever-pressing and an assessment of bioproduction capabilities in space is a crucial step for sustaining long-term missions. The Lunar Explorer Instrument for space biology Applications (LEIA) aims to investigate the cell’s response to the lunar environment and test strategies for enhancing cellular tolerance on the Moon, utilizing the budding yeast Saccharomyces cerevisiae as a biosensor. S. cerevisiae serves as an excellent model organism due to its widespread use as an analog for human cells, well-characterized set of genetic tools, and ability to survive desiccation. However, these cells must remain desiccated for up to one year prior to launch and the radiation-sensitive strains designed for LEIA will likely be more vulnerable to desiccation effects. In this work, we introduce tardigrade proteins known for their protective mechanisms into potential LEIA strains to test genetic strategies for improving desiccation and radiation tolerance. Tardigrades are extremotolerant animals well-known for their ability to survive in harsh environments. Notably, they can survive extreme desiccation and radiation due to the upregulation of intrinsically disordered proteins (IDPs). It has been shown that integration of these tardigrade IDPs, such as cytosolic abundant heat-soluble (CAHS) proteins, into yeast can confer increased desiccation tolerance in synergy with trehalose. Our ongoing testing and assessment of bioengineered radiation and desiccation tolerance will help us gain insight into optimizing strain design and selection for protecting LEIA’s biological payload as well as genetic engineering strategies for enhancing bioproduction potential for future missions beyond low Earth orbit.

Genetics↗
NASA NTRS2024

Synthetic Biology to Support Human Exploration of Deep Space

The International Space Station (ISS) has enabled a continuous human presence in space since November 2000. The ISS is in low Earth orbit, facilitating regular resupply missions to deliver air, water, food, spare parts, and science experiments. NASA’s Moon to Mars campaign seeks to return humans to the Moon and prepare for crewed missions to Mars. Increased distance from Earth poses logistical challenges to provide all resources needed by humans for deep space missions. NASA Ames Research Center is conducting a series of synthetic biology projects to test the use of microbes for on-demand biosynthesis of human micronutrients. The BioNutrients spaceflight experiments test an implementation concept to produce fermented food products in which microbe growth enhances micronutrient content. On-demand production of carotenoids was engineered into two yeast (Saccharomyces cerevisiae) strains that have been tested in nearly 5 years of storage. One of the BioNutrients strains will be incorporated into the Lunar Explorer Instrument for Space Biology Applications (LEIA) project. LEIA is developing an instrument suite to be delivered to the lunar south pole region by the Commercial Lunar Payload Services (CLPS) program. The LEIA instrument suite will be used to measure multiple yeast strains for growth, metabolic activity, and synthetic biology-enabled production of carotenoids, while taking real time measurements of biologically relevant radiation exposure on the lunar surface. LEIA data will be used to assess the impact of lunar surface radiation and reduced gravity on the production of engineered traits.

Synthetic Biology↗
NASA NTRSDate not supplied

A Heritage BioSensor for Lunar Biology Experiments

Introduction: Automated biological experiments on small spacecraft missions have gained prominence over the past decade due to their simplicity, accessibility, and small mass, volume, and power needs. Most recently, the BioSensor microfluidic CubeSat payload aboard BioSentinel used an automated microfluidic cell culture system to study the effects of environmental stressors like deep space radiation and microgravity on yeast growth and metabolism. BioSentinel’s successor, the Lunar Explorer Instrument for space biology Applications (LEIA), will study the effects of lunar gravity and radiation using an improved version of the BioSensor microfluidic platform. The BioSensor payload has great adaptability to host a diverse range of biological experiments with single- and multi-celled organisms in both crewed and uncrewed missions, making it a compelling candidate for future space biology studies in a lunar surface environment. BioSensor Instrumentation on BioSentinel: The first spaceflight mission with the BioSensor, BioSentinel’s biology experiments occurred at three locations -- deep space, ISS and ground. The payload contained 18 microfluidic cards, each featuring 16 growth wells (a total of 288 growth wells). Each well was loaded before launch with desiccated yeast. In space, liquid culture medium (nutrients) was automatically introduced to batches of wells at a time to initiate a series of biology experiments. Temperature was maintained by thin film heaters on both sides of each card. Each well was equipped with three LEDs emitting at 570 nm, 630 nm, and 850 nm, paired with photodetectors to measure cell concentration and the alamarBlue (metabolic indicator dye) color transition from blue to pink. Phenotypic parameters like cell viability, metabolic rate, and generation time can be derived from these measurements. The sequence and timing of fluid fills, optical measurements, and thermal control were stored onboard, but could be updated asynchronously via ground communication. LEIA: LEIA is slated for launch no earlier than 2026 on a CLPS lander. BioSentinel’s BioSensor has been modified for use in LEIA. These improvements include: (a) storage for multiple culture medium types, (b) additional LED color (465 nm) for a new biological assay for antioxidant (carotenoid) production, (c) housing modifications for later biology load before launch, (d) improved isolation between electronic and fluidic components, and (e) improved humidity control for prolonged organism viability in case of post-load launch delay. Future Prospects: The consistent and successful demonstration of complex fluidics platforms alongside reliable instrument operations in a space environment is poised to create strong momentum for BioSensor-based biological experiment payloads. Planned future developments with the BioSensor include extending compatibility to a broader range of organisms and assays. Preliminary work has already demonstrated successful growth of Arabidopsis seedlings in fluidic cards. With a few modifications to the optical assembly, the setup could easily measure photosynthetic traits in plants and cyanobacteria. The addition of fluorescence measurements and generation of novel luminescent assays will elevate BioSensor’s functionality further. Beyond the BioSensor’s potential uses on free-flyer missions, ISS and Gateway, and CLPS landers, deploying the BioSensor to the lunar surface or in an artificial habitat on crewed missions could enable pioneering research on both how life responds to lunar conditions and future bioproduction capabilities making the BioSensor an indispensable tool for future space biology research.

Chinmayee Govinda Raj↗
NASA NTRSDate not supplied

Development of Genetic Countermeasures for Enhancing Cellular Stress Tolerance on a Lunar Surface Mission

The Lunar Explorer Instrument for space biology Applications (LEIA) LEIA investigates the response to partial gravity and ionizing radiation of: Different DNA damage and stress response pathways and Bioproduction of antioxidants LEIA utilizes: Various strains of the yeast Saccharomyces cerevisiae, which will be desiccated in fluidic cards and rehydrated on the lunar surface LEIA develops: Genetic countermeasures to improve tolerance to the desiccation process and the constraints associated with long duration missions beyond low Earth orbit (LEO)

Neha Lingam↗