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NASA NTRS2025

Generalized Reference Targeting for Spaceflight

For spaceflight programs to achieve some of the aggressive exploration initiatives such as visiting and landing on other celestial bodies, rendezvousing with other orbiting vehicles, and ultimately returning crew safely to Earth, an assortment of targeting algorithms to compute the necessary burns to strategically maneuver a spacecraft to a variety of destinations are required. Numerous examples exist, but rather than creating and implementing multiple targeting solutions, is it possible to have a general targeting model that can accommodate a variety of applications? Originally motivated for mission design and analysis purposes, this paper outlines a generalized reference targeting algorithm for spaceflight that may also have applications in on-orbit flight software. It accommodates arbitrary flight dynamics and both impulsive and finite burns for either absolute or relative targeting applications. It also allows for an arbitrary number of targeting design parameters such as multiple discrete correction burns or finite thrust parameters to satisfy numerous combinations of targeting constraints that can have fixed or variable time epochs. Given a reference trajectory, this targeting technique provides a general framework to quickly solve an assortment of targeting problems that may be well-defined, over-determined, or under-determined while naturally producing metrics providing insight into the controllability for a given problem formulation. Due to the derivation, speed, and accuracy of the algorithm, it lends to supporting rapid linear covariance analysis and robust trajectory design applications for a variety of flight phases such as rendezvous and docking, cislunar transfer, interplanetary flight, orbit maintenance, de-orbit, and powered descent and landing.

Targeting↗

NASA NTRSDate not supplied

Detect-and-Avoid Maneuver Planning: Benefits of Including Route Recapture

Development of Detect-and-avoid (DAA) systems’ maneuver guidance requirements at RTCA has centered on tactical maneuvers away from the intruder. Recapturing the flight-plan route the initial maneuver is not directly taken into account by most DAA maneuver guidance algorithms. This work demonstrates potential challenges and inefficiencies that can arise in recapturing the route after resolving a conflict. Horizontal resolution trajectories for a test matrix of encounters with varying aircraft speeds and geometries are computed by minimizing either flight time or deviation to serve as a baseline. Turn directions computed from a reference DAA algorithm coupled with a pilot selection model and a second algorithm called Autoresolver (AR) are compared to these baseline resolution trajectories. Baseline results show that turning into the intruder yields favorable cost for most encounters. Additional analysis of pilot response data shows that only three-fourths of pilots’ horizontal maneuvers turn into intruders, a percentage much lower than the baseline results. Improvement to a DAA guidance algorithm based on findings in this work is discussed.

Route Recapture↗

NASA NTRS1987

Utilization of Global Reference Atmosphere Model (GRAM) for shuttle entry

At high latitudes, dispersions in values of density for the middle atmosphere from the Global Reference Atmosphere Model (GRAM) are observed to be large, particularly in the winter. Trajectories have been run from 28.5 deg to 98 deg. The critical part of the atmosphere for reentry is 250,000 to 270,000 ft. 250,000 ft is the altitude where the shuttle trajectory levels out. For ascending passes the critical region occurs near the equator. For descending entries the critical region is in northern latitudes. The computed trajectory is input to the GRAM, which computes means and deviations of atmospheric parameters at each point along the trajectory. There is little latitude dispersion for the ascending passes; the strongest source of deviations is seasonal; however, very wide seasonal and latitudinal deviations are exhibited for the descending passes at all orbital inclinations. For shuttle operations the problem is control to maintain the correct entry corridor and avoid either aerodynamic skipping or excessive heat loads.

Joosten, Kent↗

DOE OSTI2025

3013 Baseline Inner Container Report: Description, Imaging, and Analysis

This report presents consolidated findings from three microscopy studies of baseline Bagless Transfer Containers (BTCs), which are the inner containers used in DOE Standard 3013-compliant plutonium storage packages. The studies were conducted by Los Alamos National Laboratory (LANL), Savannah River National Laboratory (SRNL), and DNV, a subcontractor specializing in corrosion science. The baseline BTCs examined had not been exposed to corrosive environments, serving as control samples for comparison with destructively examined (DE) containers. Additionally, the Background section provides an overview of the BTC design, including manufacturing methods, material properties, and container configurations that may influence corrosion behavior. The primary objective of the LANL and SRNL studies was to establish reference characteristics of BTCs to distinguish manufacturing artifacts from corrosion-induced features observed in DE containers. Both laboratories used advanced microscopy techniques, including Wide Area Microscopy (WAMS), Laser Confocal Microscopy (LCM), Scanning Electron Microscopy (SEM), and Focused Ion Beam (FIB), to reveal that most surface anomalies were shallow and attributable to flow-forming processes, not corrosion. Subsurface impurities, identified as non-metallic inclusions, were also observed. DNV’s investigation focused on characterizing the mechanical and chemical properties of the BTC material to support crack growth modeling. Their findings indicate higher hardness and strength but reduced ductility—consistent with flow-formed 304L stainless steel and a higher-than-expected martensite content, in contrast to manufacturer claims. Ongoing and future studies, including Electron Backscatter Diffraction (EBSD), aim to further understand the microstructural factors influencing crack initiation and propagation. The combined studies provide a crucial baseline for distinguishing corrosion effects in DE containers, contributing to the safe and reliable long-term storage of plutonium-bearing materials.

36 MATERIALS SCIENCE↗

NASA NTRS2003

Experimental Evaluation of Verification and Validation Tools on Martian Rover Software

We report on a study to determine the maturity of different verification and validation technologies (V&V) on a representative example of NASA flight software. The study consisted of a controlled experiment where three technologies (static analysis, runtime analysis and model checking) were compared to traditional testing with respect to their ability to find seeded errors in a prototype Mars Rover. What makes this study unique is that it is the first (to the best of our knowledge) to do a controlled experiment to compare formal methods based tools to testing on a realistic industrial-size example where the emphasis was on collecting as much data on the performance of the tools and the participants as possible. The paper includes a description of the Rover code that was analyzed, the tools used as well as a detailed description of the experimental setup and the results. Due to the complexity of setting up the experiment, our results can not be generalized, but we believe it can still serve as a valuable point of reference for future studies of this kind. It did confirm the belief we had that advanced tools can outperform testing when trying to locate concurrency errors. Furthermore the results of the experiment inspired a novel framework for testing the next generation of the Rover.

Brat, Guillaume↗

NASA NTRS1976

Viking telecommunication effects of GEOS satellite interference based on testing at the Madrid deep space station

In support of the ongoing NASA-European Space Agency (ESA) effort to understand and control possible interference between missions, testing was conducted at the Madrid Deep Space Station from July 1975 to February 1976 to characterize the effect on Viking 1975 telecommunication link performance of Geodetic Earth-Orbiting Satellite (GEOS) downlink signals. The prime use of the data was to develop a capability to predict GEOS interference effects for evaluation of Viking 1975 mission impacts and possible temporary GEOS shutdown. Also, the data would serve as a basis for assessment of the GEOS impact on missions other than Viking as well as for more general interference applications. Performances of the reference receiver, telemetry, and planetary ranging were measured in the presence of various types of GEOS-related interference, including an unmodulated GEOS carrier and simulation of the actual spectrum by an ESA-supplied GEOS suitcase model.

Stuhr, F. V.↗

NASA NTRS2005

Mapping From an Instrumented Glove to a Robot Hand

An algorithm has been developed to solve the problem of mapping from (1) a glove instrumented with joint-angle sensors to (2) an anthropomorphic robot hand. Such a mapping is needed to generate control signals to make the robot hand mimic the configuration of the hand of a human attempting to control the robot. The mapping problem is complicated by uncertainties in sensor locations caused by variations in sizes and shapes of hands and variations in the fit of the glove. The present mapping algorithm is robust in the face of these uncertainties, largely because it includes a calibration sub-algorithm that inherently adapts the mapping to the specific hand and glove, without need for measuring the hand and without regard for goodness of fit. The algorithm utilizes a forward-kinematics model of the glove derived from documentation provided by the manufacturer of the glove. In this case, forward-kinematics model signifies a mathematical model of the glove fingertip positions as functions of the sensor readings. More specifically, given the sensor readings, the forward-kinematics model calculates the glove fingertip positions in a Cartesian reference frame nominally attached to the palm. The algorithm also utilizes an inverse-kinematics model of the robot hand. In this case, inverse-kinematics model signifies a mathematical model of the robot finger-joint angles as functions of the robot fingertip positions. Again, more specifically, the inverse-kinematics model calculates the finger-joint commands needed to place the fingertips at specified positions in a Cartesian reference frame that is attached to the palm of the robot hand and that nominally corresponds to the Cartesian reference frame attached to the palm of the glove. Initially, because of the aforementioned uncertainties, the glove fingertip positions calculated by the forwardkinematics model in the glove Cartesian reference frame cannot be expected to match the robot fingertip positions in the robot-hand Cartesian reference frame. A calibration must be performed to make the glove and robot-hand fingertip positions correspond more precisely. The calibration procedure involves a few simple hand poses designed to provide well-defined fingertip positions. One of the poses is a fist. In each of the other poses, a finger touches the thumb. The calibration subalgorithm uses the sensor readings from these poses to modify the kinematical models to make the two sets of fingertip positions agree more closely.

Goza, Michael↗

DOE OSTI2024

Zero Power Reactor Database (ZPRD) Development Plan

Past sodium-cooled fast reactors (SFR) were built with an active experimental program in place to support the design and development work. Most of the experimental facilities in the United States that were important for SFR design were shutdown in the 1980s and 1990s. Reactor licensing and construction requires any reactor design to be verified against existing reactor facilities or experimental measurements. With the absence of those experimental facilities, modern SFR projects must rely on historical measurements to demonstrate that the engineering modeling software and data being used for the new reactor design work are reliable. There has been a considerable push in the last 6 years by both DOE and commercial companies to obtain historical experimental measurements that are relevant for SFRs, in particular those with features that are important for the new reactor designs of interest. The zero power reactor experiments carried out at Argonne National Laboratory’s critical facilities (ZPR-3, ZPR-6, ZPR-9, and ZPPR) from the 1950s to the 1980s are some of the best reactor physics experiments on SFR technology that are available today. Of particular interest today are the ZPPR-15 measurements done at the ZPPR facility for the Integral Fast Reactor project in the 1980s as they are in line with most commercial and DOE interests today. In the past 10 years, the measurements done on ZPPR-15 have been processed into both Monte Carlo (MCNP) and deterministic models (MC2-3 and DIF3D) useable for validating the engineering modeling software for key parts of the SFR design work. To achieve this, a detailed model description must be created for the experiment and the experimental measurement that the engineering modeling software is to reproduce. Then, an assessment of the uncertainty on the measured quantity which considers all of the sources of uncertainty in defining the model must be obtained and documented. The models created for ZPPR-15 provide the best validation basis available today for neutronics modeling software. Reference 2 is a good resource to understand how these models were built and how the uncertainties on the measured quantities were derived. The intention of the Zero Power Reactor Database (ZPRD), hosted at frdb.ne.anl.gov, is to make available the experimental measurements and models that have been constructed to-date. Though ZPPR-15 measurements are the primary data requested for validation needs, other measurements on ZPPR, ZPR-6, and ZPR-9 in support of the Clinch River Breeder Reactor (CRBR) and Fast Test Reactor (FFTF) should also be considered important for future software validation needs. In this manuscript, the details of available measurements on ZPR-3, ZPR-6, ZPR-9, and ZPPR facilities are summarized, and a general organization of the web interface is displayed. Many of the documents associated with the measurements are export controlled information so access to the database will also have to be controlled.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗

NASA NTRS1989

Measurement of hand dynamics in a microsurgery environment: Preliminary data in the design of a bimanual telemicro-operation test bed

Data describing the microsurgeon's hand dynamics was recorded and analyzed in order to provide an accurate model for the telemicrosurgery application of the Bimanual Telemicro-operation Test Bed. The model, in turn, will guide the development of algorithms for the control of robotic systems in bimanual telemicro-operation tasks. Measurements were made at the hand-tool interface and include position, acceleration and force between the tool-finger interface. Position information was captured using an orthogonal pulsed magnetic field positioning system resulting in measurements in all six degrees-of-freedom (DOF). Acceleration data at the hands was obtained using accelerometers positioned in a triaxial arrangement on the back of the hand allowing measurements in all three cartesian-coordinate axes. Force data was obtained by using miniature load cells positioned between the tool and the finger and included those forces experienced perpendicular to the tool shaft and those transferred from the tool-tissue site. Position data will provide a minimum/maximum reference frame for the robotic system's work space or envelope. Acceleration data will define the response times needed by the robotic system in order to emulate and subsequently outperform the human operator's tool movements. The force measurements will aid in designing a force-reflective, force-scaling system as well as defining the range of forces the robotic system will encounter. All analog data was acquired by a 16-channel analog-to-digital conversion system residing in a IBM PC/AT-compatible computer at the Center's laboratory. The same system was also used to analyze and present the data.

Charles, Steve↗

DOE OSTI2025

Validation and Calibration of Energy Models with Real Vehicle Data from Chassis Dynamometer Experiments

Accurate estimation of vehicle fuel consumption typically requires detailed modeling of complex internal powertrain dynamics, often resulting in computationally intensive simulations. However, many transportation applications-such as traffic flow modeling, optimization, and control-require simplified models that are fast, interpretable, and easy to implement, while still maintaining fidelity to physical energy behavior. This work builds upon a recently developed model reduction pipeline that derives physics-like energy models from high-fidelity Autonomie vehicle simulations. These reduced models preserve essential vehicle dynamics, enabling realistic fuel consumption estimation with minimal computational overhead. While the reduced models have demonstrated strong agreement with their Autonomie counterparts, previous validation efforts have been confined to simulation environments. This study extends the validation by comparing the reduced energy model's outputs against real-world vehicle data. Focusing on the MidSUV category, we tune the baseline Autonomie model to closely replicate the characteristics of a Toyota RAV4. We then assess the accuracy of the resulting reduced model in estimating fuel consumption under actual drive conditions. Our findings suggest that, when the reference Autonomie model is properly calibrated, the simplified model produced by the reduction pipeline can provide reliable, semi-principled fuel rate estimates suitable for large-scale transportation applications.

42 ENGINEERING↗

DOE OSTI2026

Enter the AHU (36th Chamber of ASHRAE): A Multi-site Field Study of ASHRAE G36

Despite being recognized as the best practice for advanced building controls, ASHRAE Guideline 36 (G36) has seen slow adoption in retrofit cases. Decisionmakers lack credible field evidence to justify the time and person-power investment. Most prior analyses have relied on software simulations, which overlook implementation challenges and fail to persuade owners to move from models to real-world deployment. This paper presents a multi-site field study of G36 performance, drawing on measured results from 17 projects across diverse building types and climate zones. The analysis disaggregates outcomes by the most widely adopted air handling unit (AHU) based G36 strategies, including trim-and-respond approaches to supply air temperature (SAT) and duct static pressure (DSP) reset, and economizer controls. Results for controls re programming implementations are encouraging, with HVAC savings ranging from 2% - 49%, with a median of 18%. The range aligns with simulation study findings showing 1% - 46% savings through these three strategies. These findings show that even without capital investment in control infrastructure upgrades, existing building owners are reaping significant benefits from updating HVAC sequences of operation to industry best-practice solutions. The results give practitioners and decisionmakers a reference point on what to expect, which strategies deliver, and how field performance may compare to simulation.

Deshpande, Reva↗

NASA NTRS2022

A Unifying View of Estimation and Control Using Belief Propagation With Application to Path Planning

The use of estimation techniques on stochastic models to solve control problems is an emerging paradigm that falls under the rubric of Active Inference (AI) and Control as Inference (CAI). In this work, we use probability propagation on factor graphs to show that various algorithms proposed in the literature can be seen as specific composition rules in a factor graph. We show how this unified approach, presented both in probability space and in log of the probability space, provides a very general framework that includes the Sum-product, the Max-product, Dynamic programming and mixed Reward/Entropy criteria-based algorithms. The framework also expands algorithmic design options that lead to new smoother or sharper policy distributions. We propose original recursions such as: a generalized Sum/Max-product algorithm, a Smooth Dynamic programming algorithm and a modified versions of the Reward/Entropy algorithm. The discussion is carried over with reference to a path planning problem where the recursions that arise from various cost functions, although they may appear similar in scope, bear noticeable differences. We provide a comprehensive table of composition rules and a comparison through simulations, first on a synthetic small grid with a single goal with obstacles, and then on a grid extrapolated from a real-world scene with multiple goals and a semantic map.

Francesco A. N. Palmieri↗

NASA NTRS2002

On Polymorphic Circuits and Their Design Using Evolutionary Algorithms

This paper introduces the concept of polymorphic electronics (polytronics) - referring to electronics with superimposed built-in functionality. A function change does not require switches/reconfiguration as in traditional approaches. Instead the change comes from modifications in the characteristics of devices involved in the circuit, in response to controls such as temperature, power supply voltage (VDD), control signals, light, etc. The paper illustrates polytronic circuits in which the control is done by temperature, morphing signals, and VDD respectively. Polytronic circuits are obtained by evolutionary design/evolvable hardware techniques. These techniques are ideal for the polytronics design, a new area that lacks design guidelines, know-how,- yet the requirements/objectives are easy to specify and test. The circuits are evolved/synthesized in two different modes. The first mode explores an unstructured space, in which transistors can be interconnected freely in any arrangement (in simulations only). The second mode uses a Field Programmable Transistor Array (FPTA) model, and the circuit topology is sought as a mapping onto a programmable architecture (these experiments are performed both in simulations and on FPTA chips). The experiments demonstrated the synthesis. of polytronic circuits by evolution. The capacity of storing/hiding "extra" functions provides for watermark/invisible functionality, thus polytronics may find uses in intelligence/security applications.

Stoica, Adrian↗

NASA NTRS2015

Static Aeroelastic Modeling of a Sub-Scale Wind Tunnel Model with Novel Flap Concept

This paper examines the static aeroelastic modeling of a flexible wind tunnel model equipped with a novel control effector known as the Variable Camber Continuous Trailing Edge Flap (VCCTEF) system. The wind tunnel model is an approximately 10 percent sub-scale version of a wing designed to be equipped on a commerical transport aircraft such as the full-scale NASA Generic Transport Model (GTM). The structure of the model is made highly flexible such that a 10 percent of semi-span wing tip deflection is expected at a design lift condition, and a representation of the VCCTEF concept is incorporated on the model. Static aeroelastic modeling is conducted by using a representative single beam structural finite-element model coupled to a vortex-lattice aerodynamic model. The resulting aeroelastic model of the flexible wind tunnel model is compared against experimental wind tunnel test results from the actual model tested at the University of Washington Aeronautical Laboratory (UWAL). Comparison cases are made using reference and VICON measurements of VCCTEF flap deflections from the wind tunnel test data, and percent errors between the lift curve parameters are presented. The results show up to a maximum percent of error of <10% with regards to lift curve slope values between the aeroelastic model and the UWAL test data. The agreement of lift curve slope values from the aeroelastic model and experimental test results serves as validation for the coupled vortex-lattice finite-element static aeroelastic model.

Aeroelasticity↗

NASA NTRSDate not supplied

Advancements in Lunar Dust Mitigation and Leveraging the Contamination Control Community

Lunar Dust Contamination: As we get closer to putting landers, rovers, payloads, and astronauts on the Moon, dust mitigation is becoming more critical than ever. To address the many challenges of dust mitigation on the surface, engineers and scientists are looking at various technologies that may have relevance for the contamination control community. This includes active, passive, architectural, and operational solutions. There have been advances in coatings for dust mitigation at NASA and through commercial partnerships (see Wiesner, Wohl, et al). There have been advancements to previous solutions as well as novel solutions to address dust mitigation on the surface and on-orbit. There have been several efforts to better understand the effectiveness of various terrestrial and dust mitigation solutions with much of this data published in recent years. In the area of dust mitigation, dust contamination or infiltration refers to the impingement or contact of planetary dust with items not normally dusty and whose operation may therefore be compromised. Recent Advancements in Dust Mitigation: Dust mitigation strategies are becoming more common for hardware heading to the Moon, with most major programs implementing lunar dust contamination requirements for human health and equipment. In the last couple of years, NASA has created more guidance to assist hardware developers with dust mitigation. This includes NASA-STD-1008 (Classifications and Requirements for Testing Systems and Hardware to be Exposed to Dust in Planetary) and NASA TP 20220018746 (Lunar Dust Mitigation Guide and Reference). Several projects are testing hardware in relevant dusty environments to understand their tolerance to lunar dust. There has been a significant increase in the amount of testing with lunar simulants, with the NASA Simulant Advisory Committee serving a resource for NASA and our partners. There are new solutions for detecting and monitoring the dust internally and externally. Learning from the contamination control community: In addition to sharing potentially relevant solutions at the workshop, the dust mitigation community is also interested in learning from the contamination control community on ways to address lunar dust. For instance, a major aspect of managing the dust is monitoring and detection. This is critical for understanding when maintenance is needed and to inform future missions. Another challenge of working with lunar dust is quantifying and characterizing the smallest of particles. There are also challenges in modeling lunar dust and its behavior. The dust mitigation community can also leverage lessons learned from the planetary protection community.

dust mitagation↗

DOE OSTI2024

Modeling powder spreadability in powder-based processes using the discrete element method

Powder-bed fusion (PBF) processes refer to a subset of Additive Manufacturing (AM) techniques where powder is spread on the build-plate before melting (by a laser or electron beam). While PBF processes are attractive due to their ability for realizing complex structures that are either difficult or impossible to create through conventional means, the parts fabricated with these techniques can exhibit defects such as pores, inclusions, and excessive surface roughness. To minimize these defects, much research has been dedicated towards process maturation by optimizing laser or electron beam parameters. However, these developmental efforts typically do not address the recoating process where achieving dense and uniform layers of powder is a necessity for ensuring process repeatability and part quality. While the recoating process can be studied through experimentation, the dynamics of particle movement are difficult to analyze experimentally. Therefore, here, in this study, powder spreading in PBF was simulated through the Discrete Element Method (DEM) to elucidate the mechanisms that control powder-bed quality. Utilizing the Buckingham Pi theorem, a dimensionless metric referred to as the spreading index is developed that combines powder-bed density, roughness, and particle size to assess the quality of powder layers. The formulated spreading index is then related to several dimensionless quantities that provide insight into the mechanisms dominating powder spreading in PBF. The DEM simulations conducted in this work focused on the scenario where powder is spread onto an existing powder bed and revealed that a reduction in the recoating velocity causes an increase in the spreading index while little to no impact on the spreading index was observed when varying layer thickness from 30 μm to 75 μm.Particle size effects on the powder-bed quality were also investigated.

36 MATERIALS SCIENCE↗

NASA NTRSDate not supplied

Development of Urban Air Mobility (UAM) Vehicles for Ease of Operation

To date the air transportation system has been developed with the in-cremental introduction of new technology and with highly experienced air transport pilots and air traffic controllers overseeing flight operations. Thus, we currently have one of the safest commercial aviation systems in the world. General Aviation (GA) in the United States, however, has not always followed the same cautious and monitored approach to implementation; consequently, the GA safety record does not meet the high standards of commercial aviation. Recently, a new system known as Urban Air Mobility (UAM), is attracting considerable interest and investment from industry and government agencies. UAM refers to a system of passenger and small-cargo air transportation vehicles within an urban area with the goal of reducing the number of times we need to use our cars, thus improving urban traffic by moving people and cargo from crowded single pas-senger vehicles on our roads to personal and on-demand air vehicles. These UAM vehicles will be small and based on electric, Vertical-Take-Off-and-Landing (eV-TOL) systems. A significant component of UAM is offloading of flight-man-agement responsibilities from human pilots to newly-developed autonomy. Cur-rently, over 100 UAM vehicles are either in development or production. Most, if not all, have a goal of fully autonomous vehicle operations, but fully autonomous flying vehicles are not expected in the near future. Therefore, we are de-veloping concepts for UAM vehicles that will be easy to fly and/or manage by operators with minimal pilot training. In this paper we will discuss our human-automation teaming approach to develop an easy-to-operate VTOL aircraft, and some of the fly-by-wire technology needed to stabilize the vehicle so that a sim-ple ecological mental model of the flying task can be implemented. We will discuss the requirements for a stability augmentation system that must be developed to support our simple pilot input model, and also present design guidelines and requirements based on a pilot input and management model. Finally, our ap-proach to vehicle development will involve considerable operator testing and evaluation: improving pilot model, inceptors, displays and also work on a plan for how a UAM vehicle can be integrated with terminal area air traffic control airspace with minimal impact on controller workload.

UAM↗

NASA NTRSDate not supplied

A Novel Bone Marrow Single Cell Atlas of Mechanotransduction in Microgravity, Normal Gravity, Exercise, and Hindlimb Unloading for WT and CDKN1A-Null Regenerative Mice

Mechanical loading of adult stem cell progenitors is a key factor in modulating their proliferation, differentiation, and tissue regenerative potential, with loading generally promoting tissue formation and unloading mediating tissue loss. To address the role of mechanotransduction in maintaining stem cell-based tissue regenerative health, we generated a single cell transcriptomic atlas mapping responses of femur bone marrow mesenchymal and hematopoietic lineages to a range of altered mechanical loading conditions, both in WT B1629SF2/J mice as well as the p21/CDKN1A-null regenerative mice. We selected the femur marrow compartment as a model because of the diversity and high numbers of stem cell progenitor stages present, and because it undergoes static and cyclic hydrostatic pressure loading associated with weight-bearing and ambulation. Our study included normally loaded mice at 1g, unloading in microgravity during spaceflight as well as tail suspension hindlimb unloading, and voluntary running wheel exercise. Basal, one-year natural aging, and habitat controls were also conducted. Overall, the atlas encompasses 18 different experimental conditions with N=3 mice per condition and includes more than 500,000 single cell expressomes. Key specific findings include: increased mature reticulocyte populations in aging and unloading mice compared to active mice; greater hematopoietic and mesenchymal differentiating progenitors cluster identification in CDKN1A-null samples, especially the exercise model; distinct pseudotime cell trajectory shifts in the hematopoietic lineage for spaceflight and unloaded mice; and shifts in monocytic cell populations toward osteoclastic bone degenerative lineages in unloading and spaceflight samples. Overall mechanical loading shows increased marrow progenitor population differentiation while unloading is associated with increased CDKN1A expression and maintenance of marrow population stemness. Deletion of CDKN1A appears to remove a negative check on progenitor lineages commitment to differentiation. Finally, the Bone Marrow Mechanotransduction Single Cell Atlas will serve as a reference tool for studying marrow regenerative responses across a range of mechanical environments as they relate to CDKN1A status.

single-cell↗