Single-cell multi-ome and immune profiles of the Inspiration4 crew reveal conserved, cell-type, and sex-specific responses to spaceflight.

Spaceflight induces an immune response in astronauts. To better characterize this effect, we generated single-cell, multi-ome, cell-free RNA (cfRNA), biochemical, and hematology data for the SpaceX Inspiration4 (I4) mission crew. We found that 18 cytokines/chemokines related to inflammation, aging, and muscle homeostasis changed after spaceflight. In I4 single-cell multi-omics data, we identified a "spaceflight signature" of gene expression characterized by enrichment in oxidative phosphorylation, UV response, immune function, and TCF21 pathways. We confirmed the presence of this signature in independent datasets, including the NASA Twins Study, the I4 skin spatial transcriptomics, and 817 NASA GeneLab mouse transcriptomes. Finally, we observed that (1) T cells showed an up-regulation of FOXP3, (2) MHC class I genes exhibited long-term suppression, and (3) infection-related immune pathways were associated with microbiome shifts. In summary, this study reveals conserved and distinct immune disruptions occurring and details a roadmap for potential countermeasures to preserve astronaut health.

The spatial-temporal dynamics of pulmonary blood flow are altered in pulmonary arterial hypertension.

Global fluctuation dispersion (FDglobal), a spatial-temporal metric derived from serial images of the pulmonary perfusion obtained with MRI-arterial spin labeling, describes temporal fluctuations in the spatial distribution of perfusion. In healthy subjects, FDglobal is increased by hyperoxia, hypoxia, and inhaled nitric oxide. We evaluated patients with pulmonary arterial hypertension (PAH, 4F, aged 47 ± 15, mean pulmonary artery pressure 48 ± 7 mmHg) and healthy controls (CON, 7F, aged 47 ± 12) to test the hypothesis that FDglobal is increased in PAH. Images were acquired at ∼4-5 s intervals during voluntary respiratory gating, inspected for quality, registered using a deformable registration algorithm, and normalized. Spatial relative dispersion (RD = SD/mean) and the percent of the lung image with no measurable perfusion signal (%NMP) were also assessed. FDglobal was significantly increased in PAH (PAH = 0.40 ± 0.17, CON = 0.17 ± 0.02, P = 0.006, a 135% increase) with no overlap in values between the two groups, consistent with altered vascular regulation. Both spatial RD and %NMP were also markedly greater in PAH vs. CON (PAH RD = 1.46 ± 0.24, CON = 0.90 ± 0.10, P = 0.0004; PAH NMP = 13.4 ± 6.1%; CON = 2.3 ± 1.4%, P = 0.001 respectively) consistent with vascular remodeling resulting in poorly perfused regions of lung and increased spatial heterogeneity. The difference in FDglobal between normal subjects and patients with PAH in this small cohort suggests that spatial-temporal imaging of perfusion may be useful in the evaluation of patients with PAH. Since this MR imaging technique uses no injected contrast agents and has no ionizing radiation it may be suitable for use in diverse patient populations.NEW & NOTEWORTHY Using proton MRI-arterial spin labeling to obtain serial images of pulmonary perfusion, we show that global fluctuation dispersion (FDglobal), a metric of temporal fluctuations in the spatial distribution of perfusion, was significantly increased in female patients with pulmonary arterial hypertension (PAH) compared with healthy controls. This potentially indicates pulmonary vascular dysregulation. Dynamic measures using proton MRI may provide new tools for evaluating individuals at risk of PAH or for monitoring therapy in patients with PAH.

Adapting Disease Prevention Protocols for Human Spaceflight During COVID-19.

BACKGROUND: The National Aeronautics and Space Administration (NASA) Flight Crew Health Stabilization Program (HSP) was historically implemented to minimize infectious disease transmission to astronauts in the immediate prelaunch period. The first ever commercial application and adaptation of the NASA HSP was implemented during the Crew Demo-2 mission in the time of the Coronavirus disease 2019 (COVID-19) pandemic. This article details and discusses the first commercial implementation and adaptation of the HSP prior to the Crew Demo-2 launch.METHODS: This is a retrospective descriptive analysis of the application of NASA disease prevention protocols for human spaceflight during the COVID-19 pandemic. In the context of the pandemic, extra precautions added to the HSP included daily symptom surveys completed by Primary Contacts of the crew, COVID-19 RT-PCR testing, and improved quarantine protocols.RESULTS: Of the 91 SpaceX Primary Contacts who completed a total of 2720 daily symptom surveys prior to launch, 22 individuals (24.2) and 198 surveys (7.3) returned positive for potential symptoms of COVID-19. Two individuals were removed due to symptoms indistinguishable from COVID-19. Through this survey, systematic quarantine, and PCR testing, the Crew Demo-2 mission was successful with no known infectious diseases transmitted.CONCLUSIONS: Overall, the commercial implementation of the NASA Health Stabilization Program by SpaceX with adjustments required during the COVID-19 pandemic was a success, with protocols allowing identification and removal of potentially infectious persons from the program. The principles of the HSP may provide an adequate infectious disease playbook for commercial spaceflight operations going forward.Petersen E, Pattarini JM, Mulcahy RA, Beger SB, Mitchell MR, Hu YD, Middleton KN, Frazier W, Mormann B, Esparza H, Asadi A, Musk ER, Alter G, Nilles E, Menon AS. Adapting disease prevention protocols for human spaceflight during COVID-19. Aerosp Med Hum Perform. 2021; 92(7):597602.

Regional pulmonary perfusion patterns in humans are not significantly altered by inspiratory hypercapnia.

Pulmonary vascular tone is known to be sensitive to both local alveolar Po2 and Pco2. Although the effects of hypoxia are well studied, the hypercapnic response is relatively less understood. We assessed changes in regional pulmonary blood flow in humans in response to hypercapnia using previously developed MRI techniques. Dynamic measures of blood flow were made in a single slice of the right lung of seven healthy volunteers following a block-stimulus paradigm (baseline, challenge, recovery), with CO2 added to inspired gas during the challenge block to effect a 7-Torr increase in end-tidal CO2. Effects of hypercapnia on blood flow were evaluated based on changes in spatiotemporal variability (fluctuation dispersion, FD) and in regional perfusion patterns in comparison to hypoxic effects previously studied. Hypercapnia increased FD 2.5% from baseline (relative to control), which was not statistically significant (P = 0.07). Regional perfusion patterns were not significantly changed as a result of increased FICO2 (P = 0.90). Reanalysis of previously collected data using a similar protocol but with the physiological challenge replaced by decreased FIO2 (FIO2 = 0.125) showed marked flow redistribution (P = 0.01) with the suggestion of a gravitational pattern, demonstrating hypoxia has the ability to affect regional change with a global stimulus. Taken together, these data indicate that hypercapnia of this magnitude does not lead to appreciable changes in the distribution of pulmonary perfusion, and that this may represent an interesting distinction between the hypoxic and hypercapnic regulatory response.NEW & NOTEWORTHY Although it is well known that the pulmonary circulation responds to local alveolar hypoxia, and that this mechanism may facilitate ventilation-perfusion matching, the relative role of CO2 is not well appreciated. This study demonstrates that an inspiratory hypercapnic stimulus is significantly less effective at inducing changes in pulmonary perfusion patterns than inspiratory hypoxia, suggesting that in these circumstances hypercapnia is not sufficient to induce substantial integrated feedback control of ventilation-perfusion mismatch across the lung.

Susceptibility to high-altitude pulmonary edema is associated with a more uniform distribution of regional specific ventilation.

High-altitude pulmonary edema (HAPE) is a potentially fatal condition affecting high-altitude sojourners. The biggest predictor of HAPE development is a history of prior HAPE. Magnetic resonance imaging (MRI) shows that HAPE-susceptible (with a history of HAPE), but not HAPE-resistant (with a history of repeated ascents without illness) individuals develop greater heterogeneity of regional pulmonary perfusion breathing hypoxic gas (O2 = 12.5%), consistent with uneven hypoxic pulmonary vasoconstriction (HPV). Why HPV is uneven in HAPE-susceptible individuals is unknown but may arise from regionally heterogeneous ventilation resulting in an uneven stimulus to HPV. We tested the hypothesis that ventilation is more heterogeneous in HAPE-susceptible subjects (n = 6) compared with HAPE-resistant controls (n = 7). MRI specific ventilation imaging (SVI) was used to measure regional specific ventilation and the relative dispersion (SD/mean) of SVI used to quantify baseline heterogeneity. Ventilation heterogeneity from conductive and respiratory airways was measured in normoxia and hypoxia (O2 = 12.5%) using multiple-breath washout and heterogeneity quantified from the indexes Scond and Sacin, respectively. Contrary to our hypothesis, HAPE-susceptible subjects had significantly lower relative dispersion of specific ventilation than the HAPE-resistant controls [susceptible = 1.33 ± 0.67 (SD), resistant = 2.36 ± 0.98, P = 0.05], and Sacin tended to be more uniform (susceptible = 0.085 ± 0.009, resistant = 0.113 ± 0.030, P = 0.07). Scond was not significantly different between groups (susceptible = 0.019 ± 0.007, resistant = 0.020 ± 0.004, P = 0.67). Sacin and Scond did not change significantly in hypoxia (P = 0.56 and 0.19, respectively). In conclusion, ventilation heterogeneity does not change with short-term hypoxia irrespective of HAPE susceptibility, and lesser rather than greater ventilation heterogeneity is observed in HAPE-susceptible subjects. This suggests that the basis for uneven HPV in HAPE involves vascular phenomena.NEW & NOTEWORTHY Uneven hypoxic pulmonary vasoconstriction (HPV) is thought to incite high-altitude pulmonary edema (HAPE). We evaluated whether greater heterogeneity of ventilation is also a feature of HAPE-susceptible subjects compared with HAPE-resistant subjects. Contrary to our hypothesis, ventilation heterogeneity was less in HAPE-susceptible subjects and unaffected by hypoxia, suggesting a vascular basis for uneven HPV.

A statistical clustering approach to discriminating perfusion from conduit vessel signal contributions in a pulmonary ASL MR image.

The measurement of pulmonary perfusion (blood delivered to the capillary bed within a voxel) using arterial spin labeling (ASL) magnetic resonance imaging is often complicated by signal artifacts from conduit vessels that carry blood destined for voxels at a distant location in the lung. One approach to dealing with conduit vessel contributions involves the application of an absolute threshold on the ASL signal. While useful for identifying a subset of the most dominant high signal conduit image features, signal thresholding cannot discriminate between perfusion and conduit vessel contributions at intermediate and low signal. As an alternative, this article discusses a data-driven statistical approach based on statistical clustering for characterizing and discriminating between capillary perfusion and conduit vessel contributions over the full signal spectrum. An ASL flow image is constructed from the difference between a pair of tagged magnetic resonance images. However, when viewed as a bivariate projection that treats the image pair as independent measures (rather than the univariate quantity that results from the subtraction of the two images), the signal associated with capillary perfusion contributions is observed to cluster independently of the signal associated with conduit vessel contributions. Analyzing the observed clusters using a Gaussian mixture model makes it possible to discriminate between conduit vessel and capillary-perfusion-dominated signal contributions over the full signal spectrum of the ASL image. As a demonstration of feasibility, this study compares the proposed clustering approach with the standard absolute signal threshold strategy in a small number of test images.

Rapid Prototyping of Inspired Gas Delivery System for Pulmonary MRI Research.

Specific ventilation imaging (SVI) is a noninvasive magnetic resonance imaging (MRI)-based method for determining the regional distribution of inspired air in the lungs, useful for the assessment of pulmonary function in medical research. This technique works by monitoring the rate of magnetic resonance signal change in response to a series of imposed step changes in inspired oxygen concentration. The current SVI technique requires a complex system of tubes, valves, and electronics that are used to supply and rapidly switch inspired gases while subjects are imaged, which makes the technique difficult to translate into the clinical setting. This report discusses the design and implementation of custom three-dimensional (3D) printed hardware that greatly simplifies SVI measurement of lung function. Several hardware prototypes were modeled using computer-aided design software and printed for evaluation. After finalization of the design, the new delivery system was evaluated based on O2 and N2 concentration step responses and validated against the current SVI protocol. The design performed rapid switching of supplied gas within 250 ms and consistently supplied the desired concentration of O2 during operation. It features a reduction in the number of commercial hardware components, from five to one, and a reduction in the number of gas lines between the operator's room and the scanner room, from four to one, as well as a substantially reduced preparation time from 25 to 5 min. 3D printing is well suited to the design of inexpensive custom MRI compatible hardware, making it particularly useful in imaging-based research.

Inhaled nitric oxide alters the distribution of blood flow in the healthy human lung, suggesting active hypoxic pulmonary vasoconstriction in normoxia.

Hypoxic pulmonary vasoconstriction (HPV) is thought to actively regulate ventilation-perfusion (V̇a/Q̇) matching, reducing perfusion in regions of alveolar hypoxia. We assessed the extent of HPV in the healthy human lung using inhaled nitric oxide (iNO) under inspired oxygen fractions (FiO2 ) of 0.125, 0.21, and 0.30 (a hyperoxic stimulus designed to abolish HPV without the development of atelectasis). Dynamic measures of blood flow were made in a single sagittal slice of the right lung of five healthy male subjects using an arterial spin labeling (ASL) MRI sequence, following a block stimulus pattern (3 × 60 breaths) with 40 ppm iNO administered in the central block. The overall spatial heterogeneity, spatiotemporal variability, and regional pattern of pulmonary blood flow was quantified as a function of condition (FiO2 × iNO state). While spatial heterogeneity did not change significantly with iNO administration or FiO2 , there were statistically significant increases in Global Fluctuation Dispersion, (a marker of spatiotemporal flow variability) when iNO was administered during hypoxia (5.4 percentage point increase, P = 0.003). iNO had an effect on regional blood flow that was FiO2 dependent (P = 0.02), with regional changes in the pattern of blood flow occurring in hypoxia (P = 0.007) and normoxia (P = 0.008) tending to increase flow to dependent lung at the expense of nondependent lung. These findings indicate that inhaled nitric oxide significantly alters the distribution of blood flow in both hypoxic and normoxic healthy subjects, and suggests that some baseline HPV may indeed be present in the normoxic lung.

Validating the distribution of specific ventilation in healthy humans measured using proton MR imaging.

Specific ventilation imaging (SVI) uses proton MRI to quantitatively map the distribution of specific ventilation (SV) in the human lung, using inhaled oxygen as a contrast agent. To validate this recent technique, we compared the quantitative measures of heterogeneity of the SV distribution in a 15-mm sagittal slice of lung obtained in 10 healthy supine subjects, (age 37 ± 10 yr, forced expiratory volume in 1 s 97 ± 7% predicted) using SVI to those obtained in the whole lung from multiple-breath nitrogen washout (MBW). Using the analysis of Lewis et al. (Lewis SM, Evans JW, Jalowayski AA. J App Physiol 44: 416-423, 1978), the most likely distribution of SV from the MBW data was computed and compared with the distribution of SV obtained from SVI, after normalizing for the difference in tidal volume. The average SV was 0.30 ± 0.10 MBW, compared with 0.36 ± 0.10 SVI (P = 0.01). The width of the distribution, a measure of the heterogeneity, obtained using both methods was comparable: 0.51 ± 0.06 and 0.47 ± 0.08 in MBW and SVI, respectively (P = 0.15). The MBW estimated width of the SV distribution was 0.05 (10.7%) higher than that estimated using SVI, and smaller than the intertest variability of the MBW estimation [inter-MBW (SD) for the width of the SV distribution was 0.08 (15.8)%]. To assess reliability, SVI was performed twice on 13 subjects showing small differences between measurements of SV heterogeneity (typical error 0.05, 12%). In conclusion, quantitative estimations of SV heterogeneity from SVI are reliable and similar to those obtained using MBW, with SVI providing spatial information that is absent in MBW.

Spatial-temporal dynamics of pulmonary blood flow in the healthy human lung in response to altered FI(O2).

The temporal dynamics of blood flow in the human lung have been largely unexplored due to the lack of appropriate technology. Using the magnetic resonance imaging method of arterial spin labeling (ASL) with subject-gated breathing, we produced a dynamic series of flow-weighted images in a single sagittal slice of the right lung with a spatial resolution of ~1 cm(3) and a temporal resolution of ~10 s. The mean flow pattern determined from a set of reference images was removed to produce a time series of blood flow fluctuations. The fluctuation dispersion (FD), defined as the spatial standard deviation of each flow fluctuation map, was used to quantify the changes in distribution of flow in six healthy subjects in response to 100 breaths of hypoxia (FI(O(2)) = 0.125) or hyperoxia (FI(O(2)) = 1.0). Two reference frames were used in calculation, one determined from the initial set of images (FD(global)), and one determined from the mean of each corresponding baseline or challenge period (FD(local)). FD(local) thus represented changes in temporal variability as a result of intervention, whereas FD(global) encompasses both FD(local) and any generalized redistribution of flow associated with switching between two steady-state patterns. Hypoxic challenge resulted in a significant increase (96%, P < 0.001) in FD(global) from the normoxic control period and in FD(local) (46%, P = 0.0048), but there was no corresponding increase in spatial relative dispersion (spatial standard deviation of the images divided by the mean; 8%, not significant). There was a smaller increase in FD(global) in response to hyperoxia (47%, P = 0.0015) for the single slice, suggestive of a more general response of the pulmonary circulation to a change from normoxia to hyperoxia. These results clearly demonstrate a temporal change in the sampled distribution of pulmonary blood flow in response to hypoxia, which is not observed when considering only the relative dispersion of the spatial distribution.