Menu

 

Task Book: Biological & Physical Sciences Division and Human Research Program
Advanced Search     

Project Title:  Effects of Space Flights on the Proteome of Astronauts' Plasma Reduce
Images: icon  Fiscal Year: FY 2021 
Division: Human Research 
Research Discipline/Element:
HRP HHC:Human Health Countermeasures
Start Date: 04/01/2016  
End Date: 12/31/2021  
Task Last Updated: 04/09/2021 
Download report in PDF pdf
Principal Investigator/Affiliation:   Rithidech, Kanokporn  Ph.D. / State University New York at Stony Brook 
Address:  Department of Pathology 
BHS T9 Health Sciences Center 
Stony Brook , NY 11794-8691 
Email: Kanokporn.Rithidech@sbumed.org 
Phone: (631) 444-3446  
Congressional District:
Web:  
Organization Type: UNIVERSITY 
Organization Name: State University New York at Stony Brook 
Joint Agency:  
Comments:  
Co-Investigator(s)
Affiliation: 
Martin, Dwight  Ph.D. State University of New York, Stony Brook 
Project Information: Grant/Contract No. NNX16AH80G 
Responsible Center: NASA JSC 
Grant Monitor: Stenger, Michael  
Center Contact: 281-483-1311 
michael.b.stenger@nasa.gov 
Solicitation / Funding Source: 2014-15 HERO NNJ14ZSA001N-MIXEDTOPICS. Appendix E: Behavioral Health & Human Health Countermeasures Topics 
Grant/Contract No.: NNX16AH80G 
Project Type: FLIGHT 
Flight Program:  
TechPort: No 
No. of Post Docs:
No. of PhD Candidates:
No. of Master's Candidates:
No. of Bachelor's Candidates:
No. of PhD Degrees:
No. of Master's Degrees:
No. of Bachelor's Degrees:
Human Research Program Elements: (1) HHC:Human Health Countermeasures
Human Research Program Risks: (1) Immune:Risk of Adverse Health Event Due to Altered Immune Response (IRP Rev F)
Human Research Program Gaps: (1) IM-101:Evaluate the effects of deep-space radiation on immune dysfunction, as an additional hazard to the effects of psychophysiological stress and weightlessness (IRP Rev L)
(2) IM-102:Evaluate the effects of altered atmospheric conditions such as elevated CO2 levels or mildly hypoxic exploration atmosphere on immune dysfunction (IRP Rev L)
(3) IM-103:Evaluate for novel endogenous viral reactivation (IRP Rev L)
(4) IM-401:Test, optimize and validate nutrition-based preventive/mitigative countermeasures (IRP Rev L)
Flight Assignment/Project Notes: Flight definition

NOTE: End date changed to 12/31/2021 per NSSC information (Ed., 12/31/20)

NOTE: End date changed to 12/31/2020 per NSSC information (Ed., 6/12/20)

NOTE: End date changed to 3/31/2020 per NSSC information (Ed., 3/25/19)

Task Description: NOTE: This is an integrated project consisting of Dr. Brian Crucian's "Functional Immune Alterations, Latent Herpesvirus Reactivation, Physiological Stress, and Clinical Incidence Onboard the International Space Station" directed research; and Dr. Richard Simpson's "The Impact of an ISS Mission on the Anti-Viral and Functional Properties of NK-cells, T-cells, B-cells and Dendritic Cells," Dr. Kanokporn Rithidech's "Effects of Space Flights on the Proteome of Astronauts' Plasma," and Dr. Honglu Wu's "DNA Damage in the ISS Astronaut's Lymphocytes and Their Association with Stress-Induced Immune Dysfunction" solicited research.

Space flight results in exposure of astronauts to several stressors, such as space radiation, microgravity, and physiological stress, that could exacerbate the risks of adverse health effects. To protect astronauts, we must improve our understanding of molecular changes that influence immunological conditions associated with increased astronaut health risks. The in vivo response to the space environment is complex, involving multiple proteins associated with various signal transduction cascades, resulting in different outcomes. Molecular mechanisms responsible for such diverse consequences are poorly understood. It is, therefore, essential to characterize the protein signatures of responses to the space environment in blood plasma samples from astronauts, collected at pre-, in-, and post-flights. Such analyses should help to reveal a particular set of proteins causing adverse immunological changes and to develop methods that help to prevent, or at least to counteract, these effects.

In this flight definition project, we will use cutting age proteomic technology to determine protein alterations, qualitatively and quantitatively, in plasma samples collected from astronauts before, during, and after space flights. Our findings will help to provide an understanding of the time course and etiology of immune changes induced by the space environment. Furthermore, since pre- and post-flight samples, in addition to the in-flight samples, will be evaluated in the same astronaut, the direct effects of the space environment can be determined. Hence, our findings will provide high-priority and highly relevant information to NASA. We will further correlate protein expression profiles to the available data on immune dysfunction detected in each astronaut. This approach makes it possible to determine potential predictive biomarkers for space-flight-induced immune dysregulation. Consequently, effective countermeasures against such harmful effects of the space environment can be identified.

Research Impact/Earth Benefits: Our findings will deliver important information that should advance our understanding of the time course and etiology of immune changes induced by the space environment. Hence, our findings will provide high-priority and highly relevant information to NASA. Importantly, we will correlate protein expression profiles to the available data on immune dysfunction detected in each astronaut. This approach makes it possible to determine potential predictive biomarkers for space-flight-induced immune dysregulation. Such knowledge is important for the assessment of health risks and will facilitate the development of countermeasures that can help astronauts, space travelers, and people on Earth with the impairment of the immune system.

Task Progress & Bibliography Information FY2021 
Task Progress: This is an integrated project consisting of Dr. Brian Crucian's "Functional Immune Alterations, Latent Herpesvirus Reactivation, Physiological Stress, and Clinical Incidence Onboard the International Space Station" directed research; and Dr. Richard Simpson's "The Impact of an ISS Mission on the Anti-Viral and Functional Properties of NK-cells, T-cells, B-cells, and Dendritic Cells," Dr. Kanokporn Rithidech's "Effects of Space Flights on the Proteome of Astronauts' Plasma," and Dr. Honglu Wu's "DNA Damage in the ISS Astronaut's Lymphocytes and Their Association with Stress-Induced Immune Dysfunction" solicited research.

Our contribution to this Flight Definition is to identify and characterize plasma proteins in the blood plasma of astronauts that can be used as predictive biomarkers of immunological dysfunction due to space flight. Specifically, we characterize the proteome of blood plasma collected from the same astronauts at different times, i.e. pre-, in-, and post-flight.

Previously, we analyzed the data by comparing pre-vs-in-flight and in-vs-post-flight separately. During the past few months, we analyzed the data by studying the temporal changes of each protein in samples collected at pre, in-, and post-flight from the same astronauts. The results from this approach not only help us to understand the temporal changes of proteins during space flights but also are highly relevant to the effects of space flight on protein expression. Using this approach, we detected new low abundance proteins with altered expression levels that have never been reported.

Our data demonstrate that there are 19 proteins with significant changes, i.e., increased or decreased, in expression levels due to space flight. Among the 19 proteins with significant changes in expression levels, there were seven proteins with significantly decreased levels. These include lumican (LUM), extracellular matrix protein 1 (ECM1), vitronectin (VTN), filaggrin-2 (FLG2), ceruloplasmin (CP), Desmoplakin (DSP), CD5 antigen-like (CD5L). These proteins are involved in cell defense responses, inflammatory responses, and cell/tissue repair. Hence, a reduction in the expression levels of these proteins may be associated with the impairment of the immune system and the repair of damaged tissues. The deficiency of filaggrin-2 (FLG2) is involved in skin inflammation. Although further studies are needed, a reduction in the expression level of FLG2 may play a role in the occurrence of skin rashes in some astronauts. It is known that lumican (LUM) is a major keratan sulfate proteoglycan of the cornea responsible for circumferential growth, corneal transparency, epithelial cell migration, and tissue repair. Hence, LUM is critical in maintaining corneal clarity. Further, a loss of LUM expression is associated with corneal inflammation. Hence, persistent decreases in the level of LUM after space flight would impair the homeostasis of the eyes. Taken together, it is plausible to hypothesize that a reduction in LUM level may be associated with vision impairment that has been observed in many astronauts after space flight. Our findings warrant further investigation of the potential role of LUM in vision impairment detected in astronauts.

The levels of the remaining 12 proteins were significantly increased. These are antithrombin-II (SERPINC1), fibronectin (FN1), protein disulfide-isomerase A (PDIA 3), titin (TTN), tropomodulin (TMOD3), zyxin (ZYN), bridging integrator 2 (BIN2), apolipoprotein A-II (APOA2), platelet factor 4 variant (PF4V1), beta-Ala-His dipeptidase (CNDP1), alpha-2-HS-glycoprotein (AHSG), pigment epithelium-derived factor (SERPINF1). The majority of these proteins are involved in inflammatory responses, cytoskeleton organization, and aging. Increased expression levels of CNDP1 may be indicative of increased oxidative stress in the brain and the muscle since CNDP1 degrades carnosine and homocarnosine (proteins with anti-oxidative activity mostly concentrated in the brain and the muscle). The protein in the SERPIN (serine protease inhibitor) family, i.e., SERPINC1, and SERPINF1, is the majority of those with increased expression levels. The SERPIN protease inhibitors comprise a large family of molecules involved in inflammation, immune response, blood clotting, hormone transport, and complement activation, dementia, and tumorigenesis. Hence, our findings suggest that dysregulation of these proteins may affect cell/tissue integrity and homeostasis, leading to late occurring health risks.

Based on these findings, we are preparing a manuscript to be submitted for possible publication in Acta Astronautica. In addition to the preparation of a manuscript, we are currently using Orbitrap Fusion Lumos Tribrid MS coupled with Thermo Ultimate 3000 HPLC system to investigate the pattern of protein expression profiles of astronauts’ plasma. This strategy helps us to achieve more in-depth coverage of the proteome than the MudPIT. However, it was not available in our laboratory in the past. We are currently analyzing the data generated by this highly sensitive and accurate mass spectrometer system. We anticipate another publication from this analysis.

Bibliography Type: Description: (Last Updated: 01/01/2021)  Show Cumulative Bibliography Listing
 
 None in FY 2021
Project Title:  Effects of Space Flights on the Proteome of Astronauts' Plasma Reduce
Images: icon  Fiscal Year: FY 2020 
Division: Human Research 
Research Discipline/Element:
HRP HHC:Human Health Countermeasures
Start Date: 04/01/2016  
End Date: 12/31/2021  
Task Last Updated: 06/29/2020 
Download report in PDF pdf
Principal Investigator/Affiliation:   Rithidech, Kanokporn  Ph.D. / State University New York at Stony Brook 
Address:  Department of Pathology 
BHS T9 Health Sciences Center 
Stony Brook , NY 11794-8691 
Email: Kanokporn.Rithidech@sbumed.org 
Phone: (631) 444-3446  
Congressional District:
Web:  
Organization Type: UNIVERSITY 
Organization Name: State University New York at Stony Brook 
Joint Agency:  
Comments:  
Co-Investigator(s)
Affiliation: 
Martin, Dwight  Ph.D. State University of New York, Stony Brook 
Project Information: Grant/Contract No. NNX16AH80G 
Responsible Center: NASA JSC 
Grant Monitor: Norsk, Peter  
Center Contact:  
Peter.norsk@nasa.gov 
Solicitation / Funding Source: 2014-15 HERO NNJ14ZSA001N-MIXEDTOPICS. Appendix E: Behavioral Health & Human Health Countermeasures Topics 
Grant/Contract No.: NNX16AH80G 
Project Type: FLIGHT 
Flight Program:  
TechPort: No 
No. of Post Docs:
No. of PhD Candidates:
No. of Master's Candidates:
No. of Bachelor's Candidates:
No. of PhD Degrees:
No. of Master's Degrees:
No. of Bachelor's Degrees:
Human Research Program Elements: (1) HHC:Human Health Countermeasures
Human Research Program Risks: (1) Immune:Risk of Adverse Health Event Due to Altered Immune Response (IRP Rev F)
Human Research Program Gaps: (1) IM-101:Evaluate the effects of deep-space radiation on immune dysfunction, as an additional hazard to the effects of psychophysiological stress and weightlessness (IRP Rev L)
(2) IM-102:Evaluate the effects of altered atmospheric conditions such as elevated CO2 levels or mildly hypoxic exploration atmosphere on immune dysfunction (IRP Rev L)
(3) IM-103:Evaluate for novel endogenous viral reactivation (IRP Rev L)
(4) IM-401:Test, optimize and validate nutrition-based preventive/mitigative countermeasures (IRP Rev L)
Flight Assignment/Project Notes: Flight definition

NOTE: End date changed to 12/31/2021 per NSSC information (Ed., 12/31/20)

NOTE: End date changed to 12/31/2020 per NSSC information (Ed., 6/12/20)

NOTE: End date changed to 3/31/2020 per NSSC information (Ed., 3/25/19)

Task Description: NOTE: This is an integrated project consisting of Dr. Brian Crucian's "Functional Immune Alterations, Latent Herpesvirus Reactivation, Physiological Stress, and Clinical Incidence Onboard the International Space Station" directed research; and Dr. Richard Simpson's "The Impact of an ISS Mission on the Anti-Viral and Functional Properties of NK-cells, T-cells, B-cells and Dendritic Cells," Dr. Kanokporn Rithidech's "Effects of Space Flights on the Proteome of Astronauts' Plasma," and Dr. Honglu Wu's "DNA Damage in the ISS Astronaut's Lymphocytes and Their Association with Stress-Induced Immune Dysfunction" solicited research.

Space flight results in exposure of astronauts to several stressors, such as space radiation, microgravity, and physiological stress, that could exacerbate the risks of adverse health effects. To protect astronauts, we must improve our understanding of molecular changes that influence immunological conditions associated with increased astronaut health risks. The in vivo response to the space environment is complex, involving multiple proteins associated with various signal transduction cascades, resulting in different outcomes. Molecular mechanisms responsible for such diverse consequences are poorly understood. It is, therefore, essential to characterize the protein signatures of responses to the space environment in blood plasma samples from astronauts, collected at pre-, in-, and post-flights. Such analyses should help to reveal a particular set of proteins causing adverse immunological changes and to develop methods that help to prevent, or at least to counteract, these effects.

In this flight definition project, we will use cutting age proteomic technology to determine protein alterations, qualitatively and quantitatively, in plasma samples collected from astronauts before, during, and after space flights. Our findings will help to provide an understanding of the time course and etiology of immune changes induced by the space environment. Furthermore, since pre- and post-flight samples, in addition to the in-flight samples, will be evaluated in the same astronaut, the direct effects of the space environment can be determined. Hence, our findings will provide high-priority and highly relevant information to NASA. We will further correlate protein expression profiles to the available data on immune dysfunction detected in each astronaut. This approach makes it possible to determine potential predictive biomarkers for space-flight-induced immune dysregulation. Consequently, effective countermeasures against such harmful effects of the space environment can be identified.

Research Impact/Earth Benefits: Our findings will deliver important information that should advance our understanding of the time course and etiology of immune changes induced by the space environment. Hence, our findings will provide high-priority and highly relevant information to NASA. Importantly, we will correlate protein expression profiles to the available data on immune dysfunction detected in each astronaut. This approach makes it possible to determine potential predictive biomarkers for space-flight-induced immune dysregulation. Such knowledge is important for assessment of health risks and will facilitate the development of appropriate countermeasures that can help astronauts, space travelers, and people on Earth with the impairment of the immune system.

Task Progress & Bibliography Information FY2020 
Task Progress: The major goal of our Flight Definition is to identify and characterize plasma proteins in the blood plasma of astronauts that can be used as predictive biomarkers of immunological dysfunction due to space flight. Specifically, we characterize the proteome of blood plasma collected from the same astronauts at different times, i.e., pre-, in-, and post-flight.

In the last annual report, we reported that a total of 453 unique and non-redundant proteins were identified at >99% confidence. Changes in protein concentrations during the pre-, in-, and post-flight timeline were determined by Students’ t-test analysis of comparisons between groups (p<0.1 is considered significant). Our data demonstrate that there are 14 proteins with significant changes, i.e., increased or decreased, in expression levels in plasma samples collected in-flight as compared to those collected pre-flight. We also reported that there were 16 proteins identified with significant changes, i.e., increased in green or decreased in red, in expression levels found astronauts’ plasma collected post-flight, in relation to those collected in-flight. Last January, we presented our results at the Annual NASA Human Research Program (HRP) workshop in Galveston.

During the past year, we use Principal Component Analysis (PCA) to analyze these 453 unique and non-redundant proteins identified by LTQ Orbitrap XL Ion trap mass spectrometer. The PCA is frequently used in the global analysis of the “omic” datasets. It provides fully unsupervised information on the dominant directions of highest variability in the data and can, therefore, be used to investigate similarities between individual samples, or the formation of clusters. The PCA is a dimensionality reduction method used to reduce the dimensionality of large data sets by transforming a large set of variables into a smaller one while preserving as much information as possible. The new variables are called the principal components and they are linear combinations of the actual variables. The first principal component is a linear combination of all the actual variables that have maintained the greatest amount of variation. The second principal component is a linear combination of the remaining variables to give the second greatest amount of variation and this can continue for third, fourth, and so on components but usually the first and second components carry the most important information. A scatter plot of the first and second components (a score plot) will often display samples sharing similar characteristics being grouped together apart from samples with different characteristics (i.e., it shows clustering based on similarity). Another plot is also generated in this analysis called a loading plot. The loading plot shows how strongly each original variable influences the principal component. Often, most points in a loading plot will be clustered around the center and the outlier points may be associated with the variables making the largest contribution to the data variation. In summary, the PCA indicates the differences in the pattern of protein expression profiles between samples collected pre- and in-flight.

Moreover, we constructed a heatmap to visualize the result of a hierarchical clustering calculation of the 14 proteins differentially expressed in samples collected at pre- or in-flight from each astronaut. The results from the heatmap show that the intensities (level of expression) of tropomodulin-3 (TMOD3), SERPINA7, SERPING1, and SERPRINC1 were higher in samples collected in-flight, in relation to those collected pre-flight. Notably, the protein in the SERPIN (serine protease inhibitor) family, i.e., SERPINA7, SERPINC1, and ISERPING1, is the majority of those with increased intensities (expression levels). However, the data also demonstrated individual variability in the intensities of these proteins. It should be noted that SERPIN protease inhibitors comprise a large family of molecules involved in inflammation, immune response, blood clotting, hormone transport, and complement activation, dementia, and tumorigenesis. Our heatmap data also showed high intensities of CP, FLG2, KRT2, F13A1, DMKN, LUM in samples collected pre-flight. Subsequently, the intensities of expression of these proteins were declined in samples collected in-flights. Likewise, the intensities of another set of proteins (i.e., CFB, FBLN1, AHSG, and ECM1) were high in samples collected pre-flight; this followed by a reduction in intensities in samples collected in-flight, with one exception of increased intensity of FBLN1 protein in sample4s collected mid-flight from one astronaut.

Overall, the heatmap of our dataset demonstrated that the intensities of not only proteins in the SERPIN family but also TMOD3 (an actin filament pointed-end capping protein with multifunctional roles, including cell proliferation, cell migration, inflammation, and carcinogenesis) are consistently high in the in-flight samples. In contrast, low intensities of lumican (LUM) have been repeatedly detected in the in-flight samples. Of note, LUM is a major keratan sulfate proteoglycan of the cornea responsible for the circumferential growth, corneal transparency, epithelial cell migration, and tissue repair. Hence, it is possible to speculate that a reduction in LUM level may be associated with vision impairment that has been observed in many astronauts after spaceflight. In summary, our findings suggest that dysregulation of the SERPIN, TMOD3, and LUM may affect cell/tissue integrity and homeostasis, leading to late occurring health risks. Thus, our results may represent a foundation for the identification of countermeasures against the harmful effects of spaceflights.

Bibliography Type: Description: (Last Updated: 01/01/2021)  Show Cumulative Bibliography Listing
 
Abstracts for Journals and Proceedings Rithidech KN, Medococsa G, Crucian B, Martin D. "Impact of space flights on plasma proteome of astronauts." Presented at the 2020 NASA Human Research Program Investigators’ Workshop, Galveston, TX, January 27-30, 2020.

Abstracts. 2020 NASA Human Research Program Investigators’ Workshop, Galveston, TX, January 27-30, 2020. , Jan-2020

Project Title:  Effects of Space Flights on the Proteome of Astronauts' Plasma Reduce
Images: icon  Fiscal Year: FY 2019 
Division: Human Research 
Research Discipline/Element:
HRP HHC:Human Health Countermeasures
Start Date: 04/01/2016  
End Date: 03/31/2020  
Task Last Updated: 01/30/2019 
Download report in PDF pdf
Principal Investigator/Affiliation:   Rithidech, Kanokporn  Ph.D. / State University New York at Stony Brook 
Address:  Department of Pathology 
BHS T9 Health Sciences Center 
Stony Brook , NY 11794-8691 
Email: Kanokporn.Rithidech@sbumed.org 
Phone: (631) 444-3446  
Congressional District:
Web:  
Organization Type: UNIVERSITY 
Organization Name: State University New York at Stony Brook 
Joint Agency:  
Comments:  
Co-Investigator(s)
Affiliation: 
Martin, Dwight  Ph.D. State University of New York, Stony Brook 
Project Information: Grant/Contract No. NNX16AH80G 
Responsible Center: NASA JSC 
Grant Monitor: Norsk, Peter  
Center Contact:  
Peter.norsk@nasa.gov 
Solicitation / Funding Source: 2014-15 HERO NNJ14ZSA001N-MIXEDTOPICS. Appendix E: Behavioral Health & Human Health Countermeasures Topics 
Grant/Contract No.: NNX16AH80G 
Project Type: FLIGHT 
Flight Program:  
TechPort: No 
No. of Post Docs:
No. of PhD Candidates:
No. of Master's Candidates:
No. of Bachelor's Candidates:
No. of PhD Degrees:
No. of Master's Degrees:
No. of Bachelor's Degrees:
Human Research Program Elements: (1) HHC:Human Health Countermeasures
Human Research Program Risks: (1) Immune:Risk of Adverse Health Event Due to Altered Immune Response (IRP Rev F)
Human Research Program Gaps: (1) IM-101:Evaluate the effects of deep-space radiation on immune dysfunction, as an additional hazard to the effects of psychophysiological stress and weightlessness (IRP Rev L)
(2) IM-102:Evaluate the effects of altered atmospheric conditions such as elevated CO2 levels or mildly hypoxic exploration atmosphere on immune dysfunction (IRP Rev L)
(3) IM-103:Evaluate for novel endogenous viral reactivation (IRP Rev L)
(4) IM-401:Test, optimize and validate nutrition-based preventive/mitigative countermeasures (IRP Rev L)
Flight Assignment/Project Notes: Flight definition

NOTE: End date changed to 3/31/2020 per NSSC information (Ed., 3/25/19)

Task Description: NOTE: This is an integrated project consisting of Dr. Brian Crucian's "Functional Immune Alterations, Latent Herpesvirus Reactivation, Physiological Stress, and Clinical Incidence Onboard the International Space Station" directed research; and Dr. Richard Simpson's "The Impact of an ISS Mission on the Anti-Viral and Functional Properties of NK-cells, T-cells, B-cells and Dendritic Cells," Dr. Kanokporn Rithidech's "Effects of Space Flights on the Proteome of Astronauts' Plasma," and Dr. Honglu Wu's "DNA Damage in the ISS Astronaut's Lymphocytes and Their Association with Stress-Induced Immune Dysfunction" solicited research.

Space flight results in exposure of astronauts to several stressors, such as space radiation, microgravity, and physiological stress, that could exacerbate the risks of adverse health effects. To protect astronauts, we must improve our understanding of molecular changes that influence immunological conditions associated with increased astronaut health risks. The in vivo response to the space environment is complex, involving multiple proteins associated with various signal transduction cascades, resulting in different outcomes. Molecular mechanisms responsible for such diverse consequences are poorly understood. It is, therefore, essential to characterize the protein signatures of responses to the space environment in blood plasma samples from astronauts, collected at pre-, in-, and post-flights. Such analyses should help to reveal a particular set of proteins causing adverse immunological changes and to develop methods that help to prevent, or at least to counteract, these effects.

In this flight definition project, we will use cutting age proteomic technology to determine protein alterations, qualitatively and quantitatively, in plasma samples collected from astronauts before, during, and after space flights. Our findings will help to provide an understanding of the time course and etiology of immune changes induced by the space environment. Furthermore, since pre- and post-flight samples, in addition to the in-flight samples, will be evaluated in the same astronaut, the direct effects of the space environment can be determined. Hence, our findings will provide high-priority and highly relevant information to NASA. We will further correlate protein expression profiles to the available data on immune dysfunction detected in each astronaut. This approach makes it possible to determine potential predictive biomarkers for space-flight-induced immune dysregulation. Consequently, effective countermeasures against such harmful effects of the space environment can be identified.

Research Impact/Earth Benefits: Our findings will deliver important information that should advance our understanding of the time course and etiology of immune changes induced by the space environment. Hence, our findings will provide high-priority and highly relevant information to NASA. Importantly, we will correlate protein expression profiles to the available data on immune dysfunction detected in each astronaut. This approach makes it possible to determine potential predictive biomarkers for space-flight-induced immune dysregulation. Such knowledge is important for assessment of health risks and will facilitate the development of appropriate countermeasures that can help astronauts, space travelers, and people on Earth with the impairment of the immune system.

Task Progress & Bibliography Information FY2019 
Task Progress: After the last Task Book report, we obtained plasma samples from four astronauts. Prior to proteomics analysis, astronaut blood plasma was depleted of albumin and IgG using affinity spin columns. The depleted plasma was reduced and alkylated and digested with trypsin. The reaction mixture containing the resultant peptide product from the digestion was desalted by reverse phase chromatography. The desalted peptides were fractionated by a 12 step multi-dimensional chromatography process which delivered the peptides to a Thermo Fisher Orbitrap XL mass spectrometer for analysis by Liquid Chromatography with tandem mass spectrometry (LC-MS-MS).

Subsequently, the raw data files were interrogated and searched against a current UniProt human proteome database using the Andromeda search engine within MaxQuant. This produced a list of proteins for each sample which was quantitated by extracted ion chromatogram intensity by the MaxQuant program. The raw data from all the samples were searched simultaneously to reduce the occurrence of homology redundancy. The data from all the samples were compiled into a spreadsheet and the protein quantity was normalized based on the fractional signal strength within each sample. The samples were grouped according to the timeline stages of collection (i.e., pre-, in-, and post-flight). A total of 453 unique and non-redundant proteins were identified at >= 99% confidence. Changes in protein concentrations during the timeline were determined by Student’s t-test analysis of comparisons between groups (p<0.1 is considered significant). Those proteins displaying statistically significant changes were subjected to interacting network analysis using the Genemania app within the Cytoscape software package.

Our data demonstrate that there are 14 proteins with significant changes, i.e., increased or decreased, in expression levels in plasma samples collected in-flight as compared to those collected pre-flight. Four proteins with significant increases were detected in plasma collected in-flight, as compared to the plasma collected pre-flight. These are antithrombin-II (SERPINC1), plasma protease C1 inhibitor (SERPING1), Tropomodulin-3 (TMOD3), thyroxine-binding globulin (SERPINA7). The levels of the remaining 10 proteins were decreased in plasma collected in-flight as compared to those collected pre-flight. Notably, the protein in the SERPIN (serine protease inhibitor) family, i.e., SERPINA7, SERPINC1, and ISERPING1, is the majority of those with increased expression levels. The SERPIN protease inhibitors comprise a large family of molecules involved in inflammation, immune response, blood clotting, hormone transport, and complement activation, dementia, and tumorigenesis. Hence, our findings suggest that dysregulation of these proteins may affect cell/tissue integrity and homeostasis, leading to late occurring health risks. It has been suggested that overexpression of tropomodulin- 3 (TMOD3, actin pointed end-capping protein) leads to decreased endothelial motility. The majority of proteins with decreased levels are those involved in immune response and cytoskeleton systems.

Further, there were 16 proteins with significant changes in expression levels in astronauts’ plasma collected post-flight, in relation to those collected in-flight. There are eight proteins with significantly increased expression levels, while another set of eight proteins showed significantly decreased levels. The majority of proteins with changes in expression are those involved in immune and cytoskeleton systems. A new set of proteins with significant changes in expression levels was found in astronauts’ plasma collected post-flight, e.g., vitronectin, ceruloplasmin, Zyxin, and tubulin beta chain. This set of proteins may be involved in the re-adaptation to the Earth environment. Importantly, our results show that a decreased level of lumican (LUM) and an increased level of antithrombin-III (SERPINC1) persisted in astronauts’ plasma collected in- and post-flight. Such a prolonged increase in the level of SERPINC1 may contribute to coagulopathy, the phenomenon observed in individuals exposed to radiation. It is known that LUM is a major keratan sulfate proteoglycan of the cornea responsible for the circumferential growth, corneal transparency, epithelial cell migration, and tissue repair. Hence, LUM has been found to be critical in maintaining corneal clarity. Further, a loss of LUM expression has been found to be associated with corneal inflammation. Hence, persistent decreases in the level of LUM after spaceflight would impair the homeostasis of the eyes. Taken together, it is plausible to hypothesize that a reduction in LUM level may be associated with vision impairment that has been observed in many astronauts after spaceflight. Our findings warrant further investigation of the potential role of LUM in vision impairment detected in astronauts.

Highlights: The highlights of the findings from our study on the proteome of astronauts’ plasma collected at pre-, in-, and post-flights are:

• The majority of proteins with altered expression levels detected in plasma samples collected in- or post-flight are those involved in immune and cytoskeleton systems.

• A new set of proteins with altered expression levels was detected in plasma collected post-flight as compared to the samples collected in-flight.

• An increased level of antithrombin-III (SERPINC1) and a decreased level of Lumican (LUM, an important protein involved in maintaining corneal clarity protein) persisted for a long time post-flight, suggesting the potential role in late-occurring health risks.

Bibliography Type: Description: (Last Updated: 01/01/2021)  Show Cumulative Bibliography Listing
 
 None in FY 2019
Project Title:  Effects of Space Flights on the Proteome of Astronauts' Plasma Reduce
Images: icon  Fiscal Year: FY 2018 
Division: Human Research 
Research Discipline/Element:
HRP HHC:Human Health Countermeasures
Start Date: 04/01/2016  
End Date: 03/31/2019  
Task Last Updated: 01/29/2018 
Download report in PDF pdf
Principal Investigator/Affiliation:   Rithidech, Kanokporn  Ph.D. / State University New York at Stony Brook 
Address:  Department of Pathology 
BHS T9 Health Sciences Center 
Stony Brook , NY 11794-8691 
Email: Kanokporn.Rithidech@sbumed.org 
Phone: (631) 444-3446  
Congressional District:
Web:  
Organization Type: UNIVERSITY 
Organization Name: State University New York at Stony Brook 
Joint Agency:  
Comments:  
Co-Investigator(s)
Affiliation: 
Martin, Dwight  Ph.D. State University of New York, Stony Brook 
Project Information: Grant/Contract No. NNX16AH80G 
Responsible Center: NASA JSC 
Grant Monitor: Norsk, Peter  
Center Contact:  
Peter.norsk@nasa.gov 
Solicitation / Funding Source: 2014-15 HERO NNJ14ZSA001N-MIXEDTOPICS. Appendix E: Behavioral Health & Human Health Countermeasures Topics 
Grant/Contract No.: NNX16AH80G 
Project Type: FLIGHT 
Flight Program:  
TechPort: No 
No. of Post Docs:
No. of PhD Candidates:
No. of Master's Candidates:
No. of Bachelor's Candidates:
No. of PhD Degrees:
No. of Master's Degrees:
No. of Bachelor's Degrees:
Human Research Program Elements: (1) HHC:Human Health Countermeasures
Human Research Program Risks: (1) Immune:Risk of Adverse Health Event Due to Altered Immune Response (IRP Rev F)
Human Research Program Gaps: (1) IM-101:Evaluate the effects of deep-space radiation on immune dysfunction, as an additional hazard to the effects of psychophysiological stress and weightlessness (IRP Rev L)
(2) IM-102:Evaluate the effects of altered atmospheric conditions such as elevated CO2 levels or mildly hypoxic exploration atmosphere on immune dysfunction (IRP Rev L)
(3) IM-103:Evaluate for novel endogenous viral reactivation (IRP Rev L)
(4) IM-401:Test, optimize and validate nutrition-based preventive/mitigative countermeasures (IRP Rev L)
Flight Assignment/Project Notes: Flight definition

Task Description: NOTE: This is an integrated project consisting of Dr. Brian Crucian's "Functional Immune Alterations, Latent Herpesvirus Reactivation, Physiological Stress, and Clinical Incidence Onboard the International Space Station" directed research; and Dr. Richard Simpson's "The Impact of an ISS Mission on the Anti-Viral and Functional Properties of NK-cells, T-cells, B-cells and Dendritic Cells," Dr. Kanokporn Rithidech's "Effects of Space Flights on the Proteome of Astronauts' Plasma," and Dr. Honglu Wu's "DNA Damage in the ISS Astronaut's Lymphocytes and Their Association with Stress-Induced Immune Dysfunction" solicited research.

Space flight results in exposure of astronauts to several stressors, such as space radiation, microgravity, and physiological stress, that could exacerbate the risks of adverse health effects. To protect astronauts, we must improve our understanding of molecular changes that influence immunological conditions associated with increased astronaut health risks. The in vivo response to the space environment is complex, involving multiple proteins associated with various signal transduction cascades, resulting in different outcomes. Molecular mechanisms responsible for such diverse consequences are poorly understood. It is, therefore, essential to characterize the protein signatures of responses to the space environment in blood plasma samples from astronauts, collected at pre-, in-, and post-flights. Such analyses should help to reveal a particular set of proteins causing adverse immunological changes and to develop methods that help to prevent, or at least to counteract, these effects.

In this flight definition project, we will use cutting age proteomic technology to determine protein alterations, qualitatively and quantitatively, in plasma samples collected from astronauts before, during, and after space flights. Our findings will help to provide an understanding of the time course and etiology of immune changes induced by the space environment. Furthermore, since pre- and post-flight samples, in addition to the in-flight samples, will be evaluated in the same astronaut, the direct effects of the space environment can be determined. Hence, our findings will provide high-priority and highly relevant information to NASA. We will further correlate protein expression profiles to the available data on immune dysfunction detected in each astronaut. This approach makes it possible to determine potential predictive biomarkers for space-flight-induced immune dysregulation. Consequently, effective countermeasures against such harmful effects of the space environment can be identified.

Research Impact/Earth Benefits: Our findings will deliver important information that should advance our understanding of the time course and etiology of immune changes induced by the space environment. Hence, our findings will provide high-priority and highly relevant information to NASA. Importantly, we will correlate protein expression profiles to the available data on immune dysfunction detected in each astronaut. This approach makes it possible to determine potential predictive biomarkers for space-flight-induced immune dysregulation. Such knowledge is important for assessment of health risks and will facilitate the development of appropriate countermeasures that can help astronauts, space travelers, and people on Earth with the impairment of the immune system.

Task Progress & Bibliography Information FY2018 
Task Progress: We have completed our experiments on stability testing of plasma samples received from NASA Functional Immunology that were kept at room temperature (RT) for different times prior to storage at -80oC for proteomic analyses, i.e., 0 (immediately) hour (hr), day (d) 1, d2, and d3. These samples were collected using two types of anticoagulants, i.e., anticoagulant citrate dextrose (ACD) and ethylenediamine tetra acetic acid (EDTA), which may create variability in proteome yield both quantitatively and qualitatively. Additional variation can be introduced by downstream due to the necessary manipulations of sample preparation. It’s important to understand the level of variability. Hence, since the last report our effort has been to focus on understanding the variability of the observed proteome within and between the samples.

Our data on protein concentrations show no effects of time in keeping samples at room temperature prior to storage at -80oC. Additionally, there was no clear difference between the ACD and EDTA samples. We also optimized the procedures for sample preparation prior to mass spectrometry analyses. Such optimal conditions will be used to study the proteome of astronauts’ plasma samples which will be the focus of our project in the next fiscal year.

Bibliography Type: Description: (Last Updated: 01/01/2021)  Show Cumulative Bibliography Listing
 
 None in FY 2018
Project Title:  Effects of Space Flights on the Proteome of Astronauts' Plasma Reduce
Images: icon  Fiscal Year: FY 2017 
Division: Human Research 
Research Discipline/Element:
HRP HHC:Human Health Countermeasures
Start Date: 04/01/2016  
End Date: 03/31/2019  
Task Last Updated: 04/29/2017 
Download report in PDF pdf
Principal Investigator/Affiliation:   Rithidech, Kanokporn  Ph.D. / State University New York at Stony Brook 
Address:  Department of Pathology 
BHS T9 Health Sciences Center 
Stony Brook , NY 11794-8691 
Email: Kanokporn.Rithidech@sbumed.org 
Phone: (631) 444-3446  
Congressional District:
Web:  
Organization Type: UNIVERSITY 
Organization Name: State University New York at Stony Brook 
Joint Agency:  
Comments:  
Co-Investigator(s)
Affiliation: 
Martin, Dwight  Ph.D. State University of New York, Stony Brook 
Project Information: Grant/Contract No. NNX16AH80G 
Responsible Center: NASA JSC 
Grant Monitor: Norsk, Peter  
Center Contact:  
Peter.norsk@nasa.gov 
Solicitation / Funding Source: 2014-15 HERO NNJ14ZSA001N-MIXEDTOPICS. Appendix E: Behavioral Health & Human Health Countermeasures Topics 
Grant/Contract No.: NNX16AH80G 
Project Type: FLIGHT 
Flight Program:  
TechPort: No 
No. of Post Docs:
No. of PhD Candidates:
No. of Master's Candidates:
No. of Bachelor's Candidates:
No. of PhD Degrees:
No. of Master's Degrees:
No. of Bachelor's Degrees:
Human Research Program Elements: (1) HHC:Human Health Countermeasures
Human Research Program Risks: (1) Immune:Risk of Adverse Health Event Due to Altered Immune Response (IRP Rev F)
Human Research Program Gaps: (1) IM-101:Evaluate the effects of deep-space radiation on immune dysfunction, as an additional hazard to the effects of psychophysiological stress and weightlessness (IRP Rev L)
(2) IM-102:Evaluate the effects of altered atmospheric conditions such as elevated CO2 levels or mildly hypoxic exploration atmosphere on immune dysfunction (IRP Rev L)
(3) IM-103:Evaluate for novel endogenous viral reactivation (IRP Rev L)
(4) IM-401:Test, optimize and validate nutrition-based preventive/mitigative countermeasures (IRP Rev L)
Flight Assignment/Project Notes: Flight definition

Task Description: NOTE: This is an integrated project consisting of Dr. Brian Crucian's "Functional Immune Alterations, Latent Herpesvirus Reactivation, Physiological Stress, and Clinical Incidence Onboard the International Space Station" directed research; and Dr. Richard Simpson's "The Impact of an ISS Mission on the Anti-Viral and Functional Properties of NK-cells, T-cells, B-cells and Dendritic Cells," Dr. Kanokporn Rithidech's "Effects of Space Flights on the Proteome of Astronauts' Plasma," and Dr. Honglu Wu's "DNA Damage in the ISS Astronaut's Lymphocytes and Their Association with Stress-Induced Immune Dysfunction" solicited research.

Space flight results in exposure of astronauts to several stressors, such as space radiation, microgravity, and physiological stress, that could exacerbate the risks of adverse health effects. To protect astronauts, we must improve our understanding of molecular changes that influence immunological conditions associated with increased astronaut health risks. The in vivo response to the space environment is complex, involving multiple proteins associated with various signal transduction cascades, resulting in different outcomes. Molecular mechanisms responsible for such diverse consequences are poorly understood. It is, therefore, essential to characterize the protein signatures of responses to the space environment in blood plasma samples from astronauts, collected at pre-, in-, and post-flights. Such analyses should help to reveal a particular set of proteins causing adverse immunological changes and to develop methods that help to prevent, or at least to counteract, these effects.

In this flight definition project, we will use cutting age proteomic technology to determine protein alterations, qualitatively and quantitatively, in plasma samples collected from astronauts before, during, and after space flights. Our findings will help to provide an understanding of the time course and etiology of immune changes induced by the space environment. Furthermore, since pre- and post-flight samples, in addition to the in-flight samples, will be evaluated in the same astronaut, the direct effects of the space environment can be determined. Hence, our findings will provide high-priority and highly relevant information to NASA. We will further correlate protein expression profiles to the available data on immune dysfunction detected in each astronaut. This approach makes it possible to determine potential predictive biomarkers for space-flight-induced immune dysregulation. Consequently, effective countermeasures against such harmful effects of the space environment can be identified.

Research Impact/Earth Benefits: Our findings will help to provide an understanding of the time course and etiology of immune changes induced by the space environment. Hence, our findings will provide high-priority and highly relevant information to NASA. We will further correlate protein expression profiles with to the available data on immune dysfunction detected in each astronaut. This approach makes it possible to determine potential predictive biomarkers for space-flight-induced immune dysregulation. Importantly, the information obtained from this project will have a significant impact on the future development of a novel mechanism-based therapeutic strategy to prevent or mitigate the acute or long-term effects on the immune system due to space flight, as well as immunological diseases found on Earth.

Task Progress & Bibliography Information FY2017 
Task Progress: Currently, we are testing the stability of plasma samples that were prepared and kept at room temperature (RT) for different times prior to storage at -80oC for proteomic analyses, i.e., 0 (immediately) hour (hr), day (d) 1, d2, and d3. Our preliminary results suggested that the duration of storage at RT prior to plasma preparation and storage at -80oC influences protein expression profiles, as compared to those immediately kept at -80oC (0 hr) after plasma collection. A reduction in the numbers of observed proteins was detected among samples stored at d1, d2, and d3 at RT prior to plasma preparation and stored at -80oC, as compared to that observed in samples immediately kept at -80oC (0 hr) after blood collection. Our data also indicated a trend of a reduction in the numbers of observed proteins among samples stored at d1, d2, or d3 at RT prior to storage at -80oC. Moreover, since two types of anti-coagulant were used in blood collection, testing the effects of anti-coagulants on proteomic analyses is underway.

Bibliography Type: Description: (Last Updated: 01/01/2021)  Show Cumulative Bibliography Listing
 
 None in FY 2017
Project Title:  Effects of Space Flights on the Proteome of Astronauts' Plasma Reduce
Images: icon  Fiscal Year: FY 2016 
Division: Human Research 
Research Discipline/Element:
HRP HHC:Human Health Countermeasures
Start Date: 04/01/2016  
End Date: 03/31/2019  
Task Last Updated: 05/18/2016 
Download report in PDF pdf
Principal Investigator/Affiliation:   Rithidech, Kanokporn  Ph.D. / State University New York at Stony Brook 
Address:  Department of Pathology 
BHS T9 Health Sciences Center 
Stony Brook , NY 11794-8691 
Email: Kanokporn.Rithidech@sbumed.org 
Phone: (631) 444-3446  
Congressional District:
Web:  
Organization Type: UNIVERSITY 
Organization Name: State University New York at Stony Brook 
Joint Agency:  
Comments:  
Co-Investigator(s)
Affiliation: 
Martin, Dwight  Ph.D. State University of New York, Stony Brook 
Project Information: Grant/Contract No. NNX16AH80G 
Responsible Center: NASA JSC 
Grant Monitor: Norsk, Peter  
Center Contact:  
Peter.norsk@nasa.gov 
Solicitation / Funding Source: 2014-15 HERO NNJ14ZSA001N-MIXEDTOPICS. Appendix E: Behavioral Health & Human Health Countermeasures Topics 
Grant/Contract No.: NNX16AH80G 
Project Type: FLIGHT 
Flight Program:  
TechPort: No 
No. of Post Docs:  
No. of PhD Candidates:  
No. of Master's Candidates:  
No. of Bachelor's Candidates:  
No. of PhD Degrees:  
No. of Master's Degrees:  
No. of Bachelor's Degrees:  
Human Research Program Elements: (1) HHC:Human Health Countermeasures
Human Research Program Risks: (1) Immune:Risk of Adverse Health Event Due to Altered Immune Response (IRP Rev F)
Human Research Program Gaps: (1) IM-101:Evaluate the effects of deep-space radiation on immune dysfunction, as an additional hazard to the effects of psychophysiological stress and weightlessness (IRP Rev L)
(2) IM-102:Evaluate the effects of altered atmospheric conditions such as elevated CO2 levels or mildly hypoxic exploration atmosphere on immune dysfunction (IRP Rev L)
(3) IM-103:Evaluate for novel endogenous viral reactivation (IRP Rev L)
(4) IM-401:Test, optimize and validate nutrition-based preventive/mitigative countermeasures (IRP Rev L)
Flight Assignment/Project Notes: Flight definition

Task Description: NOTE: This is an integrated project consisting of Dr. Brian Crucian's "Functional Immune Alterations, Latent Herpesvirus Reactivation, Physiological Stress, and Clinical Incidence Onboard the International Space Station" directed research; and Dr. Richard Simpson's "The Impact of an ISS Mission on the Anti-Viral and Functional Properties of NK-cells, T-cells, B-cells and Dendritic Cells," Dr. Kanokporn Rithidech's "Effects of Space Flights on the Proteome of Astronauts' Plasma," and Dr. Honglu Wu's "DNA Damage in the ISS Astronaut's Lymphocytes and Their Association with Stress-Induced Immune Dysfunction" solicited research.

Space flight results in exposure of astronauts to several stressors, such as space radiation, microgravity, and physiological stress, that could exacerbate the risks of adverse health effects. To protect astronauts, we must improve our understanding of molecular changes that influence immunological conditions associated with increased astronaut health risks. The in vivo response to the space environment is complex, involving multiple proteins associated with various signal transduction cascades, resulting in different outcomes. Molecular mechanisms responsible for such diverse consequences are poorly understood. It is, therefore, essential to characterize the protein signatures of responses to the space environment in blood plasma samples from astronauts, collected at pre-, in-, and post-flights. Such analyses should help to reveal a particular set of proteins causing adverse immunological changes and to develop methods that help to prevent, or at least to counteract, these effects.

In this flight definition project, we will use cutting age proteomic technology to determine protein alterations, qualitatively and quantitatively, in plasma samples collected from astronauts before, during, and after space flights. Our findings will help to provide an understanding of the time course and etiology of immune changes induced by the space environment. Furthermore, since pre- and post-flight samples, in addition to the in-flight samples, will be evaluated in the same astronaut, the direct effects of the space environment can be determined. Hence, our findings will provide high-priority and highly relevant information to NASA. We will further correlate protein expression profiles to the available data on immune dysfunction detected in each astronaut. This approach makes it possible to determine potential predictive biomarkers for space-flight-induced immune dysregulation. Consequently, effective countermeasures against such harmful effects of the space environment can be identified.

Research Impact/Earth Benefits:

Task Progress & Bibliography Information FY2016 
Task Progress: New project for FY2016.

Bibliography Type: Description: (Last Updated: 01/01/2021)  Show Cumulative Bibliography Listing
 
 None in FY 2016