{"project":{"acronym":"","projectId":23235,"title":"In-flight Blood Analysis Technology for Astronaut Health Monitoring","primaryTaxonomyNodes":[{"taxonomyNodeId":10694,"taxonomyRootId":8816,"parentNodeId":10693,"level":3,"code":"TX06.3.1","title":"Medical Diagnosis and Prognosis","definition":"This functional area provides a suite of medical technologies, knowledge, and procedures that reduce the likelihood and/or consequence of both nominal and off-nominal medical events during exploration missions.","exampleTechnologies":"Emerging screening technologies, preventative countermeasures, low resource imaging modalities, laboratory analysis platforms and assays, sterile fluid generation, medication packaging options and long-term medication storage, medical equipment re-use and in-situ manufacturing, integrated medical equipment and software suite, autonomous clinical care and decision support","hasChildren":false,"hasInteriorContent":true}],"startTrl":4,"currentTrl":6,"endTrl":6,"benefits":"
This project developed assays and instrumentation (i.e., hardwares, and softwares) that provide new ways of WBC count and subtype analysis. This project also proved that these new methods are as good as, if not better, as currently available commercial methods on Earth. Therefore, for the first time, this project provided the capability for NASA to do blood cell analysis in space, although further improvement needs to be done over our prototype for space qualification. In addition, both the developed assays and instrument can be used on Earth, too, and the technology has been licensed to a company, i.e., LeukoDx Inc., for the development of a point of care sepsis monitoring system initially targeted for the detection and monitoring of neonatal sepsis.
","description":"Medical events happened frequently to astronauts in space. For example, even the Space Shuttle Program alone reported 1867 incidences 1981-1998. Moreover, some events were serious viral/bacterial diseases such as urinary tract, conjunctivitis, acute respiratory, dental, and Varicella Zoster virus infections. Ideally, treatment on astronauts should be based on precise medical information. Meanwhile, blood is one of the most important body fluids related to health and there's tremendous information in blood. Blood analysis, if possible, should be the first important step of health monitoring for sick and healthy astronauts. Blood analysis can also be a powerful technique to monitor bone loss and radiation effects. Therefore, NASA should have an in-space, real-time blood analysis capability. However, NASA still doesn't have blood analysis capability other than blood gas and electrolyte analysis. This proposal is specifically to develop an in-box blood analysis technology for NASA. As a whole, we believe that the lab-on-a-chip technology is the best choice for multiple blood analysis in space. Therefore, our long-term objective is to develop blood analysis in-a-box using lab-on-a-chip technology specifically for space applications, emphasizing small form factor, lightweight, and autonomous operation to accommodate Crew Exploration Vehicle (CEV) and International Space Station (ISS) size requirement for medical kits.
The specific aims for this project period are to develop space technologies for (a) 5-part WBC (white blood cell) differential, (b) analysis of WBC subtypes (e.g., CD4+ T helper and natural killer cells). Our approach to achieve the goal is to develop the capability of minimally diluted micro flow cytometer to enable a comprehensive WBC differential, and allow detection of fluorescent labels attached to ligands used for cell surface marker for WBC subtype analysis. Embedded in the two specific aims is a research component on the data analysis software. This software has been developed in Matlab to facilitate both quantitative assessment of fluorescence detection and cell and analyte recognition and quantitation.
For the last funding years, we worked extensively on searching for a new staining method and optimizing the previously proposed Acridine Orange staining. We successfully developed a series of assays including a 4-part differential assay (i.e., Lymphocyte, Monocyte, Neutrophil, and Eosinophil) with a cocktail staining of fluorescent dyes fluorescein isothiocyanate (FITC) and propidium iodide (PI), a 5-part differential assay (i.e., Lymphocyte, Monocyte, Neutrophil, Eosinophil, and Basophil) with a cocktail staining of fluorescent dyes fluorescein isothiocyanate (FITC), propidium iodide (PI), and Basic Orange 21, and a specific assay for the rare cell type basophil differential using fluorescent dye Basic Orange 21. The differential assays were investigated in a correlation study with the commercial hematology analyzer, and further verified with the purified WBC types. For the Acridine Orange assay, the differential capability is also extended from 2-part (Lymphocyte and Neutrophil) into 4-part (Lymphocyte, Monocyte, Neutrophil, and Eosinophil). The time and temperature dependence of the Acridine Orange staining are also investigated.
Within the project period, we have also explored the possibility of improving the (box) platform in terms of spectroscopic detection. Two different approaches have been implemented; one uses a commercial mini-spectrometer and the other approach uses a 8-channel PMT (photomultiplier) module. Measurement of fluorescent emission spectrum from blood cells stained with the dye assay has been successfully demonstrated on the spectroscopic approach. Single cell fluorescence emission spectrum has been measured on the mini-spectrometer prototype. Distinct spectrums were measured from lymphocyte, neutrophil, and eosinophil cells. Besides, multicolor fluorescent beads have been successfully measured on the 8-color reader. Those two approaches enable detection of multiple fluorophore simultaneously from WBC subtype immune-staining. The additional spectral information should provide better discrimination between multiple fluorophores used simultaneously. It may also provide additional information about the intracellular environment in which Acridine Orange fluorescence occurs, leading to efficient WBC subtype discrimination. For WBC subtypes analysis, we have also successfully demonstrated assays that identified and counted CD4 and CD8 WBCs. In addition, we also developed synthesized peptides specifically targeted for leukocyte cells. The binding peptides were custom-synthesized with a fluorescein fluorophore attached to their n-terminus for binding quantification. A library of 72 potential peptide candidates has been tested using a modified protocol for leukocytes utilized in our studies. Among the results, 4 peptides from our initial library exhibited a 2-3x higher binding strength to the B-cells compared to the other peptides, which confirmed the capability of this approach for WBC subtype analysis.
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2. Verification of the differential assays with purified WBC types. A procedure of preparing purified WBC individual types (Lymphocyte, Monocyte, Neutrophil, Eosinophil, or Basophil) has been developed. The differential capability of the 5-part assay (PI, FITC, BO21) and the Basophil specific assay (BO21) was verified with the purified WBC types. The staining pattern observed from the purified WBC types also provided a useful tool to study new assays.
3. Spectrum analysis capability. One unit of the prototype has been upgraded from two-color detection to spectrum analysis with a commercial mini-spectrometer. Fluorescence spectrum measurment of dye (Acridine Orange) stained white blood cells were successfully demonstrated on the microfluidic chip. Distinct spectrums were measured from the Lymphocyte, Neutrophil, and Eosinophil cells. In addition, the detection of lymphocyte subtype cells were also demonstrated with the spectrum measurement system, which paved the way for simultaneous measurement of multiple subtype cells.
4. Planning for the new generation cartridge. Components of the next generation cartridge were successfully demonstrated. In the current cartridge, manual handling was involved to process the blood sample before test, and an external pump and waste collection tube were need for the fluidic operation. In the next generation cartridge, the whole test will be integrated into a 1cm x 1cm x 3mm chip without external fluidic connection. We successfully demonstrated the on-chip staining of blood sample with fluorescent dyes on the microchip. Besides, basic components of on-chip pump, on-chip valve, and long term reagent storage capability were also demonstrated.
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HRP research focuses on reducing crew health and performance risks for exploration missions. In addition, HRP research gathers the data necessary to understand and mitigate the long-term health risks to the crew, to allow the update of specific crew health standards for each mission scenario, to support crew selection, and to address any rehabilitation requirements. The OCHMO owns and sets the standards upon which the HRP research efforts are based. The Transition to Medical Practice process defined by the OCHMO is used to review the HRP deliverable countermeasures and technologies prior to their operational use.
HRP technology development advances medical care and countermeasure systems for exploration and vehicle development programs’ missions. The HRP also develops and matures operational concepts to inform requirements for the design and operation of space vehicles and habitats needed for exploration. This includes requirements for displays and controls, internal environments, operations planning, habitability, and methodologies for maintaining crew physical and mental health as well as physical and cognitive capabilities.
The HRP is managed at the Johnson Space Center (JSC) and comprised of six research and technology development projects. These projects provide the program knowledge and capabilities to conduct research addressing the human health and performance risks as well as advancing the readiness levels of technology and countermeasures to the point of transfer to the customer programs and organizations. The six projects within the HRP are referred to as Program Elements throughout this document. Each Element is managed at the JSC with research and technology development expertise provided by JSC, Ames Research Center (ARC), Glenn Research Center (GRC), the Langley Research Center (LaRC), and the Kennedy Space Center (KSC), as well as other Agencies, institutions and organizations identified in the following Element descriptions. The six Elements are:
1) Space Radiation (SR) Element – The SR Element performs investigations to develop the scientific basis to accurately predict and mitigate health risks from the space radiation environment. This knowledge yields recommendations to permissible exposure limits, assessment/projection tools/models of crew risk from radiation exposure, and models/tools to assess vehicle design for radiation protection. The SR Element conducts research using accelerator-based simulation of space radiation. The SR Element explores and develops countermeasures to the deleterious effects of radiation on human health. The LaRC and ARC contribute to the SR Element.
2) Behavioral Health and Performance (BHP) Element – The BHP Element identifies and characterizes the behavioral and performance risks associated with training, living and working in space, and returning to Earth. The BHP Element develops strategies, tools, and technologies to mitigate these risks.
3) Exploration Medical Capability (ExMC) Element – The ExMC Element is responsible for defining requirements for crew health maintenance during exploration missions, developing treatment scenarios, extrapolating from the scenarios to health management modalities, and evaluating the feasibility of those modalities for use during exploration missions. The ExMC Element is also responsible for the technology and informatics development that will enable the availability of medical care and decision systems for exploration missions. GRC, LaRC and ARC contribute technology development and clinical care expertise to the ExMC Element.
4) Space Human Factors and Habitability (SHFH) Element – The SHFH Element is focused on the human system in space environments: how do humans interface with spacecraft systems, and what environmental and habitation factors are essential to maintain crew health and performance? The SHFH Element has three main focus areas: space human factors engineering, advanced environmental health, and advanced food technology. The ARC contributes to the SHFH Element.
5) Human Health Countermeasures (HHC) Element – The HHC Element is responsible for understanding the physiological effects of spaceflight and developing countermeasure strategies and procedures. The Element provides the biomedical expertise for the development and assessment of medical standards and vehicle and spacesuit requirements dictated by human physiological needs. In addition, the HHC Element develops a validated and integrated suite of countermeasures for exploration missions to ensure the maintenance of crew health during all mission phases. The ARC and GRC contribute to the HHC Element as well as international agencies cooperating on joint flight proposals, reduced gravity studies, and collaborative bed rest studies.
6) International Space Station Medical Projects (ISSMP) Element – The ISSMP Element is responsible for managing all ISS and ground analog human research activities, including those integrated with operational medical support of the crews, and to ensure research tasks are completed. The ISSMP is responsible for all planning, integration, and implementation services for HRP research tasks and evaluation activities requiring access to space or related flight resources on the ISS, Soyuz, Progress, Multi-Purpose Crew Vehicle (MPCV), commercial vehicles and ground-based spaceflight analogs. This includes support to related pre- and postflight activities. The ARC contributes to the ISSMP with technical support to experiment management, hardware development, and international partner integration. KSC provides support for baseline data collection requirements development for future crew vehicles.
The work performed within the six Elements is supported by numerous collaborative efforts with academia and international agencies. Relationships with the ISS Program, the National Space Biomedical Research Institute (NSBRI), the Brookhaven National Laboratory (BNL), and the University of Texas Medical Branch (UTMB) are critical to the HRP successfully meeting its objectives. The HRP also maintains collaborative relationships with the International Partners through various working groups. These relationships enhance the research capabilities and provide synergy between the research and technology efforts of different countries.
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