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Dr Doug Boreham

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Dr Doug Boreham 12/13/2019 1 12/13/2019 We are going 2024 Artemis – Greek Goddess of the Hunt, the Moon, and Virginity Twin of Apollo Man-made vs natural radiation Occupational Fallout Nuclear fuel cycle 0.5% 1% 1% Consumer products 6.5% Sources of Radiation Exposure Nuclear medicine/x-rays Man-made Natural 91% radiation background 52% radiation 48% Mettler 2007 Brooks 2010 2 12/13/2019 Interplanetary space radiation What are we going to encounter beyond LEO? Limits of life in space, as Ionizing radiation that studied to date: will affect biology: Galactic Cosmic 12.5 days on a lunar round trip Radiation (GCR) 1.5 years in low Earth Solar Particle Events orbit on ISS (SPEs) Galactic Cosmic Radiation - GCR 3 12/13/2019 Deep Space Radiation • Galactic cosmic rays (GCR)—highly-ionizing radiation (Linear Energy Transfer - LET) • Comprised of high-energy protons and heavy ions • Solar particle events (SPEs): primarily energetic protons (85%), helium (14%) and high-energy (E) and high-charge (Z) ions called HZE particles (1%) 8 4 12/13/2019 DNA Damage Credit: NASA 5 12/13/2019 Human missions will need: water, food, oxygen fuel avoid solar particle events (SPEs). Deep space missions will require significant countermeasures, likely both technological and biomedical, to protect the crew from the effects of chronic radiation exposure. 11 Autonomous Biosensors for Deep Space Organisms that can survive for extended periods with minimal life support technology, and must function reliably with intermittent communication with Earth. 12 6 12/13/2019 13 7 12/13/2019 Saccharomyces cerevisiae Saccharo – Sugar Myces – Fungus Cerevisiae – Beer (Sugar fungus that makes beer) • Winemaking, Baking and Brewing • Eukaryotic • Highly studied and understood • Genome has been sequenced • Genetics are easily manipulated • Very easy to maintain in the lab and in space 15 BioSentinel Mission NASA Ames Research Center Northern Ontario School of Medicine SNOLAB Sudbury Cubesatb NASA Ames Research Center 8 12/13/2019 NASA Ames Collaboration • Collaboration with NASA’s BioSentinel research group • Understand effects of ionizing radiation in deep space (high dose) as well as deep underground in the absence of background radiation (low dose) 17 What is BioSentinel? • Yeast cells are similar Ionizing radiation • Deep space radiation to human cells environment cannot including in terms of be precisely mimicked DNA damage and on Earth repair DNA double strand breaks S. cerevisiae (budding yeast) Yeast cells will act as biosensors to measure the effects of deep space radiation 9 12/13/2019 BioSentinel mission Main objective: develop a tool with autonomous life support technologies to study the biological effects of the space radiation environment at different orbits Launch • First biological study beyond low Earth orbit (LEO) in almost 50 years • First biological 6U CubeSat to fly beyond LEO • Far beyond the protection of Earth’s magnetosphere • Will compare the biological effects of different radiation environments (EM-1, ISS…) Space Launch System (SLS) rocket. Lunar Distance to ISS: ~ 350 km Lunar Transfer transit Distance to the Moon: ~385,000 km & Fly-by (3-7 days) Distance at 6 months: ~40,000,000 km BioSentinel escapes into a heliocentric orbit Launch Secondary payload deployment (L+4- Launch of Exploration 5 hrs) Mission - 1 Methodology Yeast Strains: • Saccharomyces cerevisiae Wild Type (YBS71-1) W303 (NASA Yeast) • Saccharomyces cerevisiae RAD51 Δ (YBS72-1) W303 (NASA Yeast) – RADiation sensitive – RAD51 assists in repair of double stranded DNA breaks 20 10 12/13/2019 NASA BioSentinel • Genetically engineered Saccharomyces cerevisiae • Goal is to study DNA damage and repair mechanisms in the deep space radiation environment beyond LEO • Simplistic colorimetric endpoint quantifies changes in metabolism due to accumulated DNA damage Long-term survival – 32 weeks RT then 64 weeks 4oC at 0% RH 22 11 Models of risk Models k s i R l Epidemiology a c i g LNT o l Threshold o i B Hormesis Radiation Dose Natural 23 Background Sub-background Radiation 12/13/2019 12 12/13/2019 Chemico-Biological Interactions Volume 301, March 2019 Tharmalingam S, Sreetharan S, Brooks AL, Boreham DR. Re-evaluation of the Linear No-threshold (LNT) Model Using New Paradigms and Modern Molecular Studies. 54-67 Zarnke AM, Tharmalingam S, Boreham, DR Brooks AL. BEIR VI Radon: The Rest of the Story. 81-87 Ricci PF, Tharmalingam S. Ionizing Radiations Epidemiology Does Not Support the LNT Model.128-140 Scott BR, Tharmalingam S. The LNT Model for Cancer Induction is Not Supported by Radiobiological Data. 34-53 Hypothesis Natural background radiation is essential for life and helps to maintain the stability of our genome Prolonged exposure to a sub-background environment will be detrimental to living systems 13 12/13/2019 SNOLAB Creighton Mine 2 km underground (6 km #9 Shaft water equivalent) Norite Class 2,000 clean room Rock 50 million times less cosmic 2 km radiation Air filtration of 50 m3 s-1 for radon reduction 6800 level SNOLAB Radiation Research SNOLAB Focus Issue Volume 188, October 2017 14 12/13/2019 REPAIR Project Researching the Effects of the Presence and Absence of Ionizing Radiation REPAIR Project Cell culture Life Sciences Laboratory 2019 Biological Safety Cabinet Microscope Cell counter Water bath Centrifuge Tissue culture incubator UPS 15 12/13/2019 Advanced Tissue Culture Incubator 31 Background Radiation Characterization Radon 150 ) 3 - 75 m 15 q -3 B Underground (123.45 ± 13.53 Bq m ) ( n -3 o Surface (3.67 ± 2.14 Bq m ) i t a 10 r -3 t Glovebox (0.79 ± 0.93 Bq m ) n e c n o c 5 n o d a R 0 0 2 4 6 8 10 12 14 16 18 20 Time (days) 16 12/13/2019 Methodology • Determining survival with Biosentinal assay and colony-forming formation • Transcriptomic analysis will be performed at the optimal time point using RNA sequencing (RNA-Seq) with next-generation sequencing (NGS) technology 34 17 12/13/2019 35 Deep Space Travel • Complex environment intrinsic to space travel will likely exacerbate radiation-induced tissue injury • Microgravity • High CO2 • 2.3 - 5.3 mm Hg vs. 0.3 mm Hg on Earth • Confined living space • Circadian disruption • 16 sunrises and sunsets/day • Altered physiological processes • Poorly understood impact on nutritional requirement 36 18 12/13/2019 Deep Space Travel - Countermeasures •Bone demineralization •Cataracts •Cancer •Cardiovascular disease •Cognitive decline •Epigenetic Modification 37 19 12/13/2019 Vineland Estate Riesling STS 118 Endeavor Doses to Astronauts 2 mSv – Apollo 11 2000 mSv - Mars 20 12/13/2019 21 12/13/2019 NASA Johnson Complex Dietary Countermeasure The Dietary Countermeasure 22 12/13/2019 NASA Johnson Complex Dietary Countermeasure Collaboration • Designed to offset key processes associated with elevated ROS – Oxidative stress – Inflammation – Impaired glucose metabolism – Membrane deterioration – Mitochondrial dysfunction Mouse Model • Transgenic Growth Hormone Mice (TGM) • Model of elevated free radical processes – Elevated superoxide radical production – Increased lipid peroxidation • Model of accelerated aging – Average lifespan: 12 months – Premature onset of senescence Model for Astronauts 23 12/13/2019 The Results • Increased lifespan by 28% in TGM • 13% in normal siblings B • Almost eliminated sarcopenia, cataracts, arthritis, alopecia • All markers of oxidative damage and/or inflammation • Increased mobility and activity levels • Protected sensory and cognitive functions • Normalized ROS and inflammatory processes • Enhanced mitochondrial function • ETC complex activity increased 200 – 400% • Eliminated metastases in P53 cancer-prone mice • Eliminated paralysis in EAE model of MS • Protected cognition in 3xTg AD model 47 Chromosome Aberrations • Spectral Karyotyping (SKY) used to examine all chromosomes for structural aberrations 24 12/13/2019 Genomic Instability • Used Spectral Karyotyping (SKY) to determine type and amount of chromosome aberrations in TGM compared to age-matched normals Chromosome Aberrations Percent Structural Chromosome Aberrations 70 0Gy 60 2Gy 50 40 30 20 Percent Structural Aberrations 10 0 Untreated Norm. Untreated TGM Suppl. Norm. Suppl. TGM Experimental Groups 25 12/13/2019 Cognition 50 45 40 35 30 25 20 15 # # of Trials to Learn 10 5 0 UT Normal UT TGM Suppl. Normal Suppl. TGM Experimental Groups Brain Cell Study • Old TGM have ~44% of brain cells remaining – No increase in apoptosis – Most acellular region corresponds with the nucleus septi medialis • one of the main origins of cholinergic fibres innervating the hippocampus • Countermeasure completely abolishes brain cell loss • Responsible for retention of learning ability? 26 12/13/2019 Brain Cell Density • 12 mo. Old normal mice Brain Cell Number in Old Female Mice do not have significant brain cell loss compared 140000 to young mice 120000 • Untreated old TGM have 100000 less than 50% of young 80000 number of brain cells 60000 • Supplemented 12 mo. old Cell Number TGM have brain cell 40000 density equivalent to 20000 young mice 0 UT Norm Suppl. Norm UT TGM Suppl. TGM Experimental Groups Glucose Metabolism in Brain TGH untreated TGH diet 27 12/13/2019 Unsupp. Supp. FDG Activity in Brain 0.4 0.35 0.3 0.25 0.2 Activity 0.15 0.1 0.05 0 Normal TGM Experimental Group Cranial Irradiation Project Time to discover peanut butter smell • 10Gy gamma dose to head only • Countermeasure treated mice • Pre-IR fed 30d prior to WBI to end of study • Post-IR fed 24h post-IR to end of study • Animals assessed for cognitive function and olfactory acuity • 30d and 120d post-irradiation
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