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Muotri. Additional support was provided by the Alzheimer's Association (AARF-25-1473595), CAPES (Finance Code 001), and CNPq (grants 311438/2022-9 and 402773/2022-5)."},{"name":"Identifiers","value":""},{"name":"Managing NASA Center","value":""},{"name":"Mission End","value":"04/30/2024"},{"name":"Mission Link","value":"https://osdr.nasa.gov/bio/repo/data/missions/SpaceX-30"},{"name":"Mission Name","value":"SpaceX-30"},{"name":"Mission Start","value":"03/21/2024"},{"name":"Project Identifier","value":"CubeLab_HSAPIENS_SpaceX-30"},{"name":"Project Link","value":""},{"name":"Project Title","value":""},{"name":"Project Type","value":"Spaceflight Study"},{"name":"Space Program","value":""},{"name":"Study Grant Number","value":""}],"description":"Spaceflight provides a unique environment to study neural responses relevant to neurodegenerative diseases. Here, human cortical organoids derived from induced pluripotent stem cells (iPSCs) that carry familial Alzheimer's disease (fAD) mutations in APP (p.V717G), PSEN1 (p.M233L), and PSEN2 (p.N141I) were cultivated at the International Space Station (ISS) alongside isogenic controls for 40 days. Multi-omic profiling, integrating transcriptomics, proteomics, and metabolic analyses, revealed a coordinated shift in neuronal and cellular states during spaceflight. AD-derived organoids showed altered glycolytic activity and altered lactate-to-malate ratios, consistent with metabolic and mitochondrial remodeling. Spaceflight was also associated with changes in amyloid-beta (AB) processing and tau phosphorylation, although the magnitude and direction of these responses varied across genetic backgrounds. Pathway analysis identified increased synaptic and neurotransmission-related signatures, particularly those associated with GABAergic and glutamatergic signaling. Integrative analysis further revealed convergent programs involving RNA processing, chromatin organization, cytoskeletal dynamics, and cellular stress responses. Proteomic and multi-omic analyses also identified changes in RNA-binding proteins, including CIRBP, consistent with altered post-transcriptional regulatory networks under spaceflight conditions. 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Cortical specification followed a semi-guided differentiation protocol.\n\nNeural induction (days 3\u20139): On day 3, mTeSR1 was replaced with Media 1: Neurobasal Medium (Thermo Fisher Scientific,  21103049) containing 1\u00d7 GlutaMAX, 1% N2 supplement, 2% Gem21 NeuroPlex (GeminiBio), 1% MEM Non-Essential Amino Acids, 1% Penicillin-Streptomycin, 10 uM SB431542, and 1 uM Dorsomorphin. Organoids were cultured in Media 1 for 6 days, with media changes every other day.\n\nNPC proliferation (days 9\u201322): On day 9, cultures were transferred to Media 2: Neurobasal with 1\u00d7 GlutaMAX, 2% Gem21 NeuroPlex, 1% NEAA, and 1% Penicillin-Streptomycin, supplemented with 20 ng/mL bFGF (PeproTech). Media was exchanged daily for 7 days. From days 16\u201322, organoids were kept in Media 2 with both 20 ng/mL bFGF and 20 ng/mL EGF (PeproTech), with media changes every other day.\n\nNeuronal differentiation (days 22\u201328): On day 22, organoids were switched to Media 3: Media 2 supplemented with 10 ng/mL BDNF, 10 ng/mL GDNF, 10 ng/mL NT-3 (all PeproTech), 200 \u03bcM L-ascorbic acid, and 1 mM dibutyryl-cAMP (both Sigma-Aldrich). Media was refreshed every other day for 6 days. From day 28 onward, organoids were kept in Media 2, with media exchanges every 3\u20134 days.\n\nQuality control: All organoid cultures were routinely tested for mycoplasma by PCR on media supernatants, and only negative cultures were used. After differentiation, the diameter of every organoid was measured using GelCount (Oxford Optronix), and cortical identity was confirmed by immunofluorescence (MAP2, SOX2) on 30-day-old organoids.","name":"growth protocol","parameters":[{"@id":"#139875369258576","parameterName":{"@id":"3Hq6ViAPdhnUQVpmPN4g5qe1Sc2CIbQALaTc3jdF","annotationValue":"","comments":[],"termAccession":"","termSource":""}}],"protocolType":{"@id":"0gMEkcMNeDUnXJOXyMiO3HWs7cSThiG9RpoYliDD","annotationValue":"growth protocol","comments":[],"termAccession":"http://www.ebi.ac.uk/efo/EFO_0003789","termSource":"EFO"},"uri":"","version":""},{"@id":"#139875369256080","comments":[],"components":[],"description":"Cortical organoids were generated from four human iPSC lines: a healthy control line (7889SA) and three isogenic lines carrying heterozygous CRISPR-engineered familial Alzheimer\u2019s disease mutations, PSEN1 (WT/M233L), PSEN2 (WT/N141I), and APP (WT/V717G). At one month of age, organoids of each genotype (Control, APP, PSEN1, PSEN2) were prepared for spaceflight, with twelve independent organoids per genotype \u00d7 environment combination. Organoids were transferred to cryovials (Thermo Fisher Scientific,\n 374088) containing ~2 mL of Media 2 (Neurobasal Medium with 1\u00d7 GlutaMAX, 2% Gem21 NeuroPlex, 1% NEAA, 1% Penicillin-Streptomycin, and 20 ng/mL bFGF). Loaded cryovials were hand-carried from UC San Diego to NASA Kennedy Space Center (KSC) and placed into a High Sample Capacity CubeLab (Space Tango Inc.), an autonomous incubator that maintained 37C and 5% CO2 throughout the mission. The flight CubeLab was integrated into the SpaceX Dragon capsule and launched to the ISS on the SpaceX CRS-30 mission on March 21, 2024. It returned to Earth on April 30, 2024. Ground control organoids were kept in a matching CubeLab at KSC labs under identical conditions: the same sealed cryovial system, CubeLab hardware, media volume, culture duration, temperature, CO2 control, and handling procedures.","name":"treatment protocol","parameters":[{"@id":"#139875369256704","parameterName":{"@id":"ZjmSSbPuCF9KW9S9qrQGc9mjnHs9TzHb1bMgnKcX","annotationValue":"duration","comments":[],"termAccession":"http://www.ebi.ac.uk/efo/EFO_0000433","termSource":"EFO"}},{"@id":"#139875372912112","parameterName":{"@id":"h9PQXMXXC1yOsFrPEWev3JtxYv4nNBpHkUvRH6wd","annotationValue":"Growth Temperature","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":"OSD"}},{"@id":"#139875372910912","parameterName":{"@id":"OqdFxo3iFGK3zWxVzb2EArzED1HinXKNpfayTxso","annotationValue":"Culture Vessel","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":"OSD"}},{"@id":"#139875372909184","parameterName":{"@id":"wo8Nyv8GLsF5cUaFVIokEOnbhWAjaExxVnwOCXB9","annotationValue":"Sample Media Information","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":"OSD"}},{"@id":"#139875372911872","parameterName":{"@id":"6igTY4thAsWlxyOsW7Ulx7TfqS4aKkH7lxmFJ9Fp","annotationValue":"Sample Media Quantity","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":"OSD"}}],"protocolType":{"@id":"PGCRAeZJDifKUY4b2TtcTGR2iEqwo3p6PKTVbSlo","annotationValue":"sample treatment protocol","comments":[],"termAccession":"http://www.ebi.ac.uk/efo/EFO_0003809","termSource":"EFO"},"uri":"","version":""},{"@id":"#139875369256752","comments":[],"components":[],"description":"Following splashdown on April 30, 2024, the CubeLab payload was returned to NASA's Space Station Processing Facility (SSPF), where it was opened and the organoid samples were retrieved during de-integration on the same day. The samples were immediately frozen on dry ice and subsequently stored at \u221280\u00b0C until further processing. For RNA sequencing, individual organoids were harvested as separate samples, and total RNA was extracted from each organoid independently. A total of 46 organoid samples were included in the final RNA-seq dataset: Control (Ground n equals 6, ISS n equals 6), APP (Ground n equals 6, ISS n equals 6), PSEN1 (Ground n equals 4, ISS n equals 6), and PSEN2 (Ground n equals 6, ISS n equals 6). One PSEN1 Ground sample was excluded due to poor RNA quality (RIN 2.6). To maintain consistency and avoid including an incomplete biological replicate, we excluded both experimental replicates associated with that biological sample. Therefore, the final dataset contains four samples representing two independent biological replicates (n equals 4).","name":"sample collection protocol","parameters":[{"@id":"#139875372910480","parameterName":{"@id":"mNsijgBSEnsCKOvqsd2bq0kKthgtK3FmpjfqpWEJ","annotationValue":"Sample Preservation Method","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":"OSD"}},{"@id":"#139875372912256","parameterName":{"@id":"f35LRInkOhIipZJMj3QqwZfzknHysHZYIWTPkIq5","annotationValue":"Sample Storage Temperature","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":"OSD"}}],"protocolType":{"@id":"8EuLrUZjLXgSuMKVI5oy9cqAEjPd73aK0xfAFloq","annotationValue":"sample collection protocol","comments":[],"termAccession":"http://www.ebi.ac.uk/efo/EFO_0005518","termSource":"EFO"},"uri":"","version":""},{"@id":"#139875369256368","comments":[],"components":[],"description":"Total RNA was extracted from individual cortical organoids using the RNeasy Plus Micro Kit Qiagen according to the manufacturer instructions. Organoids were lysed and homogenized, and genomic DNA was removed using gDNA eliminator columns included in the kit. RNA was purified through silica membrane spin columns and eluted in RNase free water. RNA concentration was measured using a Qubit fluorometric assay and RNA integrity was assessed using an Agilent Tapestation system. Only samples with RNA integrity number greater than 8 were used for downstream library preparation. For RNA sequencing n 3 biological replicates per genotype per condition total n 23 samples were processed.","name":"Nucleic Acid Extraction","parameters":[{"@id":"#139875371118256","parameterName":{"@id":"3Hq6ViAPdhnUQVpmPN4g5qe1Sc2CIbQALaTc3jdF","annotationValue":"","comments":[],"termAccession":"","termSource":""}}],"protocolType":{"@id":"SUvQ2YF4kYqggIqs6nZbO3jPYZOOa6kg99LuQwua","annotationValue":"Nucleic Acid Extraction","comments":[],"termAccession":"","termSource":"EFO"},"uri":"","version":""},{"@id":"#139875369256944","comments":[],"components":[],"description":"RNA sequencing libraries were prepared from high quality total RNA using the Illumina Stranded mRNA Prep kit according to the manufacturer instructions. Briefly polyadenylated RNA was enriched from total RNA and fragmented followed by first strand cDNA synthesis and second strand synthesis to generate double stranded cDNA. End repair and adapter ligation were performed and libraries were amplified by PCR. Library quality and concentration were assessed prior to sequencing. For RNA sequencing, n equals 3 biological replicates (individual organoids) per genotype per condition were used. Two independent technical-duplicate libraries were prepared from each RNA sample (48 libraries in total) and sequenced on separate lanes (L005 and L006) of the same NovaSeq 6000 run. One PSEN1 Ground biological replicate (GROUND_PSEN1_C) was excluded due to low RNA quality (RIN equals 2.6), and both of its libraries were removed, resulting in 46 libraries from 23 biological replicates in the final dataset.","name":"Library Construction","parameters":[{"@id":"#139875371116672","parameterName":{"@id":"gtKoOB4pXqmeWeZUfaA9fbez5HqlqkExIy7Egxfe","annotationValue":"Library Kit","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":"OSD"}},{"@id":"#139875371114656","parameterName":{"@id":"cSxJXkwH0s91Jd9yfYm2td3pbM1gGx6f4P9bIE8o","annotationValue":"stranded","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":"OSD"}},{"@id":"#139875371115424","parameterName":{"@id":"aqfZL5hvAkuf2Y5iTljXjMYxxUQ0V6TMjltaRpJa","annotationValue":"Library Selection","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":"OSD"}},{"@id":"#139875371117584","parameterName":{"@id":"JpyIKQUUZnStfVHj5N4P2R1Jkk72xPk7RosQz16j","annotationValue":"Library Layout","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":"OSD"}}],"protocolType":{"@id":"XwjyzSmjqwin5GREdbnjSVPuFEUzn18nShdNZvvS","annotationValue":"Library Construction","comments":[],"termAccession":"","termSource":"EFO"},"uri":"","version":""},{"@id":"#139875372912064","comments":[],"components":[],"description":"RNA sequencing libraries were multiplexed and sequenced on an Illumina NovaSeq 6000 platform using paired end reads of 151 base pairs. Sequencing was performed to achieve an average depth of approximately 25 million reads per sample. Base calling and demultiplexing were carried out using Illumina bcl2fastq software to generate FASTQ files for downstream analysis. Two independent technical-duplicate libraries were prepared from each RNA sample (48 libraries in total) and sequenced on separate lanes (L005 and L006) of the same NovaSeq 6000 run. One PSEN1 Ground biological replicate (GROUND_PSEN1_C) was excluded due to low RNA quality (RIN equals 2.6), and both of its libraries were removed, resulting in 46 libraries from 23 biological replicates in the final dataset.","name":"Nucleic Acid Sequencing","parameters":[{"@id":"#139875371117920","parameterName":{"@id":"ii6C5hK77k3RXUNA60xnDBRuoitkNvkXwZqxF9QJ","annotationValue":"Sequencing Instrument","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":"OSD"}},{"@id":"#139875371115040","parameterName":{"@id":"DOWz9RyvfMoltYGyC7An2WqtHdVXvLLJU32JiRbM","annotationValue":"Read Length","comments":[],"termAccession":"http://ncicb.nci.nih.gov/xml/owl/EVS/Thesaurus.owl","termSource":"NCIT"}},{"@id":"#139875371115088","parameterName":{"@id":"RNSQEM8tFAWy1LmmWB8QihODuaWzTWSvhVPzm8XT","annotationValue":"Base Caller","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":"OSD"}}],"protocolType":{"@id":"PsdoouSqq1TS1MOsEV6KSaZ1z6HqOAcEOC3cEEVX","annotationValue":"Nucleic Acid Sequencing","comments":[],"termAccession":"","termSource":"EFO"},"uri":"","version":""},{"@id":"#139875372909616","comments":[],"components":[],"description":"Fastq format was checked with fastq_utils v0.25.1. Quality assessment of reads was performed with FastQC v0.11.9 and reports were combined with MultiQC v1.12. rRNA percentages of reads were estimated with the 'hts_SeqScreener' program of htstream v1.4.1 screened against a fasta file of H-sapiens rRNA sequences retrieved from NCBI on 08-Jan-2026.","name":"GeneLab raw data processing protocol","parameters":[{"@id":"#139875371114704","parameterName":{"@id":"Ohs1BbENFJo4CWnwszKmpZ4g1nZJNqqJv60k0wyP","annotationValue":"Read Depth","comments":[],"termAccession":"http://ncicb.nci.nih.gov/xml/owl/EVS/Thesaurus.owl","termSource":"NCIT"}},{"@id":"#139875371118160","parameterName":{"@id":"ZzenbtS4P7ghaybWjqC3aI8kCjQrrZviCO3NL21k","annotationValue":"rRNA Contamination","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":"OSD"}},{"@id":"#139875371114992","parameterName":{"@id":"F7hQ9tMR9H2PMbQLZu0rqx36vnG0kIkup3vnJ6MU","annotationValue":"MultiQC File Names","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":"OSD"}}],"protocolType":{"@id":"gHRCpX8ZCZrvMb1jv0PpSihU4DurynDUUnkOSfet","annotationValue":"data transformation","comments":[],"termAccession":"http://purl.obolibrary.org/obo/OBI_0200000","termSource":"OBI"},"uri":"","version":""}],"publicReleaseDate":"26-Oct-2026","publications":[{"authorList":"Natalia C. S. Moreira, Luisa B. V. Coelho, Luiz P. Petroski, Stephanie S. Almeida, Luiza P. M. Gaudio, Livia L. Nascimento, Jeong-Hee Lee, Stevens Rehen, Blake Tsu, Juliana de S da G Fischer, Paulo C Carvalho, Aline M. A. Martins, Roberto H. Herai,      Alysson R. Muotri","comments":[],"doi":"","pubMedID":"","status":{"@id":"ewNKMJAJks5cjHqe1a4mkZdGRWwY30e2kbI5RBWb","annotationValue":"in preparation","comments":[],"termAccession":"http://www.ebi.ac.uk/efo/EFO_0001795","termSource":"EFO"},"title":"Spaceflight accelerates Alzheimer\u2019s disease\u2013associated molecular programs in human brain organoids"}],"studyDesignDescriptors":[{"@id":"q38Ii93JkgpqCZo21nbBgMqVw8pRvdAaUO0M8Ckf","annotationValue":"Alzheimer's disease","comments":[],"termAccession":"http://purl.obolibrary.org/obo/DOID_10652","termSource":"RBO"},{"@id":"XjiB5oWCcOxiqas5uTnohFkhOc0ZoQF7IybJuFMa","annotationValue":"Spaceflight","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":"OSD"},{"@id":"gVSq21JQZc5WN0DTTnnJZ8bOo201SFe5qvbMUGZ3","annotationValue":"Multi-omics analysis","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":"OSD"},{"@id":"LYJGWVPUQ5LcOAG9sP9lo8N4Obs9v9av0jZqAmRB","annotationValue":"Organoids","comments":[],"termAccession":"http://purl.bioontology.org/ontology/MESH/D009940","termSource":"MESH"},{"@id":"k8rppZHuvd3QhkjZ7dYmbh6mJcuFjH5YjOrZK3MB","annotationValue":"Weightlessness","comments":[],"termAccession":"http://purl.bioontology.org/ontology/MESH/D014893","termSource":"MESH"},{"@id":"w8gE63MhRE2jPbKwIDLmJc8Ur44kBpN33MAsibxa","annotationValue":"brain","comments":[],"termAccession":"http://purl.obolibrary.org/obo/UBERON_0000955","termSource":"OBI"}],"submissionDate":"19-Nov-2025","title":"Spaceflight reshapes Alzheimer's disease\u2013associated molecular programs in human brain organoids","unitCategories":[{"@id":"0egjOe4tupLenTJ2RAdBhRffOpl8xZWv0FRLbF9W","annotationValue":"day","comments":[],"termAccession":"http://purl.obolibrary.org/obo/UO_0000033","termSource":""},{"@id":"ixhZXi79C7bGPbpBgn0nhYS3WcKUHFxvfrXat1ju","annotationValue":"degree Celsius","comments":[],"termAccession":"http://purl.obolibrary.org/obo/UO_0000027","termSource":""},{"@id":"u5hUzYE0N0xDfj0iUQCW3tdbfFRfHT1ccYPDERfY","annotationValue":"milliliter","comments":[],"termAccession":"http://purl.obolibrary.org/obo/UO_0000098","termSource":""}]}],"version":1}},"success":true}