Whole Blood Measurements from the SpaceX Inspiration4 Mission (RNA-seq, epitranscriptome, WGS, clonal hematopoiesis, CBC)
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The SpaceX Inspiration4 mission was a 3-day mission with four private astronauts to low Earth orbit that occurred in September 2021. The crew collected biospecimen samples before, during, and after flight. One of these biospecimen collections included whole blood collected via venipuncture into PaxGene Blood RNA tubes (BD Biosciences, #762165), collected pre-flight (L-92, L-44, L-3) and post-flight (R+1, R+45, R+82, R+194). Total RNA was purified from each tube and mRNA was sequenced using the Oxford Nanopore Technologies direct RNA-sequencing kit on the PromethION. Data was used to generate differential gene expression profiles and annotate sites of m6A modification. Additionally, whole genome sequencing was performed on crew members at L-44 and R+45 and a clonal hematopoiesis panel was performed on a subset of 9 genes (DNMT3A, TET2, ASXL1, TP53, CHEK2, JAK2, SRSF2, SF3B1, PPM1D). For these, DNA was isolated from the pellet of a cfDNA blood collection tube (Streck, cat #230470) (L-44, L-3, R+1, R+45, R+82, R+194). Finally, whole blood collected in K2 EDTA tubes (BD Biosciences, cat# 367844) were submitted to Quest Diagnostics for a complete blood count at all ground timepoints (L-92, L-44, L-3, R+1, R+45, R+82, R+194).
SpaceX Inspiration4任务是2021年9月开展的一项为期3天的近地轨道(low Earth orbit)飞行任务,搭载四名私人宇航员。任务机组人员在飞行前、飞行中及飞行后采集了生物标本(biospecimen samples)。其中一类生物标本采集采用静脉穿刺(venipuncture)法获取全血,将其注入PaxGene血液RNA管(PaxGene Blood RNA tubes,碧迪生物(BD Biosciences),货号#762165)中,分别于飞行前(L-92、L-44、L-3)与飞行后(R+1、R+45、R+82、R+194)节点完成采样。随后从每支试管中纯化总RNA(total RNA),使用牛津纳米孔科技(Oxford Nanopore Technologies)的直接RNA测序试剂盒(direct RNA-sequencing kit)在PromethION测序平台上完成信使RNA(mRNA)测序。所得测序数据用于构建差异基因表达谱(differential gene expression profiles),并注释m6A修饰(m6A modification)位点。此外,研究人员于L-44与R+45两个时间点对机组人员开展全基因组测序(whole genome sequencing),并针对包含DNMT3A、TET2、ASXL1、TP53、CHEK2、JAK2、SRSF2、SF3B1、PPM1D在内的9个基因组成的克隆性造血基因面板(clonal hematopoiesis panel)进行检测。上述检测所用的DNA均从循环游离DNA(cfDNA)血液采集管(Streck,货号#230470)的沉淀物中分离,采样时间点涵盖L-44、L-3、R+1、R+45、R+82、R+194。最后,采集于K2 EDTA管(碧迪生物(BD Biosciences),货号#367844)中的全血样本,在所有地面采样时间点(L-92、L-44、L-3、R+1、R+45、R+82、R+194)均被送至奎斯特诊断(Quest Diagnostics)完成全血细胞计数(complete blood count)检测。




