Transcriptomic atlas reveals organ-specific disease tolerance in sickle cell mice. Dataset for HbAA lung injected or not with heme

<div> <div> <p>The objective of this experiment was to explore the transcriptome&nbsp;of the&nbsp;HbSS Townes&nbsp;mouse model&nbsp;of&nbsp;sickle cell disease.Townes model mice carry several human hemoglobin&nbsp;knock-in&nbsp;genes replacing the endogenous mouse genes and may be useful in studying&nbsp;sickle cell disease.All mice were&nbsp;genotyped, age- and sex-matched littermates. All&nbsp;HbAA&nbsp;(control, normal human hemoglobin) vs HbSS (sickle cell disease, mutated human hemoglobin) mice were used for experimentations at 6-8 weeks of age, to limit intra-group&nbsp;heterogeneity.&nbsp;Hemin&nbsp;(Ferriprotoporphyrin IX) was purchased from&nbsp;Frontiers Scientific&nbsp;and injected&nbsp;intravenously&nbsp;(iv.) in a retroorbital sinus at a concentration of 24 &micro;mol/kg. Control mice received&nbsp;PBS&nbsp;instead. Mice were anesthetized with&nbsp;isoflurane&nbsp;2-3% for injections, blood collection and sacrifice. All mice were sacrificed by cervical dislocation, 4 hours after injection.</p> <p>The results of lung (indicated poumon) from HbAA mice injected or not with&nbsp;heme&nbsp;are presented here .&nbsp;</p> <p>The results of lung (indicated poumon) from HbSS mice injected or not with heme can be found at number 10.5281/zenodo.10966094</p> <p>Thirty &mu;m-thick frozen tissue sections of lungs were cut as above and homogenized in 200&mu;L of 1-Thioglycerol/Homogenization Solution (Maxwell&reg; 16 LEV simplyRNA Tissue Kit Promega&nbsp;AS1280).&nbsp;The quality and quantity of mRNA were evaluated using a 2100&nbsp;bioanalyzer with TNA 6000 NanoKits (all&nbsp;Agilent Technologies,&nbsp;Palo Alto, CA,&nbsp;USA). RNA Integrity Numbers superior to 7 were eligible for subsequent&nbsp;reverse transcription&nbsp;into&nbsp;cDNA.&nbsp;RNAseq&nbsp;was performed at the GenomIC plateform&nbsp;Cochin&nbsp;Institute INSERM U1016. After&nbsp;RNA extraction, RNA quality (RNA integrity number) was estimated. 1&mu;g of high-quality total RNA sample (RIN &amp;gt;7) was processed to build up the libraries, using TruSeq Stranded mRNA kit (Illumina) according to manufacturer instructions. Briefly, purified&nbsp;poly-A&nbsp;containing mRNA molecules were fragmented and&nbsp;reverse-transcribed&nbsp;using random&nbsp;primers. Replacement of dTTP by dUTP during second strand synthesis allowed us to achieve strand specificity. Addition of a single A base to the&nbsp;cDNA&nbsp;was followed by&nbsp;ligation&nbsp;of&nbsp;Illumina&nbsp;adapters.<br>Libraries were quantified by&nbsp;qPCR&nbsp;using&nbsp;KAPA Library Quantification&nbsp;Kits for&nbsp;Illumina&nbsp;Libraries (KapaBiosystems,&nbsp;Wilmington, MA). Library profiles were assessed using DNA High Sensitivity LabChip kits on an&nbsp;Agilent&nbsp;Bioanalyzer. Libraries were sequenced on an&nbsp;Illumina&nbsp;Nextseq 500 instrument using 75 base-lengths read V2 chemistry in a&nbsp;paired-end&nbsp;mode. After sequencing, primary analysis based on AOZAN software (ENS,&nbsp;Paris), was applied to&nbsp;demultiplex&nbsp;and control the quality of the&nbsp;raw data&nbsp;(based of FastQC modules / version 0.11.5).</p> <p>The dataset here represents 4 groups of mice, 4 mice per group as follows: HbAA&nbsp;PBS, HbAA&nbsp;heme, HbSS&nbsp;PBS,&nbsp;HbSS&nbsp;heme.&nbsp;</p> </div> </div>