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

<div> <p>The objective of this experiment was to explore the&nbsp;<span>transcriptome</span>&nbsp;of the&nbsp;<span>HbSS Townes</span>&nbsp;<span>mouse model</span>&nbsp;of&nbsp;<span>sickle cell disease</span>.Townes model mice carry several human hemoglobin&nbsp;<span>knock-in</span>&nbsp;genes replacing the endogenous mouse genes and may be useful in studying&nbsp;<span>sickle cell disease</span>.All mice were&nbsp;<span>genotyped</span>, age- and sex-matched littermates. All&nbsp;<span>HbAA</span>&nbsp;(control, normal human hemoglobin) vs HbSS (<span>sickle cell disease</span>, mutated human hemoglobin) mice were used for experimentations at 6-8 weeks of age, to limit intra-group&nbsp;<span>heterogeneity</span>.&nbsp;<span>Hemin</span>&nbsp;(<span>Ferriprotoporphyrin IX</span>) was purchased from&nbsp;<span>Frontiers Scientific</span>&nbsp;and injected&nbsp;<span>intravenously</span>&nbsp;(iv.) in a retroorbital sinus at a concentration of 24 &micro;mol/kg. Control mice received&nbsp;<span>PBS</span>&nbsp;instead. Mice were anesthetized with&nbsp;<span>isoflurane</span>&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 heart (indicated coeur) from HbSS mice injected or not with <span>heme</span>&nbsp;are presented here .&nbsp;</p> <p>The results of heart (indicated coeur) from&nbsp;<span>HbAA</span>&nbsp;mice injected or not with&nbsp;<span>heme</span> can be found at number 10.5281/zenodo.10964042</p> <p>Thirty &mu;m-thick frozen tissue sections of kidneys&nbsp;were cut as above and homogenized in 200&mu;L of 1-Thioglycerol/Homogenization Solution (Maxwell&reg; 16 LEV simplyRNA Tissue Kit&nbsp;<span>Promega</span>&nbsp;AS1280).&nbsp;The quality and quantity of mRNA were evaluated using a 2100&nbsp;bioanalyzer with TNA 6000 NanoKits (all&nbsp;<span>Agilent Technologies</span>,&nbsp;<span>Palo Alto, CA</span>,&nbsp;<span>USA</span>). RNA Integrity Numbers superior to 7 were eligible for subsequent&nbsp;<span>reverse transcription</span>&nbsp;into&nbsp;<span>cDNA</span>.&nbsp;<span>RNAseq</span>&nbsp;was performed at the GenomIC plateform&nbsp;<span>Cochin</span>&nbsp;Institute INSERM U1016. After&nbsp;<span>RNA extraction</span>, RNA quality (<span>RNA integrity number</span>) 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 (<span>Illumina</span>) according to manufacturer instructions. Briefly, purified&nbsp;<span>poly-A</span>&nbsp;containing mRNA molecules were fragmented and&nbsp;<span>reverse-transcribed</span>&nbsp;using random&nbsp;<span>primers</span>. Replacement of dTTP by dUTP during second strand synthesis allowed us to achieve strand specificity. Addition of a single A base to the&nbsp;<span>cDNA</span>&nbsp;was followed by&nbsp;<span>ligation</span>&nbsp;of&nbsp;<span>Illumina</span>&nbsp;adapters.<br>Libraries were quantified by&nbsp;<span>qPCR</span>&nbsp;using&nbsp;<span>KAPA Library Quantification</span>&nbsp;Kits for&nbsp;<span>Illumina</span>&nbsp;Libraries (KapaBiosystems,&nbsp;<span>Wilmington</span>, MA). Library profiles were assessed using DNA High Sensitivity LabChip kits on an&nbsp;<span>Agilent</span>&nbsp;Bioanalyzer. Libraries were sequenced on an&nbsp;<span>Illumina</span>&nbsp;Nextseq 500 instrument using 75 base-lengths read V2 chemistry in a&nbsp;<span>paired-end</span>&nbsp;mode. After sequencing, primary analysis based on AOZAN software (ENS,&nbsp;<span>Paris</span>), was applied to&nbsp;<span>demultiplex</span>&nbsp;and control the quality of the&nbsp;<span>raw data</span>&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;<span>PBS</span>, HbAA&nbsp;<span>heme</span>, HbSS&nbsp;<span>PBS</span>,&nbsp;<span>HbSS</span>&nbsp;<span>heme</span>.&nbsp;</p> </div>