HDAC inhibitors and IgH locus transcription. Mini 3'RR and IgH locus transcription

Numerous B-cell lymphomas feature translocations linking oncogenes with the IgH locus and epigenetic drugs such as histone deacetylase inhibitors (HDACi) have been approved to treat some of them. In this study we investigated IgH locus transcription in B-cell splenocytes stimulated with LPS and the HDACi SAHA. B-cell development is spatially and temporally regulated with the 3'RR enhancer of the IgH locus as a conductor. 3'RR is composed of 4 enhancer elements with a palindromic structure of great significance. We investigated the role of this palindrome with KOKI mice where the 30Kb structure of the 3'RR has been deleted of its palindromic structure. Overall design: Femoral B-cells from KOKI mice (in a RAG2-deficient and RAG2/gamma c-deficient background) and RAG2/gamma c -deficient mice were recovered. RNA was extracted using miRNeasy kit from QIAGEN, according to the manufacturer's instructions. Two pooled RNAs (each with 3 mice samples) were obtained for the three genotypes. CD43- splenocytes were obtained from KOKI mice and stimulated 2 days (1x10^6 cells/ml in RPMI 1640 with 10% FCS) with 5µg/ml LPS. RNA was extracted using miRNeasy kit from QIAGEN, according to the manufacturer's instructions. CD43- splenocytes were obtained from wt mice and stimulated 2 days (1x10^6 cells/ml in RPMI 1640 with 10% FCS) with 5µg/ml LPS ± 200 ng/ml SAHA. RNA was extracted using miRNeasy kit from QIAGEN. There are two replicates per condition.