Harnessing endogenous miR-181a to segregate transgenic antigen receptor expression in developing versus post-thymic T cells in murine hematopoietic chimeras Journal Article

   

Authors:	Papapetrou, E. P.; Kovalovsky, D.; Beloeil, L.; Sant'angelo, D.; Sadelain, M.
Article Title:	Harnessing endogenous miR-181a to segregate transgenic antigen receptor expression in developing versus post-thymic T cells in murine hematopoietic chimeras
Abstract:	MicroRNAs (miRNAs) are small, noncoding RNAs that regulate gene expression by targeting complementary sequences, referred to as miRNA recognition elements (MREs), typically located in the 3′ untranslated region of mRNAs. miR-181a is highly expressed in developing thymocytes and markedly downregulated in post-thymic T cells. We investigated whether endogenous miR-181a can be harnessed to segregate expression of chimeric antigen receptors (CARs) and TCRs between developing and mature T cells. Lentiviral-encoded antigen receptors were tagged with a miR-181a-specific MRE and transduced into mouse BM cells that were used to generate hematopoietic chimeras. Expression of a CAR specific for human CD19 (hCD19) was selectively suppressed in late double-negative and double-positive thymocytes, coinciding with the peak in endogenous miR-181a expression. Receptor expression was fully restored in post-thymic resting and activated T cells, affording protection against a subsequent challenge with hCD19<sup>+</sup> tumors. Hematopoietic mouse chimeras engrafted with a conalbumin-specific TCR prone to thymic clonal deletion acquired peptide-specific T cell responsiveness only when the vector-encoded TCR transcript was similarly engineered to be subject to regulation by miR-181a. These results demonstrate the potential of miRNA-regulated transgene expression in stem cell-based therapies, including cancer immunotherapy.
Keywords:	unclassified drug; human cell; genetics; nonhuman; antigen expression; t lymphocyte; t-lymphocytes; animal cell; mouse; animal; cytology; metabolism; animals; mice; bone marrow cells; cancer immunotherapy; microrna; cell maturation; animal experiment; stem cell transplantation; physiology; cell lineage; transgenic mouse; mice, transgenic; stem cell; chimera; immunological tolerance; lymphocyte activation; engraftment; hybrid protein; recombinant fusion proteins; thymus; receptors, antigen, t-cell; thymus gland; hematopoietic cell; transgene; hematopoietic stem cells; down regulation; bone marrow cell; hematopoietic stem cell; micrornas; cell protection; thymocyte; lymphocyte antigen receptor; receptors, antigen; transgenes; microrna 181a; ovotransferrin; mirn181 microrna, mouse; lentivirinae
Journal Title:	Journal of Clinical Investigation
Volume:	119
Issue:	1
ISSN:	0021-9738
Publisher:	American Society for Clinical Investigation
Date Published:	2009-01-05
Start Page:	157
End Page:	168
Language:	English
DOI:	10.1172/jci37216
PUBMED:	19033646
PROVIDER:	scopus
PMCID:	PMC2613472
DOI/URL:	http://www.scopus.com/inward/record.url?eid=2-s2.0-61749100637&partnerID=40&md5=25c9f7ea796e3cde4ae83656e3e453a4
Notes:	--- - "Cited By (since 1996): 11" - "Export Date: 30 November 2010" - "CODEN: JCINA" - "Source: Scopus"

https://synapse.mskcc.org/synapse/works/has_related_data_chk/1193