Mus musculus
Protocols: C57BL/6 female mice (8 weeks old) were irradiated and transplanted with mouse hematopoietic progenitor/stem cells transduced with a control mTie2-GFP-mirT LV. At 20 weeks of age mice were sacrificed. Spleen tissue was dissociated and cells subjected to FACS sorting. F4/80+ macrophages were recovered. IFN treatment was performed by reconstituting lethally irradiated mice with hematopoietic stem and progenitor cells (HSPCs) that were previously transduced with a lentiviral vector (LV) containing the cDNA sequence of the IFN-alpha gene under the control of the Tie2 promoter/enhancer sequences. Among differentiated hematopoietic cells, Tie2 is selectively expressed in a population of tumor-infiltrating monocytes (TEMs), thus resulting in tumor-specific IFN-alpha delivery. The LV design also incorporates into the IFN-alpha transgene multiple miR-126 and miR-130a target sequences to suppress post-transcriptionally transgene expression in HSPCs limiting potential transgene in this compartment. C57BL/6 female mice (8 weeks old) were irradiated and transplanted with mouse HSPC transduced with a mTie2-IFN-mirT LV. At 20 weeks of age mice were sacrificed. Spleen tissue was dissociated and cells subjected to FACS sorting. F4/80+ macrophages were recovered. C57BL/6 female mice (8 weeks old) were irradiated and transplanted with mouse HSPC transduced with a control mTie2-IFN-mirT LV. At 16 weeks of age mice were injected intravenously with 10-100 thousand B-ALL cells. At 20 weeks of age mice were sacrificed. Spleen tissue was dissociated and cells subjected to FACS sorting. F4/80+ macrophages were recovered. ReliaPrep RNA MiniPrep System (Promega) RNA-Seq libraries were prepared with the SMART-Seq2 protocol, with minor modification. Briefly, RNA (1-5 ng) was reverse transcribed using custom oligodT and template-switching LNA oligos (sequences), followed by PCR amplification and clean-up (Ampure XP beads, Beckman Coulter). The resulting cDNA (0,5- 1 ng) was tagmented at 55°C for 30 minutes and final RNA-Seq libraries generated using reagents from the Nextera XT DNA Library Prep Kit (Illumina).
ENA-FIRST-PUBLIC
2018-07-26T17:02:32Z
ENA-LAST-UPDATE
2018-05-29T14:42:41Z
External Id
SAMEA4696493
INSDC center name
San Raffaele Telethon Institute for Gene Therapy.
INSDC first public
2018-07-26T17:02:32Z
INSDC last update
2018-05-29T14:42:41Z
INSDC status
public
Submitter Id
E-MTAB-6482:ALL_r2
age
20
broker name
ArrayExpress
cell type
macrophage
common name
house mouse
disease
B-cell acute lymphoblastic leukemia
genotype
lentiviral expression of mTie2-GFP-mirT
organism part
spleen
phenotype
F4/80 positive
sample name
E-MTAB-6482:ALL_r2
scientific_name
Mus musculus
sex
female
stimulus
intravenous injection of 10-100 thousand B-ALL cells
strain
C57BL/6