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GEO help: Mouse over screen elements for information. |
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Status |
Public on Oct 08, 2024 |
Title |
Expansion of in vivo HDAd-transduced HSPCs by constitutive expression of tHMGA2 without additional treatment allows for therapeutically relevant transgene expression in HSPC progeny cells without clonal dominance. |
Organism |
Mus musculus |
Experiment type |
Other
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Summary |
We developed an in vivo hematopoietic stem and progenitor cell (HSPC) gene therapy approach that does not require HSC cell transplantation. To achieve therapeutically relevant numbers of corrected cells, we tested truncated HMGA2 instead of chemo-selection for enrichment of transduced HSPCs. We constructed HSC-tropic HDAd5/35++ vectors expressing a 3’UTR HMGA2 truncated gene and a GFP reporter gene. A SB100x transposase vector was used to mediate random integration of the tHMGA2/GFP transgene cassette. Mice were mobilized by subcutaneous injections of G-CSF and AMD3100/Plerixafor and intravenously injected with the integrating tHMGA2/GFP vector. This resulted in slow but progressive competitive expansion of GFP+ PBMCs, reaching about 50% by week 44 with further expansion in secondary recipients. Expansion occurred at the level of HSCs as well as at the level of myeloid, lymphoid, and erythroid progenitors within the bone marrow and spleen. Importantly, based on genome-wide integration site analyses, expansion was polyclonal, without any signs of clonal dominance. The results were validated in humanized mice. This is the first demonstration that simple subcutaneous and intravenous injections of mobilization drugs and HDAd vectors can trigger autocatalytic, self-limiting expansion of in vivo transduced HSPCs resulting in marking rates that, theoretically, are curative for hemoglobinopathies.
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Overall design |
CD46 transgenic mice were mobilized by subcutaneous (SC) injections of human recombinant G-CSF (250 mg/kg/mouse/day, 4 days) followed by an SC injection of AMD3100 (5mg/kg) on day 5. In addition, animals received Dexamethasone (10 mg/kg, IP) 16 h and 2 h before virus injection to blunt innate toxicity associated with intravenous HDAd injection. Forty-five minutes after AMD3100, animals were intravenously injected with HDAd virus vectors through the retro-orbital plexus (4x1010 viral particles per mouse). Group 1 (N=10) were injected with HDAd-mgmt/GFP + HDAd-SB). Group 2 received HDAd-tHMGA2/GFP + HDAd-SB. DNA from bone marrow MNCs of week 44 primary mice with different rates of GFP marking in PBMCs (ranging from 25% to 65% ) were subjected to genome-wide transgene integration site analysis.
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Contributor(s) |
Wang H, Georgakopoulou A, Eluère R, Mok KW, Policastro RA, Lieber A |
Citation(s) |
39282078 |
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Submission date |
May 30, 2024 |
Last update date |
Oct 09, 2024 |
Contact name |
Robert Policastro |
Organization name |
Ensoma Inc.
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Street address |
451 D St Unit #600
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City |
Boston |
State/province |
Massachusetts |
ZIP/Postal code |
02210 |
Country |
USA |
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Platforms (1) |
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Samples (18)
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Relations |
BioProject |
PRJNA1118248 |
Supplementary file |
Size |
Download |
File type/resource |
GSE268678_HDAd_tHMGA2_GFP.fasta.gz |
9.1 Kb |
(ftp)(http) |
FASTA |
GSE268678_RAW.tar |
40.0 Kb |
(http)(custom) |
TAR (of BED) |
GSE268678_annotated_insertion_sites.tsv.gz |
84.6 Kb |
(ftp)(http) |
TSV |
SRA Run Selector |
Raw data are available in SRA |
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