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GEO help: Mouse over screen elements for information. |
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| Status |
Public on Sep 18, 2024 |
| Title |
c-Myc alone is enough to reprogram fibroblasts into functional macrophages [scRNA-seq] |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Background Cell therapy based on macrophages is promising in solid tumors, but effective acquisition of macrophages is a challenge. Induced pluripotent stem cell (iPSC)-induced macrophages are a good source but are time-consuming and costly. Macrophages derived from somatic cells by reprogramming technologies have the potential to enable the development of cell-based therapies for numerous malignant diseases.
Methods The composition of CD45+ myeloid-like cell complex (MCC) was analyzed by flow cytometry and single-cell sequencing. The engraftment capacity of CD45+ MCC was evaluated by two transplantation assays. Regulation of c-myc on MafB was evaluated by ChIP-qPCR and promoter reporter and dual luciferase assays. The phenotype and phagocytosis of induced macrophages were explored by flow cytometry and immunofluorescence. Leukemia, breast cancer, and patient-derived tumor xenograft models were used to explore the anti-tumor function of induced macrophages.
Results Here we report on the establishment of a novel methodology allowing for reprogramming fibroblasts into functional macrophages with phagocytic activity by c-Myc overexpression. Fibroblasts with ectopic expression of c-Myc in iPSC medium rapidly generated CD45+ MCC intermediates with engraftment capacity as well as the repopulation of distinct hematopoietic compartments. MCC intermediates were stably maintained in suspension culture and continuously generated functional and highly pure-induced macrophage (iMac) just by M-CSF cytokine stimulation. Single-cell transcriptomic analysis of MCC intermediates revealed that c-Myc up-regulates the expression of MafB, a major regulator of macrophage differentiation, to promote macrophage differentiation. Characterization of the iMac activity showed NF-κB signaling activation and a pro-inflammatory phenotype. iMac cells display significantly increased in vivo persistence and inhibition of tumor progression in leukemia, breast cancer, and patient-derived tumor xenograft models.
Conclusions Our findings demonstrate that c-Myc alone is enough to reprogram fibroblasts into functional macrophages, supporting that c-Myc reprogramming strategy of fibroblasts could help circumvent long-standing obstacles to gaining “off-the-shelf” macrophages for anti-cancer immunotherapy.
Keywords c-Myc, fibroblast, macrophage, immunotherapy, reprogramming
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| Overall design |
A single-cell RNA sequencing of c-Myc-induced macrophage (iMac) was performed
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| Contributor(s) |
Li Y, Chen G |
| Citation(s) |
39267119 |
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| Submission date |
Aug 22, 2024 |
| Last update date |
Sep 18, 2024 |
| Contact name |
Guoyu Chen |
| E-mail(s) |
chenguoyu@live.com
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| Phone |
+8618621591690
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| Organization name |
PICB
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| Street address |
Xuhui District Yueyang Road 320
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| City |
Shanghai |
| ZIP/Postal code |
200031 |
| Country |
China |
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| Platforms (1) |
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| Samples (2) |
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| Relations |
| BioProject |
PRJNA1150967 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE275457_barcodes.tsv.gz |
17.7 Kb |
(ftp)(http) |
TSV |
| GSE275457_features.tsv.gz |
75.1 Kb |
(ftp)(http) |
TSV |
| GSE275457_matrix.mtx.gz |
61.1 Mb |
(ftp)(http) |
MTX |
SRA Run Selector |
| Raw data are available in SRA |
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