Single-cell transcriptional profiling informs efficient reprogramming of human somatic cells to cross-presenting dendritic cells
(2022) In Science Immunology 7(69). p.1-18- Abstract
Type 1 conventional dendritic cells (cDC1s) are rare immune cells critical for the induction of antigen-specific cytotoxic CD8+ T cells, although the genetic program driving human cDC1 specification remains largely unexplored. We previously identified PU.1, IRF8, and BATF3 transcription factors as sufficient to induce cDC1 fate in mouse fibroblasts, but reprogramming of human somatic cells was limited by low efficiency. Here, we investigated single-cell transcriptional dynamics during human cDC1 reprogramming. Human induced cDC1s (hiDC1s) generated from embryonic fibroblasts gradually acquired a global cDC1 transcriptional profile and expressed antigen presentation signatures, whereas other DC subsets were not induced at the single-cell... (More)
Type 1 conventional dendritic cells (cDC1s) are rare immune cells critical for the induction of antigen-specific cytotoxic CD8+ T cells, although the genetic program driving human cDC1 specification remains largely unexplored. We previously identified PU.1, IRF8, and BATF3 transcription factors as sufficient to induce cDC1 fate in mouse fibroblasts, but reprogramming of human somatic cells was limited by low efficiency. Here, we investigated single-cell transcriptional dynamics during human cDC1 reprogramming. Human induced cDC1s (hiDC1s) generated from embryonic fibroblasts gradually acquired a global cDC1 transcriptional profile and expressed antigen presentation signatures, whereas other DC subsets were not induced at the single-cell level during the reprogramming process. We extracted gene modules associated with successful reprogramming and identified inflammatory signaling and the cDC1-inducing transcription factor network as key drivers of the process. Combining IFN-γ, IFN-β, and TNF-α with constitutive expression of cDC1-inducing transcription factors led to improvement of reprogramming efficiency by 190-fold. hiDC1s engulfed dead cells, secreted inflammatory cytokines, and performed antigen cross-presentation, key cDC1 functions. This approach allowed efficient hiDC1 generation from adult fibroblasts and mesenchymal stromal cells. Mechanistically, PU.1 showed dominant and independent chromatin targeting at early phases of reprogramming, recruiting IRF8 and BATF3 to shared binding sites. The cooperative binding at open enhancers and promoters led to silencing of fibroblast genes and activation of a cDC1 program. These findings provide mechanistic insights into human cDC1 specification and reprogramming and represent a platform for generating patient-tailored cDC1s, a long-sought DC subset for vaccination strategies in cancer immunotherapy.
(Less)
- author
- organization
-
- LUCC: Lund University Cancer Centre
- Cell Reprogramming in Hematopoiesis and Immunity (research group)
- StemTherapy: National Initiative on Stem Cells for Regenerative Therapy
- WCMM-Wallenberg Centre for Molecular Medicine
- Bone marrow stem cells and cellular therapies (research group)
- Hematopoiesis and Gene Therapy (research group)
- publishing date
- 2022
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Science Immunology
- volume
- 7
- issue
- 69
- article number
- eabg5539
- pages
- 1 - 18
- publisher
- American Association for the Advancement of Science (AAAS)
- external identifiers
-
- pmid:35245086
- scopus:85125875360
- ISSN
- 2470-9468
- DOI
- 10.1126/sciimmunol.abg5539
- project
- Generating Dendritic Cells by Direct Cell Reprograming
- Elucidating the Mechanisms of Dendritic cell reprogramming
- language
- English
- LU publication?
- yes
- id
- b45ab2fd-8511-4aed-986f-a22de14a636e
- date added to LUP
- 2022-03-17 14:52:27
- date last changed
- 2024-10-03 23:23:02
@article{b45ab2fd-8511-4aed-986f-a22de14a636e, abstract = {{<p>Type 1 conventional dendritic cells (cDC1s) are rare immune cells critical for the induction of antigen-specific cytotoxic CD8+ T cells, although the genetic program driving human cDC1 specification remains largely unexplored. We previously identified PU.1, IRF8, and BATF3 transcription factors as sufficient to induce cDC1 fate in mouse fibroblasts, but reprogramming of human somatic cells was limited by low efficiency. Here, we investigated single-cell transcriptional dynamics during human cDC1 reprogramming. Human induced cDC1s (hiDC1s) generated from embryonic fibroblasts gradually acquired a global cDC1 transcriptional profile and expressed antigen presentation signatures, whereas other DC subsets were not induced at the single-cell level during the reprogramming process. We extracted gene modules associated with successful reprogramming and identified inflammatory signaling and the cDC1-inducing transcription factor network as key drivers of the process. Combining IFN-γ, IFN-β, and TNF-α with constitutive expression of cDC1-inducing transcription factors led to improvement of reprogramming efficiency by 190-fold. hiDC1s engulfed dead cells, secreted inflammatory cytokines, and performed antigen cross-presentation, key cDC1 functions. This approach allowed efficient hiDC1 generation from adult fibroblasts and mesenchymal stromal cells. Mechanistically, PU.1 showed dominant and independent chromatin targeting at early phases of reprogramming, recruiting IRF8 and BATF3 to shared binding sites. The cooperative binding at open enhancers and promoters led to silencing of fibroblast genes and activation of a cDC1 program. These findings provide mechanistic insights into human cDC1 specification and reprogramming and represent a platform for generating patient-tailored cDC1s, a long-sought DC subset for vaccination strategies in cancer immunotherapy.</p>}}, author = {{Rosa, Fábio F and Pires, Cristiana F and Kurochkin, Ilia and Halitzki, Evelyn and Zahan, Tasnim and Arh, Nejc and Zimmermannová, Olga and Ferreira, Alexandra G and Li, Hongzhe and Karlsson, Stefan and Scheding, Stefan and Pereira, Carlos-Filipe}}, issn = {{2470-9468}}, language = {{eng}}, number = {{69}}, pages = {{1--18}}, publisher = {{American Association for the Advancement of Science (AAAS)}}, series = {{Science Immunology}}, title = {{Single-cell transcriptional profiling informs efficient reprogramming of human somatic cells to cross-presenting dendritic cells}}, url = {{http://dx.doi.org/10.1126/sciimmunol.abg5539}}, doi = {{10.1126/sciimmunol.abg5539}}, volume = {{7}}, year = {{2022}}, }