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Direct Reprogramming of Mouse and Human Fibroblasts into Conventional Dendritic Cells Type 1

Fiúza Rosa, Fábio LU ; Pires, Cristiana LU ; Kurochkin, Ilia LU ; Ferreira, Alexandra G. LU orcid ; Schulz, Oliver ; Reis e Sousa, Caetano and Pereira, Carlos-Filipe LU orcid (2022) In Molecular Immunology 150. p.22-22
Abstract
Cell fate reprogramming of adult cells towards pluripotency or unrelated somatic cell-types has been explored in the context of regenerative medicine. Dendritic cells (DCs) are professional antigen presenting cells (APCs) specialized in the recognition, processing and presentation of antigens to T-cells, inducing adaptive immunity. In particular, the mouse conventional DCs type 1 (cDC1) subset or DC1 human equivalent excel on the ability to perform antigen cross-presentation, a critical step for inducing cytotoxic responses. We hypothesized that the unique properties of cDC1s could be induced in unrelated cell-types, allowing the direct control of immune responses with cell reprogramming.

Here, the requirements to induce cDC1s... (More)
Cell fate reprogramming of adult cells towards pluripotency or unrelated somatic cell-types has been explored in the context of regenerative medicine. Dendritic cells (DCs) are professional antigen presenting cells (APCs) specialized in the recognition, processing and presentation of antigens to T-cells, inducing adaptive immunity. In particular, the mouse conventional DCs type 1 (cDC1) subset or DC1 human equivalent excel on the ability to perform antigen cross-presentation, a critical step for inducing cytotoxic responses. We hypothesized that the unique properties of cDC1s could be induced in unrelated cell-types, allowing the direct control of immune responses with cell reprogramming.

Here, the requirements to induce cDC1s were investigated using combinatorial overexpression of Transcription Factors (TFs) in Clec9a-tdTomato mouse fibroblasts. In the hematopoietic system, Clec9a specifically marks the DC lineage, including all conventional dendritic cells type 1 (cDC1). We have identified PU.1, IRF8 and BATF3 (PIB) as sufficient and necessary to induce Clec9a reporter activation, establish DC morphology and activate a cDC1 transcriptional program in mouse fibroblasts. The over- expression of PIB ignites the expression of DC markers including CD103, XCR1, MHC-I, MHC-II and co-stimulatory molecules. Functionally, Induced DCs (iDCs) secrete inflammatory cytokines and engulf, process, present and cross-present antigens to CD4+ and CD8+ T cells, respectively. Additionally, we have demonstrated that combined expression of PIB factors induces DC1 reprogramming in human fibroblasts. Human iDC1s acquire DC morphology, express DC1 markers, including Clec9a, CD141 and the co-stimulatory molecules CD40, CD80 and CD86, and acquire a DC1 transcriptional signature at the single cell level. Interestingly, DC1 reprogramming efficiency can be enhanced 70-fold by supplementing culture media with inflammatory cytokines, suggesting a regulatory role of inflammation during DC1 reprogramming.

Hence, we provide evidence that antigen presentation and cross-presentation can be dynamically programmed by a small combination of TFs. These findings provide insights into cDC1 specification and a platform for developing cancer immunotherapies based on cell reprogramming. (Less)
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author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Molecular Immunology
volume
150
pages
22 - 22
publisher
Pergamon Press Ltd.
ISSN
1872-9142
DOI
10.1016/j.molimm.2022.05.076
language
English
LU publication?
yes
id
db11ebbf-47cd-4820-8024-6f7f1f4b7c57
date added to LUP
2023-08-10 21:15:57
date last changed
2023-08-11 11:06:08
@misc{db11ebbf-47cd-4820-8024-6f7f1f4b7c57,
  abstract     = {{Cell fate reprogramming of adult cells towards pluripotency or unrelated somatic cell-types has been explored in the context of regenerative medicine. Dendritic cells (DCs) are professional antigen presenting cells (APCs) specialized in the recognition, processing and presentation of antigens to T-cells, inducing adaptive immunity. In particular, the mouse conventional DCs type 1 (cDC1) subset or DC1 human equivalent excel on the ability to perform antigen cross-presentation, a critical step for inducing cytotoxic responses. We hypothesized that the unique properties of cDC1s could be induced in unrelated cell-types, allowing the direct control of immune responses with cell reprogramming.<br/><br/>Here, the requirements to induce cDC1s were investigated using combinatorial overexpression of Transcription Factors (TFs) in Clec9a-tdTomato mouse fibroblasts. In the hematopoietic system, Clec9a specifically marks the DC lineage, including all conventional dendritic cells type 1 (cDC1). We have identified PU.1, IRF8 and BATF3 (PIB) as sufficient and necessary to induce Clec9a reporter activation, establish DC morphology and activate a cDC1 transcriptional program in mouse fibroblasts. The over- expression of PIB ignites the expression of DC markers including CD103, XCR1, MHC-I, MHC-II and co-stimulatory molecules. Functionally, Induced DCs (iDCs) secrete inflammatory cytokines and engulf, process, present and cross-present antigens to CD4+ and CD8+ T cells, respectively. Additionally, we have demonstrated that combined expression of PIB factors induces DC1 reprogramming in human fibroblasts. Human iDC1s acquire DC morphology, express DC1 markers, including Clec9a, CD141 and the co-stimulatory molecules CD40, CD80 and CD86, and acquire a DC1 transcriptional signature at the single cell level. Interestingly, DC1 reprogramming efficiency can be enhanced 70-fold by supplementing culture media with inflammatory cytokines, suggesting a regulatory role of inflammation during DC1 reprogramming.<br/><br/>Hence, we provide evidence that antigen presentation and cross-presentation can be dynamically programmed by a small combination of TFs. These findings provide insights into cDC1 specification and a platform for developing cancer immunotherapies based on cell reprogramming.}},
  author       = {{Fiúza Rosa, Fábio and Pires, Cristiana and Kurochkin, Ilia and Ferreira, Alexandra G. and Schulz, Oliver and Reis e Sousa, Caetano and Pereira, Carlos-Filipe}},
  issn         = {{1872-9142}},
  language     = {{eng}},
  month        = {{10}},
  note         = {{Conference Abstract}},
  pages        = {{22--22}},
  publisher    = {{Pergamon Press Ltd.}},
  series       = {{Molecular Immunology}},
  title        = {{Direct Reprogramming of Mouse and Human Fibroblasts into Conventional Dendritic Cells Type 1}},
  url          = {{http://dx.doi.org/10.1016/j.molimm.2022.05.076}},
  doi          = {{10.1016/j.molimm.2022.05.076}},
  volume       = {{150}},
  year         = {{2022}},
}