Pereira Lab

#KSMYELOID26 #Immunology #CellReprogramming #CancerImmunotherapy

Looking forward to my talk today, where I will present an update on the conversion of cancer cells into dendritic cells to reinstate anti-tumor immunity, and discuss how cDC1 reprogramming can be used as a model to study tertiary lymphoid structure formation.

Currently on the slopes of Keystone, Colorado attending the for the Symposium Myeloid Cells: Functional Heterogeneity with Therapeutic Promise. It has been a fantastic meeting so far; inspiring science, great discussions on the slopes, and exciting new perspectives on myeloid cells.

Excited to welcome our new postdocs to the lab! Looking forward to exciting science, collaborations, and discoveries together. Welcome aboard!✨
Read more about them on our website: pereiralab.com

Huge congrats to Ervin and thanks to Lund University and the LUCC for awarding the potential of dendritic cell reprogramming for cancer immunotherapy.
@lundsuniversitet.bsky.social @lundstem.bsky.social

We are happy to announce that Ervin Ascic was awarded the Best Cancer Thesis of 2025 at Lund University!

Read full paper here : sciencedirect.com/science/articl…

#CellSystems #CellIdentity #ImmuneReprogramming #CoverArt

The artwork depicts a field of blooming tulips symbolizing immune cell diversity rooted in transcription factor combinations that define immune cell fates. Our combinatorial TF screening platform maps landscape to enable immune cell reprogramming. Illustration: Lilith Lawrence

We’re excited to share that our recent paper, “A combinatorial transcription factor screening platform for immune cell reprogramming”, has been selected for the cover of the January issue of Cell Systems!✨

New from @cellreprolab.bsky.social! The REPROcode library is a collection of over 400 immune-related human transcription factors in barcoded lenti vectors. Explore the set and find your favorite or mix-and-match: www.addgene.org/browse/artic...

🙏🏻 A big thank you to all the funding agencies for making this possible: @ERC_Research, @HorizonEU, @Cancerfonden, @Vetenskapsradet, @novonordiskfond, @fct_pt, #NextgenerationEU, @CancerResearch, WCMM Lund, @Lunduniversity, @Lund_Stem, LUCC, @CNC_UC and Region Skåne.

14) 👥 We are grateful to our collaborators @AsgardThx, @FRosa93, @_CristianaPires, Branko Cirovic, @fabian_theis, @mariia_minaeva, @Dominik1Klein, Ewa Sitnicka, Markus Ringnér and Marcel Martin. We thank everyone for their fantastic contributions!

13) 📖 This study was only possible because of the great effort from current and past Pereira Lab members: @Ilya_ibc, @AbigailRAltman, Inês Caiado, @Halitzki, Diogo Cabral, @luisfholiveira, @nmalavika05 and Daniel Oliveira.

12) Take-home message: REPROcode is a versatile, single-cell-resolved platform to decode TF logic and engineer immune cells, from DC subsets to NK-like cells. REPROcode has broad future applications in cancer, autoimmunity, and tissue repair.

11) Finally, we experimentally validated an NK reprogramming combination: TBX21, ETS1, NFIL3, and EOMES generated NK-like cells expressing CD56, NKG7, NCR1, GZMB, with functional capacity for degranulation and cytokine release.

10) A decision tree trained on barcode counts and single-cell transcriptomes pinpointed key identity drivers: PU.1 at the root of reprogrammed cells, with KLF4 as a major early branch point, revealing a hierarchical TF map to guide immune cell reprogramming.

9) Beyond cDC1: We uncovered TF modules for cDC2, pDC, macrophage, monocyte and NK trajectories. This highlighted the potential of REPROcode to generate immune heterogeneity.

8) Programming immune diversity: Screening 48 TFs enriched in three DC subsets generated multiple immune phenotypes including myeloid and lymphoid, and extinguished fibroblast gene signatures. These patterns were reproduced across three experiments.

7) Beyond cell fate, we could also use REPROcode to identify factors that modify cDC1 cell states when co-expressed with PIB:
• IκB-α/β (NFKBIA/B) → immature cDC1
• MXD1, ID2 → immunostimulatory/mature cDC1
• BCL6, IRF7 → CCR7⁺ migratory cDC1

6) The levels of identified co-factors also matter: GATA2 increased fidelity in a narrow window, as higher levels reduced reprogramming efficiency and XCR1 expression. GFI1B boosted efficiency at higher levels. This reveals distinct mechanisms of cooperation of GATA2 and GFI1B with PIB.

5) We showed that stoichiometry matters: high PU.1 + high BATF3 + intermediate IRF8 = best cDC1 induction from human fibroblasts or multiple cancer cells.

4) Barcode recovery was robust and evenly distributed in reprogrammed cells. On the optimization side, we observed that more TFs, lower MOI, and higher cell recovery sharpened inference from large pools. Our meta-analysis shows this boosts accuracy for identifying instructive TFs.

3) By applying multiplexed TF libraries (9, 22, and 42 factors), we reconstructed PIB-mediated (PU.1, IRF8, BATF3) cDC1 reprogramming, enabling resolution of minimal TF combinations and stoichiometries and revealing GATA2 and GFI1B as key enhancers of lineage fidelity.

2) What's REPROcode? A barcoded, arrayed lentiviral library of 408 immune TFs (now available through Addgene - www.addgene.org/Filipe_Perei...) + scRNA-seq that links which reprogramming TFs a cell receives to the identity it acquires. With no extra barcode PCR steps!

1) Cell reprogramming holds great promise for next-generation immunotherapies, but combinatorial logic of TF network remains a challenge. We developed REPROcode to systematically interrogate TF combinations at single-cell resolution while resolving cell identity and state.

Here is a summary of what we discovered 🧵:

In a game of Mancala, each move strategically redistributes pieces mirroring how TF combinations direct immune cell fate. We present REPROcode to uncover these combinatorial rules providing a roadmap to reprogram cells into defined immune identities. Illustration: Lilith Lawrence

A combinatorial transcription factor screening platform for immune cell reprogramming Direct reprogramming of immune cells holds promise for immunotherapy but is constrained by limited knowledge of transcription factor (TF) networks. He…

📢 New study from out lab published in @CellSystemsCP: REPROcode — a combinatorial single-cell screening platform to discover the transcription-factor (TF) recipes that reprogram immune cells.
Read it here: www.sciencedirect.com/science/arti...

The Pereira Lab was awarded a Discovery grant from the American Brain Tumor Association (ABTA) - Pereira Lab The project “In vivo Dendritic Cell Reprogramming as a New Immunotherapy for Glioblastoma” was awarded a Discovery grant from the ABTA, a non-profit organization that supports breakthrough research id...

3/3 In collaboration with Dr. Johan Bengzon, Dr. Anna Darabi, and Dr. Peter Siesjö (Lund University), as well as Asgard Therapeutics, we seek to change the landscape of treatment for aggressive GBM tumors.
For more details: pereiralab.com/lab-news/the...

#ABTA #Glioblastoma #Immunotherapy

2/3 Here, we will reprogram tumor cells into type 1 conventional dendritic cells (cDC1s) by intra-tumoral delivery of cDC1-inducing transcription factors in glioblastoma (GBM). Since GBM is highly immunosuppressive, cDC1 reprogramming offers a strategy to restore immunogenicity!