Natural Killer cells drive immunotherapy efficacy in Triple-Negative Breast Cancer patients by modulating their transcriptional plasticity — ASN Events
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Natural Killer cells drive immunotherapy efficacy in Triple-Negative Breast Cancer patients by modulating their transcriptional plasticity (#166)

Elisa Salviato 1 , Veronica Manicardi 1 , Gloria Manzotti 1 , Anna Rita Redavid 1 , Federica Torricelli 1 , Emanuele Vitale 1 , Elisa Gasparini 2 , Moira Ragazzi 3 4 , Carmine Pinto 2 , Alessia Ciarrocchi 1 , Francesca Reggiani 1
  1. Translational Research, Azienda USL-IRCCS of Reggio Emilia, Reggio Emilia, Emilia Romagna, Italia
  2. Oncology Unit, Azienda USL-IRCCS of Reggio Emilia, Reggio Emilia, Emilia Romagna, Italia
  3. Medical and Surgical Sciences for Children and Adults, University of Modena and Reggio Emilia, Modena, Emilia Romagna, Italy
  4. Pathology Unit, Azienda USL-IRCCS of Reggio Emilia, Reggio Emilia, Emilia Romagna, Italia

Background

Immunotherapy in combination with neoadjuvant chemotherapy (NAT) is changing the therapeutic perspective for Triple-Negative Breast Cancer (TNBC) patients, as demonstrated by the KEYNOTE522 trial. We previously showed by morphology-guided digital spatial transcriptome profiling, that Natural Killer (NK) infiltration in diagnostic biopsies of TNBC patients was associated with increased NAT success, suggesting a role of NKs in promoting NAT efficacy1.

Aim

Since NKs are characterized by high transcriptional plasticity associated with their activation2, we aimed to define the transcriptional landscape and crucial transcription factors (TFs) controlling NK effector functions and immunotherapy efficacy in TNBC.

Results

Through single-cell RNA-sequencing (scRNA-seq) and flow cytometry, we explored the intracellular dynamics leading to NK activation in TNBC patients undergoing KEYNOTE-522 NAT. We prospectively enrolled 45 patients and collected peripheral blood at diagnosis (T0) and after NAT completion before surgery (T1). We did not observe a significant difference in NK number or sub-clustering before therapy (T0) when comparing responders to partial/no responders. However, after NAT conclusion (T1), NKs from partial responders displayed an immune-tolerant profile characterized by down-regulation of cytotoxicity-related genes.

A 300-gene signature associated with NK activation was obtained by merging RNA-seq profiles of activated NK92 cells2 and TNBC patient-derived NKs. Gene promoters of this signature were in silico scanned with the FIMO algorithm to predict upstream transcriptional regulators. Among top-scoring TFs, we selected VEZF1, RXRA, and MAZ. Predicted genes recognized in NKs by these TFs were largely overlapped, suggesting a mutual transcriptional regulation and cooperation, further confirmed by ChIP-sequencing. In TNBC patients, differential TF expression was associated with distinct NAT response, confirming their implication in regulating NK functions and therapy outcomes.

Conclusions

Collectively, our data unveil the driving role of NKs in triggering immunotherapy efficacy in TNBC patients and that the differential expression of crucial TFs can improve NK effector functions in TNBC.

  1. Donati B, Reggiani F, Torricelli F, Santandrea G, Rossi T, Bisagni A, Gasparini E, Neri A, Cortesi L, Ferrari G, Bisagni G, Ragazzi M, Ciarrocchi A. Spatial Distribution of Immune Cells Drives Resistance to Neoadjuvant Chemotherapy in Triple-Negative Breast Cancer. Cancer Immunol Res. 2024 Jan 3;12(1):120-134. doi: 10.1158/2326-6066.CIR-23-0076. PMID: 37856875.
  2. Reggiani F, Talarico G, Gobbi G, Sauta E, Torricelli F, Manicardi V, Zanetti E, Orecchioni S, Falvo P, Piana S, Lococo F, Paci M, Bertolini F, Ciarrocchi A, Sancisi V. BET inhibitors drive Natural Killer activation in non-small cell lung cancer via BRD4 and SMAD3. Nat Commun. 2024 Mar 22;15(1):2567. doi: 10.1038/s41467-024-46778-8. PMID: 38519469; PMCID: PMC10960013.