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Poster Display session 3

2316 - A 3D co-culture platform of breast cancer and patient derived immune cells to analyse the response to chemotherapy and immunotherapies


30 Sep 2019


Poster Display session 3


Translational Research

Tumour Site

Breast Cancer


Diana Saraiva


Annals of Oncology (2019) 30 (suppl_5): v760-v796. 10.1093/annonc/mdz268


D.P. Saraiva1, A. Jacinto1, S. Braga2, M.G. Cabral1

Author affiliations

  • 1 Cedoc, NOVA Medical School, 1150-347 - Lisbon/PT
  • 2 Instituto Cuf Oncologia, J Mello Saude, 2790-072 - Lisbon/PT


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Abstract 2316


Neoadjuvant chemotherapy (NACT) is the treatment option for locally advanced breast cancer (BC). However, more than half of the patients have no response. Thus, there is an urgent need to find biomarkers of NACT response and personalized therapies for non-responders. Recently, we described a biomarker of NACT response, assessed both in biopsy and blood - HLA-DR expressing cytotoxic T cells (CTLs) with a sensitivity and specificity of 94.12% and 100%, respectively. HLA-DR is associated with CTLs’ cytotoxicity, and it was only observed in NACT-responders, suggesting that functional CTLs are required in the tumor for NACT success. Now, we are validating these results in an independent cohort. Simultaneously, we are establishing 3D spheroid co-culture platforms to tackle new therapeutic options for NACT non-responders.


MCF7 BC cell line was used to spontaneously form a spheroid in agarose coated plates. Peripheral blood mononuclear cells (PBMCs) from NACT-responders or non-responders were added after spheroid formation and they were able to infiltrate the structure. Doxorubicin (doxo), anti-PD-1 and anti-PD-L1 were added in different combinations.


The validation of HLA-DR level in CTLs to predict NACT response has been in agreement with the first cohort. Regarding the BC spheroids, doxo alone or in combination with NACT non-responders PBMCs (low HLA-DR in CTLs) did not affect them. However, when PBMCs from NACT responders (high HLA-DR in CTLs) were added in combination with doxo, the viability and proliferation of MCF7 cells reduced significantly. Notably, when PBMCs from NACT non-responders were stimulated ex vivo, to increase CTLs’ HLA-DR level and IFN-γ production (p = 0.002), MCF7 cells viability decreased (p = 0.0007). Considering that in biopsies of non-responders, the low HLA-DR level in CTLs was negatively correlated with the expression of tumor PD-L1, we are also exploring this platform to inhibit the PD-1/PD-L1 axis in order to stimulate the PBMCs and decrease BC cells viability.


Besides determining if BC patients will respond to NACT, by analysing HLA-DR levels in CTLs, we will now be able to define the best therapy to NACT non-responders, hopefully improving the BC treatment.

Clinical trial identification

Editorial acknowledgement

Legal entity responsible for the study

The authors.


Fundação para a Ciência e Tecnologia (FCT) - PD/BD/114023/2015 and PTDC/BBB-BMD/4497/2014. Liga Portuguesa Contra o Cancro - LPCC-NRS/Terry Fox 2019/2020.


All authors have declared no conflicts of interest.

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