A high CD8 to FOXP3 ratio in the tumor stroma and expression of PTEN in tumor cells are associated with improved survival in non-metastatic triple-negative breast carcinoma

- A high CD8 to FOXP3 ratio in the tumor stroma and expression of PTEN in tumor cells are associated with improved survival in non-metastatic triple-negative breast carcinoma.
- However, it is still unclear how each diverse cell of the immune system contributes to the prognosis of patients with breast cancer.
- PD-L1 expression in the stroma was associated with the percentage of TILs ( p = 0.018) as, PD-L1 expression in the tumor was associated with the percentage of TILs ( p = 0.049).
- We found a correlation between TILs and PD-L1 expression in stroma cells ( p = 0.020) and in tumor cells ( p = 0.027).
- The CD8 to FOXP3 ratio and the CD4 to FOXP3 ratio were associated with better OS as well, however, only the CD8 to FOXP3 ratio had its prognostic impact confirmed in the METABRIC TNBC cohort.
- There was no association between PD-L1 expression and OS..
- The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material.
- If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.
- FOXP3 expression and the CD8 to FOXP3 ratio in the tumor stroma as well as the loss of PTEN expression in tumor cells are prognostic factors in non-metastatic TNBC..
- Triple-negative breast cancer (TNBC) refers to breast carcinomas that lack the expression of hormone recep- tors (estrogen and progesterone receptors), and that do not express the human epidermal growth factor receptor 2 (HER2) and do not have amplification of the corre- sponding gene (ERBB2).
- Activation of PD-1 by PD-L1 or PD-L2 decreases T cell activity, reduces cytokine production and induces antigen tolerance [12].
- the inhibition of co-inhibitory molecules of the immune system (such as PD-1/PD-L1), has demon- strated high tumor response rates in several tumors, and PD-L1 expression is correlated with response to those treatments [13 – 16].
- In our study we evaluated the prognostic value of the different immune cell subtypes and, as well as the associ- ation between some immunophenotypic immunohisto- chemical biomarkers in the inflammatory infiltrate of early triple-negative breast cancer..
- Exclusion criteria were in situ ductal carcinoma and a previous invasive cancer in the last 5 years preceding study entry.
- The expression of CD20 (B lymphocyte B), CD4 and CD8 (T lymphocyte), FOXP3 (regulatory T-lymphocyte- Treg), CD68 (macrophages), CD163 (M2 response mac- rophages), pSTAT1 and PD-1 was evaluated in inflam- matory cells located in the tumor stroma.
- PD-L1 and PD-L2 expression was evaluated both in the tumor cells and in the inflammatory infiltrate in the tumor stroma..
- Immunohistochemical reactions used an automated protocol and ready-to-use reagents and were performed in the Benchmark Ultra (Ventana Medical Systems, Inc.) equipment following standardized dilutions and reaction conditions for each antibody (Table S1)..
- PD-L1 expression analysis.
- PD-L1 expression was evaluated with the SP263 clone ( Spring Bioscience) and was analyzed by direct optical microscopy by a pathologist (M.M.P.) with specialized training.
- PD-L1 expression was counted as the percent- age of cells with positive membrane staining and was evaluated separately in tumor cells and in the stroma..
- For each sample, the average value obtained from the duplicates was used in the statistical analysis.
- When only one core was evaluable, the value obtained from this core was employed in the analysis..
- Automated counting of the total number of positive cells in the tumor stroma from each core to each biomarker studied (except for PD-L1) was performed using the Aperio AT2 (Leica Biosystems) equipment and the man- ufacturer ’ s suggested protocols for membrane (Aperio Membrane Algorithm) and cytoplasmic (Positive Pixel- Count v9) staining.
- without knowledge of the clinical data.
- In each of the technical replicas in the TMA, an area was delimited, and the number of positive cells for each marker was calculated as cells/mm2.
- Optimal cutoff points for the biomarkers associated with survival were determined by the maximization of the log-rank test method described by LAUSEN e SCHU- MACHER 1992.
- Initially, 379 patients identified as TNBC in the A.
- From this group, 166 patients with HE slides available in the Pathology Department archive were se- lected for analysis of TILs, and among them, 76 patients also had paraffin blocks available, which were used to construct the TMA for immunohistochemical analysis (Fig.
- Clinical-pathological features and outcomes of patients included in the TMA analysis.
- The mean age of the 76 women whose tumors were in- cluded in the TMA was 48.4 years old.
- Analysis of the immune cell infiltrate composition.
- The mean percentage of TILs was 25.78%, and the mean percentage of cells expressing PD-L1 in the tumor and stroma was 14.48 and 20.85%, respectively.
- Repre- sentative microphotographs for each parameter investi- gated are depicted in the Supplementary Online Material (Figures S1 to S9)..
- PD-L1 expression in the stroma was associated with the percentage of TILs ( p = 0.018).
- PD-L1 expression in tumor cells was associated with the percentage of TILs ( p = 0.049) and with N staging ( p = 0.046).
- We found a positive and statistically significant, al- though weak, correlation between the percentage of TILs in the stroma and the expression of PD-L1 in stromal cells (p = 0.020) and in tumor cells.
- CD163 with CD8, CD68, FOXP3, PD-L1 in the stroma and absence of PTEN expression.
- and stromal PDL-1 with tumor PD-L1 and absence of PTEN expression.
- The number of FOXP3 positive cells were also inversely correlated with the expression of PD- L1 in the tumor (Table 4)..
- 1 Flowchart of patients included in the study.
- Although the median survival was numerically dif- ferent (77 and 120 months, respectively for PD-L1 ex- pression in tumor>.
- p = 0.224) for patients whose tumors had high PD-L1 expression, this was not statistically significant (Figure S10)..
- Similarly, PD-L1 expression in the stroma and the number of cells positive for CD8, CD4, CD20, pSTAT-1, CD68, CD163 or PD-1 in the stroma were not associated with overall survival (Figures S11 to S13 and S15 to S19)..
- Impact of CD8A to FOXP3 ratio on the survival of TNBC in the METABRIC cohort.
- We calculated the overall survival of non-metastatic, triple-negative breast cancer patients included in the METABRIC according to the ratio between the ex- pression of CD8A and FOXP3 genes (CD8A/FOXP3 Table 1 Clinical-pathological features and outcomes of patients included in the TMA analysis ( N = 76).
- We could not validate the prognostic impact of PTEN in the METABRIC dataset since the gene was not annotated..
- Although we observed a high number of CD8 + and CD4 + cells in the stroma of TNBC, these cells were directly correlated with increased numbers of CD68 + and CD163 + cells, as well as FOXP3 + cells.
- Be- sides, the number of CD8 + cells also correlated with a higher number of PD-L1+ cells in the stroma.
- We hypothesize that, although TN breast tumors may effect- ively attract T lymphocytes at some point in their pro- gression, these lymphocytes probably have their effector activity suppressed by the co-recruitment of immuno- suppressive cells (M2 macrophages and regulatory lym- phocytes) and the acquisition of an exhausted phenotype due to the PD-L1 expression, nevertheless this needs to be further validated..
- The description of the frequency of these cells in the intratumoral or stromal compartments is very heteroge- neous in the literature, as well as the determination of its clinical significance [21].
- evaluated in the tumor cells or in the tumor stroma.
- PD-L1 Tumor 62 4.53.
- PD-L1 Estroma 48 12.17.
- Table 3 Association between clinical-pathological characteristics and the expression of PD-1, PD-L1 in tumor cells and in stromal cells.
- Median was used to dichotomize the expression of PD-L1 and PD1.
- of the immunophenotype of the cells in the tumor im- mune infiltrate by flow or mass cytometry would be very informative..
- We also found a positive and statistically signifi- cant correlation between the percentage of cells ex- pressing PD-L1 in the stroma (p = 0.020) and PD-L1 in the tumor (p = 0.027), reinforcing some data from the literature suggesting that the higher the.
- inflammatory tumor infiltrate the higher the expres- sion levels of PD-L1 [22].
- Nevertheless, PD-L1 ex- pression, both in the stroma and in the tumor was not associated with OS.
- [17] demon- strated that PD-L1 expression is a negative prognos- tic factor for OS, independent of the histological tumor subtype.
- Table 4 Correlation of the percentage of TILs and the expression of the studied biomarkers among themselves.
- Markers CD20 CD4 CD163 CD8 CD68 FOXP3 PD-L1 Stroma PD-L1 Tumor PD1 PTEN Absent pSTAT1 TIL*.
- PD-L1 Stroma.
- PD-L1 Tumor.
- In some samples analyzed for PD-L1 expression, both cores in the TMA were unevaluable because the tissue detached from the slide, in other cases, the core represented a tumor with high cellularity and did not contain stroma.
- imbalance in the proportion of informative cases in each scenario.
- Lymphocytes infiltrated into the tumor are an import- ant immune component of the cancer response..
- The only variables associated with overall survival were the lack of PTEN expression in tumor cells and the number of FOXP3+ cells in the stroma.
- Together, these data suggest that the T lympho- cytes in the tumor microenvironment may exhibit antitumor activity and promote growth control, as opposed to events favoring immunosuppression (regulatory lymphocyte infiltration).
- This data is rep- licated in the TNBC cohort of METABRIC for those patients whose tumors have a high CD8A/FOXP3 ratio..
- On the other hand, a high CTL/Treg ratio in the tissue surrounding the tumor was signifi- cantly associated with improved OS and PFS [9]..
- 4 Overall Survival of triple-negative breast cancer patients in the METABRIC cohort according to the ratio between the expression of CD8A and FOXP3.
- Our data suggest that the presence of exuberant in- flammatory infiltrate in TNBC is associated with PD-L1 expression both in the tumor itself and in the tumor stroma, as well as with enrichment for T lymphocytes (CD8+ and CD4.
- which may make these tumors im- portant targets for treatment with PD-1 or PD-L1 inhibi- tors.
- Furthermore, macrophages and regulatory T lymphocytes probably represent important counter- regulatory mechanisms that the tumor recruits to allow its escape of the adaptive immune response and a high CD8 to FOXP3 ratio in the tumor stroma is a predictor of improved survival in non-metastatic TNBC (Fig.
- In the future, it would be interesting to evaluate the role of strategies that deplete FOXP3+ cells in the tumor microenvironment..
- Microscopic analysis of the tumor infiltrating lymphocytes.
- PD-L1 expression on stromal and tumor cells.
- A - PD- L1 membrane staining in stromal cells (macrophages).
- B - PD-L1 membrane staining in tumor cells.
- Brown coloration indi- cates PD-L1 expression (immunohistochemistry with DAB).
- Overall survival curves according to labeling for PD-L1 in the stroma (>.
- The vertical strokes in the curves represent censored cases.
- Overall survival curves according to PD-L1 expression in tumor cells.
- A more exuberant mononuclear infiltrate is associated with increased overall survival, as well as with increased PD-L1 expression in both the stroma and tumor cells.
- Overall survival curves according to the ratio of the number of CD4 and FOXP3 positive cells.
- The vertical strokes in the curves represent cen- sored cases.
- Overall Sur- vival of triple-negative breast cancer patients in the METABRIC cohort ac- cording to the expression of FOXP3 .
- Overall Survival of triple-negative breast cancer patients in the METABRIC cohort according to the ratio between the expression of CD4 to FOXP3 .
- Macedo analyzed the expression of PD-L1.
- https://doi.org .
- doi.org .
- https://doi.org/10.1007/s .
- Targeting immune co-stimulatory effects of PD-L1 and PD-L2 might represent an effective therapeutic strategy in stroke.
- Safety and activity of anti-PD-L1 antibody in patients with advanced cancer..
- Association of PD- 1, PD-1 ligands, and other features of the tumor immune.
- https://doi.org/10.1 016/S .
- High PD-L1 expression is closely associated with tumor-infiltrating lymphocytes and leads to good clinical outcomes in Chinese triple negative breast cancer patients.
- Variation in the incidence and magnitude of tumor-infiltrating lymphocytes in breast cancer subtypes: a systematic review.
- https://doi.org/10.1186/s

Xem thử không khả dụng, vui lòng xem tại trang nguồn
hoặc xem Tóm tắt