Breast Cancer Research最新文献

Background: The insulin-like growth factor (IGF) pathway is implicated in a naturally occurring process of tissue remodeling during which cells acquire stem cell-like characteristics. We examined associations of circulating IGF-1 and IGF binding protein-3 (IGFBP-3) with expression of CD44, CD24, and ALDH1A1 stem cell markers in benign breast biopsies.

Methods: This study included 151 cancer-free women with incident biopsy-confirmed benign breast disease and blood samples within the Nurses' Health Study II. The data on reproductive and other BCa risk factors were obtained from biennial questionnaires. Immunohistochemistry (IHC) was done on tissue microarrays. For each core, the IHC expression was assessed using QuPath, and expressed as % of cells that stain positively for a specific marker out of the total cell count. Generalized linear regression was used to examine the associations of plasma IGF-I and IGFBP-3 (continuous log-transformed and quartiles) with log-transformed expression of each marker (in epithelium and stroma), adjusted for BCa risk factors.

Results: In multivariate analysis, continuous circulating IGF-1 and IGFBP-3 measures were not associated with the continuous expression of any of the markers in the epithelium or stroma. Women whose IGFBP-3 levels were in the top quartile appeared to have lower expression of stromal CD24 compared to those in the lowest quartile (β = - 0.38, 95% CI - 0.69, - 0.08, p-trend = 0.06).

Conclusions: Higher circulating IGFBP-3 levels were associated with lower stromal CD24 expression in benign breast tissue. Our findings provide indirect evidence of the inducing effect of IGF pathway on epithelial-to-mesenchymal transitions and stem cell activity in the breast.

Background: Early prediction of treatment response to neoadjuvant therapy (NAT) in breast cancer patients can facilitate timely adjustment of treatment regimens. We aimed to develop and validate a MRI-based enhanced self-attention network (MESN) for predicting pathological complete response (pCR) based on longitudinal images at the early stage of NAT.

Methods: Two imaging datasets were utilized: a subset from the ACRIN 6698 trial (dataset A, n = 227) and a prospective collection from a Chinese hospital (dataset B, n = 245). These datasets were divided into three cohorts: an ACRIN 6698 training cohort (n = 153) from dataset A, an ACRIN 6698 test cohort (n = 74) from dataset A, and an external test cohort (n = 245) from dataset B. The proposed MESN allowed for the integration of multiple timepoint features and extraction of dynamic information from longitudinal MR images before and after early-NAT. We also constructed the Pre model based on pre-NAT MRI features. Clinicopathological characteristics were added to these image-based models to create integrated models (MESN-C and Pre-C), and their performance was evaluated and compared.

Results: The MESN-C yielded area under the receiver operating characteristic curve (AUC) values of 0.944 (95% CI: 0.906 - 0.973), 0.903 (95%CI: 0.815 - 0.965), and 0.861 (95%CI: 0.811 - 0.906) in the ACRIN 6698 training, ACRIN 6698 test and external test cohorts, respectively, which were significantly higher than those of the clinical model (AUC: 0.720 [95%CI: 0.587 - 0.842], 0.738 [95%CI: 0.669 - 0.796] for the two test cohorts, respectively; p < 0.05) and Pre-C (AUC: 0.697 [95%CI: 0.554 - 0.819], 0.726 [95%CI: 0.666 - 0.797] for the two test cohorts, respectively; p < 0.05). High AUCs of the MESN-C maintained in the ACRIN 6698 standard (AUC = 0.853 [95%CI: 0.676 - 1.000]) and experimental (AUC = 0.905 [95%CI: 0.817 - 0.993]) subcohorts, and the interracial and external subcohort (AUC = 0.861 [95%CI: 0.811 - 0.906]). Moreover, the MESN-C increased the positive predictive value from 48.6 to 71.3% compared with Pre-C model, and maintained a high negative predictive value (80.4-86.7%).

Conclusion: The MESN-C using longitudinal multiparametric MRI after a short-term therapy achieved favorable performance for predicting pCR, which could facilitate timely adjustment of treatment regimens, increasing the rates of pCR and avoiding toxic effects.

Trial registration: Trial registration at https://www.chictr.org.cn/ .

Registration number: ChiCTR2000038578, registered September 24, 2020.

Neoadjuvant checkpoint blockade immunotherapy (NATI) significantly prolonged outcomes for triple-negative breast cancer (TNBC). Residual tumor cells that survive NATI represent high-risk cell populations with metastatic potential and usually evade immunosurveillance by NK cells. Using an 82-protein panel, we here profiled single-cell membrane proteomics of CD56+ (NCAM1+) NK cells from tumor, peri-cancerous tissue, as well as peripheral blood from 28 TNBC patients post-NATI of residual cancer burden II/III. Unsupervised clustering resulted in several distinct clusters: 2 tumor-infiltrating NK (TINK) clusters with divergent functions of immune activation (TNFRSF7+) and suppression (SELL+); 2 immuno-suppressive peri-cancerous clusters; and 1 periphery-specific cluster. Considering the contradiction of the 2 TINK clusters, we further tested cytokine functions of SELL + and TNFRSF7 + TINKs by single-cell secreting proteomics using a 32-cytokine panel. Consistently, SELL + TINK clusters were characterized by immuno-suppressive secretion patterns (IL10+). A low proportion of SELL + TINK cluster and low proportion of IL10 + secreting SELL + TINK cluster (single-cell secreting proteomics) were both associated with better progression-free survival time. These findings were validated in an independent cohort of 15 patients during 16-month follow-up. Overall, we identified a distinct immuno-suppressive TINK cell group, featuring IL10 + secreting and SELL expression with a strong relation to poor survival prognosis in TNBC patients post-NATI.

Background: The role of T cell immunity during antineoplastic therapy is poorly understood. In the BEGYN-1 study, patients with breast cancer underwent quarterly assessments prior to and during antineoplastic therapy over a period of 12 months.

Methods: We used flow cytometry and multiplex immunoassays to quantify 25 T cell subpopulations and seven T cell associated plasma cytokines in peripheral blood from 92 non-metastatic breast cancer patients, respectively. In addition, the association between T cell dynamics and the outcome of patients undergoing neoadjuvant chemotherapy was investigated.

Results: In patients undergoing chemotherapy, a significant reduction in T helper (Th) cells, particularly naïve central and effector cells and thymus positive Th cells, was observed over time. Interestingly, Th1 immune response-associated cytokines (IL-12, TNF, IFN-γ) declined while Th2 cells and cytotoxic T cells increased over time.

Conclusions: We conclude that in breast cancer patients, chemotherapy is associated with a transition from a Th1 immune response towards Th2 and an increase in cytotoxic T cells, whereas in patients without chemotherapy, these alterations were less pronounced. Future studies should clarify whether patterns of T cell subsets or plasma cytokines can be used as biomarkers to monitor or even improve therapeutic interventions.

Background: Trastuzumab emtansine (T-DM1) is an antibody-drug conjugate that is an effective therapy for HER2-positive breast cancer; however, its efficacy is limited by drug resistance. While multiple mechanisms of resistance have been proposed, these are not yet well understood. Greater understanding of T-DM1 sensitivity and resistance could provide new combination strategies to overcome resistance or predictive biomarkers to guide therapy.

Methods: We have conducted CRISPR/Cas9 functional genomics modifier screens in HER2-positive breast cancer cell lines to allow for unbiased discovery of T-DM1 sensitivity and resistance genes. Whole-genome knockout screens were carried out in MDA-MB-361 and MDA-MB-453 cells treated with T-DM1 and its payload cytotoxin DM1. Hits were validated in secondary T-DM1 screens using a focused single-guide RNA (sgRNA) library and subsequently by individual gene knockout.

Results: The whole-genome CRISPR screens with T-DM1 and DM1 identified 599 genes as potential modifiers of T-DM1 sensitivity and resistance. Of these, 17 genes were significantly enriched and 3 genes depleted at P < 0.001 in either or both MDA-MB-361 and MDA-MB-453 libraries in the secondary screens. Among the top hits, were known T-DM1 sensitivity genes ERBB2 and SLC46A3, in addition to negative regulators of mTOR complex 1: TSC1 and TSC2. MDA-MB-453 clones with knockout of TSC1 or partial knockout of TSC2 were more resistant to T-DM1 than wild type cells in competition growth assays and to T-DM1 and other HER2 targeting therapies (T-DXd, lapatinib and neratinib) in growth inhibition assays, and had increased internalisation of T-DM1 at 6 h. T-DM1 and the mTOR inhibitor everolimus demonstrated synergistic activity at inhibiting cell proliferation at multiple T-DM1 concentrations across four HER2-positive breast cancer cell lines.

Conclusions: Our CRISPR screening approach with T-DM1 in HER2-positive breast cancer cell lines identified genes not previously implicated in T-DM1 sensitivity or resistance, including TSC1 and TSC2. These genes may inform new strategies to enhance T-DM1 therapy in the clinic.

Breast cancer is the most prevalent cancer in women worldwide. Aberrant epigenetic reprogramming such as dysregulation of histone acetylation has been associated with the development of breast cancer. Histone acetylation modulators have been targeted as potential treatments for breast cancer. This review comprehensively discusses the roles of these modulators and the effects of their inhibitors on breast cancer. In addition, epigenetic reprogramming not only affects breast cancer cells but also the immunosuppressive myeloid cells, which can facilitate breast cancer progression. Therefore, the review also highlights the roles of these immunosuppressive myeloid cells and summarizes how histone acetylation modulators affect their functions and phenotypes. This review provides insights into histone acetylation modulators as potential therapeutic targets for breast cancer.

Background: Triple negative breast cancer (TNBC) is an aggressive breast cancer subtype characterized with poor prognosis and high metastatic potential. Although traditional chemotherapy, radiation, and surgical resection remain the standard treatment options for TNBC, bispecific antibody-based immunotherapy is emerging as new strategy in TNBC treatment. Here, we found that the receptor tyrosine kinase-like Orphan Receptor 1 (ROR1) was highly expressed in TNBC but minimally expressed in normal tissue. A bispecific ROR1-targeted CD3 T cell engager (TCE) was designed in IgG-based format with extended half-life.

Method: The expression of ROR1 in TNBC was detected by RT-qPCR and immunohistology analysis. The killing of ROR1/CD3 antibody on TNBC cells was determined by the in vitro cytotoxicity assay and in vivo PBMC reconstituted mouse model. The activation of ROR1/CD3 on T cells was analyzed by the flow cytometry and ELISA assay. Pharmacokinetics study of ROR1/CD3 was performed in mouse.

Results: The ROR1/CD3 TCE triggered T cell activation and proliferation, which showed potent and specific killing to TNBC cells in ROR1-depedent manner. In vivo mouse model indicated that ROR1/CD3 TCE redirected the cytotoxic activity of T cells to lyse TNBC cells and induced significant tumor regression. Additionally, the ROR1/CD3 bispecific antibody exhibited an extended half-life in mouse, which may enable intermittent administration in clinic.

Conclusions: Collectively, these results demonstrated that ROR1/CD3 TCE has a promising efficacy profile in preclinical studies, which suggested it as a possible option for the treatment of ROR1-expressing TNBC.

Background: In HR+/HER2- early breast cancer (EBC) patients, approximately one-third of stage II and 50% of stage III patients experience recurrence, with poor outcomes after recurrence. Given that these patients commonly undergo adjuvant chemo-endocrine therapy (C-ET), accurately predicting the recurrence risk is crucial for optimizing treatment strategies and improving patient outcomes.

Methods: We collected postoperative histopathological slides from 1095 HR+/HER2- EBC who received C-ET and were followed for more than five years at West China Hospital, Sichuan University. Two deep learning pipelines were developed and validated: ACMIL-based and CLAM-based. Both pipelines, designed to predict recurrence risk post-treatment, were based on pretrained feature encoders and multi-instance learning with attention mechanisms. Model performance was evaluated using a five-fold cross-validation approach and externally validated on HR+/HER2- EBC patients from the TCGA cohort.

Results: Both ACMIL-based and CLAM-based pipelines performed well in predicting recurrence risk, with UNI-ACMIL demonstrating superior performance across multiple metrics. The average area under the curve (AUC) for the UNI-ACMIL pipeline in the five-fold cross-validation test set was 0.86 ± 0.02, and 0.80 ± 0.04 in the TCGA cohort. In the five-fold cross-validation test sets, effectively stratified patients into high-risk and low-risk groups, demonstrating significant prognostic differences. Hazard ratios for recurrence-free survival (RFS) ranged from 5.32 (95% CI 1.86-15.12) to 15.16 (95% CI 3.61-63.56). Moreover, among six different multimodal recurrence risk models, the WSI-based risk score was identified as the most significant contributor.

Conclusion: Our multimodal recurrence risk prediction model is a practical and reliable tool that enhances the predictive power of existing systems relying solely on clinicopathological parameters. It offers improved recurrence risk prediction for HR+/HER2- EBC patients following adjuvant C-ET, supporting personalized treatment and better patient outcomes.

Background: Blood DNA methylation (DNAm) profiles have been used to show that changes in circulating leukocyte composition occur during breast cancer development, suggesting that peripheral immune system alterations are markers of breast cancer risk. Blood DNAm profiles have recently been used to predict plasma protein concentrations ("Protein EpiScores"), but their associations with breast cancer risk have not been examined in detail.

Methods: Whole blood DNAm profiles were obtained for a case-cohort sample of participants in the Sister Study and used to calculate 109 Protein EpiScores. Of the 4,479 women included, 2,151 (48%) were diagnosed with breast cancer within 15 years of their baseline blood draw (median time to diagnosis: 8.6 years; 1,673 invasive cancer and 478 ductal carcinomas in situ). Protein EpiScores associations with breast cancer incidence were estimated using weighted Cox regression models, overall and stratified by time and participant characteristics.

Results: Protein EpiScores for RARRES2, IGFBP4, and CCL21 were positively associated with invasive breast cancer risk (hazard ratios from 1.17 to 1.24), while those for F7, SELL, CXCL9, CD48, and IL19 were inversely associated (hazard ratios from 0.82 to 0.86) (all FDR < 0.10). Eight immune response-related Protein EpiScores (CXCL9, CD48, FCGR3B, CXCL11, CCL21, CRTAM, VCAM1, GZMA) were associated with invasive cancers diagnosed within five years of enrollment. Protein EpiScore associations were consistently stronger for estrogen receptor-negative tumors.

Conclusions: Several Protein EpiScores, including many related to immune response, were associated with breast cancer risk, highlighting novel changes to the peripheral immune system that occur during breast cancer development.

Background: Accurately assessing HER2-low (immunohistochemistry [IHC] 1 + and IHC 2+/in situ hybridization [ISH]-) and HER2-ultralow (IHC > 0 < 1+) is essential given the emergence of novel therapies. Thorough understanding of the reproducibility of rescoring IHC stained slides or re-staining archived tissue slides is essential.

Methods: 2,869 breast cancer patients diagnosed between July 2021 and July 2022 from 10 hospitals in China were included in this multicentre study. The prevalence of different HER2 expression levels and distribution of HER2 IHC scores were assessed by HER2 status determination from rescored historical slides. Concordance was evaluated across historical results versus rescored results, historical results versus re-stained results, and leading center results versus local site results. Clinicopathological characteristics were retrospectively analyzed as well.

Results: HER2 IHC 0, IHC 1+, IHC 2+, and IHC 3 + were identified in 682 (23.8%), 871 (30.4%), 801 (27.9%), and 515 (18.0%) cases, respectively. HER2-positive, HER2-low, and HER2 IHC 0 (HER2-ultralow and IHC null) were identified in 21.7%, 54.5%, and 23.8% of cases, respectively. The prevalence of HER2-ultralow and IHC null was 10.6% and 13.2%, respectively. The concordance for HER2-ultralow was 43.3%; 30% of cases that were scored as HER2-ultralow at local sites were rescored as HER2-null and 26.7% of cases were rescored as IHC 1 + at the leading site. Overall, there was substantial agreement (83.1%) between rescored and historical IHC results. A high concordance rate of 91.7% was observed for HER2-low classification.

Conclusions: This is the first multicenter study to determine the prevalence of HER2-low and HER2-ultralow based on rescored results in the Chinese breast cancer population. The concordance analysis carries important implications for the diagnosis of HER2-low and HER2-ultralow cases in clinical practice. The relatively low concordance in identifying HER2-ultralow suggested that the reproducibility of scoring HER2-ultralow needed to be improved through training.

Trial registration: ClinicalTrials.gov identifier NCT05203458.