Background:The GenesWell BCT is a prognostic assay that integrates the expression of six prognostic and three reference genes with clinicopathological factors (tumor size and nodal status) to generate a BCT Score. The BCT Score is used to predict the risk of recurrence in early breast cancer. We excluded clinical variables and developed a gene-only version of the BCT Gene Score to evaluate its independent prognostic performance. Methods:We assessed the prognostic value of the BCT Gene Score in a cohort of patients with ER+/HER2- early breast cancer using available Oncotype DX Recurrence Score (RS) data. The primary endpoint was concordance between the assays and recurrence-free survival (RFS). Results:We analyzed 759 samples from five Korean institutions. Risk classification concordance between the BCT Gene Score and the BCT Score was 81.2%, whereas concordance with the Oncotype DX RS was 74.2%, indicating moderate agreement. All three scores were significantly associated with RFS (p < 0.05). Combining the BCT and BCT Gene Scores improved prognostic stratification compared with either score alone. Patients classified as high risk by both scores exhibited the poorest prognosis, whereas those classified as low risk by both scores exhibited the most favorable outcomes. Among patients with low RS, those reclassified as high risk by the BCT Gene Score exhibited significantly worse RFS. Conclusions:The BCT Gene Score exhibits independent prognostic utility and complements the original BCT Score. Risk stratification may be enhanced by incorporating both scores, ultimately guiding more precise treatment decisions in ER+/HER2- early breast cancer.

Purpose:Triple-negative breast cancer (TNBC) is an aggressive subtype with poor prognosis. CD103+ tissue-resident memory T (TRM) cells are crucial for anti-tumor immunity in TNBC. We investigated whether their spatial interactions with other T-cells influence clinical outcomes, particularly following neoadjuvant chemotherapy (NAC). Materials and Methods:This retrospective study analyzed 182 TNBC patients (98 NAC-treated; 84 non-NAC). Using Opal™ multiplex immunohistochemistry data and the Spatial Image Analysis of Tissues (SPIAT) R package, we analysed spatial interactions between CD103+ cells and other T cell subsets (CD45RO, CD8, CD4, PD-1) in central/peripheral tumor regions. Normalized mixing score (NMS) quantified spatial interactions. Results:NMS-based clustering revealed two distinct CD103+ cell interaction patterns-Cluster 1 (low NMS) characterized by weaker and Cluster 2 (high NMS) by stronger spatial interactions between CD103+ and other T cell subsets. In the NAC group, Cluster 2 in the tumor periphery was associated with lower pathologic stage (p=0.002), higher stromal tumor-infiltrating lymphocyte level (p=0.031), and significantly improved recurrence-free survival (p=0.028) and overall survival (p=0.018) compared to Cluster 1. Central tumor region clustering patterns had no association with prognosis. No significant survival-related differences were observed in the non-NAC group according to NMS-based clustering. Conclusion:Spatial interaction patterns between CD103+ and other T cell subsets in the tumor periphery influence clinical outcomes in NAC-treated TNBC patients. Analysing such spatial relationships between T cells, rather than their presence alone, may provide additional prognostic information for patients undergoing NAC.

Background:Activating mutations in ESR1 gene are a known mechanism of endocrine resistance. We analyzed ESR1 mutations in cell-free DNA (cfDNA) from serially collected blood from hormone receptor-positive, human epidermal growth factor receptor 2-negative metastatic breast cancer patients to investigate their impact on outcomes. Methods:A total of 268 cfDNA samples serially collected from 25 patients who underwent palliative endocrine therapy were examined. Seven ESR1 hotspot mutations were tested in cfDNA using droplet digital polymerase chain reaction. Progression-free survival (PFS) was analyzed using the Kaplan-Meier method. Blood ESR1 mutation (bESR1) results were correlated with clinical response. Results:Among 25 patients, baseline bESR1 mutation was negative for all patients (0/4) and 64.0% (16/25) was bESR1-positive at any time. bESR1 positivity did not affect PFS. Those whose bESR1 cleared in subsequent cfDNA exhibited longer PFS. Among patients with clinical progression, 57.1% (8/14) were bESR1-positive, and four had bESR1 detected before clinical progression. Conclusions:We observed worse outcomes in patients with persistent bESR1 detection during first-line endocrine therapy and then sustained positive. bESR1 was detected prior to clinical progression in half of patients. Our study suggests the benefit of bESR1 monitoring during palliative endocrine therapy.

AIM:To understand whether the worldwide implementation of PD-L1 testing in triple-negative breast cancer (TNBC) can be achieved in routine clinical practice. METHODS AND RESULTS:The multicentre retrospective observational VANESSA study consecutively and uniformly enrolled patients treated with systemic therapy for early or metastatic (e/m)TNBC diagnosed between 2014 and 2017. PD-L1 status was retrospectively assessed locally and centrally using the VENTANA PD-L1 (SP142) Assay (PD-L1 expression on tumour-infiltrating immune cells covering ≥1% of the tumour area). The primary objective was to determine the prevalence of PD-L1 positivity assessed locally on primary and/or metastatic tumour tissue. Concordance between local and central testing was a secondary endpoint. PD-L1-positive prevalence was 38% in eTNBC (728/1902) and 20% in mTNBC (30/152) and was higher in submitted tissue size >5 versus <5 mm diameter (eTNBC: 43% versus 16%; mTNBC: 24% versus 13%). Among 1967 samples tested both centrally and locally, concordance was 75% (Cohen's κ coefficient 0.52, 95% CI 0.48-0.55) and was similar regardless of cohort (eTNBC versus mTNBC), sample collection method (biopsy versus resection) or sample origin (primary versus metastatic). PD-L1-positive prevalence was higher by central versus local assessment (eTNBC: 55% versus 39%; mTNBC: 26% versus 20%). CONCLUSION:In this real-world study, PD-L1-positive prevalence was lower than in prospective trials assessing PD-L1 status centrally, lower in mTNBC than eTNBC, lower in smaller than larger tissue samples and lower by local than central assessment. These findings underline the importance of central PD-L1 testing on sufficiently large samples to ensure optimal selection for therapies targeting PD-(L)1 in mTNBC.

Background:This study aimed to determine the prevalence of human epidermal growth factor receptor 2 (HER2)-ultralow breast cancer among cases initially classified as HER2 immunohistochemistry (IHC) 0 and assess interobserver variability in interpreting low-level HER2 expression. Methods: In this multicenter retrospective study, all invasive breast cancer cases diagnosed between January and December 2022 across 10 Korean institutions were retrieved. Institutional pathologists reexamined HER2 IHC slides originally reported as IHC 0 according to the 2018 American Society of Clinical Oncology/College of American Pathologists guidelines and reclassified them as HER2-null (0), HER2-ultralow (0+), or HER2-low (1+). Slides from 10% of HER2-null and HER2-ultralow cases were digitized for central review and independently assessed by two pathologists, with discrepancies resolved by consensus. Results: Among 8,026 cases, 2,836 cases (35.5%) were initially reported as IHC 0. Upon re-review, 1,673 (59.0%), 1,139 (40.2%), and 24 (0.8%) cases were reclassified as HER2-null, HER2-ultralow, and HER2-low, respectively. The prevalence of HER2-ultralow breast cancer varied considerably across institutions (23.7%-78.1%). Central review of 268 digitized cases showed concordance in 193 cases (72.0%). Among the 75 discordant cases, 54 tumors (72.0%) were upgraded from HER2-null to HER2-ultralow, and 18 (24.0%) tumors were upgraded from HER2-ultralow to HER2-low. Furthermore, two tumors (2.7%) were downgraded from HER2-ultralow to HER2-null. Conclusions: Approximately 40% of cases initially categorized as IHC 0 were reclassified as HER2-ultralow. The substantial inter-institutional variability observed in interpreting low-level HER2 expression highlights the need for standardized training and quality assurance to ensure accurate identification of patients eligible for HER2-targeted antibody-drug conjugates.

Human epidermal growth factor receptor type 2 (HER2) is overexpressed in 15–20

BACKGROUND:The recent approval of trastuzumab deruxtecan for human epidermal growth factor receptor 2 (HER2)-low and HER2-ultralow breast cancer mandates an adequate assessment of these categories. METHODS:Seven breast pathologists from the Breast Pathology Study Group of the Korean Society of Pathologists held an on-site expert consensus meeting. Fifteen sets of virtual whole slide images (WSI) of hematoxylin and eosin stain and HER2 immunohistochemistry were provided. The pathologists were given 60 minutes to submit their diagnosis of HER2 expression into null, ultralow, 1+, 2+, or 3+. Afterwards, in-depth discussion and consensus diagnoses were made by real-time visualization of the WSI. RESULTS:After the consensus meeting, unanimous 100% agreements were seen only in five (33.3%) of the examined cases, which consisted of three 1+ cases and two 2+ cases. Two cases (13.3%) had mild disagreement, with only one pathologist's disagreement. Of note, eight cases (53.3%) showed significant disagreement, defined by more than two pathologists' disagreement. All HER2-null cases were reclassified as ultralow after consensus review, suggesting potential widespread underclassification of ultralow cases in clinical practice. CONCLUSIONS:Experts had significant discrepancies in interpreting HER2-low/ultralow status. It is important to assess if the distinction between HER2-low and ultralow is strictly required and if HER2-null breast cancer exists in reality.

PURPOSE:Based on the KEYNOTE-522 study, neoadjuvant pembrolizumab plus chemotherapy has become the standard treatment for early-stage triple-negative breast cancer (TNBC). This study evaluated the real-world efficacy, safety, and predictors of pathologic complete response (pCR) in Korean patients. METHODS:We conducted a retrospective cohort study of 174 patients with early-stage TNBC who received the KEYNOTE-522 regimen (neoadjuvant pembrolizumab plus paclitaxel and carboplatin, followed by doxorubicin and cyclophosphamide) at a tertiary cancer center between August 2022 and July 2024. We assessed the primary endpoints, including pCR rate and event-free survival (EFS). We performed univariable and multivariable logistic regression analyses to identify independent predictors of pCR. RESULTS:The median patient age was 50 years (range, 24-74 years). The clinical stages were II and III in 79.3% and 20.1% of patients, respectively, and 10.9% had clinical N3 disease. The overall pCR rate was 62.1%, and the N3 subgroup had a pCR rate of 47.4%. On multivariable analysis, high baseline Ki-67 expression (≥ median, 75%) was significantly associated with pCR (odds ratio, 2.84; 95% confidence interval, 1.45 to 5.66; p = 0.002). At a median follow-up of 18.4 months, the 12-month EFS rate was 97.4%, with significantly superior outcomes observed in patients who achieved pCR compared with those who did not achieve pCR (100% vs. 93.1%, p = 0.007). The treatment completion rate was 92.0%, and immune-related adverse events occurred in 13.8% of patients. CONCLUSION:In this real-world analysis of one of the largest Asian cohorts of patients with early-stage TNBC treated with neoadjuvant pembrolizumab, the KEYNOTE-522 regimen demonstrated substantial efficacy and manageable toxicity, consistent with the original trial findings.

Biomarker testing is central to precision oncology, enabling effective use of targeted therapies in hormone receptor-positive (HR+), human epidermal growth factor receptor 2 (HER2)-negative metastatic breast cancer (mBC). Recommendations for current and future biomarker testing and diagnostic workflows were developed using a three-step approach: pre-meeting survey, descriptive analysis, and steering committee discussion. A panel of 14 experts from Latin America, Asia Pacific, the Middle East, and Africa developed recommendations using an expert opinion-based approach without a formal consensus methodology. Recommendations with >90% agreement were considered panel recommendations. Some recommendations are expert-driven and not yet standard-of-care. High agreement was observed for biomarker testing at diagnosis for estrogen receptor (ER), progesterone receptor (PR), HER2, and BReast CAncer gene (BRCA)1/2. For endocrine-resistant patients, phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) testing is recommended at diagnosis; otherwise, during or after first-line treatment. AKT serine/threonine kinase 1 (AKT1) and phosphatase and tensin homolog (PTEN) testing can occur either during first-line treatment or at progression, while estrogen receptor 1 (ESR1) testing is advised at progression. Tumor tissue (preferably from a re-biopsy) is the preferred sample for PIK3CA, AKT1, and PTEN testing; if unavailable, primary tissue or circulating tumor DNA (ctDNA) may be considered. Blood is recommended for germline BRCA1/2 or PALB2 testing, while ctDNA is preferred for ESR1 detection. Next-generation sequencing (NGS) is the preferred method for most biomarkers, with polymerase chain reaction as an alternative where NGS is unavailable; PTEN may also be assessed using immunohistochemistry. Above 90% of experts proposed future biomarker testing at diagnosis for ER, PR, HER2, germline BRCA1/2, PALB2, PIK3CA, AKT1, PTEN, mismatch repair, microsatellite instability, tumor mutational burden, and neurotrophic tyrosine receptor kinase in all patients with HR+, HER2-negative mBC. This article offers a practical guide for biomarker testing in HR+, HER2-negative mBC.

OBJECTIVE:Cyclin-dependent kinase 4/6 (CDK4/6) inhibitors combined with endocrine therapy (ET) constitute the standard systemic treatment for estrogen receptor-positive, human epidermal growth factor 2-negative (ER+/HER2-) metastatic breast cancer (MBC). However, treatment responses remain heterogeneous, highlighting the need for reliable prognostic markers. This study aimed to evaluate the prognostic significance of ¹⁸F-fluorodeoxyglucose (FDG) PET/CT findings in this setting. MATERIALS AND METHODS:This retrospective single-center cohort study included patients with ER+/HER2- MBC who underwent ¹⁸F-FDG PET/CT before initiating CDK4/6 inhibitors plus ET between 2018 and 2023. Maximum standardized uptake value (SUVmax), whole-body metabolic tumor volume (MTV), and total lesion glycolysis (TLG) were measured. Progression-free survival (PFS) and overall survival (OS) were evaluated as the primary and secondary outcomes, respectively, using multivariable Cox models. PET parameters (SUVmax, MTV, and TLG) were analyzed as both continuous and dichotomized variables based on median values, adjusting for relevant clinical covariates. RESULTS:Among the 374 patients, 82 (21.9%) presented with de novo metastatic disease, and 357 (95.5%) received CDK4/6 inhibitors as first-line therapy. In multivariable Cox analysis, all continuous PET parameters were independently associated with PFS (adjusted hazard ratio for SUVmax 1.05 [95% confidence interval 1.02-1.08]; log-transformed MTV 1.16 [1.08-1.25]; and log-transformed TLG 1.14 [1.07-1.23]) and OS (SUVmax 1.08 [1.04-1.11]; log-transformed MTV 1.24 [1.12-1.38]; and log-transformed TLG 1.22 [1.11-1.34]) with all P < 0.001. Results based on dichotomized PET parameters were similar to those obtained with continuous values: PFS (adjusted hazard ratio for SUVmax ≥ 7.6, 1.41 [1.08-1.85]; MTV ≥ 21.2 cm³, 1.41 [1.08-1.86]; and TLG ≥ 78.9, 1.51 [1.14-1.99]) with P ≤ 0.013 and OS (1.43 [1.01-2.04]; 1.84 [1.28-2.66]; and 1.73 [1.20-2.50], respectively) with P ≤ 0.046. CONCLUSION:Pretreatment ¹⁸F-FDG PET/CT parameters are independent prognostic markers in patients with ER+/HER2- MBC receiving CDK4/6 inhibitors with ET, supporting their potential utility in risk stratification.

BACKGROUND:Cytological diagnosis of pleural effusion plays an important role in the early detection and diagnosis of lung cancers. Recently, attempts have been made to overcome low diagnostic accuracy and interobserver variability using artificial intelligence-based image analysis. However, such analysis is primarily performed at the image-patch level and not at the whole-slide image (WSI) level. This study aims to develop a WSI-level classification of malignant effusions in metastatic lung cancer based on pleural fluid cytology using a quality-controlled, nationwide dataset. METHODS:The dataset was collected by a consortium research group that included three major university hospitals and the Quality Assurance Program Committee of the Korean Society of Cytopathology. It contains 576 normal and 309 cancer WSIs from pleural fluids. A clustering-constrained attention multiple-instance learning (CLAM) model was used for WSI-level classification. RESULTS:The CLAM model achieved a high accuracy of 97%, with an area under the curve of 0.97, representing a 13% improvement over the image patch classification model-based WSI classification. It also significantly reduced the analysis time and computing resources compared to those required during image patch-level classification and heat map generation on the WSIs. CONCLUSION:The CLAM model successfully demonstrated high performance in differentiating malignant effusion at the WSI level using a large, quality-controlled, nationwide dataset. Further external validation is required to ensure generalizability.

PURPOSE:Although human epidermal growth factor receptor 2 (HER2) positivity is prevalent in microinvasive breast cancer (MIBC), data focused on HER2-positive MIBC are limited. We investigated the clinical course and long-term outcomes of HER2-positive MIBC and evaluated the role of adjuvant chemotherapy. MATERIALS AND METHODS:The study included patients with curatively resected pT1mi pN0 HER2-positive breast cancer between January 2000 and January 2020. Treatments and survival outcomes, including invasive breast cancer-free survival (IBCFS), distant recurrence-free survival (DRFS), and overall survival (OS) were analyzed. RESULTS:The analysis included 799 female patients. The median age was 51 years (range, 23 to 79 years), and 51.6% (n=412) were premenopausal. Multifocality was confirmed in 17.3% (n=138), and estrogen receptor (ER) positivity in 29.8% (n=238). Adjuvant chemotherapy was administered to 17.5% (n=140), with doxifluridine in 96.4% of cases. One patient (0.1%) received trastuzumab. With a median follow-up of 119.0 months (95% confidence interval [CI], 114.0 to 127.0), the 8-year IBCFS, DRFS, and OS were 91.2% (95% CI, 89.1 to 93.3), 97.5% (95% CI, 96.4 to 98.7), and 98.8% (95% CI, 98.0 to 99.6), respectively. No significant differences were observed between patients with and without adjuvant chemotherapy. The lack of differences in IBCFS by chemotherapy was consistent across subgroups, including pre-/postmenopausal patients, grade 1-2/3 tumors, and ER-negative disease. CONCLUSION:A clinically meaningful proportion of HER2-positive MIBC patients experience IBCFS events with long-term follow-up. Adjuvant chemotherapy did not improve survival, potentially due to the use of an outdated, ineffective regimen. The role of modern adjuvant regimens, particularly those incorporating HER2-targeted therapy, warrants further exploration.

BACKGROUND:In early triple-negative breast cancer (TNBC), surgery following neoadjuvant chemotherapy (NAC) is standard. Adjuvant capecitabine has shown survival benefits in patients with residual disease; however, data remain limited regarding the subgroups that benefit most, particularly when stratified by residual cancer burden (RCB), a key prognostic indicator post-NAC. METHODS:Patients with early TNBC and residual disease following NAC (Feb 2008-Dec 2021) were retrospectively included. After propensity score matching to balance baseline characteristics, recurrence-free survival (RFS) was compared between patients who received adjuvant capecitabine and those who did not. RESULTS:Of 828 eligible patients, 631 were included in the final propensity score matched analysis (222 capecitabine; 409 observation). Median ages were 48 years (range, 40-56) and 50 years (range, 41-56) in the observation and capecitabine groups. RCB distribution was comparable between groups: RCB-I (10% vs. 11%), RCB-II (68% in both), and RCB-III (22% vs. 21%). RCB was prognostic across the cohort, with 3-year RFS rates of 92.2%, 73.6%, and 43.3% for RCB-I, -II, and -III, respectively. Adjuvant capecitabine improved RFS in RCB-III (HR, 0.6; 95% CI, 0.3-0.9; P = .02), but not in RCB-I (HR, 1.1; 95% CI, 0.2-6.5; P = .93) or RCB-II (HR, 0.9; 95% CI, 0.7-1.4; P = .84). In RCB-III, distant metastasis occurred less frequently in the capecitabine group compared with the observation group (34.0% vs. 62.9%). CONCLUSION:Adjuvant capecitabine was most beneficial in patients with RCB-III disease, primarily through a reduction in distant metastasis. These findings support its selective use in high-risk TNBC populations.

PURPOSE:This study analyzed the pathological complete response (pCR) rates, long-term outcomes, and biological features of human epidermal growth factor receptor 2 (HER2)-zero, HER2-low, and HER2-positive breast cancer patients undergoing neoadjuvant treatment. METHODS:This single-center study included 1,667 patients who underwent neoadjuvant chemotherapy from 2008 to 2014. Patients were categorized by HER2 status, and their clinicopathological characteristics, chemotherapy responses, and recurrence-free survival (RFS) rates were analyzed. RESULTS:Patients with HER2-low tumors were more likely to be older (p = 0.081), have a lower histological grade (p < 0.001), and have hormone receptor (HorR)-positive tumors (p < 0.001). The HER2-positive group exhibited the highest pCR rate (23.3%), followed by the HER2-zero (15.5%) and HER2-low (10.9%) groups. However, the pCR rate did not differ between HER2-low and HER2-zero tumors in the HorR-positive or HorR-negative subgroups. The 5-year RFS rates increased in the following order: HER2-low, HER2-positive, and HER2-zero (80.0%, 77.5%, and 74.5%, respectively) (log-rank test p = 0.017). A significant survival difference between patients with HER2-low and HER2-zero tumors was only identified in HorR-negative tumors (5-year RFS for HER2-low, 74.5% vs. HER2-zero, 66.0%; log-rank test p-value = 0.04). Multivariate survival analysis revealed that achieving a pCR was the most significant factor associated with improved survival (hazard ratio [HR], 4.279; p < 0.001). Compared with HER2-zero, the HRs for HER2-low and HER2-positive tumors were 0.787 (p = 0.042) and 0.728 (p = 0.005), respectively. After excluding patients who received HER2-targeted therapy, patients with HER2-low tumors exhibited better RFS than those with HER2-zero (HR 0.784, p = 0.04), whereas those with HER2-positive tumors exhibited no significant difference compared with those with HER2-low tumors (HR, 0.975; p = 0.953). CONCLUSION:Patients with HER2-low tumors had no significant difference in pCR rate compared to HER2-zero but showed better survival, especially in HorR-negative tumors. Further investigation into biological differences is warranted.

Adoptive cell therapy (ACT) is a promising method for treating cancer and viral infections. Identifying antigen-specific T cells (ASTs) is critical to ACT. We investigated biomarkers for identifying ASTs. Peripheral blood mononuclear cells from healthy donors underwent staining with carboxyfluorescein succinimidyl ester (CFSE) to detect proliferating ASTs. Following exposure to CMV pp65 peptide and interleukin-2 for T-cell expansion, CD3+/CD8+ T cells were isolated at varying time points, revealing distinct populations. TCR repertoire analysis unveiled nine major clones in CFSE-/CD3+/CD8+ T cells on day 7, constituting 93.9 % of total cells. Contrarily, CFSE+/CD3+/CD8+ T cells exhibited minimal overlap with major TCR clones. Combined single-cell RNA-seq analyses highlighted upregulated genes associated with cell cycle proliferation and T-cell cytotoxicity. Engineered T cells expressing dominant clones effectively engaged CMV pp65 peptide, triggering T-cell activation and interferon-γ production. A set of seven upregulated genes in CFSE-/CD3+/CD8+ T cells on day 7, indicative of proliferating ASTs, was used to identify antigen-specific CD3+/CD8+ T cells on days 2-3, exhibiting 93.52 % accuracy. These markers predicted CFSE-/CD3+/CD8+ T cells with dominant TCR clones following exposure to EBV peptide with 74.59 % accuracy. In conclusion, we identified new markers facilitating the early isolation of viral antigen-responsive T cells.

Estrogen receptor (ER) expression and heterogeneity affect endocrine therapy efficacy. 18F-fluoroestradiol (18F-FES) PET/CT is an effective non-invasive method to analyze systemic ER expression. This study aimed to examine the predictive/prognostic value of 18F-FES PET/CT for patients treated with endocrine therapy plus cyclin-dependent kinase 4/6 (CDK4/6) inhibitors. Patients were identified from a prospective cohort for post-marketing surveillance of 18F-FES enrolled between April 2021 and April 2023 at Asan Medical Center. In this retrospective analysis, patients with ER-positive, HER2-negative recurrent/metastatic breast cancer who underwent CDK4/6 inhibitor-based endocrine therapy and pre-treatment 18F-FES PET/CT were included. A total of 127 women were included. The endocrine therapy used was aromatase inhibitors in 96 patients (76

Adoptive cell therapy (ACT) utilizing tumor-infiltrating lymphocytes (TILs) has emerged as a successful treatment modality for various malignancies. However, TILs cultured from colorectal cancer (CRC) liver metastasis remain underexplored. Fifteen CRC liver metastasis tissues underwent initial expansion (IE) of TILs and rapid expansion (REP). Histologic examination including the level of stromal TILs and Klintrup-Mäkinen score, were assessed by pathologists and deep learning-derived spatial analysis. We performed correlation analysis between expanded TILs and histopathologic factors. All cases exhibited successful IE, with a mean IE TIL count per fragment and total IE TIL per case of 2.59 ± 2.79e5 cells and 167.79 ± 126.97e5 cells, respectively. Five cases underwent REP, with a median fold change of 3,610 (range, 1,136–4,925). The median CD4+/CD8 + ratio in IE TILs and REP TILs were 3.66 and 0.68, respectively. A significant correlation was observed between the mean number of expanded TILs per fragment and KM score (p = 0.022). Successful expansion of TILs from CRC liver metastasis was achieved. Assessment of KM score may serve as a predictive tool for the obtainable TILs before IE. These findings lay the groundwork for future studies to establish effective ACT in patients with metastatic CRC.

Recently, a strategy involving the engineering of chemokine receptors on immune cells was developed to optimize adoptive cell therapy (ACT) for solid tumors. Given the variability in chemokine secretion among different tumor types, identifying and modulating the appropriate chemokine receptors is crucial. In this study, we utilized extensive RNA sequencing data from both tumor tissues from The Cancer Genome Atlas and normal tissues from Genotype-Tissue Expression to investigate the expression profiles of chemokines. Through analysis, we identified eight chemokine receptors associated with increased chemokine levels in tumor tissues compared to normal tissues, making them promising candidates for enhancing ACT. Subsequent examination of tumor-infiltrating lymphocytes and chimeric antigen receptor-T cells revealed that five out of the eight candidate chemokine receptors did not exhibit elevated expression in T cells. Among five candidates, we demonstrated that CXCR5 is a particularly promising candidate for enhancing cell migration without compromising cell viability or cytotoxicity. In conclusion, our study underscores the potential of five chemokine receptors (CCR6, CCR9, CXCR1, CXCR5, and XCR1) as valuable targets for modulating ACT to enhance cell trafficking and potentially improve cancer therapy outcomes.

Background:The 21-gene recurrence score (RS)-guided decision-making for adjuvant treatment has been utilized as a standard of care for early ER+HER2- breast cancer. We investigated a prognostic value of the Breast Cancer Test (BCT) score, a multigene assay incorporating clinical risk, in estrogen receptor (ER)+HER2- breast cancer patients receiving RS-guided adjuvant treatments, specifically focusing on chemotherapy-untreated patients with low RS. Methods:This multicenter cohort study included 759 patients who received RS-guided adjuvant treatment. The primary endpoint was recurrence-free survival (RFS), and the secondary endpoint was distant recurrence-free survival (DRFS). Results:At a median follow up of 85 months, the 7-year RFS was 92.9% (95% CI, 90.9%-94.9%). Among the 592 chemotherapy-untreated patients with low RS, the RFS differed significantly according to the BCT score (P=.014); the 7-year RFS was 95.5% (95% CI, 93.4%-97.7%) in the BCT-low group, while it was 89.9% (95% CI, 84.9%-95.1%) in the BCT-high group. The BCT score was an independent prognostic factor for both RFS and DRFS. In addition, the RFS of the low-BCT score group was superior to that of the high-BCT group in women aged 50 years or younger, with an RS of 16 to 25. Conclusions:Our study suggests the utility of the BCT score in stratifying the relapse risk among chemotherapy-untreated patients with a low RS, particularly in young women with an RS of 16-25 who are at risk for long-term recurrence.