Metastatic brain disease occurs in up to 30% of patients with lung, melanoma and breast cancers, and the median survival time remains less than a year. Treating these patients is a challenge because surgical approaches are limited and most chemotherapeutic drugs and immunotherapies are ineffective at crossing the blood-brain barrier (BBB). Given the unique abilities of macrophages to cross the BBB and exert their phagocytic function on tumour cells, we genetically engineer macrophages that express a chimaeric antigen receptor (CAR) targeting mesothelin (MSLN). To specifically target metastatic brain tumours, we fused the cells with the immune signalling molecule MyD88. This chimaeric antigen receptor macrophage (CARMA) penetrates the BBB and decreases brain metastasis growth in a humanized mouse model. MSLN-CARMA shows antigen-specific phagocytosis activity against tumour cells and exhibits a bystander effect by releasing TNF to act on surrounding tumour cells lacking the tumour antigen. These features of CARMA represent advantages over other immune therapies and CARMA may serve as a promising therapeutic tool for the treatment of brain metastasis.

Glioblastoma (GBM) is an extremely aggressive and incurable primary tumor of the brain. GBM is characterized by interpatient and intratumoral heterogeneity, making this cancer particularly resistant to therapy and likely to recur. Mapping the complex dynamics that underpin the development and evolution of gliomas with human-based in vitro models is difficult. This study aimed to generate 3D glioma patient-derived tumor constructs (PTCs) using a clinically relevant, Matrigel-free, hyaluronic acid system, evaluate their suitability in drug screening assays, and determine the stability of their genetic profiles compared to originating tumors. In this study, we utilized a synthetically modified hyaluronic acid and gelatin hydrogel system to generate tumor constructs containing cells from clinical glioma biospecimens. PTCs were characterized phenotypically, after which they were deployed in chemotherapy drug screens using temozolomide (TMZ) and a P53 activator compound. Drug responses of these 3D cultures were compared with 2D cultures, as well as PTCs that were generated after passaging in 2D. RNA sequencing was used to evaluate genetic parity between PTCs or 2D cultures with originating tumor tissues, using The Cancer Genome Atlas (TCGA) GBM subpopulations for subcategorizing. PTCs were created successfully from five World Health Organization (WHO) grade 4, two grade 3, and two grade 2 gliomas. PTCs were maintained with high viability. Chemotherapy drug screens demonstrated that expected TMZ responses were observed for Isocitrate dehydrogenase (IDH) mutant diffuse gliomas while drug response was variable for IDH wildtype GBM PTCs. PTCs demonstrated stable drug response over time, while 2D passaging resulted in significant shifts in drug sensitivity. RNA sequencing revealed maintenance of subpopulation signatures for PTCs which clustered with their originating patient tumor tissue. In contrast, 2D cultures largely clustered together regardless of the patient. Our PTC approach utilizes a defined hydrogel biomaterial system that maintains the genotypic and drug response characteristics of patient tumors making this an ideal ex vivo model for translational applications.

Harnessing patient immune cells via adoptive cellular therapy is a promising cancer therapeutic strategy. However, major challenges remain for advanced solid malignancies, including difficulty isolating sufficient tumor infiltrating lymphocytes (TILs) and limited targeting of diverse neoantigens in heterogenous tumors. To address this, we have developed a tumor-on-a-chip platform with co-cultured patient-derived tumor cells, autologous peripheral blood mononuclear cells (PBMCs), and lymphoid tissue-derived antigen presenting cells. This approach generates organoid interacting lymphocytes (OILs) with enhanced anti-tumor activity. In peritoneal malignancies, OIL-induced cytotoxicity of patient-matched tumor cells surpasses both TILs and static-expanded PBMCs. We find that this improved performance was linked to increased CD8 + T and NK cells among OILs, and increased effector cytokine polyfunctionality, particularly Granzyme A. Our platform represents a versatile and scalable approach to generate patient-specific therapeutic lymphocytes even when TILs are insufficient, offering a promising avenue to treat diverse solid tumors associated with poor outcomes under current immunotherapies. Teaser:A tumor-on-a-chip device primes patient immune cells with tumor recognition for personalized immunotherapy applications.

BACKGROUND:Triple negative breast cancer (require) new treatment strategies due to poor responses to current therapies. While myeloid SIRPα mediates immunosuppression, its cancer intrinsic role remains poorly understood. METHODS:Human breast cancer scRNAseq profiles were used to examine SIRPα expression across different cell populations and subtypes. TNBC brain-tropic cells were injected into the mouse mammary fat pad for the orthotopic tumor model, and intracardiac-injected for brain metastasis models. Bulk RNA sequencing was used to determine SIRPα-regulated pathway. Stably SIRPα overexpressed and knockout TNBC cell lines were established to determine SIRPα intracellular regulation. Digital spatial profiling was utilized to investigate the orthotopic and brain metastasis tumor immune microenvironment. RESULTS:Human single-cell data showed that SIRPα levels increased in malignant TNBC epithelial cells. We observed that SIRPα is upregulated in patient breast-to-brain metastatic lesions. SIRPα is overexpressed in TNBC brain-tropic cells compared to parental cells. Bulk RNA-Seq showed that targeting SIRPα affects genes involved in mitochondrial dynamics, and that SIRPα upregulates mitochondrial fission and induces metastasis through the SHP2/Erk/Drp1 signaling pathway. In vivo, overexpression of SIRPα in cancer cells significantly increases TNBC systemic metastasis. Next, spatial proteomics revealed changes in the immune microenvironment associated with the SIRPα-regulated ECM protein fibronectin. Fibronectin induces microglial tolerance by impairing inflammatory signaling and metabolic reprogramming, allowing cancer to escape microglial immunosurveillance. Most importantly, SIRPα inhibition reduced TNBC brain metastatic lesions in mouse metastasis models. CONCLUSION:: Cancer-intrinsic SIRPα promotes TNBC brain metastasis through increased mitochondria fission and triggering microglia tolerance, and targeting SIRPα reduces brain metastasis.

Abstract Background: Mucinous Appendiceal Cancer (mAC) is a rare and poorly characterized malignancy with no known molecular targets and is treated using colorectal cancer protocols. In this study, we employed whole-exome sequencing (WES) on 3D cultures to investigate how mAC-specific oncogenic programming associates with patient clinical variables and therapeutic responses. Methods: Patient-derived tumor organoids (PTOs) were generated from multiple mAC lesions from patients undergoing cytoreductive surgery and hyperthermic intraperitoneal chemotherapy under IRB approval. PTOs were treated with FOLFOX or FOLFIRI for 72 hours with response measured by Cell TiterGlo to determine a response phenotype. A subset of PTOs from 10 patients and 31 tumor deposits were treated with DMSO vehicle, FOLFOX, or FOLFIRI, after which genomic DNA was isolated and sequenced on an Illumina NextSeq 6000. GATK and DRAGEN pipelines were used for mutation calling, and alterations associated with drug response were analyzed by Ingenuity Pathway Analysis (IPA), with stratification informed by relevant patient clinical variables. Results: Across the 10 mAC cases profiled, WES identified a total of 28,245 mutations, including therapeutically actionable KRAS G12D variants in 40% of cases. No targetable alterations were detected in BRAF, EGFR, or ALK. When pooling all pre-treated PTOs and averaging the variant allele frequency (VAF) across all 31 lesions, with a focus on mutations with an average VAF >=10% and present in at least 2/10 patients, individual gene mutations that were undetectable (i.e. VAF=0%) after FOLFIRI treatment included TAX1BP1, PDXK, ZFPM2, and H1FX-AS1. The lone genetic mutation undetectable after FOLFOX treatment was LINC01197, with mutations in SORD and ZBTB24 associated with a complete clonal loss after either FOLFIRI or FOLFOX treatment. Of note, SORD overexpression is implicated in the migration of CRC via upregulation of the mevalonate pathway. IPA canonical pathway analysis of patients progressing within 2 years revealed alterations in D-Myo-Inositol regulation associated pathways, whereas progression-free patients displayed alterations in pathways related to insulin-like growth factor uptake and iron regulation. Further pathway analysis was performed to discover mutations that differentiate pathways by sex, race, tumor grade, TNM scores, obesity status, among other clinical variables. Conclusions: Comprehensive mutational profiling of mAC using paired PTO models via WES is feasible and clinically informative, enabling the detection of actionable oncogenic drivers and chemotherapy-sensitive clonal populations. These data provide the first integrated framework linking mAC-specific genomic alterations to patient outcomes and therapeutic responses and offers new biological insight into the molecular underpinnings of mAC. Citation Format: Daniel J. Gironda, Cecilia R. Schaaf, Steven D. Forsythe, R. Andrew Erali, Tiefu Liu, Ming Leung, Ashok K. Pullikuth, Helen R. Bernardirathgeb, Eleftherios Makris, Shay Soker, Edward A. Levine, Konstantinos I. Votanopoulos, Lance D. Miller. Mutational profiling of patient tumor organoids for clinical variable association and markers of therapeutic response in mucinous appendiceal cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 4854.

Abstract Background: Triple-negative breast cancer (TNBC) is an aggressive subtype with no targeted therapies and limited response to immunotherapy. TNBC evades immune surveillance by repressing antigen presentation machinery (APM) genes, reducing MHC Class I expression. Using bioinformatics, we identified SOX11 as a potential transcriptional regulator associated with APM suppression and immune evasion in TNBC. Methods: We analyzed gene expression data from basal breast cancer (BRCA) tumors (n = 189) in TCGA and cancer cell lines (n = 1,189) in DepMap. Spearman correlation coefficients were used to evaluate associations between a T-cell infiltration signature and an APM signature. SOX11 was prioritized as it was strongly negatively correlated with both signatures. To identify downstream effectors, we performed RNA-seq analysis on SOX11 knockdown (KD) CAL-148 cells (high endogenous SOX11) and integrated these results with publicly available SOX11-overexpression and KD datasets. To validate key targets, SOX11 KD and MEX3A KD were performed using siRNA, followed by flow cytometry to measure surface MHC Class I (HLA-ABC) expression. Results: SOX11 was strongly inversely correlated (rho < -0.4, adjusted p < 0.01) with APM and T-cell infiltration signatures in basal BRCA and DepMap datasets. In basal BRCA tumors, SOX11 expression was enriched in T-cell cold tumors (p < 0.01). Analysis of our SOX11 KD RNA-seq data, integrated with public SOX11-modulated datasets, revealed a highly reproducible SOX11-driven gene signature. This signature was notably enriched for known suppressors of innate immunity. Among the most consistent SOX11 targets were genes implicated in destabilizing HLA-A (MEX3B), degrading innate immune sensors (MEX3A), activating immunosuppressive signaling (SBK1), and facilitating epigenetic silencing of interferon pathways and APM (RCOR2, NELL2). Consistent with this, SOX11 KD in CAL-148 cells significantly increased surface HLA-ABC expression. Furthermore, targeted knockdown of its effector, MEX3A, also resulted in a significant increase in APM gene expression. Conclusions: SOX11 suppresses HLA-A/B/C expression and contributes to immune evasion in TNBC. Its high expression in T-cell cold tumors and its regulation of multiple APM-suppressive targets highlight its potential as a therapeutic target. We hypothesize that SOX11-mediated APM repression is not driven by a single gene but by the concerted activity of this multi-effector network. Targeting SOX11 or its key effectors could restore antigen presentation, promote immune recognition, and enhance immunotherapy responses in TNBC. Citation Format: Salomat Abdulkhuseynova, Ashok Pullikuth, Lance D. Miller. From correlation to causation: SOX11 drives a coordinated immune-suppressive network that represses antigen presentation in triple-negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 7009.

Cancer-induced bone pain (CIBP) is among the most common and debilitating symptoms in patients with bone metastasis. Current treatments are somewhat effective but have severe side effects. For the future development of safer CIBP treatments, in this study, we sought to investigate the mechanisms whereby the nerve-cancer interaction controls CIBP. We found that c-Kit, a receptor tyrosine kinase, was activated in the dorsal root ganglia (DRG) sensory neurons of mice with CIBP and that c-Kit’s sole ligand, stem cell factor (SCF), was enhanced in the bone marrow with bone metastasis. When DRGs were treated with SCF or conditioned medium from high SCF-expressing cancer cells, in vitro nerve sprouting was enhanced, and this effect was abolished with c-Kit inhibitors. Mice inoculated intrafemorally with cancer cells that had varying levels of SCF expression developed CIBP and enhanced peripheral nerve sprouting in an SCF-dependent manner. Downstream proteomic analysis revealed that SCF upregulated and activated fibroblast growth factor 1 (FGF1) in DRGs. When FGF1 was knocked down in DRGs, SCF-mediated nerve sprouting was prevented. Taken together, our studies demonstrate the importance of the SCF/c-Kit axis in CIBP and nerve sprouting and identify the SCF/c-Kit/FGF1 pathway as a potential therapeutic target for CIBP.

Chemotherapy-induced peripheral neuropathy (CIPN) is the most prevalent and limiting side effect of paclitaxel treatment in patients with cancer. CIPN affects sensory neurons through neuroinflammatory mechanisms, but how immune cells sense and interpret systemic paclitaxel exposure during treatment is unclear. Here, we found that paclitaxel administration activated the endoplasmic reticulum (ER) stress sensor inositol-requiring enzyme 1α (IRE1α) in circulating and dorsal root ganglion-resident myeloid cells, engendering an inflammatory milieu that promotes CIPN. Mechanistically, paclitaxel induced the overproduction of mitochondria-derived reactive oxygen species (ROS) that provoked ER stress and IRE1α hyperactivation in macrophages. This process reprogrammed macrophages toward an inflammatory state characterized by IRE1α-dependent production of TNF-α, IL-1β, PGE2, IL-6, IL-5, GM-CSF, MCP-1, and MIP-2. Ablation of IRE1α in leukocytes, or treatment with a selective IRE1α pharmacological inhibitor, prevented dorsal root ganglion neuroinflammation and CIPN-related pain behaviors in mice. Furthermore, the development and severity of CIPN in patients with gynecological cancer were associated with the status of IRE1α activation in their circulating leukocytes. Our study uncovers leukocyte-intrinsic IRE1α as a key mediator of CIPN and suggests that targeting its dysregulated activation could help mitigate CIPN in patients with cancer who are receiving paclitaxel.

Background/Objectives: Appendiceal cancer (AC) is a rare and understudied malignancy with limited genomic data available to guide clinical interventions. Historically treated as a subtype of colorectal cancer, AC is now recognized as a distinct disease with unique histologic subtypes and molecular features. This review aims to consolidate current genomic data across AC subtypes and explore the clinical relevance of recurrent mutations. Methods: A systematic literature review was performed in accordance with general Preferred Reporting Items for Systemic Reviews and Meta-Analyses (PRISMA) guidelines. Using search engines such as PubMed and Web of Science, we selected studies based on relevance to AC genomics using search terms such as “appendix cancer”, “appendiceal cancer”, “pseudomyxoma peritonei”, “sequencing”, “mutation”, and “genotype”. Results: AC comprises five major histologic subtypes—appendiceal neuroendocrine neoplasms (ANENs), mucinous appendiceal neoplasms (MANs), goblet cell adenocarcinomas (GCAs), colonic-type adenocarcinomas (CTAs) and signet ring cell adenocarcinomas (SRCs)—each with unique clinical behaviors and mutational profiles. Low-grade tumors, such as ANENs and MANs, frequently harbor KRAS and GNAS mutations, while high-grade subtypes, such as CTAs and SRCs, are enriched for TP53, APC, and SMAD gene alterations. GCA tumors exhibit a distinct mutational spectrum involving chromatin remodeling genes such as ARID1A and KMT2D. Compared to colorectal cancer, AC demonstrates lower frequencies of APC and TP53 mutations and a higher prevalence of GNAS mutations, consistent with a pathological divergence from CRC. Conclusions: The genomic heterogeneity of AC is commensurate with its histological complexity and has important implications for diagnosis, prognosis and treatment. While certain actionable mutations are present in a subset of tumors, large-scale genomic characterization efforts and development of subtype-specific models will be essential for advancing precision medicine in AC.

BACKGROUND:Intrabuccal administration of amplitude-modulated 27.12 MHz radiofrequency electromagnetic fields (AM RF EMF) resulting in the systemic delivery of low and safe levels of AM RF EMF has shown activity in several forms of cancer. METHODS:Glioblastoma (GB) cell lines were exposed to GB-specific AM RF EMF (GBMF) three hours per day at a level of exposure identical to patients during treatment. Cellular assays and agnostic genomic approaches were used to characterize the mechanism-of-action. One patient with therapy refractory GB received compassionate use treatment with GBMF as well as a second patient with refractory oligodendroglioma. RESULTS:Treatment with GBMF inhibited the proliferation of several GB cell lines. CACNA1H mediates the effect of GBMF. GBMF modulates the "Mitotic Roles of Polo-Like Kinase" pathway resulting in the disruption of GB mitotic spindle. There was evidence of clinical and radiological benefit in a 38-year-old patient with recurrent GB and evidence of safety and feasibility in a 47-year-old patient with oligodendroglioma. CONCLUSIONS:This is the first report showing in vitro antitumor activity, disruption of the mitotic spindle, activation of the Mitotic Roles of Polo-like kinase pathway in GB. This is also the first report showing feasibility and clinical activity in patients with brain tumor.

Small cell lung cancer (SCLC) is a highly proliferative and aggressive type of lung cancer, accounting for 10–15% of all lung cancer cases. SCLC is characterized by early metastasis, with more than half of the cases presenting with metastases at the time of diagnosis. While comprehensive genomic studies have been performed for SCLC, metastasis-associated mutations have not been fully characterized. In this study, 57 SCLC samples from the Wake Forest Baptist Comprehensive Cancer Center (WFBCCC), profiled by either FoundationOne CDx or Guardant360 CDx between 2013 and 2022, were analyzed. The most common metastatic sites were identified as the brain (54.3%), liver (45.6%), and bone (33.3%). Associations between clinical features and distant metastases were examined, with no significant differences observed in age, sex, race, or smoking status among the metastatic groups. To explore the relationship between gene mutation rates and distant metastatic sites, we collected somatically altered genes from these patients. Stratified by metastatic site, Cox proportional hazards regression analysis was performed to identify mutations associated with the progression of metastases. BRCA2 and APC alterations were significantly associated with brain metastases, while PDGFRA mutations were linked to liver metastases. No mutations were found to be associated with bone metastases. Our findings suggest the potential efficacy of PARP inhibitors for treating brain metastases and PDGFRA inhibitors for addressing liver metastases in SCLC. Validation in SCLC animal model is under investigation. This study reveals specific mutations associated with metastatic progression to the brain and liver. These findings underscore the importance of precision medicine approaches in tailoring treatments based on the genetic profile of tumors, offering the potential to improve outcomes for patients with this aggressive cancer. Citation Format: Yin Liu, Yuezhu Wang, Ralph D’Agostino, Jimmy Ruiz, Lance D Miller, Wencheng Li, Michael D Chan, Liang Liu, Fei Xing. Mutation analysis of SCLC patients with distant metastases [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Functional and Genomic Precision Medicine in Cancer: Different Perspectives, Common Goals; 2025 Mar 11-13; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2025;85(5 Suppl):Abstract nr A012.

Cholesterol is an integral sterol lipid that influences the homeostatic maintenance of cellular structure, hormone synthesis, and metabolic regulation within the cell. Accumulation of intracellular cholesterol has shown to drive the progression of multiple human cancers. Here we report that mutation or transcriptional repression of the cholesterol exporter, ABCA1, is a common phenomenon conserved in bowel cancers with novel mechanistic and therapeutic implications. Whole-exome sequence analysis was conducted on 24 appendiceal cancers (AC) from consented patients. ABCA1 coding mutations and expression levels were analyzed in tumors from The Cancer Genome Atlas (TCGA) and Broad DepMap databases. One AC cell line (Naxorini) was created in house and four commercial CRC cell lines were used in ABCA1 functional studies. Combination treatment of 22-(R)-hydroxycholesterol (OHC) and 9-cis-retinoic acid (CRA) was used to induce ABCA1. ABCA1 knockdown (KD), measures of cellular viability, cell death, apoptosis, and free cholesterol were quantified. Protein coding mutations in the ABCA1 gene were identified in 7/24 (29%) AC specimens. Mutational analysis of other gastrointestinal cancers shows that ABCA1 is preferentially mutated in mucinous CRCs and mucinous stomach adenocarcinomas compared to their non-mucinous counterparts (X2 p-value <0.001 and 0.047). ABCA1 mRNA expression levels were significantly lower in CRC and gastric tumors as compared to 28 different tumor types (ANOVA p<0.001), and significantly lower in bowel cancer cell lines as compared to 34 different cancer lineages (ANOVA p<0.001). 10uM OHC+CRA treatment robustly induced ABCA1 protein expression in ABCA1 wildtype (WT) cell lines (Naxorini, MDST8, COLO201), but not in ABCA1 mutant lines (HCT15, SNU81), significantly induced cell death in ABCA1 WT cell lines (all, p<0.001), and decreased free cholesterol accumulation (MDST8, p=0.010; Naxorini, p=0.007). Importantly, KD of ABCA1 rescued the viability of OHC+CRA treated cells (MDST8, p<0.001) demonstrating that OHC+CRA cytotoxicity is dependent on the induction of ABCA1. Furthermore, the KD of ABCA1 in untreated MDST8 cells significantly enhanced cell viability (p=0.004) consistent with a growth advantage owing to reduced activity of ABCA1. Analysis of ABCA1-dependent, OHC+CRA induced cell death found alterations in protein levels of Bad, Bcl-2 and HtrA2/Omi (all, p<0.05), associating ABCA1 induction with mitochondrial apoptosis. Dysregulation of cellular cholesterol due to dysfunctional ABCA1, whether by somatic mutation or transcriptional downregulation, may represent an oncogenic addiction conserved in bowel cancers. These findings warrant further investigation into the therapeutic potential of ABCA1 and cholesterol targeting strategies for bowel cancers. Daniel J. Gironda, Calvin J. Wagner, Ashok K. Pullikuth, Lance D. Miller. Dysfunction in ATP-binding cassette 1 (ABCA1): a driver of intracellular cholesterol accumulation and potential therapeutic vulnerability of bowel cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 5565.

Introduction: Triple-negative breast cancer (TNBC) is group of heterogeneity caner. Despite majority of them had the unfavorable prognosis, a subset of patients who do not receive chemotherapy exhibit a good prognosis. Biomarkers are required to recognize these group of pateints and improve the current therapeutic strategies for them. Methods: A retrospective analysis of 997 patients with TNBC from three datasets including 188 case of Peking Union Medical College Hospital (PUMCH) and two TNBC datasets from published cohort studies(279 case of Affy-set, 530 case of GSE set) was conducted. Intrinsic subtypes (basal-like, immune-enhanced, human epidermal growth factor receptor-2 [HER2]-enriched and luminal A/B) and tumor environmental immunity were evaluated using expression profiles of a 72-gene panel. Association of intrinsic subtype and immunity score with distant metastasis-free survival (DMFS) and overall survival (OS) was analyzed. Results: Five intrinsic subtypes were identified in the patients with TNBC, comprising 64 % basal-like, 19 % immune-enhanced, 11 % HER2-enriched, 5 % luminal A, and 2 % luminal B. In the absence of adjuvant chemotherapy (ACT), Luminal A and immune-enhanced subtypes showed better DMFS than basal-like, HER2-enriched, and luminal B subtypes(P = 0.35). Significantly good OS was observed in luminal A and immune-enhanced subtypes compared to basal-like and HER2-enriched subtypes (P < 0.05). So two subtype groups were further classified as low-risk subtypes, including luminal A and immune-enhanced, and high-risk subtypes, including basal-like, HER2-enriched, and luminal B. Except for the immune-enhanced subtype, each subtype was further sorted and grouped according to immunity score, istrong and iweak. Significant improvements in both DMFS and OS were observed in patients with istrong compared with those with iweak(P = 0.01 and 0.0051 respectively). When combining intrinsic subtype and immunity status to predict the benefit from ACT, all high-risk subtype patients demonstrated improved DMFS(P = 0.075) and OS(P < 0.0001), with istrong patients exhibiting greater benefit; low-risk subtype plus iweak patients showed marginal benefit, whereas low-risk subtype plus istrong patients demonstrated least benefit from ACT. Conclusion: Intrinsic subtype and immunity score is good prognostic biomarkers for patients with TNBC in the absence of chemotherapy. Combined intrinsic subtype and immunity evaluation could identify patients with TNBC who do not benefit from chemotherapy.