Purpose High-risk neuroblastoma presents a serious clinical challenge with survival rates below 50%. Disease relapse most commonly occurs at distant metastatic sites and remains the primary driver of poor outcomes, emphasizing the need for therapies to target drivers of relapse. Experimental design This study identified DNA-PKcs as a critical determinant of poor survival and metastatic relapse in neuroblastoma patients. We evaluated which therapeutic modality—chemotherapy or radiotherapy—when combined with DNA-PKcs inhibition, more effectively reduces metastatic burden and prevents recurrence. Results Colony-forming assays revealed that established neuroblastoma colonies resist doxorubicin alone and require high-dose doxorubicin paired with DNA-PKcs inhibition to suppress progression. In contrast, low-dose radiotherapy in combination with DNA-PKcs inhibition effectively controlled colony progression. Maximal synergy between radiotherapy and DNA-PKcs inhibition was achieved when the inhibitor was administered within 4 h post-irradiation. Chronic co-exposure to doxorubicin and peposertib encouraged emergence of therapy-resistant cells, whereas chronic co-exposure to radiotherapy combined with peposertib disrupted neuroblastoma cells self-renewal and prevented long-term colony maintenance. In neuroblastoma metastases, adding DNA-PKcs inhibition to doxorubicin improved efficacy but induced gastrointestinal side effects and failed to eradicate tumors; pairing it with low-dose, fractionated radiotherapy resulted in total lesion regression, impaired tumor self-renewal, and prevented systemic adverse effects. Conclusions Our findings correlate elevated DNA-PKcs levels with poor patient prognosis and show that low-dose radiotherapy combined with peposertib effectively abrogates neuroblastoma self-renewal compared to chemotherapy-based regimens, thereby implicating DNA-PKcs as a key mediator of metastatic relapse and supporting radiotherapy plus DNA-PKcs inhibition as a compelling therapeutic strategy for relapsed or refractory high-risk neuroblastoma.

Immune checkpoint blockade can elicit durable antitumor responses, yet tumor heterogeneity and adaptive resistance often necessitate combination strategies that increase systemic toxicity. In vivo genome editing offers a programmable route to durable immunomodulation but remains difficult to spatially confine in solid tumors. Here we develop a magnetically gated genome-editing platform that enables spatially confined immunomodulation. The system integrates a non-replicating baculoviral vector with magnetic nanoparticles (MBV), in which magnetic activation restores viral transduction despite complement-mediated inactivation, thereby confining CRISPR activity to tumor regions. Baculoviral transduction engages antiviral innate programs that promote chemokine signaling and antigen presentation in tumors. In a syngeneic colon cancer model, MBV-mediated disruption of Pdl1 restricts checkpoint loss to tumor tissue while preserving immune activation, enhancing immune infiltration and suppressing tumor growth. Local Pdl1 editing synergizes with CTLA-4 blockade, extending survival without overt toxicity. These findings define MBV as a controllable genome-editing architecture for localized combination immunotherapy.

MAPK inhibitors (MAPKi) are important treatment options for some patients with metastatic melanoma; however, resistance inevitably develops in the vast majority of cases (>70%). Here, using immune-proficient mouse models, single-cell RNA sequencing, flow cytometry, and datasets from patients treated with MAPKi, we demonstrate that ABL kinases (ABL1/2) promote MAPKi resistance not only by impacting intracellular signaling but also by driving secretion of chemokines that suppress the immune microenvironment. Targeting ABL1/2 reduces chemokine secretion by melanoma cells, prevents immunosuppressive myeloid-derived suppressor cell (MDSC) infiltration, and promotes accumulation of cytotoxic CD8+ T cells. Depletion of MDSCs prevents resistance, and depletion of CD8+ T cells or re-expression of chemokines blocks ABL inhibitors from preventing resistance. Importantly, CXCR2 receptor ligand expression and ABL1/2 activity are reduced in patients who respond to treatment but increased during resistance. Thus, the dual role of ABL1/2 in proliferation/survival and immune suppression makes them attractive targets for potential treatments. Although MAP kinase inhibitors (MAPKi) can initially reduce tumour burden in metastatic melanoma, resistance to treatment often occurs. Here, the authors demonstrate that concomitant treatment with ABL kinase inhibitors overcomes tumour resistance to MAPKi by promoting the accumulation of cytotoxic T lymphocytes and inhibiting the infiltration of immunosuppressive cells.

The presence of BRAFV600E mutations is associated with poor prognosis in colorectal cancer (CRC). Although the FDA-approved combination of encorafenib and cetuximab provides clinical benefit in this population, only 22% of patients respond and most eventually develop resistance. This study investigated the mechanisms of resistance to PLX8394, a second-generation BRAF inhibitor. Using primary and established BRAFV600E CRC cells, we show that the development of resistance to PLX8394 results in cross-resistance of cells to encorafenib. Moreover, the acquired resistance is associated with increased proliferation, invasion, and upregulation of lipid metabolism, including increased expression of fatty acid synthase (FASN), a key enzyme of lipid synthesis. Yet, the combination of PLX8394 and FASN inhibitor TVB3664 has a synergistic effect on cell viability and colony formation in parental CRC cells, but not in PLX-resistant cells. Importantly, we demonstrate that addition of TVB3664 to the PLX8394 or encorafenib regimen significantly postpones development of resistance to BRAF-targeted therapy by inhibiting the cell cycle progression via a decrease in pRb (Ser780) and downregulation of E2F transcription factor and Cyclin D1 expression. Consistently, clinical data show that patients with BRAFV600E CRC who have high FASN expression in tumor tissues have higher expression of cell cycle-associated genes, including CDKs, E2F, CCDN1 (Cyclin D1), survivin, and MKI67. Collectively, these findings identify FASN-driven lipid metabolism as a critical mediator of resistance to BRAF-targeted therapy and suggest that incorporation of FASN inhibitors may enhance therapeutic efficacy and delay acquired resistance in BRAFV600E CRC.

Metastatic castration-resistant prostate cancer (CRPC) remains a clinical challenge, and epithelial-mesenchymal transition (EMT) contributes to metastatic progression and reduced response to therapy. However, the upstream epigenetic mechanisms that sustain EMT programs in advanced prostate cancer (PCa) are not fully defined. We identify the histone H3 lysine 9 (H3K9) methyltransferase SET domain bifurcated 1 (SETDB1) as a key regulator of EMT and metastasis through direct repression of RhoB, the small GTPase. SETDB1 is genomically amplified and transcriptionally upregulated in metastatic CRPC, and SETDB1 depletion reduces cell migration, invasion, and metastatic dissemination. Integrated chromatin profiling and transcriptomic analyses demonstrate that SETDB1 occupies the RhoB promoter and mediates its transcriptional silencing through H3K9 methylation. Restoration of RhoB reverses EMT gene expression and suppresses invasive behavior, whereas RhoB knockdown rescues the effects of SETDB1 depletion, establishing RhoB as a critical downstream effector of SETDB1 function. Androgen signaling inhibitor-resistant PCa models exhibit RhoB loss and EMT activation, linking this axis to therapy-resistant phenotypes. Finally, antisense oligonucleotide-mediated SETDB1 silencing restores RhoB expression and suppresses EMT and invasion in CRPC cell models. Together, these findings define a SETDB1-RhoB epigenetic pathway that promotes EMT and metastatic progression in PCa and may be therapeutically targeted in advanced disease.

Programmed cell death 4 (Pdcd4) is a well-established tumor suppressor as well as an inhibitor of protein translation. Although Pdcd4-mediated translational repression contributes to tumor suppression, emerging evidence suggests that Pdcd4 also exerts translation-independent functions. In this study, we found that Pdcd4 suppresses tumorigenesis through disrupting mTORC2 complex formation by binding with the rapamycin-insensitive companion of mTOR (Rictor), a core component of the mTORC2 complex. Using deletion mapping and site-directed mutagenesis, we defined the Rictor-binding domain of Pdcd4 and identified three critical residues, R105, K108, and R110, for this interaction. Co-immunoprecipitation and in vitro kinase assays demonstrated that Pdcd4 binding to Rictor disrupted mTORC2 complex assembly and inhibited its kinase activity. Reverse phase protein array analysis revealed that 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3), a key regulator of glycolysis, was markedly upregulated in Pdcd4-knockdown cells. Restoration of wild-type Pdcd4, but not a Rictor-binding–deficient mutant, reduced PFKFB3 protein abundance by promoting ubiquitin–proteasome–mediated degradation. Functionally, Pdcd4–Rictor interaction suppressed glycolytic activity and inhibited tumor cell proliferation in cultured cells and xenograft models. Consistent with these findings, non–small cell lung cancer (NSCLC) tissues exhibited significantly elevated protein levels of Rictor and PFKFB3 compared with adjacent normal tissues, with a positive correlation between their expression. Collectively, these results demonstrate that Pdcd4-Rictor interaction disrupts mTORC2 signaling and downregulates PFKFB3, which plays a critical role in suppressing NSCLC growth and glycolysis.

Immune checkpoint blockade can elicit durable antitumor responses, yet tumor immune heterogeneity and adaptive resistance often necessitate combination strategies that may cause systemic toxicity. Here we develop a magnetically activatable nanosystem that integrates a non-replicating insect viral vector with magnetic nanoparticles (MBV) to enable spatially confined genome editing within tumors. Magnetic activation functions as a physical on-switch that restores viral entry under complement pressure, enabling localized CRISPR-mediated disruption of immune checkpoints. Concurrently, baculoviral transduction engages antiviral programs, reshaping the tumor immune microenvironment. In a syngeneic colon cancer model, MBV-mediated disruption of Pdl1 confines checkpoint loss to tumor tissue while preserving virus-induced immune activation, resulting in increased immune infiltration and tumor growth suppression. MBV-Pdl1 further synergizes with systemic CTLA-4 blockade, extending survival without overt toxicity. These results establish MBV as a controllable genome-editing platform that couples magnetic precision with virus-encoded immune priming to enable localized, combination immunomodulation in solid tumors. ### Competing Interest Statement X.Y. and S.T. are inventors on a pending, unlicensed patent application assigned to the University of Kentucky and related to the technologies described in this work. National Institute of Biomedical Imaging and Bioengineering, https://ror.org/00372qc85, R01EB026893

Abstract In recent decades, there has been an unprecedented rise in gastric cancer among younger individuals, contrasting with a decline among older individuals. However, the biological underpinnings of gastric cancer in younger individuals remain poorly understood. We described the clinicopathologic and genomic characteristics of early-onset gastric cancer (EOGC) compared to average-onset gastric cancer (AOGC). We analyzed 311 patients using the multi-institutional prospective Oncology Research Information Exchange Network (ORIEN) database to compare demographic, clinicopathologic, genomic, and survival outcomes between EOGC (<50 years; N=72) vs AOGC (>50 years N=239). Genomic, germline, and RNA sequencing data were analyzed and compared between the cohorts. Mutational and immune signatures were also processed. EOGC patients exhibited significantly higher rates of pain at diagnosis (53% vs 30% p=0.001), but not anemia or reflux. EOGC patients were more likely to present with stage III/IV disease (70% vs 45% p=0.006) and diffuse/signet ring histology (47% vs 16% p=0.002). Consequently, OS was decreased in the younger cohort (HR 1.52; p=0.03). Somatic mutational load was decreased in young patients. Significantly mutated genes in the entire cohort included CDH1, ARID1A, TP53, PIK3CA, but CDH1 was more frequently mutated in the EOGC cohort (40% vs 18% p=0.09). Significant differences in RNA expression were observed, with upregulated epithelial mesenchymal transition (EMT), myogenesis, and apical junction. Immune deconvolution revealed a predominance of M2 macrophages, mast cells, and CD4+ T cell subsets, but no differences between cohorts. Early-onset gastric cancer has unique clinical and genomic features. Pathway dysregulation in EOGC may contribute to tumorigenesis and therapy resistance. This study underscores the necessity for further research into novel therapies, biomarker discovery, and early detection methodologies in younger individuals. Citation Format: Hannah McDonald, Lilia Turcios, Neelima Hosamani, Abu Saleh Mosa Faisal, Chi Wang, Joseph Kim, Mautin Barry-Hundeyin.. Real-world clinicogenomic comparison of early- and average- onset gastric 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 5347.

Glioblastoma (GBM) is the most aggressive primary brain tumor in adults. Cognitive impairment is a common sequela in glioblastoma survivors, yet the underlying mechanisms remain poorly understood. Extracellular vesicles (EVs) derived from glioblastoma are established mediators of intercellular signaling within the tumor microenvironment. Here, we investigated whether GBM-derived EVs released after radiation treatment (RT-EVs) regulate cognitive function. Treatment with RT-EVs was associated with cognitive deficits and neuroinflammatory responses in vivo. In vitro, RT-EVs activated the NFκB pathway and induced the release of neurotoxic H 2 O 2 . Importantly, NFκB p50 knockdown abolished the H 2 O 2 release previously triggered by RT-EVs, demonstrating mechanistic dependence on NFκB signaling. Collectively, these findings identify GBM-derived RT-EVs as critical mediators of cognitive impairment through NFκB-dependent redox imbalance. EV-driven redox dysregulation may therefore represent a therapeutic target to mitigate GBM-associated cognitive dysfunction. Highlights:Radiation induces the release of glioblastoma-derived EVs that are biologically different from those released under non-irradiated conditions.EVs released from glioblastoma after radiation are sufficient to impair cognition EVs from irradiated glioblastoma can activate microglia via NFκB and induce production of neurotoxic H 2 O 2.

e16080 Background: In recent decades, there has been an unprecedented rise in gastric cancer among younger individuals, contrasting with a decline among older individuals. However, the biological underpinnings of gastric cancer in younger individuals remain poorly understood. We described the clinicopathologic and genomic characteristics of early-onset gastric cancer (EOGC) compared to average-onset gastric cancer (AOGC). Methods: We analyzed 311 patients using the multi-institutional prospective Oncology Research Information Exchange Network (ORIEN) database to compare demographic, clinicopathologic, genomic, and survival outcomes between EOGC ( < 50 years; N = 72) vs AOGC ( > 50 years N = 239). Genomic, germline, and RNA sequencing data were analyzed and compared between the cohorts. Mutational and immune signatures were also processed. Results: EOGC patients exhibited significantly higher rates of pain at diagnosis (53% vs 30% p = 0.001 ), but not anemia or reflux. EOGC patients were more likely to present with stage III/IV disease (70% vs 45% p = 0.006 ) and diffuse/signet ring histology (47% vs 16% p = 0.002 ). Consequently, OS was decreased in the younger cohort (HR 1.52; p = 0.03). Somatic mutational load was decreased in young patients. Significantly mutated genes in the entire cohort included CDH1 , ARID1A , TP53 , PIK3CA, but CDH1 was more frequently mutated in the EOGC cohort (40% vs 18% p = 0.006 ). Significant differences in RNA expression were observed, with upregulated epithelial mesenchymal transition (EMT), myogenesis, and apical junction. Immune deconvolution revealed a predominance of M2 macrophages, mast cells, and CD4 + T cell subsets, but no differences between cohorts. Conclusions: Early-onset gastric cancer has unique clinical and genomic features. Pathway dysregulation in EOGC may contribute to tumorigenesis and therapy resistance. This study underscores the necessity for further research into novel therapies, biomarker discovery, and early detection methodologies in younger individuals.

Sarcoidosis is a chronic granulomatous disease marked by persistent inflammation and immune cell aggregation, yet its molecular underpinnings remain incompletely understood, hindering the development of effective targeted therapies. Here, we report that deletion of TSC1 or TSC2 in mice using a Fsp1-Cre leads to spontaneous formation of sarcoid-like granulomas, driven by hyperactivation of the mTORC1 pathway in fibroblasts and interstitial macrophages. Through inflammatory cytokine/chemokine array, we identified CCL24, a chemokine ligand for CCR3, as a key immunoregulatory molecule downregulated in both our murine model and sarcoid cohort plasma. Mechanistically, mTORC1 suppresses CCL24 expression via aberrant STAT3 signaling in fibroblasts and promotes CCR3 expression in interstitial macrophages, uncovering a novel regulatory axis in granuloma formation and maintenance. Pharmacological inhibition using rapamycin and azithromycin markedly attenuated granuloma burden and normalized CCL24-CCR3 signaling, underscoring the therapeutic relevance of this axis. Together, our study establishes a mechanistic link between mTORC1 activation, CCL24-CCR3 dysregulation, and granuloma persistence, offering not only a new insight into molecular mechanisms in sarcoidosis but also identifying promising targets for clinical intervention.

Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with limited therapeutic options. Here, we investigated how integrin-dependent signaling pathways regulate tumor metabolism and therapeutic vulnerability in TNBC. Pharmacological inhibition of the integrin/FAK axis and/or BRD4 induced cell cycle arrest, autophagy, and senescence in highly proliferative cells, consistent with a metabolic stress phenotype. Metabolomic analyses using [U-¹³C]-glucose revealed a marked suppression of glycolytic carbon flux, accompanied by an approximately 30-47% reduction in intracellular NAD⁺ levels and coordinated alterations in NADH and tricarboxylic acid (TCA) cycle intermediate α-ketoglutarate. Mechanistically, we identified nicotinamide phosphoribosyltransferase (NAMPT), the rate-limiting enzyme in NAD⁺ biosynthesis, as a central metabolic node integrating signaling/function of the two axes. NAMPT expression/activity was sustained transcriptionally or post-translationally, including sirtuin-associated deacetylation and neddylation-dependent proteasomal turnover. In BRCA1/2-deficient TNBC, integrin-FAK and NAMPT/NAD+ pathways converged on Wnt/β-catenin signaling to regulate DNA repair, and response to PARP1/2 inhibitors. Co-inhibiting FAK and NAMPT synergistically suppressed tumor growth by approximately 80%. Elevated stromal NAMPT expression was associated with a trend toward favorable clinical outcomes. Collectively, these findings uncover a previously unrecognized crosstalk between integrin/FAK and NAMPT/NAD⁺ pathways in TNBC and identify a synthetic lethal-like therapeutic vulnerability that warrants further evaluation in clinically relevant models.

Cyclin-dependent kinase 1 (CDK1), a serine/threonine kinase essential for cell cycle progression, also plays critical roles in DNA damage response, gene expression regulation, and therapeutic resistance. In this study, we identify a previously unrecognized regulatory link between CDK1 and apurinic/apyrimidinic endonuclease 1 (APE1) in non-small cell lung cancer (NSCLC). Both CDK1 and APE1 are highly expressed in lung tumors and exhibit an inverse correlation in protein expression levels across multiple lung cancer cell lines. Mechanistically, CDK1 phosphorylates APE1 at Ser54 to promote its degradation, whereas inhibition of CDK1 stabilizes APE1 protein levels and reduces the efficacy of APE1-targeted therapy. Importantly, activation of CDK1 through WEE1 inhibition using MK1775 enhances APE1 degradation and markedly sensitizes NSCLC cells to APE1 inhibition, inducing synthetic lethality. CDK1-mediated APE1 degradation mimics APE1 loss, leading to genome instability, cell-cycle dysregulation, and altered metastatic behavior. Notably, the effect of CDK1-mediated APE1 phosphorylation on cell motility is p53-dependent: in p53-proficient cells, phosphorylated APE1 activates p53-dependent programs that suppress migration and invasion, whereas in p53-deficient cells the same modification enhances metastatic potential. Furthermore, further investigation revealed that APE1 deficiency activates the cGAS–STING pathway and induces PD-L1 expression through the MAPK–ERK axis, thereby reshaping the tumor immune microenvironment and enhancing sensitivity to immune checkpoint blockade. Given the dual roles of CDK1 in APE1 regulation and immune modulation, we propose that a sequential or combinatorial strategy in which WEE1 inhibition activates CDK1 to promote APE1 degradation, priming cancer cells for APE1 inhibition and inducing synthetic lethality, while minimizing toxicity to normal cells. This approach simultaneously disrupts APE1’s endonuclease and homologous recombination repair functions and exploits CDK1-driven immune reprogramming to potentiate anti–PD-L1 immunotherapy. Collectively, our findings establish CDK1-mediated APE1 phosphorylation as a key node linking DNA repair, immune evasion, and therapeutic response, and providing a mechanistic rationale for combined CDK1–APE1–PD-L1-targeted therapy in NSCLC.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is the fastest-growing etiology of hepatocellular carcinoma (HCC). Understanding the metabolic heterogeneity of MASLD-driven tumors is crucial to inform strategies for future treatment options. Paired tumor (n = 8) and adjacent non-tumor tissue (n = 8) were collected from patients with steatohepatitic HCC at the University of Kentucky Markey Cancer Center. Hematoxylin and eosin (H&E) staining was used for pathological determination of tumor and adjacent nontumor tissue by a board-certified pathologist. Lipidomic, metabolomic, and transcriptomic analyses were performed, and data were integrated across platforms to identify novel relationships across tumor and adjacent nontumor tissue. Paired transcriptomic analyses were validated in 424 human samples from The Cancer Genome Atlas-Liver Hepatocellular Carcinoma (TCGA-LIHC). Histological analysis by H&E showed significant lipid vacuole accumulation and inflammatory foci in HCC tumors relative to nontumor tissue. Across omics platforms, we identified 1,679 genes, 1,696 metabolites, and 292 lipids that were significantly (padj < 0.01) increased or decreased across all paired (tumor vs. nontumor) patient samples. We identified significant reductions in ceramides and linoleic acid-enriched lipids, and increases in fatty acyl chain saturation in tumor tissue. Metabolites involved in purine and fatty acid catabolism were commonly decreased in tumors relative to nontumor tissue across paired samples. We also identified a total of 303 highly significant and novel transcript-metabolite associations (117 gene-metabolite; 186 gene-lipid) across tumor and nontumor tissue. Taken together, this integrative analysis reveals novel relationships between steady-state gene transcripts and specific metabolites in steatohepatitic tumors, thereby identifying new pharmacological targets that may be exploited for therapeutic benefit.

Abstract Background: Cancer-associated fibroblasts (CAFs) are the major sources of secreted cytokines and chemokines contributing to the shaping of immunosuppressive tumor microenvironment (TME). Dysregulation of glycogen metabolism is integral to cancer cell proliferation and metastasis. The purpose of this study was to identify whether targeting glycogen metabolism in CAFs could alleviate immunosuppression and inhibit colorectal cancer (CRC) growth. Methods: Human CAFs (hCAFs) were isolated from CRC patient samples. APCf/f; CDX2-Cre/ERT2 mice were injected with tamoxifen to induce adenoma formation. Mouse CAFs (mCAFs) were isolated and cultured. The expression of glycogen phosphorylase liver form (PYGL), phospho-PYGL, hexokinase 2 (HK2), GLUT1, IL6, CLCF1, LIF, and CXCL6 was determined by either real time (RT)-PCR or western blot. Glycogen levels were determined using a Glycogen Analysis Kit. Cell proliferation was determined using WST-1. Extracellular acidification rate (ECAR) was determined utilizing an Agilent Seahorse XFe96 extracellular flux analyzer. To determine whether targeting PYGL would inhibit tumor growth, APCf/f; CDX2-Cre/ERT2 and MC38 mouse tumor models were used. Results: (i) Elevated glucose metabolism in CAFs was found compared to normal fibroblasts. (ii) Treatment of hCAFs with either conditioned medium (CM) derived from HCT116 CRC cells or TGFβ resulted in activated glycogen metabolism as noted by increased expression of p-PYGL/PYGL and the reduced level of glycogen in hCAFs. Moreover, CM increased the expression of IL6 family cytokines such as IL6, CLCF1, LIF and CXCL6 in hCAFs. (iii) Inhibition of glycogen metabolism by knockdown of PYGL or treatment with the PYG inhibitor, CP-91149, significantly repressed hCAFs proliferation and reduced the expression of these cytokines in hCAFs. Knockdown of PYGL decreased glycolytic activity as noted by decreased ECAR. (iv) Treatment with CP-91149 significantly inhibited tumor growth. In addition, mCAF cells co-implanted with MC38 cells increased MC38 tumor growth; this increase was attenuated by knockdown of PYGL in mCAFs. Conclusion: Our results demonstrate that glycogen metabolism is crucial for promoting CAF immunosuppressive functions. Importantly, our findings suggest that targeting glycogen metabolism in CAFs alleviates immunosuppression and inhibits CRC malignancy. Citation Format: Xudong Zhu, Yuning Zhou, Haoxiang Zhang, Yinping Jiang, Jinpeng Liu, Chi Wang, B. Mark Evers, Qingding Wang. Targeting glycogen metabolism in cancer-associated fibroblasts alleviates immunosuppression and inhibits colorectal cancer malignancy [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 4713.

Hematopoietic stem cells (HSCs) have the ability to self-renew and differentiate to all blood cell types. HSCs and their differentiated progeny show sex/gender differences. The fundamental mechanisms remain largely unexplored. We previously reported that latexin (Lxn) deletion increased HSC survival and repopulation capacity in female mice. Here, we find no differences in HSC function and hematopoiesis in Lxn knockout (Lxn-/-) male mice under physiologic and myelosuppressive conditions. We further find that Thbs1, a downstream target gene of Lxn in female HSCs, is repressed in male HSCs. Male-specific high expression of microRNA 98-3p (miR98-3p) contributes to Thbs1 suppression in male HSCs, thus abrogating the functional effect of Lxn in male HSCs and hematopoiesis. These findings uncover a regulatory mechanism involving a sex-chromosome-related microRNA and its differential control of Lxn-Thbs1 signaling in hematopoiesis and shed light on the process underlying sex dimorphism in both normal and malignant hematopoiesis.

OBJECTIVE:We investigated neurotensin receptor 1 (NTSR1) as a potential mediator of the mechanical immune barrier that contributes to T-cell exclusion. SUMMARY BACKGROUND DATA:The efficacy of immunotherapy in advanced colorectal cancer (CRC) is limited by primary resistance, frequently characterized by an "immune-excluded" phenotype where physical and biological barriers prevent effector T-cell infiltration. Through its high-affinity receptor, NTSR1, neurotensin signaling has been implicated in tumor progression, yet its role in shaping the immune landscape of CRC remains unclear. METHODS:We integrated transcriptomic and clinical data from The Cancer Genome Atlas discovery cohort with an independent surgical validation cohort from the University of Kentucky. Random Forest machine learning identified molecular features associated with NTS/NTSR1 signaling, and a 3D tumor spheroid co-culture model was used to evaluate the impact of NTS/NTSR1 signaling on spatial T-cell distribution in vitro . RESULTS:Dual-cohort analyses revealed high NTSR1 expression as an independent risk factor for progression-free interval in the discovery cohort and overall survival in the validation cohort. Mechanistically, the NTS/NTSR1 axis upregulates a mechano-structural network centered on PLXNB3, FLNC, and AHNAK2, while simultaneously driving epithelial dedifferentiation through downregulation of core transcription factors (CDX2, SATB2). Together, these transcriptional programs are associated with the remodeling of the tumor microenvironment into an immune-cold, suppressive state. Moreover, 3D modeling indicated that ligand-activated NTS/NTSR1 signaling restricts T-cell infiltration, which was significantly reversed by the NTSR1 antagonist SR48692, providing functional support for a mechanical immune barrier proposed based on transcriptomic remodeling. CONCLUSIONS:Targeting the NTS/NTSR1 axis may attenuate biological and physical barriers underlying immune exclusion, providing a preclinical rationale for converting immune-excluded CRCs into immune-responsive tumors.