PURPOSE:AXL, is a receptor tyrosine kinase implicated in EGFR resistance. The primary objective of this study was to test the hypothesis, generated in preclinical studies, that low AXL correlates with clinical response to cetuximab in patients with head and neck cancer (HNC). The secondary objectives were to further describe the feasibility and safety of preoperative administration of cetuximab. METHODS:We performed a single-site, single arm, open label, window-of-opportunity study utilizing preoperative cetuximab in 15 patients with HNC. Patients received two doses of cetuximab and underwent resection of their tumor. Biopsies were used to assess AXL expression, change in Ki67 staining, and to establish patient-derived xenografts. scRNA-seq of treated xenograft tissue was used to assess differences between sensitive and resistant tumors. RESULTS:Fifteen subjects were registered for treatment on the study and fourteen completed treatment with cetuximab prior to surgical resection. No correlation between AXL expression at baseline and change in tumor size in response to cetuximab was seen. Preoperative cetuximab did not delay surgical resection and no unexpected treatment-related adverse events were seen. scRNA-seq identified that cetuximab sensitivity is marked by transcriptional remodeling, while resistance is associated with stable HER signaling and elevated TAM activity. CONCLUSION:Delivery of preoperative cetuximab is safe, and a short course can be utilized to identify patients with HNC responding to single-agent cetuximab treatment.
PURPOSE:PF-07062119 is an anti-GUCY2C/anti-CD3ε bispecific Fc antibody designed to elicit systemic anti-tumor immunity. This phase 1, first-in-human study evaluated the safety, tolerability, pharmacokinetics (PK), pharmacodynamics, immunogenicity, and anti-tumor activity of PF-07062119 as monotherapy and in combination with sasanlimab or bevacizumab in patients with advanced gastrointestinal cancers expressing GUCY2C. PATIENTS AND METHODS:Patients received escalating doses of PF-07062119 (45-3700 μg subcutaneously [SC]) in Part 1A. In Part 1B, PF-07062119 was administered with either sasanlimab (300 mg SC) or bevacizumab (5 mg/kg intravenously). Primary endpoints were safety and tolerability; secondary endpoints included PK, immunogenicity and efficacy. RESULTS:A total of 79 patients were enrolled. In Part 1A, maximum tolerated dose without a priming dose was 800 μg. Dose-limiting toxicities observed at this level were grade 3 cytokine release syndrome (CRS), colitis, and diarrhea. A 400 μg priming dose with premedication reduced CRS incidence and recommended dose for expansion was 2100 μg. Treatment-related grade 3/4 treatment-emergent adverse events (TEAEs) were observed in 27 (34.2%) patients, with no grade 5 treatment-related TEAEs. Diarrhea remained the most clinically significant toxicity in Part 1A. Combination therapy in Part 1B did not result in significant additional toxicity. PK analyses demonstrated dose-proportional increases in exposure. The overall response rate was 2.5% across all cohorts. CONCLUSIONS:PF-07062119 monotherapy demonstrated a generally tolerable safety profile within the context of this early-phase study with evidence of clinical activity in patients with advanced/metastatic gastrointestinal cancers. CLINICAL TRIAL INFORMATION:NCT04171141.
At least 15% of children with cancer have a pathogenic germline variant in a cancer predisposition gene. Studies of germline cancer predisposition, however, have focused primarily on single nucleotide and copy number variants, leaving other classes such as transposable elements (TEs) largely unexplored. Although rare pathogenic TE insertions have been implicated in inherited cancer, their contribution to pediatric cancer predisposition remains unknown, partly because these repetitive sequences often require whole-genome sequencing for detection. We characterized the germline TE insertion landscape using non-tumor whole genome sequencing data from 2,334 pediatric cancer and 3,447 controls. Across 5,781 genomes, we identified 96,484 TE insertions, most of which were rare and located in intergenic or intronic regions. While global TE burden did not differ between cases and controls, rare TE insertions were significantly enriched in cancer genes in patients with solid tumors, particularly within 3' untranslated regions (p<0.02). Gene-phenotype concordance analysis identified 19 insertions in genes with established dominant cancer predisposition, representing ~0.8% of cases. Integration of RNA-seq data revealed transcriptional impact for a subset of insertions. Notably, a 3' UTR L1 insertion in the tumor suppressor PTEN disrupted alternative polyadenylation, whereas an SVA insertion in a STIM1 intron induced exonization, generating a novel transcript containing SVA sequence. These findings demonstrate that rare germline TE insertions in cancer predisposition genes can have functional consequences at the RNA level. Incorporating TE detection into genomic workflows may improve identification of cancer predisposition syndromes and expand understanding of noncoding contributions to pediatric cancer susceptibility.
Immune checkpoint blockade (ICB) is an effective treatment for microsatellite instability-high (MSI-H) colorectal cancers (CRCs) that are highly infiltrated by CD8+ T cells. Microsatellite stable (MSS) CRCs are unresponsive to ICB, at least in part, due to the paucity of intratumoral CD8+ T cells. We recently identified Discoidin Domain Receptor 1 (DDR1) as one of four genes associated with CD8+ T-cell exclusion in MSS CRC. There are conflicting reports about the presence and role of the cleaved ectodomain (cECD) of DDR1 in mouse models of breast and pancreatic cancer. To explore the role of the DDR1 cECD in human CRC, we developed Collagen Alignment and Spatial Transcriptomics Analysis (CASTA), which revealed that genes involved in fibroblast contractility were associated with both DDR1 tumor expression and collagen alignment as determined by label-free second harmonic generation (2HG) imaging of the tumor collagen. Using 3D collagen co-cultures of CRC spheroids and fibroblasts, we show that DDR1 promotes collagen alignment and CD8+ T-cell exclusion. We found large amounts of DDR1 cECD in MSS CRC supermeres, 25-35 nm secreted amembranous nanoparticles. Supermeres containing DDR1 cECD were sufficient to induce contraction of human colonic fibroblasts. We propose a model in which supermeres containing DDR1 cECD promote the contraction of stromal fibroblasts in MSS CRC, leading to collagen alignment and CD8+ T-cell exclusion. These results support DDR1 cECD as an attractive therapeutic target in MSS CRC.
The development of liver metastases in prostate cancer is associated with aggressive disease and poor prognosis. Because hepatocytes exhibit high metabolic activity with unique secretory profiles, we hypothesized that hepatocyte-to-tumor cell signaling plays a role in promoting liver metastasis. We evaluated single-cell transcriptomic data and metastatic tissue from patients with castration-resistant prostate cancer spanning androgen receptor (AR)-positive and AR-negative pathologies. Despite extensive intra- and inter-sample heterogeneity, communication analysis predicted vitronectin engagement of tumor integrins as a common feature. Vitronectin-positive hepatocytes were observed in prostate tumors with vitronectin accumulation in sinusoidal patches. Consistent with integrin activation, vitronectin treatment of AR-positive and AR-negative prostate cancer cells significantly promoted tumor cell adhesion, inhibited hypodiploid accumulation consistent with survival effects, and stimulated FAK-dependent phosphorylation of ERK and AKT. FAK inhibition with defactinib abrogated vitronectin- and serum-mediated adhesion in a cell-specific manner and mitigated VTN-driven depletion of hypodiploid populations. These findings support a model where dense intra-sinusoidal vitronectin deposits might capture metastatic prostate tumor cells in the liver and biochemically activate tumorigenic signaling, promoting tumor aggressiveness.
Non-small-cell lung cancer (NSCLC) is responsible for the majority of cancer-related mortality worldwide. Lung adenocarcinoma (LUAD) is the most common NSCLC subtype. Despite advances in targeted therapies, treatment resistance remains a critical challenge. Ribonucleotide reductase (RNR), a crucial enzyme in deoxyribonucleotide triphosphate (dNTP) biosynthesis, is frequently upregulated in cancer, contributing to genomic instability and poor prognosis in multiple malignancies. However, the role of the RNR complex in driving tumorigenesis is not fully understood in oncogenic-driven LUAD. Transcriptomic analysis of more than 27,000 real-world NSCLC patient samples revealed that RNR subunits (RRM1 and RRM2) are significantly upregulated in TP53 mutated NSCLC and correlated with significantly poor prognosis in multiple oncogene-driven LUAD. Using pharmacologic and genetic approaches to inhibit RNR in LUAD models, we assessed functional consequences through molecular, biochemical, and imaging techniques. RNR inhibition induced appreciable replication stress and triggered DNA damage, leading to cell death in LUAD cells. Notably, we uncovered that RNR suppression preferentially induced ferroptosis, an iron-dependent cell death driven by lipid peroxidation. This represents a previously unrecognized mechanism of RNR-mediated cell death by which mutant LUAD cells can be selectively targeted. Our study establishes RNR inhibition as a potent strategy to selectively induce ferroptosis in oncogenic addicted LUAD, offering a new therapeutic avenue for genetically defined patient subgroups. Targeting nucleotide metabolism could serve as an effective approach to overcome treatment resistance and improve clinical outcomes for patients with high-risk LUAD.
Fibroblast growth factor receptor 1 (FGFR1) amplification is frequently observed in ER⁺/HER2⁻ breast cancer and has been linked to poor response to endocrine therapy. While FGFR1 has been implicated in therapeutic resistance, its role in regulating interferon (IFN) response in the context of endocrine resistance remains unclear. We investigated whether FGFR1 modulates the IFN response through the GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway in tamoxifen-resistant breast cancer. We employed tamoxifen-resistant ER⁺ breast cancer cell lines and a tamoxifen-sensitive FGFR1-amplified patient-derived xenograft (PDX) model to examine the impact of FGFR1 inhibition on the IFN response through RNA sequencing, FGFR1 knockdown or pharmacologic inhibition, and functional assays. Clinical relevance was evaluated in breast cancer cohorts. Tamoxifen-resistant cells exhibited increased cytosolic, cGAS-positive dsDNA foci alongside elevated p-IRF3, indicating constitutive engagement of the cGAS-STING pathway. FGFR1 inhibition re-sensitized resistant cells to tamoxifen and induced IFN response gene expression, which was further amplified by tamoxifen co-treatment. FGFR1 inhibition similarly potentiated tamoxifen-induced IFN response gene expression in vivo in a FGFR1-amplified PDX model. Mechanistically, FGFR1 knockdown enhanced STING-mediated IFNB1 expression in response to cytosolic DNA. In the METABRIC breast cancer cohort, FGFR1 amplification was associated with poor prognosis specifically in tumors with high STING1 expression. These findings identify FGFR1 as a suppressor of cGAS-STING-mediated interferon response, driven by tamoxifen in ER⁺ breast cancer. Attenuation of this response may represent a previously unrecognized contribution of FGFR1 to tamoxifen resistance.
High grade serous carcinoma (HGSC) of the ovary, fallopian tube and peritoneum has an immunosuppressive tumor immune microenvironment (TIME), mediated in part by infiltrating innate immune cells such as macrophages, neutrophils, and myeloid derived suppressor cells (MDSCs). We hypothesize that manipulating these immunoinhibitory cells will improve both response to treatment and outcomes for patients with HGSC. Using publicly available databases, we demonstrate that higher expression of CXCR2, a chemotactic receptor for MDSCs and neutrophils, is correlated with decreased overall survival and increased infiltration of neutrophils, monocytes, and M2-like macrophages in HGSC. We targeted the migration of MDSCs and neutrophils to the TIME in the ID8-p53null mouse model of HGSC with the CXCR2-selective inhibitor SB-225002 (CXCR2i) alone or in combination with cisplatin. Tumor weight and dissemination were decreased in mice treated with single-agent cisplatin or CXCR2i. In cisplatin treated tumors compared with vehicle treated tumors, there was an increase in suppressive myeloid cells measured by flow cytometry and multispectral immunohistochemistry. Moreover, cisplatin-mediated modulation of tumor immune cell populations was abrogated by the addition of CXCR2i to cisplatin treatment. These findings demonstrate that inhibition of CXCR2 can slow tumor progression and decrease the proportion of immunosuppressive myeloid immune cells in the TIME. Furthermore, these data reveal new insights into detrimental chemotherapy-induced remodeling of the TIME, which should be investigated as mechanistic targets to improve outcomes in HGSC. Overall, our data demonstrate the clinical relevance and therapeutic potential of targeting CXCR2 to prevent their recruitment of tumor-promoting innate immune cells to the TIME.
Abstract Acute lymphoblastic leukemia (ALL) shows persistent outcome disparities among socioeconomically disadvantaged and Hispanic children. Neighborhood socioeconomic status (nSES) and residence in Hispanic enclaves may contribute to these disparities, yet their impact on pediatric ALL is unclear. We examined associations of nSES and Hispanic enclave residence with ALL treatment outcomes. We analyzed children (0–24 years) diagnosed with ALL between 2005 and 2017 and treated at REducing Disparities In Acute Leukemia (REDIAL) Consortium centers in Texas. Census-tract nSES (Yost) and Hispanic enclave indices were computed from US Census and American Community Survey (ACS) data, classifying tracts into low/high nSES and enclave/no enclave residence. Outcomes included end-of-induction (EOI) minimal residual disease (MRD) positivity (MRD ≥ 0.01%), relapse, event-free survival (EFS), and overall survival (OS). Associations were evaluated using multivariable logistic and Cox regression models adjusting for sex, age at diagnosis, race/ethnicity, EOI MRD, cytogenetic features, National Cancer Institute (NCI) risk group at diagnosis, ALL immunophenotype, and treating institutions. Among 1,348 patients, 41% resided in low-nSES tracts and 42% in Hispanic enclaves. Neither enclave residence nor low nSES was associated with EOI MRD positivity; similar null associations were observed for relapse, EFS, and OS. Although unadjusted analyses indicated slightly poorer 5-year EFS for low-nSES areas (81% vs. 85%, P = 0.04), this difference attenuated after adjustment. In this large, multicenter, and diverse cohort of children with ALL, neighborhood socioeconomic disadvantage and Hispanic enclave residence were not independently associated with treatment outcomes after accounting for clinical and cytogenetic features. Future studies should examine other social determinants and neighborhood influences on disease characteristics, survivorship, and late effects. Significance: In this multicenter study, neither neighborhood socioeconomic disadvantage nor residence in a Hispanic/Latino enclave was associated with inferior outcomes among children with ALL after controlling for clinical and cytogenetic disease features. Disparities affecting economically disadvantaged or minoritized populations may be mediated by other factors.
Abstract Trabectedin, prototype of the ecteinascidin class of drugs, is a known second- or later-line therapeutic option for advanced soft tissue sarcoma (STS). We evaluated the antitumor activity of two novel synthetic trabectedin derivatives, ecubectedin and PM54, in selected patient-derived xenograft (PDX) STS models. In total, 364 Naval Medical Research Institutenu/nu mice were transplanted bilaterally with two leiomyosarcoma (LMS), two dedifferentiated liposarcoma (DDLPS), one synovial sarcoma (SynSa), and one CIC-rearranged sarcoma (CRS) PDX models. Mice were randomized to six groups and treated via tail vein injection with (i) vehicle, (ii) doxorubicin, (iii) trabectedin, (iv) lurbinectedin, (v) ecubectedin, or (vi) PM54. Treatment was given on days 1, 8, and 15, and mice were sacrificed on day 16. The SynSa experiment included extra mice to investigate posttreatment xenograft evolution. Antitumor activity was assessed by tumor volume measurement, histopathologic, and immunohistochemical analyses. In all LMS and DDLPS models, ecubectedin and PM54 led to tumor growth delay compared with trabectedin. Histopathologic evaluation of treated tumors showed moderately increased antitumor activity of novel ecteinascidins compared with trabectedin. The CRS model showed tumor shrinkage in response to ecubectedin (63% regression from baseline) and PM54 (24% regression from baseline), while the SynSa model showed tumor volume stabilization. Both translocation-related models showed significant antitumor effect on histopathologic evaluation in response to the novel drugs compared with trabectedin. Ecubectedin and PM54 have modest antitumor activity in STS PDX models, with the strongest effects seen in the translocation-related sarcoma models, showing the potential of this class of drugs in the treatment of STS. Significance: Advanced STS remains without truly effective therapeutic options. Translocation-related sarcomas demonstrate higher vulnerability to next-generation ecteinascidin treatment compared with more complex sarcoma subtypes. This vulnerability builds upon known literature and could be exploited in future clinical trials using the next-generation ecteinascidins.
Pathologic complete response (pCR) after cisplatin-based neoadjuvant chemotherapy (NAC) is associated with improved outcomes in muscle-invasive bladder cancer (MIBC), but genomic predictors of benefit remain unvalidated. Based on observations in lung cancer, we hypothesized that patients with MIBC with high tumor mutational burden (TMB) would be unlikely to benefit from NAC, indicated by residual disease after cystectomy. Pretreatment tumor specimens from patients treated with cisplatin-based NAC were analyzed using Tempus xT/xR platforms. Sample size was prespecified by statistical design. High TMB was defined as the upper TMB quintile within the cohort. Correlation of TMB values and genomic variants with outcomes was performed using logistic regression, Cox proportional hazards models, and Kaplan-Meier techniques. Ninety-one patients were included, with a median follow-up of 63.6 months. pCR occurred in 31.9%. Median TMB was 8.9 mutations per megabase (Mut/Mb). Contrary to the prespecified hypothesis, higher TMB was associated with more favorable outcomes. Patients in the upper TMB quintile had a numerically higher pCR rate (47.4% vs. 27.8%; P = 0.165) and improved relapse-free survival (RFS)/overall survival [OS; hazard ratio (HR), 0.456; P = 0.08 and HR, 0.495; P = 0.16]. Using a clinically relevant cutoff, TMB ≥ 10 Mut/Mb was associated with higher pCR [43.6% vs. 23.1%; odds ratio (OR), 0.39; P = 0.044] and improved RFS (HR, 0.330; P < 0.001) and OS (HR, 0.388; P = 0.013). Exploratory analyses showed a trend toward enrichment of mismatch repair alterations (OR, 3.57; P = 0.087) in TMB-high tumors. Baseline TMB ≥ 10 Mut/Mb was associated with improved pathologic response and survival in MIBC treated with cisplatin-based NAC and warrants prospective evaluation in contemporary perioperative regimens. SIGNIFICANCE:In MIBC treated with cisplatin-based neoadjuvant chemotherapy, we found that baseline TMB ≥10 Mut/Mb was associated with higher pCR rates and improved RFS and OS. These findings support the prospective study of TMB as a prognostic biomarker in contemporary perioperative treatment regimens.
Pancreatic ductal adenocarcinoma (PDAC) remains a formidable clinical challenge. Next-generation protein arginine methyltransferase 5 (PRMT5) inhibitors show promising clinical results in a subset of PDACs with codeletion of the tumor-suppressor CDKN2A and the methylthioadenosine phosphorylase (MTAP) gene, but resistance limits their efficacy. Our study suggests that compensatory spliceosomal reprogramming contributes to adaptation to PRMT5 inhibition. Through comprehensive molecular profiling, we demonstrate that PRMT5 inhibitors induce upregulation of RNA-binding proteins, including RNA-binding protein 39 (RBM39). We investigated whether this response could be therapeutically leveraged by combining PRMT5 inhibition with indisulam-mediated RBM39 degradation, which yielded synergistic activity in cellular model systems. The combination strategy significantly enhanced apoptotic cell death and suppressed tumor outgrowth in resistance assays compared with single-agent treatments. Multiomics analysis revealed concomitant suppression of DNA repair and metabolic pathways. Collectively, our work support spliceosomal rewiring as a candidate adaptive response to PRMT5 inhibition and nominates RBM39 as a candidate therapeutic vulnerability, thereby supporting further evaluation of dual targeting of the splicing machinery. SIGNIFICANCE:Our study suggests that compensatory spliceosomal reprogramming occurs in response to PRMT5 inhibition. We investigated this vulnerability by combining PRMT5 inhibition with indisulam-mediated RBM39 degradation, which yielded synergistic antitumor activity in selected cellular PDAC models.
Abstract Purpose: Despite widespread use of targeted therapies, little is known about how tumors and their microenvironment respond in the earliest phases of treatment and whether these early changes reveal new therapeutic opportunities. In this study, we investigated how short-term epidermal growth factor receptor (EGFR) inhibition reshapes tumor biology and the tumor microenvironment using comprehensive multiregion and spatial molecular profiling. Patients and Methods: Patients with stage IA–IIIA EGFR-mutated non–small cell lung cancer (NSCLC) received at least 4 weeks of gefitinib (250 mg daily) before surgery. Multiregion whole-exome and RNA sequencing of resected tumors were compared with a purity-matched treatment-naïve cohort. Results: Thirteen patients received neoadjuvant gefitinib for a median of 1.4 months, achieving objective response and disease control rates of 62% and 100%, respectively. Pathologic downstaging occurred in 46% of patients, with major pathologic response in 8%. Deep genomic and transcriptomic profiling revealed rapid tumor and microenvironmental adaptation to EGFR inhibition without emergence of common acquired resistance mutations such as EGFR T790M. Compared with treatment-naïve tumors, on-treatment tumors showed reduced tumor purity, decreased EGFR amplification, increased immunoregulatory and inflammatory gene expression, and a metabolic shift from glycolysis to oxidative phosphorylation, spatially validated. Conclusions: These findings show that early EGFR inhibition triggers coordinated remodeling of tumor cells and the microenvironment, uncovering new therapeutic vulnerabilities and opening a window for rational combination strategies to improve outcomes in EGFR-mutated NSCLC. Significance: This study demonstrated that neoadjuvant gefitinib is safe, feasible, and effective. Multiregion and spatial genomic and transcriptomic sequencing provided unique insight into adaptive response in the tumor and microenvironment. This window-of-opportunity study uncovered the potential for rational combination approaches to improve therapeutic efficacy in EGFR-mutated NSCLC.
How mitochondrial DNA (mtDNA) polymorphisms influence complex phenotypes remains poorly understood. Using mitochondrial-nuclear exchange mice, we previously showed that mtDNA single-nucleotide polymorphisms (SNP) modify metastasis, cardiovascular disease, and epigenetic marks independently of metabolic differences. The only mtDNA SNP correlating with these phenotypes resides in the gene encoding mitochondrial transfer RNA (tRNA)-arginine [mt-tRNAArg (UCG), mt-TR], suggesting a role for non-protein-coding loci. In this study, we identify and preliminarily characterize previously undescribed tRNA-derived fragments (tRF) generated from mt-TRs. Northern blotting revealed distinct tRF that are differentially expressed among mtDNA SNPs, between lung and liver, and between sexes. Surprisingly, small RNA sequencing of untreated RNA did not detect the same tRFs in high abundance. However, demethylating and restoring 5'-OH and 3'-PO4 termini allowed detection of sequences consistent with the northern blot bands. Enforcing exact matching to the mitochondrial genome and normalizing to their parental molecule revealed putative tRF sequences with shared cleavage sites. Based on connections among mtDNA SNPs, the resulting SNP-dependent tRF, and SNP-metastasis correlation, we propose that these tRFs may function as metastasis modifiers. These data also expand the functional output of the mitochondrial genome that can contribute to phenotype modification. SIGNIFICANCE:This study describes identification and initial characterization of mitochondrial tRNA fragments that are suspected to regulate metastasis efficiency.
PURPOSE:A pilot clinical trial (NCT03519308) was conducted to evaluate perioperative chemotherapy [nab-paclitaxel, gemcitabine, and cisplatin (NGC)] plus nivolumab, with or without synthetic vitamin D (paricalcitol) in resectable pancreatic ductal adenocarcinoma (PDAC). Preclinical studies were designed with statistical power to match the experimental regimen and compare NGC only with NGC plus synthetic vitamin D [calcipotriol (Cal)] or losartan (Losa; an antihypertensive medication). MATERIALS AND METHODS:Molecular subtyping was applied to enroll patients with epithelial subtype PDAC. Genetically engineered PDAC models (KrasG12D;Trp53R172H;Pdx1-Cre mice, N = 140) were randomized to NGC only and combined therapies. Multiparametric MRI and molecular profiling were integrated to assess treatment-induced effects on stroma, including capillary perfusion and density, matrix collagen content, fibroblast activation protein (FAP) levels and transcriptome changes. RESULTS:The clinical trial had limited enrollment (N = 3) but confirmed the safety and feasibility of weekly paricalcitol infusion in combination with multiagent chemotherapy. NGC chemotherapy had a profound impact on stroma, inducing a remarkable reduction in FAP levels accompanied by a significant increase in matrix collagen content; chemotherapy also activated epithelial-mesenchymal transition (EMT) and significantly reduced blood perfusion and capillary density, resulting in features of chemoresistance. Combining NGC with stromal-targeting agents-Cal or Losa-diminished EMT and partially restored perfusion through distinct mechanisms: Cal activates the vitamin D receptor to reprise a quiescent stroma, whereas Losa suppresses TGFβ to increase lymphocyte infiltration. CONCLUSIONS:These findings support further investigations of stromal-directed combination strategies to overcome chemotherapy-induced resistance of PDAC. Dynamic contrast-enhanced MRI may provide a valuable platform to monitor stromal adaptations to therapy in vivo. SIGNIFICANCE:Results from co-clinical trials integrating MRI and molecular profiling provide a translational rationale for investigating stromal-directed combination strategies in PDAC. These approaches may help preserve chemosensitivity, counteract chemotherapy-induced stromal and vascular remodeling, and overcome adaptive resistance mechanisms that limit durable treatment response.
Biliary tract cancers (BTC) pose clinical challenges due to poor chemotherapy response and aggressive disease course. We evaluated patient-derived tumor organoid-based drug sensitivity testing as a tool to guide therapy. In this multicenter study, 26 tumor organoids were successfully derived from 43 patients with BTC and tested with an average of 50 cancer-directed therapies using the Clinical Laboratory Improvement Amendments-certified PARIS assay. Despite most organoids being from late-stage disease, 24/26 (92.3%) exhibited strong sensitivity to one or more targeted agents. Active drugs included inhibitors of EGFR/HER2, MEK, ERK, BCR-ABL and SRC family, mTOR, PI3K, MDM2, BCL2, and BET. Drug sensitivities aligned with known genetic biomarkers but were also observed in cultures lacking them, indicating ex vivo testing can expand actionability beyond genomics. In five cases, results guided therapy; one patient with an FGFR-BICC1 fusion refractory to FGFR inhibitors responded to dasatinib, achieving symptomatic improvement, stable disease, and >8-month survival. SIGNIFICANCE:Ex vivo drug testing of tumor-derived organoids is clinically feasible and can be used to identify personalized treatment options for patients with BTC, to evaluate the functional relevance of genomic biomarkers, and to guide treatment in real time.
Abstract Classic Hodgkin lymphoma (CHL) commonly affects adolescents and young adults (AYA), including service members (SM). The impact of deployment on CHL outcomes remains unknown. TRICARE beneficiaries diagnosed with CHL from 2001 to 2022 were identified through the Department of War cancer registry. Deployment history came from the Defense Manpower Data Center, and mortality data came from the National Death Index. Mortality was compared between SMs with and without post-9/11 deployment, and associations with deployment-specific variables were evaluated using Cox models adjusted for demographic and clinical factors. An exploratory analysis was performed to assess the survival of AYA SMs by race and to compare mortality with the Surveillance, Epidemiology, and End Results (SEER) database. Among 752 SMs identified, 369 (49%) had deployed after 9/11. Deployment was not associated with overall mortality [adjusted hazard ratio (aHR), 1.64; 95% confidence interval (CI), 0.84–3.19] or disease-specific mortality (aHR, 0.67; 95% CI, 0.13–3.55). No deployment-specific variable meaningfully influenced mortality. There were 21 out of 417 (5%) all-cause deaths in White AYA SMs compared with 0 out of 89 in Black AYA SMs (log-rank test P = 0.0198). Compared with SEER, SMs had lower all-cause mortality (aHR, 0.52; 95% CI, 0.38–0.72) and disease-specific mortality (aHR, 0.29; 95% CI, 0.15–0.59). Post-9/11 deployment did not adversely affect survival among SMs with CHL, highlighting the need for exposure-specific and survivorship-focused research. Survival disparities by race were not appreciated, and mortality in SMs was lower than SEER, highlighting access to care within the Military Health System (MHS) and the healthy soldier effect. Significance: In this national cohort of 752 U.S. SMs with CHL, post-9/11 deployment and deployment-related variables did not adversely affect overall or disease-specific survival. These findings suggest that deployment exposures or occupational demands do not meaningfully alter CHL outcomes. Lower mortality compared with SEER underscores the healthy soldier effect and highlights the advantages of universal access to timely, coordinated cancer care within the MHS.
Abstract As Helicobacter pylori prevalence declines, understanding the contribution of modifiable systemic factors to gastric cancer risk becomes increasingly important. We evaluated the association of metabolic syndrome (MetS), its individual components, and anatomic subsites with incident gastric cancer in 471,540 UK Biobank participants. Multivariable Cox proportional hazards models were used to estimate the risk of overall, cardia, and non-cardia gastric cancers, and restricted cubic splines were used for exploratory nonlinear analyses. Over a mean follow-up of 6.5 years, 332 incident gastric cancer cases were identified. In the fully adjusted model, baseline MetS was associated with increased gastric cancer risk [HR = 1.36; 95% confidence interval (CI), 1.08–1.71]. A positive trend was observed with the accumulation of metabolic components (P for trend = 0.033). Subsite analyses showed a directionally positive association for cardia gastric cancer (HR = 1.48; 95% CI, 1.03–2.11) and a nonsignificant positive association for non-cardia gastric cancer (HR = 1.28; 95% CI, 0.95–1.73); however, formal testing found no statistical evidence of subsite heterogeneity (P for heterogeneity = 0.547). Exploratory spline analysis suggested a possible nonlinear association for systolic blood pressure. Overall, MetS and the accumulation of its components were associated with a moderately increased risk of gastric cancer. Waist circumference showed the most consistent component-level signal, and further studies are needed to clarify the role of metabolic health in gastric cancer risk assessment. Significance: In this prospective analysis of 471,540 UK Biobank participants, MetS was associated with a moderately increased risk of incident gastric cancer. A positive trend was observed with cumulative metabolic component burden, and waist circumference showed the most consistent component-level signal. Subsite analyses suggested a stronger point estimate for cardia gastric cancer, but formal testing found no statistical evidence of heterogeneity between cardia and non-cardia tumors.
Abstract Understanding complex cellular niches, such as tertiary lymphoid structures, requires spatially resolved, multi-omics approaches that link transcriptional states and protein expression levels within individual cells. Here, we present an integrated spatial multi-omics workflow that enables sequential mapping of hundreds of genes via the Xenium In Situ platform and more than 40 protein markers via imaging mass cytometry technology on a single formalin-fixed, paraffin-embedded (FFPE) tissue section. We applied this multi-modal approach to colorectal liver metastases and matched adjacent normal liver tissues. Our results demonstrate that the sequential application of these technologies maintains tissue morphology and the assay’s technical sensitivity. Furthermore, high-dimensional data integration was performed through optimized computational coregistration at a single-cell level. Through this approach, we observed a correlation between β-catenin proteomic levels and malignant cell states, characterized immune cell phenotypes in lymphoid aggregates, and identified discrepancies between RNA and protein levels for key checkpoint molecules (PD-L1, TIM-3, and IDO). Overall, this technical framework enables robust profiling of functional cellular states, spatial mapping of chemokine expression, and more sensitive detection of clinically relevant targets missing in single-modality methods. Significance: We present an integrated spatial multi-omics workflow that enables direct comparison of transcript and protein levels within a single FFPE section. This enables detailed profiling of the cellular microenvironment, along with relevant protein biomarkers, for clinical decision-making.
Abstract Late distant recurrence (DR) remains a persistent risk in hormone receptor–positive (HR+) early breast cancer after completion of 5 years of endocrine therapy (ET). We developed and validated a multimodal artificial intelligence (AI) model to improve long-term risk stratification and to explore heterogeneity in benefit from extended letrozole therapy (ELT). The deep learning model integrating digitized hematoxylin and eosin whole-slide images with clinicopathologic variables was developed using 2,271 patients from the National Surgical Adjuvant Breast and Bowel Project (NSABP) B-42 trial with five-fold cross-validation and externally validated in 4,300 patients from the TAILORx trial who were disease-free at 5 years from the initial diagnosis. Prognostic performance was evaluated using hazard ratios (HR) and absolute risk differences. Exploratory analyses assessed ELT benefit across model-defined risk groups. In NSABP B-42, the model stratified patients into groups with markedly different outcomes, with a 10-year absolute DR risk difference of 7.95% between high- and low-risk groups [HR, 5.71; 95% confidence interval (CI), 3.5–9.317; P < 0.001]. High-risk patients derived greater absolute benefit from ELT (4.09%) than low-risk patients (0.49%). External validation in the independent TAILORx cohort confirmed prognostic performance, with MI Clarity multimodal–multitask identifying patients with significantly different late DR outcomes (HR, 1.893; 95% CI, 1.413–2.534; P < 0.001). This multimodal AI approach using routine pathology and clinical data enables robust and generalizable stratification of late DR risk in HR+ breast cancer. This scalable strategy may complement existing genomic assays and support more individualized decisions about extended ET. Significance: Late DR is a major cause of death in HR+ breast cancer, and deciding who needs longer hormone therapy is challenging. Current genomic tests are useful but have limitations. This study shows that an AI model using pathology and clinical data identifies patients at high or low risk of late recurrence. High-risk patients may benefit more from extended hormone therapy, offering an accessible way to guide treatment decisions.
