Protective Effect Research Articles

Abstract 6221: Gastric atrophy is associated with increased risk of esophageal squamous cell carcinoma in Lilongwe, Malawi

Background: Esophageal cancer (EC) is the 7th leading cause of cancer worldwide, with nearly 500,000 annual deaths. Esophageal squamous cell carcinoma (ESCC), the most prevalent subtype, has high incidence in East Asia, South and East Africa. Malawi has the highest EC incidence rate, almost exclusively ESCC, with risk factors including tobacco use, hot beverages, fuel source, moldy grain consumption, geophagia, poor oral health, and possibly HIV. H. pylori’s role in ESCC is unclear, with adverse, null, and protective effects across different populations. Gastric atrophy, a potential result of H. pylori infection, may increase ESCC risk by promoting gastrointestinal bacterial overgrowth that exposes esophageal mucosa to carcinogens (e.g. nitrosamines). HIV, common in Malawi, may interfere with H. pylori antibody tests. Limited data, especially from African populations, leaves a gap in understanding the roles of H. pylori and gastric atrophy in ESCC. Methods: We enrolled 300 ESCC cases and 300 controls, matched by age, sex and region, at two Lilongwe, Malawi hospitals (2017-2020) and collected demographic, health, and behavioral data. We measured H. pylori antibodies and gastric atrophy (pepsinogen I (PGI), pepsinogen II (PGII)) via serology. Atrophy was defined as PGI:II <3 or PGI <30μg/L per kit specifications. We adjusted logistic regression models for design variables, known and potential confounders. Results: H. pylori seropositivity was 68% in cases, 80% in controls and significantly associated with lower ESCC risk (aOR 0.54; 95% CI 0.32-0.92). High HIV positivity in cases (21%) may obscure the association due to assay interference. Having a PGI <30μg/L was significantly associated with higher ESCC risk (aOR 4.92; 95% CI 2.04-11.87) while having a PGI:II <3 was marginally associated (aOR 2.14; 95% CI 0.97-4.70). Conclusion: Serologic gastric atrophy was associated with strongly increased ESCC risk. H. pylori's protective effect may be due to measurement interference. Citation Format: Kadiatou Traore, Yingxi Chen, Yukiko Yano, Paul S. Albert, Sung Duk Kim, Sanford M. Dawsey, Shiraz Khan, Christian C. Abnet. Gastric atrophy is associated with increased risk of esophageal squamous cell carcinoma in Lilongwe, Malawi [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 6221.

Exploring the role of gut microbiota in intervertebral disc degeneration: insights from bidirectional Mendelian randomization analysis.

Although previous studies have indicated a potential association between the gut microbiota and intervertebral disc degeneration (IVDD), the precise nature of this relationship remains unclear. The objective of this study is to further explore the potential causal relationship between gut microbiota and IVDD using a bidirectional Mendelian randomization approach, with the aim of identifying potential microbial characteristics associated with IVDD. Using the data from genome-wide association studies (GWAS) involving 412 gut microbiota species and 227,388 controls and 29,508 cases of IVDD. Inverse variance weighted (IVW) was used as the primary Mendelian randomization (MR) analysis, complemented by weighted median, MR-Egger regression, weighted mode and simple mode methods. Extensive sensitivity analyses were performed to confirm the robustness of the results and to assess heterogeneity and horizontal pleiotropy. This study revealed a positive genetic predisposition between 6 types of gut microbiota and IVDD through the IVW method, indicating that increased levels of these microbiota may lead to a higher risk of IVDD. Conversely, 6 types of gut microbiota were found to have negative effects on IVDD, suggesting that increased levels of these microbiota may have a protective effect against IVDD. Reverse MR analysis results revealed such possibilities as 1 positive and 5 negative causal relationships between IVDD and gut microbiota. The results of Cochran's Q test, MR-Egger regression, and MR-PRESSO analysis from the bidirectional Mendelian randomization all yielded p-values greater than 0.05, indicating that there is no significant heterogeneity or pleiotropy in the genetic effect analysis between gut microbiota and IVDD. We used a bidirectional Mendelian randomization approach to identify various gut microbiota associated with IVDD. Our findings lay the foundation for further exploration of the pathogenesis and treatment strategies of gut microbiota and IVDD, and provide new possibilities for research on biomarkers of IVDD-related metabolic microbiota.

Exosomes secreted from adipose-derived stem cells inhibit M1 macrophage polarization ameliorate chronic endometritis by regulating SIRT2/NLRP3.

Chronic endometritis (CE) is a key factor in adverse pregnancy outcomes such as miscarriage and infertility. Macrophages are an important immune cell type that secrete pro-inflammatory and anti-inflammatory cytokines that are essential for maintaining endometrial function. This study aimed to investigate the key mechanisms by which exosomes derived from adipose-derived mesenchymal stem cells (ADSCs) regulate macrophage polarization through the sirtuin 2 (SIRT2)/NOD-like receptor pyrin containing 3 (NLRP3) axis and exert a protective effect on CE. Exosomes were obtained from ADSCs (ADSCs-exo) using the classical ultracentrifugation method and characterized using transmission electron microscopy, nanoparticle tracking analysis, and western blotting. ADSCs-exo protective effects on CE mice and RAW 264.7 cells and its related molecular mechanisms were investigated using real-time quantitative polymerase chain reaction, western blotting, enzyme-linked immunosorbent assay, flow cytometry, immunofluorescence, immunoprecipitation, hematoxylin and eosin staining, and immunohistochemistry. ADSCs-exo significantly inhibited M1 macrophage polarization, as evidenced by a 54% reduction in tumor necrosis factor alfa (TNF-α), a 46% reduction in interleukin 1β (IL-1β), and a 36% reduction in interleukin 6 (IL-6) levels in LPS-induced RAW264.7 cells. In vivo, ADSCs-exo treatment reduced the expression of TNF-α by 50%, IL-1β by 58%, and IL-6 by 49% in the uterine tissues of CE mice. Moreover, ADSCs-exo upregulated the expression of SIRT2, promoted the deacetylation modification of NLRP3 to inhibit NLRP3 inflammasome activation, and further suppressed M1 macrophage polarization. However, these trends were reversed after SIRT2 silencing. Our experimental results demonstrate that ADSCs-exo alleviate CE by regulating the SIRT2/NLRP3 axis to inhibit M1 macrophage polarization. This provides a potential theoretical basis for the therapeutic role of stem cells in CE.

Protective Effect of Enterococcus faecium Against Alcohol-Induced Acute Liver Injury Via Extracellular Vesicles in Rats.

The beneficial effects of probiotics on alcohol-induced liver injury have been studied, but the mechanisms by which Enterococcus regulates liver function are still under investigation. In this study, we examined Enterococcus faecium (Efm) and E. faecium-derived extracellular vesicles (EfmEVs) to provide a protective effect against ethanol-induced liver injury in rats. We evaluated the impact of EfmEVs on liver histological lesions, antioxidative function, alanine aminotransferase (ALT), aspartate aminotransferase (AST) activities, and serum ALT, AST, blood alcohol concentration. The results demonstrated that pretreatment with Efm significantly ameliorated ethanol-induced liver injury. Efm pretreatment mitigated the decline in ethanol-induced liver antioxidant indicators (malondialdehyde, superoxide dismutase, and glutathione peroxidase. Additionally, Efm pretreatment significantly reduced ethanol-induced ALT activities in the liver and serum, potentially by lowering blood ethanol concentration. Further, functional studies on three bioactive components (inactivated Efm, EfmEVs, and EVs-free supernatants) from the bacterial culture revealed that EVs were primarily responsible for the liver-protective effect. Moreover, EVs secretion contributed to the overall liver-protective effect of Efm. In summary, EfmEVs mediated the protective effect of Efm against ethanol-induced liver injury, potentially by improving antioxidative function and lowering blood ethanol concentration. These findings suggest that EfmEVs could serve as a potential antioxidative strategy to alleviate alcohol-induced acute liver injury.

Ecdysteroid-enriched fraction of Cyathula officinalis suppresses synovial proliferation and inflammation to ameliorate RA by inhibiting the AKT/PI3K/mTOR signaling pathway.

This study investigated the chemical compositions isolated from Cyathula officinalis (CES) and elucidated its anti-RA effects and potential mechanisms. In vitro results revealed that CES suppressed interleukin (IL)-1β-stimulated proliferation, migration, and invasion of MH7A cells in a concentration-dependent manner. In vivo results indicated that CES could reduce the arthritis score, paw swelling, and improve histopathological deterioration. The levels of inflammatory factors, matrix metalloproteinases, and proteins expressed in rat synovial tissue were suppressed after treatment with CES. These findings illustrate that CES is involved in the PI3K/AKT/mTOR/SREBP1/SCD-1/GPX4 axis-mediated ferroptosis and anti-inflammation, exerting its protective effects against the progression of RA.

Abstract 1334: FLASH proton radiotherapy mitigates corneal damage and stimulates wound healing in mice

Eye cancers are rare but highly aggressive, with significant risk to vision and quality of life. Worldwide, thousands are affected by ocular cancers, which are challenging to treat due to their proximity to sensitive structures. Proton radiotherapy (PRT) is the current gold standard for intraocular tumors due to its precision in limiting RT exposure to surrounding healthy tissue. However, the delicate location restricts the PRT dose that can be safely administered without risking damage to essential orbital structures. FLASH (FR), a novel approach delivering PRT at ultrahigh dose rates (>40 Gy/s) compared to standard dose rate PRT (SR, <0.003 Gy/s), has shown reduced toxicity in normal tissues like the brain and skin without compromising tumor control. However, its protective effects on eye structures remain unexplored. This study aims to investigate the potential of FR in preserving ocular tissues in a mouse model. Female C57BL/6J mice received 24 Gy of either FR or SR or remained untreated (control) using a "shoot-through" approach, in which both eyes received the same dose. Retinal and corneal damage were monitored using optical coherence tomography and confocal scanning laser ophthalmoscopy, with retinal vascular permeability evaluated by fluorescein angiography and retinal function by electroretinography (ERG). Corneal wound healing was assessed using corneal micro puncture. Evaluations occurred at one week, one, three, and five months post-PRT. Mice treated with SR showed significant corneal thickening and ulceration, leading to visual impairment, while those treated with FR did not. SR mice also showed declining ERG responses, suggesting diminished response to light stimuli, while FR mice demonstrated significantly less ERG abnormalities. Additionally, mice treated with SR showed over 55% reduction in eye surface area compared to both FR and control mice. Both SR and FR reduced retinal thickness, with no significant difference between the PRT treatment groups. Fluoresceine angiography showed no leakage in the retinal microvasculature. To further characterize corneal repair following PRT, we performed a corneal wound healing assay. Notably, FR demonstrated an enhanced wound healing response, with faster recovery of corneal incisions compared to both SR and control mice. This study presents a novel model for evaluating PRT-induced ocular damage, highlighting FR’s potential to protect healthy tissues. FR was particularly effective in preserving corneal integrity and promoting wound healing, which are crucial for maintaining both visual function and cosmetic outcomes. Ongoing studies aim to quantify the preservation of stem cells in the corneoscleral junction as a potential mechanism for the observed tissue-sparing effects. Citation Format: Elias El Hoyek, Uri Amit, Brent A. Bell, Vivian Lee, Joshua L. Dunaief, Costantinos Koumenis. FLASH proton radiotherapy mitigates corneal damage and stimulates wound healing in mice [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 1334.

Abstract 2844: A cytosolic DNA/Ca2+-dependent signaling pathway for genome protection during replication stress

Abstract Protecting the replication fork structure, especially under replication stress, is essential for maintaining genome integrity and stability. Defects in fork protection, often linked to mutations in tumor suppressor genes like BRCA1, BRCA2 and FANCD2, can lead to genomic instability and consequently, cancer progression. Such defects may also influence cancer progression by altering the tumor microenvironment through the generation of cytosolic DNA fragments that activate the innate immune response. Intriguingly, our recent findings indicate that cytosolic DNA, produced after replication stress, unexpectedly also supports replication fork protection. This protective effect is mediated through a complex signaling pathway that involves the activation of the cGAS/cGAMP/STING axis, Ca2+ release from the endoplasmic reticulum via the TRPV2 ion channel, activation of CaMKK2 and AMPK protein kinases, and the inhibition of the EXO1 nuclease. Disruption of this pathway results in excessive fork processing, chromosomal instability and heightend sensitivity to replication stress inducers. The newly uncovered links between innate immune factors, intracellular Ca2+ and genome maintenance in this novel signaling pathway may open new avenues for cancer treatment. (Li et al., Mol Cell 2019, 74:1123-1137; Li, Kong et al., Mol Cell 2023, 85:556-573; Kong, Cheng et al., Nat Commun 2024, 15:4609) Citation Format: Zhongsheng You. A cytosolic DNA/Ca2+-dependent signaling pathway for genome protection during replication stress [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 2844.

Abstract 1348: Investigation of sex disparity in hepatocarcinogenesis using a hydrodynamic tail vein injection mouse model

Abstract The incidence of hepatocellular carcinoma (HCC), the major form of liver cancer, is two to three times higher in men than in women. Understanding the gender disparity in HCC incidence is challenging due to HCC development being multifactorial, including variations in risk factors, behaviors, and hormones. The liver is a sexually dimorphic organ, and estrogen has been proven to have a protective effect against the development of HCC, although the mechanism is not fully understood. We used the hydrodynamic tail vein injection (HTVI) mouse model of HCC to investigate potential mechanisms underlying the sex disparity of HCC incidence. This model eliminates social and behavioral aspects of gender disparity and allows us to determine if there is biological sex disparity and what role estrogen and androgens play in tumor development. In our preliminary study, the expression vectors of oncogenes, ΔN90-β-catenin and c-MET, to be integrated into the hepatocyte genome via transposons, were co-injected with a Sleeping Beauty (SB) transposase to induce HCC in FVB/NJ mice. Based on plasma alpha-fetoprotein (AFP), an HCC blood biomarker, male FVB/NJ mice had a significantly higher mean AFP trend in comparison to females, indicating that the males developed liver tumors at a faster rate than females. ΔN90-β-catenin and c-MET gene copy number were confirmed to be equally transfected and integrated in both male and female FVB/NJ tumors by quantitative PCR (qPCR). The expression of ΔN90-β-catenin and c-MET transgenes was also shown to be similar between male and female mouse HCC tumors by RT-qPCR and RNA-seq. We plan to repeat the HTVI of ΔN90-β-cat, c-MET, and SB to validate that the males will develop liver tumors at a faster rate than female mice. We will also Western Blot to determine if there is sex disparity in the abundance of the transfected oncogene proteins. In addition, we will compare the abundance of various types of immune cells present in the tumor immune microenvironment (TIME) to interpret potential sex-dependent immune cell differences in the tumor liver. Our research has the potential to uncover mechanisms underlying the sex-dependent disparity in HCC development and progression and the contribution of immune cells to the sex disparity. Citation Format: Araceli Selena Bernal, Morgana McLaughlin, Ankur Tiwari, Brittany Barre, Francisco Cigarroa, LuZhe Sun. Investigation of sex disparity in hepatocarcinogenesis using a hydrodynamic tail vein injection mouse model [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 1348.

Abstract 3562: Associations between premature aging and cancer among middle-class Black and White women in the US: A case-control NHANES analysis

Abstract Although overall cancer mortality rates have decreased, Black women in the U.S. continue to have higher mortality rates from cancer than their White counterparts. Recent studies reveal that both genetic and environmental factors combine to contribute to these racial disparities in cancer, and may be modulated by differences in tumor biology. With cancer being an aging-related disease, it has previously been hypothesized that racial disparities in cancer outcomes are exacerbated by prolonged exposures to social and economic stressors that lead to premature aging. Our study aims to explore these effects on cancer status among U.S. Black and White women with annual household incomes of > $75K/year. Using data from the National Health and Nutrition Examination Survey (NHANES) from 1999 to 2021, we quantified social and economic stressors using three factors: (1) allostatic load (AL) score to predict confounding stress factors on an individual’s health, (2) telomere length to predict premature aging, and (3) validated epigenetic aging markers from NHANES methylation data. We calculated the total AL score by summing the individual components for a score ranging from 0 to 9. Participants with high AL have a score ≥3. Mean telomere length was measured continuously. All aging metrics represent different aspects of accelerated epigenetic aging (Horvath Age and Hannum Age), phenotypic age (PhenoAge), pacing of aging (DunedinPoAm), and predicted age of mortality (GrimAge2Mort). Adjusted regression models were used to identify associations between these metrics, any cancer status, and race. We restricted our study sample to women with annual household incomes >$75K/year, resulting in a final NHANES cohort of 3, 690 women (cancer cases, n=302; controls, n=3, 388). In this cross-sectional study of Black (n=850) and White (n=2, 840) women, increasing allostatic load was associated with decreased odds of cancer case status overall (odds ratio (OR)=0.89, 95%CI=0.81, 0.98, p=0.02) and specifically for White women (OR=0.89, 95%CI=0.80, 0.99, p=0.03). Increasing Horvath Age was associated with a 20% increased odds of cancer case status overall (OR=1.20, 95%CI=1.01, 1.43, p=0.04) and significantly for White women (OR=1.36, 95%CI=1.11, 1.166, p=0.003), but not Black women. We also found that AL and telomere length were both associated with several accelerated aging metrics in cases and controls. Our results suggest that increasing allostatic load had a protective effect in this cohort of high-earning women, specifically among White women. Increasing epigenetic aging metrics correlated to odds of a cancer diagnosis overall and with race specific implications. Some differences by race may be attributed to the low sampling of Black women in this cohort with corresponding telomere and methylation data, emphasizing the need for future epigenetic studies to include more Black populations. Citation Format: Kayla S. Ingram, Brittany Jenkins-Lord, Kassandra L. Alcaraz, Avonne E. Connor. Associations between premature aging and cancer among middle-class Black and White women in the US: A case-control NHANES analysis [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 3562.

Peri-traumatic consumption of classic psychedelics is associated with lower anxiety and post-traumatic responses 3 weeks after exposure.

Emerging evidence indicates the therapeutic potential of psychedelic compounds for post-traumatic stress, yet the mechanisms mediating their effects remain unclear. Delineating the effect of psychedelics on traumatic memory formation could shed light on target therapeutic mechanisms. Here, we report on 343 adult survivors of a single, large-scale terrorist attack taking place during a festival in which different psychedelic compounds were consumed, in whom levels of anxiety and post-traumatic symptoms were assessed 3 weeks following the attack. Findings indicated that those who were under the influence of classic psychedelics during the attack reported significantly lower levels of anxiety and post-traumatic responses compared to those who were under the influence of 3,4-methylenedioxymethamphetamine and those who consumed no psychedelics. Furthermore, the protective effects of classic psychedelics for post-traumatic responses manifested more strongly among participants who did not consume additional recreational substances alongside psychedelics. These findings suggest that pharmacologic targets of classic psychedelics may modulate the formation of enduring trauma memories and confer a protective effect against the development of post-traumatic stress and anxiety responses.

MOB1 deletion in murine mature adipocytes ameliorates obesity and diabetes

There is currently a global epidemic of obesity and obesity-related diseases such as type 2 diabetes due to decreased physical activity, excessive food intake, and/or genetic predisposition. The Hippo-YAP1 pathway has attracted attention as a potential therapeutic target because YAP1/TAZ activation in murine immature adipocytes in vitro suppresses their differentiation by inhibiting PPARγ activity. However, the role of YAP1 activation in mature adipocytes in vivo remains unclear. MOB1, whose expression is increased in obesity, is the hub of the Hippo core molecule complex and negatively regulates YAP1/TAZ activation. Therefore, we generated aMob1DKO mutant mice, which feature deficiency of Mob1a/b specifically in mature adipocytes. Compared to controls, aMob1DKO mice subjected to a high-fat diet showed beneficial changes consistent with resistance to diet-induced obesity. The mutants exhibited increases in basal lipolysis, “beiging,” and energy expenditure, as well as suppression of ROS production and inflammation in white adipose tissue. Insulin sensitivity and glucose tolerance were improved, and ectopic fat accumulation was reduced. Most of these changes were dependent on the YAP1 activation observed in mature white adipose tissue of aMob1DKO mice. FGF21, which improves lipid metabolism, was upregulated directly via YAP1 activation, and many of the phenotypes seen in aMob1DKO mice were also dependent on FGF21. Thus, the aMob1DKO mouse is an interesting model for the study of the metabolic effects of diet-induced obesity and protection against diabetes. Our work suggests that a YAP1-FGF21 axis exists in adipocytes that may be a potential therapeutic target for obesity.

Green synthesis of silver nanoparticles using guava leaves: an effective strategy to control chilli fruit rot disease

Abstract Background Anthracnose, caused by Colletotrichum capsici, is a significant fungal disease affecting chilli crops, leading to yield losses of 10–25%. Traditional control methods, primarily chemical fungicides, not only pose risks to the environment and soil health but also threaten public safety. In contrast, nanotechnology presents a promising eco-friendly alternative, leveraging the unique properties of nanoparticles, such as their small size and high surface-to-volume ratio, to effectively manage fungal infections with minimal environmental impact. Results This study investigates the synthesis, characterization, and antifungal activity of silver nanoparticles (Ag-NPs) synthesized from guava leaf extract against chilli fruit rot. UV-Vis spectroscopy confirmed the synthesis of Ag-NPs with a peak absorption at 431 nm. X-ray diffraction (XRD) analysis revealed a crystalline structure with an average particle size of 42.5 nm, while scanning electron microscopy (SEM) showed spherical nanoparticles with sizes ranging from 30.5 nm to 50.3 nm across different samples. Fourier transform infrared spectroscopy (FTIR) identified functional groups involved in silver ion reduction. Zeta size analysis confirmed particle sizes of 500.1 nm, 1.0 nm, 62.4 nm, 262.8 nm, and 178.8 nm for samples S1 through S5, respectively. In antifungal assays, S1 at 50 ppm exhibited the highest mycelial growth inhibition (47.9%), with significant protective (87%) and curative (93%) effects. Additionally, in in-vitro leaflet assays, S1 demonstrated 86% inhibition of C. capsici at 50 ppm, highlighting its potential as an effective agent for managing chilli fruit rot. Conclusions This study presents a rapid, eco-friendly method for synthesizing Ag-NPs using guava leaf extract, showing their potential in managing chilli fruit rot caused by C. capsici. The results highlight their effectiveness in both protective and curative applications, offering a sustainable alternative to chemical fungicides. Future research should focus on scaling up the synthesis process for industrial applications, exploring the long-term environmental impact, and assessing the broader applicability of Ag-NPs in managing other phytopathogenic diseases across various crops. Summary Silver nanoparticles (Ag-NPs) synthesized from guava leaf extract effectively inhibit Colletotrichum capsici, with 86% antifungal activity at 50 ppm. This study highlights an eco-friendly, rapid synthesis method for Ag-NPs as a promising alternative to chemical fungicides in managing chilli fruit rot disease. Clinical trial number Not applicable. Graphical Abstract

Abstract 2316: Young-onset cancer incidence trends: Cohort effects on onset versus period effects on diagnosis

Background: The disproportionate increase in the diagnosis of certain cancers among younger individuals is generating intense concern. Many studies attribute the increase in so-called “young-onset” cancer to an etiologic cause but questions have also arisen about the role of earlier diagnosis. Methods: We simulate incidence trends from a natural history model that includes healthy, preclinical, and clinical disease states, where the clinical state represents diagnosis. We superimpose birth cohort effects on the rate of disease onset and period effects on the rate of transitioning to the clinical state. We examine patterns of relative incidence by age group and five-year calendar interval from 2000-2019 for six “young-onset” cancers (colon, rectum, female breast, gastric, pancreas, kidney), and identify cohort and period effects that lead to matching relative incidence patterns. Results: Two types of effects are broadly consistent with the trends in younger individuals: (1) A birth-cohort effect on disease onset that begins around 1970 and becomes more pronounced with later birth years, or (2) A progressive reduction over time in the duration of preclinical disease. An earlier, protective birth cohort effect is consistent with recent trends in incidence in older individuals that are the reverse of those in younger individuals for colon, rectal, and gastric cancers. Discussion: A disease model provides clues about the possible drivers of cancer incidence trends, suggests constraints on the patterns of exposures that might be implicated etiologically, and indicates that consideration of the role of diagnostic changes is warranted alongside potential etiologic explanations. Citation Format: Lukas Owens, Jonathan Shuhendler, Marc D. Ryser, Ruth D. Etzioni. Young-onset cancer incidence trends: Cohort effects on onset versus period effects on diagnosis [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 2316.

Investigating the causal association between coffee and migraine using Mendelian randomization analysis

ABSTRACT Background: Migraine is a chronic neurological disorder influenced by various risk factors, including inadequate medication, anxiety, sleep disorders, depression, and stress-inducing events. Coffee is well-known for its diverse bioactive properties and may help relieve acute migraine symptoms; however, stopping long-term coffee consumption can unexpectedly trigger migraines. While some migraine sufferers view caffeine as a potential trigger, research suggests it may also have preventive effects. The complex physiological and pharmacological mechanisms behind coffee and its active components are not fully understood. Objective: This study aims to clarify the relationship between coffee consumption and its components and the risk of developing migraines. Methods: We conducted a Mendelian randomization (MR) analysis, using genetic variants as proxies for exposure to reduce confounding factors and biases. Results: Our results show a significant inverse relationship between coffee intake and migraine risk, consistent with previous epidemiological studies that indicate coffee may protect against headache disorders. The MR analysis also found that 7-methylxanthine is linked to a lower risk of migraines, while caffeic acid sulfate is associated with a higher risk. Sensitivity analyses showed no heterogeneity in SNP selection for all components, except for trigonelline, and both MR-Egger and MR-PRESSO tests did not indicate horizontal pleiotropy or outliers. Our research highlights the significant impact of coffee and its components on migraine risk, providing valuable dietary recommendations. The protective effect of coffee may be closely related to adenosine receptor antagonism, which warrants further investigation to better understand the underlying mechanisms.

Abstract 3315: Unveiling a protective role of B-cells in classical Hodgkin lymphoma progression through gene expression signatures

Background: Classical Hodgkin Lymphoma (cHL) is considered highly curable, but identifying patients at risk of relapse or progression after initial treatment remains a challenge. Disease progression may involve biological factors not accounted for by current prognostic models, which could potentially be revealed by molecular analysis. In this study, we performed comprehensive gene expression profiling to identify molecular markers that could help to develop new prognostic tools in cHL patients. Patients and Methods: We conducted a retrospective review of local clinical records to identify patients with confirmed cHL diagnosed between 2004 and 2019, aged 18-65, at any disease stage, and treated with systemic chemotherapy. Gene expression analysis using nCounter Nanostring Technology with the PanCancer Immune Profiling Panel was performed on baseline diagnostic biopsies. Genomic data were correlated with clinical, laboratory, and radiomic information. Progression-Free Survival (PFS) was the primary endpoint. Immunohistochemistry was used for validation. Results: We identified 185 patients with classical Hodgkin lymphoma (cHL), of whom 155 had available formalin-fixed paraffin-embedded (FFPE) tissue samples. Of these, 32% were aged over 45 years, 46% had stage III-IV disease, and 10% presented with bulky disease. After a median follow-up of 72 months (range, 6-171 months), 31 PFS events occurred, with a 4-year PFS rate of 80.4% (95% CI 74.1-87.3). Through Cox Proportional Hazards modeling, we identified 66 genes that were significantly associated with PFS (p<0.05). Of these, 41 genes were positively associated with better PFS, suggesting a protective effect, while 25 genes were linked to a reduced probability of survival. Through correlation analysis and gene ontology, we were able to identify a 7-gene signature associated with B-cell signaling pathways. An unsupervised analysis on the entire cohort based on the expression of the identified B cell signature has allowed us to define two main patient clusters: low- B cell and high- B cell. The former cluster displayed a significant increase in the incidence of clinical events (p=0.03) and worse PFS compared to the high B-cell cluster (p=0.007). Additionally, elevated expression of PAX5, a key regulator of B-cell differentiation, was significantly associated with better PFS, with a 4-year PFS rate of 90% (95% CI 82.7-97.3) compared to 71% (95% CI 61.3-82.4) in patients with lower expression levels. Immunohistochemical evaluation of PAX5 in the tumor microenvironment further supported its potential as a prognostic marker. Conclusion: These results suggest that gene expression analysis may assist in the early detection of disease relapse and highlight the immune-modulatory role of B-cells in cHL progression. Citation Format: Stefano Pozzi, Francesca Reggiani, Tanja Lazic, Nizzoli Maria Elena, Alberto Bavieri, Benedetta Donati, Durmo Rexhep, Riccardo Valli, Attilio Gennaro, Cristian Ascione, Alessia Ruffini, Annibale Versari, Francesco Merli, Massimo Dominici, Alessia Ciarrocchi, Stefano Luminari. Unveiling a protective role of B-cells in classical Hodgkin lymphoma progression through gene expression signatures [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 3315.

Abstract 6266: Spatial profiling of prognostic differences in the tumor microenvironment between primary and metastatic ovarian cancer using multiplexed protein imaging

Abstract Background: The tumor microenvironment (TME) plays a pivotal role in ovarian cancer (OC) progression and prognosis, while also contributing to biological differences between primary and metastatic tumors. This study examines OC TMEs to uncover associations with clinical prognosis and tumor biology by analyzing paired primary and metastatic tumors from the same patients. Methods: This study focused on 57 stage-IIIC/IV OC patients treated at the Mayo Clinic, all of whom underwent surgical debulking and adjuvant therapies. All patients provided informed consent, and the study received IRB approval. A tissue microarray (TMA) was specifically designed and constructed, containing paired primary and metastatic tissue cores from the same patients. 147 tissue cores from the constructed TMA were profiled using a 35-antibody panel, with 17 antibodies for cell phenotyping and 18 for functional state characterization. Profiling was performed using Cell DIVE, a multiplexed imaging solution at the single-cell level developed by Leica Microsystems. After cell segmentations and classifications, detailed spatial niche and regional analyses were performed at the single-cell level. Immune cell populations, as well as their TME distributions, were characterized to conduct prognostic associations and contrast primary and metastasis differences. Results: A total of 1,292,988 total cells (after excluding low-density signals and ambiguous classifications) were analyzed, with a median of 8,033 cells per core. Using spatial neighborhood analysis, 12 distinct spatial niches were identified with specific cell types and/or unique functional states. Among these, a macrophage- and fibroblast-enriched niche at the tumor-stroma interface was associated with improved overall survival (p=0.008). Another rare Treg-enriched niche, along with other immune cells, demonstrated protective effects, particularly in patients with residual macroscopic tumors after debulking surgery (p=0.05), while no significant effects were observed in those without visible residual disease (p=0.8). When comparing primary and metastatic tumors, metastatic lesions exhibited significantly increased tumor mass (p=2e-4) and a notable reduction in helper T cells (p=0.04) in tumor areas, as well as significantly increased fibroblast cells in tumor-stroma interfacing regions (p=0.002), reflecting distinct immunosuppressive and tumor-promoting shifts. Conclusion: This study reveals spatial and cellular differences in primary and metastatic OC TMEs, distribution differences linked to prognosis and surgical outcomes. These findings emphasize the potential of spatially resolved TME profiling to refine adjuvant and neoadjuvant therapeutic strategies. Citation Format: {Authors}. {Abstract title} [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 6266.

Abstract 7039: Synergistic antitumor activity of the combined PARP1 inhibition and alkylating DNA damage in malignant rhabdoid tumors

Abstract Background: Malignant rhabdoid tumors (MRTs) are among the most aggressive and treatment-resistant malignancies affecting infants, originating in the kidney, brain, liver, and soft tissues. The 5-year event-free survival rate for these cancers is a mere 20%. In nearly all cases of MRT, the SMARCB1 gene (occasionally SMARCA4) - a pivotal component of the SWI/SNF chromatin remodeling complex - is homozygously deleted, although the precise etiology of these tumors remains unknown. While young patients with localized MRT generally show improved outcomes, especially those who are older and have early-stage disease, the overall prognosis remains poor despite optimal standard treatments. This highlights the urgent need for more effective treatment strategies. PARP1 is a widely targeted molecule in cancer treatment and, beyond its role in DNA repair, it participates in transcriptional regulation by recruiting chromatin remodeling complexes to modulate DNA accessibility for RNA polymerases. Methods: We evaluated the antitumor activity of the PARP1 inhibitor talazoparib (TLZ) in combination with the DNA alkylating agent temozolomide (TMZ) across six MRT xenograft models in CB-17 scid mice. The dosing schedule for this combination followed a previously reported protocol (iScience 2021; 25(2):103725). Time-to-event was defined as 4-fold increase in tumor xenograft volume from the treatment initiation date. Event-free survival (EFS) between treated and control groups was analyzed using Kaplan-Meier method. Objective response categories including partial (PR), complete (CR), and maintained complete responses (MCR), were determined based on previously established criteria (Ped Blood Cancer 2007;49:928). To widen the therapeutic window of the drug combination, we employed PEGylated TLZ (PEG-TLZ), which has been reported to reduce systemic toxicity through slow drug release. Results: Remarkably, our findings indicate that five out of six MRT xenografts exhibited objective response to PEG-TLZ+TMZ therapy. Notably, SMARCB1 loss was found to confer a protective effect, correlating with elevated expression of DNA damage and repair proteins in SMARCB1-deficient MRT cells. Additionally, MGMT emerged as a potential biomarker predictive of in vivo MRT response to PEG-TLZ+TMZ treatment. In addition, our analysis revealed alterations in signaling pathways associated with the observed antitumor activity. Conclusions: This study presents a novel and efficacious therapeutic approach for MRT, along with a promising candidate biomarker for predicting tumor response. Citation Format: Elena Mironova, Sebastian Molinas, Vanessa Del Pozo, Abhik Bandyopadhyay, Zhao Lai, Dias Kurmashev, Eric L. Schneider, Daniel V. Santi, Yidong Chen, Raushan Kurmasheva. Synergistic antitumor activity of the combined PARP1 inhibition and alkylating DNA damage in malignant rhabdoid tumors [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 7039.

Abstract 6231: Associations of serum concentrations of α- and γ-tocopherols with cancer mortality

Background: While tocopherols’ antioxidant and anti-inflammatory properties suggest their potential anti-cancer effects, little is known about the association between circulating tocopherols and mortality outcomes, particularly among cancer survivors. This study aimed to evaluate whether serum concentrations of α- and γ-tocopherols were associated with the risk of death from all causes, cancer, and cardiovascular disease (CVD). Methods: We incorporated 16,696 US adults aged ≥20 (1,466 cancer survivors and 15,230 cancer-free individuals) from the National Health and Nutrition Examination Survey, 1999-2006. Serum α- and γ-tocopherols were quantified by high-performance liquid chromatography and categorized into quartiles. Mortality and cause of death were ascertained by linkage to National Death Index records through December 31, 2019 (median follow-up of 16.1 years). Cox proportional hazards regression was used to estimate hazard ratios (HRs) and 95% CIs for all-cause, cancer, and CVD mortality after adjustment for comprehensive covariates, including serum cholesterol levels, sociodemographics, lifestyle (e.g., diet, physical activity), and comorbidities. Restricted cubic splines were used to evaluate the potential non-linearity of the associations. Results: Cancer survivors in the highest quartile of serum α-tocopherol demonstrated a 50% lower risk of cancer mortality compared to those in the lowest quartile (HR=0.50, 95% CI:0.32-0.80, p-trend=0.009). Similar protective associations were observed in cancer-free individuals, with those in the highest quartile of serum α-tocopherol showing a 31% reduction in cancer mortality risk (HR=0.69, 95% CI: 0.46-1.02, p-trend=0.05). The association between α-tocopherol and cancer mortality was non-linear (p<0.001), exhibiting the strongest protective effects at concentrations ranging from 35 μmol/L to 50 μmol/L. All-cause mortality risk was also lower among those with higher α-tocopherol in cancer survivors (HR=0.75, 95% CI: 0.54-1.03, p-trend=0.02) and cancer-free individuals (HR=0.86, 95% CI: 0.74-1.00, p-trend=0.03). No significant associations were found for γ-tocopherols. Conclusions: In this nationally representative sample of US adults, higher circulating concentrations of α-tocopherol were associated with a reduced risk of cancer mortality among both cancer survivors and cancer-free individuals. Our findings indicate the potential health benefits of circulating α-tocopherol in reducing preventable deaths and provide population-based evidence for the role of α-tocopherol in cancer survivorship care. Citation Format: Yoo-Min Koh, Jae Jeong Yang, Young-Rock Hong, Shama Karanth, Dejana Braithwaite, Hyung-Suk Yoon. Associations of serum concentrations of α- and γ-tocopherols with cancer mortality [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 6231.

Abstract 4696: Targeting CSF1R+ macrophages to synergize immune checkpoint therapy in SCCA-mediated radioresistant cervical cancer

Abstract Tumor-associated factors released by cancer cells can promote immune suppression and tolerance to radiation or immune therapy. Squamous cell carcinoma antigen (SCCA) is a tumor-associated antigen released by cancer cells. In cervical cancer, elevated pre-treatment SCCA in the serum is seen in 28-88% of patients and these patients are more likely to experience treatment resistance and recurrence. The poor prognosis for patients with elevated SCCA shows the need for improve treatment strategies. Here, we aim to understand the mechanism of action of circulating SCCA to tumor progression and response to radiotherapy. Using sequencing analysis of paired pre- versus on-treatment biopsies, we found that patients with elevated serum SCCA had increased macrophages expressing immunosuppressive features and T-cell immune checkpoints, inversely correlated with inflammatory signaling and radiation-induced interferon-stimulated gene expression. The supplement of SCCA protein (rSCCA) to the media during in vitro macrophage differentiation promotes anti-inflammatory cytokines and gene expression. Peritoneal macrophages isolated from mice treated with intra-peritoneal rSCCA expressed high M2-associated markers and functional inhibition of T cell activation. Moreover, we found that mouse tumors engineered to express SCCA with a signal peptide promoting extracellular secretion displayed increased CSF1R+ TAM infiltration and decreased T cell activation, associated with reduced response to combined radiation and anti-PDL1 treatment. Giving the findings, we hypothesized that targeting TAMs in tumors with elevated SCCA is imperative. We further demonstrated that tumor-bearing mice showed improved response to radiation and immune checkpoint therapy when receiving concurrent CSF1R blockade, abolishing the protective effect of high circulating SCCA. Our results suggest that blockade of CSF1R signaling or other myeloid-targeting therapeutics will optimize response to radiation and PD-L1 blockade in cervical cancer, particularly those with high circulating SCCA. Citation Format: Liyun Chen, Eric Liu, Victoria Shi, Marlene L. Campos Guerrero, Jasmin Yang, Abhay Singh, Matthew Inkman, Jin Zhang, Julie Schwarz, Stephanie Markovina. Targeting CSF1R+ macrophages to synergize immune checkpoint therapy in SCCA-mediated radioresistant cervical cancer [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 4696.

Abstract 2042: ROS1 single nucleotide variants predict favorable survival outcomes on immunotherapy regimens in non-small cell lung cancer

Abstract Background: Only a fraction of non-small cell lung cancer patients who receive immune checkpoint inhibitor (ICI) treatment will respond, largely because current FDA-approved biomarkers still leave room for improvement to identify more responders. We hypothesized that modeling the time-dependent interaction between treatment and tumor genetics may reveal novel associations. We further hypothesized that real-world data (RWD), as opposed to clinical trial data, may contain these associations, as RWD possesses a potentially more diverse patient population and time-to-event treatment history across the diverse regimens used in routine clinical practice. Methods: We examined the relation between tumor genetics, treatment, and real-world progression-free survival (PFS) in two RWD cohorts (Tempus discovery cohort, n = 1458; Dana Farber Cancer Institute Profile independent validation cohort, n = 466) by utilizing a Cox proportional hazards model with a time-dependent covariate for whether a patient was receiving an ICI-containing treatment regimen. Specifically, we included an indicator variable that denoted whether an individual was receiving an ICI regimen (=1) or not (=0), and enumerated the time intervals associated with each treatment status. We performed a univariate analysis for each mutated gene from tumor sequencing, and included a gene-treatment interaction term to permit identification of genes that were predictive (i.e. specific to a treatment) rather than prognostic. Our model included additional covariates to control for known predictive (tumor mutation burden [TMB]) and prognostic factors (age, sex). Results: In the discovery cohort, we identified a significant association between ROS1 single nucleotide variants (SNV; prevalence = 5%) and favorable PFS on ICI therapy (interaction HR = 0.34, 95% CI = 0.18-0.64, p = 7.8e-4, q = 0.08) versus non-ICI treatment. This association was not explained by ROS1 rearrangement or ROS1 TKI therapy. ROS1 SNV associated with worse PFS when not on ICI treatment (marginal HR = 2.3, 95% CI = 1.4-3.6, p = 4e-4, q = 0.044). This ROS1 SNV association was replicated in the validation cohort, which exhibited a favorable interaction with ICI treatment (interaction HR = 0.11, 95% CI = 0.04-0.33, p = 7.46e-5), and poor PFS on non-ICI treatment (marginal HR = 5.39, 95% CI = 2.5-11.6, p = 1.96e-5). ROS1 SNV treatment interaction effect was also not explained by TMB or PD-L1 status. Finally, we observed the ROS1 SNV ICI-specific protective effect was specific to patients ICI-treated in the first line setting. Conclusions: We identified an association between ROS1 SNVs and favorable PFS specific to ICI treatment, independent of common pre-existing predictive biomarkers for ICIs. Because this association was specific to the first line ICI setting, our finding may clarify ICI usage where clinical practice for ICI treatment is still evolving. Citation Format: Yue Wu, Jaclyn LoPiccolo, Sebastian Carrasco Pro, Krishna C. Bulusu, Etai Jacob, Alexander Gusev, Gerald J. Sun. ROS1 single nucleotide variants predict favorable survival outcomes on immunotherapy regimens in non-small cell lung cancer [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 2042.