Nieters, Alexandra

Abstract Introduction: With over 80, 000 new cases of non-Hodgkin's lymphoma (NHL) diagnosed annually, modifiable risk factors remain unclear. Circadian disruption, linked to cancers like breast and prostate, affects immune cells (e.g., natural killer cells and T-helper cells) by altering their trafficking and proliferation. Some studies (PMID: 30566672, 27611440) suggest long sleep may increase overall NHL risk, including a Mendelian randomization (MR) study (PMID: 32895918). However, no research has examined the causal relationship between sleep traits, including long sleep, and NHL risk, by subtypes. We hypothesize that sleep traits are causally associated with NHL subtypes in European populations, testing this using MR with data from the International Lymphoma Epidemiology Consortium (InterLymph). Methods: We conducted two-sample MR using genome-wide association studies (GWAS) summary data for sleep traits and NHL subtypes. Only independent, genome-wide significant (p < 5×10-8) Single Nucleotide Polymorphisms (SNPs) were selected as valid instruments, sourced from the UK Biobank. Sleep traits included chronotype (153 SNPs), insomnia (48 SNPs), sleep duration (78 SNPs), excessive daytime sleepiness (37 SNPs), short sleep (27 SNPs), and long sleep (8 SNPs). NHL subtype data were from InterLymph: follicular lymphoma (6, 508 cases / 64, 183 controls), mantle cell lymphoma (1, 169 cases / 61, 603 controls), Waldenstrom macroglobulinemia/lymphoplasmacytic lymphoma (WM/LPL: 1, 697 cases / 59, 333 controls), and chronic lymphocytic leukemia (8, 522 cases / 67, 653 controls). The primary analysis used inverse-variance weighted (IVW) random-effects, with MR-Egger for pleiotropy adjustment. Results: Using the IVW method, we did not identify any statistically significant causal association between sleep traits and the risk of NHL subtypes, but did yield several intriguing trends. Long sleep (sleep duration >= 9 h per night) showed a marginally increased NHL risk [Odds ratio (OR) range: 1.51 - 247.9] across each subtype. Short sleep (sleep duration < 7 h per night) showed a similar trend [OR range: 1.25 - 1.81], except for WM/LPL, where we noticed a negative trend. The confidence limit for long sleep was imprecise due to the small number of variants. We did not notice any significant directional horizontal pleiotropy or weak instrument bias. Conclusions: We found no conclusive evidence of a relationship between sleep traits and four NHL subtypes. However, we see suggestive trends of long and short sleep associated with higher risk of some NHL subtypes. These findings align with the previous studies, which showed long sleep was associated with higher risk of NHL overall. Future directions include expanding our analysis to additional NHL subtypes, generating polygenic risk scores, and conducting stratified analyses. Citation Format: Pankhil Shah, Brittany Crawford, Anwar Merchant, Brenda Birmann, Angelica Macauda, Michelle A. Hildebrandt, Aaron Norman, Neil E. Caporaso, Meredith Yeager, Michael Dean, Immaculata De Vivo, Lynn Goldin, Nicola J. Camp, Rosalie Griffin, Delphine Casabonne, Federico Canzian, Pelin Unal, Elad Ziv, Catherine R. Marinac, Alexandra Nieters, Stephen J. Chanock, Mitchell J. Machiela, Michael Conry, Charlie Zhong, Hanla A. Park, Simon Cheah, Jonathan N. Hofmann, Elizabeth E. Brown, Celine Vachon, Susan Slager, Sonja Berndt, Sophia S. Wang, Vijai Joseph, James McKay, Henrik Hjalgrim, Lara Sucheston-Campbell, Karl Smith-Byrne, Alyssa Clay-Gilmour. Association between sleep traits and risk of non-Hodgkin's lymphoma subtypes: a mendelian randomization study in the International Lymphoma Epidemiology (InterLymph) Consortium [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 7415.

Introduction: Follicular lymphoma (FL) is the most common indolent subtype of non-Hodgkin lymphoma. Family history is a risk factor. Genome-wide association studies (GWAS) have identified several germline variants, especially within the HLA region, to be associated with an increased risk of FL, but these variants only explain a fraction (16%) of heritability. Here, we present novel findings from GWAS of FL uncovering multiple germline genetic variants contributing to disease susceptibility. Methods: A GWAS was performed comprised of a discovery analysis in 6,508 FL cases and 64,183 controls of European ancestry followed by a replication of the most significant variants (P<5x10-7) in 2,745 FL cases and 782,226 controls from the biobanks of UK, Copenhagen Hospital, FinnGen and Mayo Clinic . At 9,253 FL cases and 846,409 FL free controls imputed genome-wide, this is the largest FL GWAS of European genetic ancestry to date. HLA alleles were imputed, and Class I and Class II signals were analysed at the amino acid level. A transcriptome wide association study (TWAS) was performed using FUSION investigate relationship to expression in GTEx and Young Finns Study in whole blood and cross-tissue . Functional annotations were done using FUMA, F-MAGMA, Open Targets and FORGE. Colocalization was conducted using CoLoC with whole blood gene expression data to understand causal variants. Mendelian randomization (MR) was performed on blood cell traits and leukocyte telomere lengths (LTL) to infer their relationships with FL genetic risk. Results: We replicated the previously published susceptibility loci within and outside the HLA region, finding two additional independent SNPs at two known loci (P < 5 x10-8) and discovered thirteen novel, genome-wide significant loci located at chromosome 2q13, 3p13, 3p24.1,3q13, 3q24.1, 4q31.21, 6p21, 6p22, 7q31, 10p14, 10q22, 10q23, 17q12, 19p13 and 20p11 with P < 5 x10-8. These SNPs were located near genes that show relevance for B-cell biology, such as BCL2L11, FOXP1, CD86, EOMES, IZKF3, PRF1, KLF2 NKX2-2 and HLA Class I and Class II genes. TWAS uncovered a role for H2AX, IGKV2-29, IMP, UBE4A and IKZF3 and provided directionality of effect for associated transcripts. Colocalization analysis corroborated a positive association with IKZF3 expression showing a robust correlation with FL risk variants (posterior probability PP4>0.7). In addition, several genome-wide significant variants in IKZF3 colocalize with eQTLs across tissues. IKZF3 (Aiolos) is part of the Ikaros family of transcription factors, critically important to the regulation of B-cell development and function. Therapeutic strategies leveraging Immunomodulatory imide drugs (IMiDs) to degrade IKZF3 have shown efficacy in multiple myeloma, and we speculate may also have potential benefits in FL by enhancing anti-tumor immunity and disrupting malignant B-cell function. Gene-set analysis showed blood and spleen to be the most relevant tissues for FL risk variants with lymphocyte activation and immune cell type differentiation genes demonstrating enrichment based on gene-ontology terms. Associated traits included allergies, respiratory diseases (e.g., asthma phenotypes), inflammatory bowel diseases, type 1 diabetes (T1D), lipid levels and commonly measured blood cell traits, with some SNPs showing evidence of colocalization with select blood traits. Interestingly, no causal association to blood traits was observed using MR; however, MR analyses did reveal a strong association with LTL. For the HLA loci, FL shows a diverse set of amino acid associations in HLA Class I and Class II genes that extends beyond what is known from autoimmune disease associations, such as rheumatoid arthritis or T1D. Our study also reveals interesting biological insights about FL susceptibility, such as a novel locus intragenic to FOXP1, an activated oncogene preferentially expressed in FL and DLBCL tumors, but rarely observed in Hodgkin's lymphoma. High levels of FOXP1 are known to result in poor survival in FL. Conclusion: This large GWAS of FL has identified multiple susceptibility variants, uncovering interesting gene candidates for further pathophysiological characterization of FL risk and B-cell development. It suggests new biological pathways and genes that could be potential leads for drug development and targeting treatment, in addition to identifying genetically high-risk populations for FL susceptibility.

Introduction: Over 80,000 new cases of Non-Hodgkin's Lymphoma (NHL) are diagnosed annually, but modifiable risk factors are largely unknown. Circadian disruption has been identified as a risk factor for several cancers, including breast and prostate cancer, and affects immune cells such as Natural Killer (NK) cells and T-helper cells. Disruptions in circadian rhythms can lead to aberrant immune cell trafficking and proliferation. Despite this biological connection, evidence linking circadian disruptions to NHL remains inconclusive. Only single Mendelian Randomization (MR) study (PMID: 32895918) has explored the causal relationship between sleep traits and overall NHL risk, without examining NHL subtypes. Studies have shown distinct risk factor profiles among NHL subtypes. Therefore, this study aims to investigate the causal relationship between sleep traits and NHL subtypes in European populations using MR. Methods: We conducted a two-sample MR analyses using genome-wide association studies (GWAS) summary data on sleep traits and NHL subtypes. The GWAS summary statistics on sleep traits were obtained from publicly available UKBiobank datasets. The sleep traits analyzed included chronotype (N=449,734), insomnia (N=237,627), sleep duration (N=446,118), daytime sleepiness (N=452,071), short sleep (N=411,934), and long sleep (N=339,926). Only independent, genome-wide significant SNPs (p < 5×10-8) were considered as valid instruments. The NHL subtype-specific summary statistics came from the InterLymph including follicular lymphoma (FL: N=6,508 cases / 64,183 controls), mantle cell lymphoma (MCL: N=1,169 cases / 61,603 controls), and Waldenström macroglobulinemia/lymphoplasmacytic lymphoma (WM/LPL: N=1,697 cases / 59,333 controls). Our primary analysis used the random-effects inverse-variance weighted (IVW) method. In addition, we employed MR-Egger to adjust for directional pleiotropy. Heterogeneity in the MR results was evaluated with Cochran's Q statistic. Results: The IVW method did not identify any statistically significant causal association between sleep traits and the risk of NHL subtypes (FL,MCL,WM/LPL), but did yield several intriguing associations. Chronotype (early rising) showed a tendency towards a marginally an increased NHL risk across subtypes [Odds ratio (OR) range: 1.07 - 2.56]. Conversely, a decreased risk was noted for long sleep (≥ 9 hours vs. 7-8 hours) [OR range: 0.19 - 0.62], insomnia [OR range: 0.72 - 0.88], and short sleep (excludes MCL) [OR range: 0.12 - 0.72]. Confidence limit for long sleep was wide due to small number of variants (N=8). The MR-Egger regression analysis did not provide evidence of significant directional horizontal pleiotropy. The F-statistics for all sleep traits were greater than 10 indicating absences of weak instrument bias. Conclusions: Our study found no statistically significant associations between sleep traits and three NHL subtypes (FL, MCL, WM/LPL) in European populations. However, we observed potential influences of chronotype, long and short sleep, and insomnia on NHL risk. These findings align with and extend previous MR study by providing subtype-specific insights. Future directions include expanding our analysis to additional NHL subtypes, generating polygenic risk scores, and conducting stratified analyses using the InterLymph population. These efforts aim to further elucidate the complex relationship between sleep characteristics and NHL risk across diverse subtypes and populations.