Presenter : Conde-Lopez Cristina (DKFZ Heidelberg)
Characterizing Sex Chromosome Dosage Differences in Head and Neck Squamous Cell Carcinoma Tumor Microenvironment
Conde-Lopez Cristina1, Marripati Divyasree3,4, Elkabets Moshe3,4, Hess Jochen1,2, Kurth Ina1
- 1German Cancer Research Center (DKFZ), Radiooncology/Radiobiology, Heidelberg, Germany
2Department of Otorhinolaryngology, Head and Neck Surgery, Heidelberg University Hospital, Heidelberg, Germany - 3The Shraga Segal Department of Microbiology, Immunology, and Genetics, Ben-Gurion University of the Negev, Beer-Sheva, Israel
4Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva, Israel
Head and neck squamous cell carcinoma (HNSCC) presents significant sex differences in incidence, prognosis, and treatment response, attributed to sex chromosome variations and hormonal expression. This study explores these sex-specific differences, particularly focusing on the genetic foundations by examining sex chromosome dosage (XX, XY, X∅) and its effects on the tumor microenvironment (TME). Utilizing omics data from the TCGA-HNSC and single- cell datasets, our analysis applies advanced analytical methods like the Seurat pipeline, scGate for cell annotation, and ProjectTILS for an in-depth examination of gene regulation, oncogenic pathways, and cell composition differences across different sex chromosome compositions. A significant aspect of this analysis is the categorization of male patients based on the expression of the Y chromosome into XY and X∅ groups and comparing these with the XX group to observe variations in TME composition, employing tools like Cellchat for detailed ligand-receptor interaction studies. The findings reveal significant intratumoral heterogeneity and a correlation with HPV status, presenting distinct cell dynamics and signaling activities in the TME across XX, XY, and X∅ groups. Particularly, the study highlights the role of fibroblasts, which exhibit high COX2/PTGS2 and AR expression, in modulating the TME. A balanced investigation into both the stromal and immune compartments and their communication with tumor cells becomes relevant, as their interplay may differ across sex chromosomal backgrounds, influencing the TME in unique ways. Our findings are expected to add valuable insights to the field of cancer research, providing new perspectives on how sex chromosomes influence cancer pathology and treatment strategies.
Presenter: Dr. James P. Cleland (DKFZ/EMBL Heidelberg)
Cell type-specific mechanisms of hepatic sexual dimorphism
James P. Cleland1,2, A. Dugourd3,4, L. Saunders1, J. van Riet1,5, E. Heidari5, B. Drotleff6, G. Rukhovich5, K. Schelzig1, M.-L. Koch1, A. Schneider1, T. Rose4, P. Sant7, J.-P. Mallm7, H. T.- K. Vu8, M. Gerstung5, J. Saez-Rodriguez3,4, E. Heard2,9 and D. T. Odom1.
- 1Division of Regulatory Genomics and Cancer Evolution, German Cancer Research Center (DKFZ), Heidelberg, Germany
2Directors' Research Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
3EMBL-EBI, Hinxton, UK - 4Institute for Computational Biomedicine, Heidelberg University, Heidelberg, Germany
- 5Division of Artificial Intelligence in Oncology, DKFZ, Heidelberg, Germany
- 6Metabolomics Core Facility, EMBL, Heidelberg, Germany
7Single-Cell Open Lab, DKFZ, Heidelberg, Germany - 8Developmental Biology Unit, EMBL, Heidelberg, Germany
- 9Collège de France, Paris, France
The adult mammalian liver is molecularly and functionally highly sexually dimorphic. Past studies suggest a role of both gonadal hormones and sex chromosome complement to hepatic sexual dimorphism; however, the precise contribution of each class of factors to the physiology of the several major hepatic cell types has not been defined. To address this question, we applied a battery of omics and imaging approaches to a large set of livers from the “Four Core Genotypes” sex reversal mouse model. Here, we present our preliminary conclusions. First, sexually dimorphic gene expression is primarily and cell type-dependently determined by gonadal hormones. Second, gonad-dependent gene expression is tightly coupled to metabolic zonation and to a lesser extent cellular ploidy. Third, global hepatic lipid abundance is also largely determined by gonadal hormones and strongly correlated with hepatocyte gonad- dependent gene expression. Fourth, testosterone-mediated repression of Pnpla3 likely causes sexually dimorphic triglyceride metabolism. Overall, these findings suggest extensive, cell type-specific and interconnected sex-dependencies across the adult liver and point toward differential gonadal hormone activity as the primary underlying mechanism.
Presenter: Leticia Rodríguez-Montes (Heidelberg University)
Sex differences in gene expression across mammalian organ development and evolution
Leticia Rodríguez-Montes1, S. Ovchinnikova2, X. Yuan1, T. Studer1, I. Sarropoulos1, S. Anders2, H. Kaessmann1*, M. Cardoso-Moreira3*.
- 1Center for Molecular Biology of Heidelberg University (ZMBH), DKFZ-ZMBH Alliance, Heidelberg, Germany.
- 2BioQuant, Heidelberg University, Heidelberg, Germany.
- 3Evolutionary Developmental Biology Laboratory, The Francis Crick Institute, London, UK.
- *Equal contribution
Sexual dimorphism is widespread in mammals, where males and females differ in their physical, physiological, and behavioural traits. These differences arise during development through sex-specific gene expression programs. Despite the prevalence of such dimorphisms, our understanding of these genetic programs is limited. Questions remain about which genes and cell types exhibit sex differences, when these differences appear during development, and how they evolve across species. Moreover, whether these sex differences are conserved across species—making conclusions derived from animal studies applicable to human biology—remains unclear. To address these questions, we used a transcriptomic resource of gene expression profiles in males and females covering the development of five organs (brain, cerebellum, heart, kidney, and liver) in five mammalian species (human, mouse, rat, rabbit, and opossum) and a bird (chicken). Our findings reveal that sex-biased gene expression is highly variable across organs and species, often tied to specific cell types. Interestingly, these sex differences emerge sharply around sexual maturity rather than gradually over time. Furthermore, the evolution of sex biased gene expression occurs rapidly at the gene level, with only a few genes showing conserved sex-biased expression across species. Despite rapid gene-level evolution, sex biased expression evolves slowly at the cellular level, where the same cell types remain sexually dimorphic across species (for example hepatocytes). These novel insights indicate that care should be taken when using nonhuman mammals to model human sex differences in individual genes, but that using model organisms might be suitable for studying sex differences at the cell type level.
Presenter: Anna E Spiering (University Medical Center Utrecht)
X escape genes in female atherosclerosis are linked to smooth muscle cell biology
Anna E Spiering1, M. Mokry1,2, E.- Diez Benavente1, H. M den Ruijter1.
- 1Laboratory of Experimental Cardiology, University Medical Center Utrecht, Utrecht University, The Netherlands
- 2Central Diagnostic Laboratory, University Medical Center Utrecht, Utrecht University, The Netherlands
Background Sex differences in atherosclerosis are well-documented, but the underlying biological mechanisms remain unclear. Genes that escape X chromosome inactivation are higher expressed in female tissues than in males and may, in part, explain the female atherosclerotic phenotype, characterized by increased fibrosis and high smooth muscle cell content. Here, we aim to identify X escape genes in female plaques and explore their role in atherosclerosis.
Methods Human end-stage carotid plaques from the Athero-Express biobank were histologically assessed and processed to obtain bulk RNA (n=632, female: 156, male: 476) and single-cell RNA (n=46, female: 20, male: 26) sequencing data. Using female-to-male differential gene expression, we defined X escape defined as 10% overexpression in females (FDR 1%).
Results We identified 171 X escape genes in plaques, showing considerable inter-individual variability across female indivduals. Women with lower X escape had a less severe indication for carotid endarterectomy while plaque histology was similar regardless of escape level. Genes higher expressed in females with low rate of X escape were predominantly expressed in endothelial and smooth muscle cells, were enriched for extracellular matrix organization and vascular development pathways and were linked to contractile smooth muscle cells gene networks.
Conclusion X escape is common in female atherosclerotic plaques and displays high variability across individuals. This variability is associated with distinct gene expression profiles, particularly in genes expressed in smooth muscle cells. This study suggests a role for X escape in the distinct female atherosclerotic molecular phenotype.
Presenter: Francesca Maranghi (Istituto Superiore di Sanità, Rome)
Rodent Models of Gender-Affirming Hormone Therapies to identify and characterize risks for Transgender People
Francesca Maranghi1, R. Tassinari1.
- 1Center for Gender-Specific Medicine, Istituto Superiore di Sanità, Rome, Italy.
Individuals whose gender identity differs from the biological sex and the social norms are defined transgender (TG); sometimes they undergo gender affirming hormone therapy (GAHT) that for TG men involves testosterone treatment and for TG women estrogen plus antiandrogens. Due—but not limited—to the lifelong lasting of GAHT, TG may show different and specific susceptibility and vulnerability compared to general population, including in the response to chemical contaminants present in daily life; among them, the endocrine disrupters (EDs) affecting hormonal and metabolic processes deserve special attention. Indeed, the endocrine system of TGs is overstimulated by GAHT and the ED exposure overlap targets with GAHT: it’s reasonable to hypothesize that TG health should be carefully evaluated in the frame of toxicological risk assessment also by setting specific animal models mimicking GAHT. Such models aim at providing robust data for hazard identification in TG, leading to a more reliable risk assessment, and at studing potential long-term consequences of GAHT. The first results are the identification of: i) suitable dose of testosterone (for defeminizing/masculinizing model) and estradiol plus cyproterone acetate (for demasculinizing/feminizing model) to be used in long-term studies; ii) high-quality biomarkers to support the success of both the HTs to obtain a reliable animal model for TG; iii) "new" (and neglected) targets affected by GAHT. The implementation of animal models mimic gender affirm HT for risk assessment is critical to support clinical studies and to filling data gap in order to ensure accurate and personalized care for TG people.
Presenter: Dr. Valentina Biasin (Medical University of Graz)
Testosterone level: an important determinant in Systemic sclerosis
P. Waked1, A. Birnhuber2, P. Kotzbeck3,4, I. Foessl4,5, D. Zabini1, L. M. Marsh2, S. Crnkovic2, A. Olschewski6, B. Obermayer-Pietsch5, G. Kwapiszewska2*, Valentina Biasin1*.
- 1Division of Physiology and Pathophysiology, Otto Loewi Research Center, Medical University of Graz, Austria.
- 2Otto Loewi Research Center, Medical University of Graz, Austria.
- 3COREMED, Joanneum Research Forschungsgesellschaft, Austria.
- 4Research Unit for Tissue Regeneration, Department of Surgery, Medical University of Graz, Austria.
- 5Department of Endocrinology, Medical University of Graz, Austria.
- 6Department of Experimental Anaesthesiology, Medical University of Graz, Austria.
- *Equal contribution
Systemic sclerosis (SSc) is an autoimmune disease which affects connective tissue leading to fibrosis in the skin and different internal organs, vascular abnormalities and strong inflammatory component. SSc has higher prevalence in females, however males with SSc often experience a more severe course of the disease. The worse prognosis of SSc male patients, is often due to a more progressive lung fibrosis and pulmonary arterial hypertension (PAH) eventually leading to respiratory failure or right heart failure respectively. The sexual dimorphism in SSc suggest a potential impact of sex hormones in SSc development and progression. In order to further elucidate the role of sexual hormones in SSc manifestations we took advantage of an established mouse model of SSc, the Fra-2 Tg mouse model; which was subjected to castration. While ovariectomy did not altered disease course in females Fra-2 Tg mice, orchiectomy clearly exacerbated the lung phenotype in males, increasing the inflammatory infiltrate, the lung fibrosis and vascular remodelling. Subsequent testosterone replacement therapy in orchiectomized Fra-2 Tg mice, resulted in a significant improvement in disease phenotype assessed by enhanced lung function, improved hemodynamic, reduced fibrosis, reduced inflammation, and alleviated vascular remodelling in the lungs. These findings showed a strong link between testosterone levels and lung phenotype severity in Fra-2 Tg mice. These results underscore the significance of sex hormones in SSc pathology, and suggest a possible causative effect of testosterone level and lung phenotype in SSc male patients.
Presenter: Dr. Ivana Jaric (University of Zurich)
Beyond SABV: Ovarian hormone fluctuations and their role in modulating brain and behavior.
Ivana Jaric1
- 1Institute of Laboratory Animal Science, University of Zurich, Zurich, Switzerland.
For decades, female subjects have been neglected in biomedical research, particularly in neuroscience, where, for every six studied male animals, only one female animal was examined. Such disparities have led to oversights in crucial sex differences, translational failures, and the unnecessary suffering and waste of animals used in inconclusive studies. In response, major funding agencies now require the inclusion of sex as a biological variable (SABV) in all basic and preclinical experiments. However, including females prompts consideration of accounting for ovarian hormone fluctuation. Despite the known impact of ovarian hormones—which vary significantly across the rodent estrous cycle, similarly to the human menstrual cycle—on a range of biological functions, this variable is still largely ignored. I will present how accounting for the estrous cycle enhances the resolution of neuroscience studies by (i) identifying previously masked sex differences and (ii) providing mechanistic insights into these differences, spanning a range of neurobehavioral outcomes. I will also present the critical role of ovarian sex hormone-driven epigenetic mechanisms, particularly chromatin organizational changes, which dynamically regulate neuronal gene expression and brain plasticity. Additionally, I will explain how fluctuations in ovarian sex hormones may prime the (epi)genome for psychopathology. Finally, I will outline future strategies for incorporating endocrine states into rodent studies, with the goal of improving data interpretability, precision, and the translational relevance of basic and preclinical research.
Presenter: Prof. Dr. Valentina Proserpio (University of Turin)
Vulvodynia at high resolution
Albano Alessia1,2, Brasi Luna1, Cianci Marco1, Griffante Gloria1,2, Loperfido Federica1, Federico Oreglia1, Monteleone Emanuele1,2, Molineris Ivan1 & Proserpio Valentina1,3.
- 1University of Turin, Torino, Italy
- 2Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.
- 3IIGM Foundation, Italian Institute for Genomic Medicine, Candiolo, Torino, Italy.
It is commonly assumed that the more severe and widespread a disease is, the more attention it will receive from the scientific community, but this is not the case for Vulvodynia (VD). VD is defined as vulvar pain without a clear identifiable cause persisting for at least 3-months. VD is a high frequency, albeit neglected, women’s health condition that affects 1 in 7 women. The average diagnostic delay is about 4 years. Our project aims at finding new diagnostic markers and, possibly, new therapeutic targets for VD. To this end, we performed global transcriptomic analysis on more than 120 VD patients and age-matched healthy controls. We also analyzed the vulvar microbiome composition by metatranscriptomic approach, considering its important pathogenic role in local immune-inflammatory responses. Differential gene expression analyses reveal altered pathways shared amongst VD patients. In particular, our preliminary analysis shows that VD patients have ~300 significantly differentially expressed Genes (DEGs) from healthy controls and that the great majority of these genes are overexpressed in VD patients. Gene ontology enrichment analysis revealed immune deregulation together with changes in adhesion and cell cycle. We also stratified patients into different clusters based on the integration between transcriptome, microbiome and patient’s metadata. This analysis has never been performed on VD patients with this omics approach and at such high resolution and holds the potential to shed light on the molecular mechanisms behind this complex disease.
Presenter: Dr. Magdalena Holze (Heidelberg University Hospital)
The role of sex and gender in pancreatic cancer: A focus on surgical outcomes
Magdalena Holze1,2, Ulf Hinz1, Christoph W. Michalski1, Martin Loos1, Rosa Klotz1,2.
- 1Department of General, Visceral and Transplantation Surgery, Heidelberg University Hospital, Heidelberg, Germany
- 2The Study Center of the German Surgical Society, Heidelberg University Hospital, Heidelberg, Germany
Background Sex and gender are increasingly recognized as modulators of health and disease, yet their impact in oncology, particularly in pancreatic cancer, remains underestimated. Recent studies found favourable outcomes for women in systemic treatment, though short-term outcomes after surgery remain controversial. Personalized perioperative strategies, including risk assessment and complication management, may improve both perioperative and oncological outcomes. This study explores how sex and gender affect the pathway of pancreatic cancer patients with a focus on the surgical setting.
Methods A literature review was conducted to assess the current evidence on the impact of biological and social sex in patients with pancreatic cancer. Based on these findings, a study concept for a retrospective analysis was developed to specifically investigate sex and gender differences in patients who underwent pancreatic cancer surgery at the surgical department of the Heidelberg University Hospital.
Results Sex differences with respect to incidence, treatment response and prognosis are described in pancreatic cancer. Sex influences both innate and adaptive immune responses, thereby affecting treatment response and survival rates. Women are less likely to receive systemic treatment and tend to wait longer for surgery but have better perioperative outcomes. Men have a higher overall postoperative mortality and develop more pancreatic-specific complications, despite women being more likely to have a soft pancreas. Female pancreatic cancer patients seem to have longer survival under treatment; however, they report a lower quality of life and higher disease burden.
Discussion In patients with pancreatic cancer, the current literature suggests relevant gender differences that may impact the treatment pathway. Thus, high-quality evidence is needed to clarify the role of sex and gender aspects in pancreatic cancer surgery and to potentially change clinical practice in terms of a precision surgical pathway.
Presenter: Dr. Raja Idris (Frankfurt University Hospital)
Sex-dependent variability of isoniazid and rifampicin serum levels in patients with tuberculosis
Raja Idris1,2, A. Z. Dayani1,2, A. M. Groh1, A. Mohr1,3 J. Koepsell1, A.-S. Zielbauer1, E. Herrmann4, M. J.G.T. Vehreschild1, T. A. Wichelhaus5, N. Wetzstein1,2,6.
- 1Goethe University Frankfurt, University Hospital, Department of Internal Medicine, Infectious Diseases, Frankfurt am Main, Germany.
- 2Goethe University Frankfurt, Mycobacterial Infection Research Unit (MIRU), Frankfurt am Main, Germany
- 3Goethe University, University Hospital, Hospital Pharmacy, Frankfurt am Main, Germany.
- 4Goethe University Frankfurt, Institute of Biostatistics and Mathematical Modeling, Frankfurt am Main, Germany.
- 5Goethe University Frankfurt, University Hospital, Institute of Medical Microbiology and Infection Control, Frankfurt am Main, Germany.
- 6Molecular and Experimental Mycobacteriology, Research Center Borstel, Borstel, Germany.
Introduction Drug-sensitive TB (DS-TB) is treated with isoniazid, rifampicin, ethambutol, and pyrazinamide. Factors like fast-metabolizing enzymes, malabsorption, and drug interactions can influence serum drug levels. Current TB treatment guidelines recommend weight -adapted dosing without considering sex differences. This study examines drug levels of isoniazid and rifampicin in TB patients treated between 2019-2023 at our center focusing on sex-specific aspects.
Methods Patients diagnosed with TB and available serum levels of isoniazid or rifampicin between 2019-2023 were retrospectively identified. Serum levels were measured using liquid chromatography–mass spectrometry and high-performance liquid chromatography. Patients were stratified by sex and a linear regression mixed effect model was used to assess predictors for different serum levels.
Results The study included 281 single therapeutic drug monitoring (TDM) measurements from 59 patients (28 women, 47.5%). Adverse effects were noted in 43.8% (42/96) of measurements in women and 29.5% (54/183) of measurements in men (p=0.03). For isoniazid, no sex-specific differences were identified. On the other hand, female sex was a significant predictor of higher rifampicin plasma levels (coefficient 4.16, 95% CI 0.74-7.59, p=0.009). Only 38.2% of rifampicin serum level measurements in male patients were within target range. Women displayed higher overall rifampicin serum levels than men (median 14.7mg/ml vs. 7.1 mg/ml, p=0.04), although weight adjusted doses were not significantly different (median 10.0 mg/kg bodyweight vs. 9.8 mg/kg, p=0.56).
Discussion Rifampicin levels were significantly lower in men compared to women, despite weight-adjusted dosing. Clinicians should consider TDM and potential sex differences when treating patients with TB.
Presenter: Evelyn Röser (Heidelberg University Hospital)
Differences between men and women in prolonged weaning
Evelyn Röser1, Julia D. Michels-Zetsche, Benjamin Neetz, Philipp Höger, Frederik Trinkmann, Michael M. Müller, Laura Klotz, Hilal Ersöz, Konstantina Kontogianni, Jana-Christina Dahlhoff, H. Winter, Sabine Krysa, Felix J.F. Herth, Franziska C. Trudzinski1.
1Thoraxklinik Heidelberg gGmbH, Heidelberg University Hospital, Heidelberg, Germany.
Background In recent years, the importance of sex as a factor influencing medical care has received increasing attention in the field of intensive care medicine. The objective of this study was to examine the influence of sex in prolonged weaning.
Methods A retrospective analysis of patients undergoing prolonged weaning at Thoraxklinik, University Hospital Heidelberg between 12/08 and 12/23 was conducted. Patients with neuromuscular diseases were excluded from the analyses. The risk factors for weaning failure in men and women were identified through stepwise Cox regression analyses.
Results A total of 785 patients were included, of whom 313 (39.9%) were women. 77.9% of the women and 75.4% of the men were successfully weaned from invasive ventilation. In group comparisons and multivariable analyses, sex was not found to be a risk factor for weaning failure. Cox regression analyses were performed separately for both sexes on the outcome of weaning failure, adjusting for relevant covariates. The results indicated that age (HR 2.38, p<0.001) and duration of mechanical ventilation before transfer (HR 1.01, p<0.001) were independent risk factors in men. In women, however, the duration of mechanical ventilation before transfer (HR 1.01, p<0.001), the presence of critical illness polyneuropathy (HR 1.82, p=0.040) and delirium (HR 2.50, p=0.017) were identified as relevant risk factors. In contrast, delirium had a favourable effect on weaning success in men (HR 0.38, p=0.020) and nosocomial pneumonia in women (HR 0.38, p=0.032).
Conclusion The analyses indicate that there are sex-based differences in the risk factors associated with weaning failure. This suggests that sex should be considered in weaning from invasive mechanical ventilation to improve outcomes.
Presenter: Dr. Anna Ruggieri (Italian National Institute of Health, Rome)
Potential sex indicators of the COVID-19 vaccination response in healthcare workers: Sex hormones and microRNAs
Anna Ruggieri1, Simona Anticoli1, Maria Dorrucci1, Elena Ortona1, Nicoletta Vonesch2, Paola Tomao2.
- 1Istituto Superiore di Sanità, Rome,Italy.
- 2INAIL, Rome, Italy.
Age and sex are the two most significant characteristics, among several, that influence the strength and effectiveness of the immune response to vaccinations, with women being more immune reactive than men. Sex-disaggregated analysis of humoral response to vaccinations, including COVID-19 vaccines, is rarely available and frequently controversial. We conducted a study to assess variations in anti-S/RBD antibody levels between male and female healthcare workers (HCWs), naïve to SARS-CoV-2 infection, at various time points following the second dose of the mRNA COVID-19 vaccine, accounting for immunization age. To the aim of identifying sex-specific markers of immune response to the vaccine we investigated microRNAs, -regulators of multiple immune functions- sex hormones—important players in the sex difference in immune response—. The potential relationship between anti-S/RBD antibody levels and these factors were statistically analyzed. Compared to male HCWs, females had a significantly stronger anti-S response; the anti-S decline, which occurred 77 days after the second vaccine dose, was more abrupt in female than in male workers, and in younger age groups than in those over 50. A role for testosterone as a sex-specific predictive biomarker of response to the COVID-19 vaccination in young men has been suggested by greater plasma levels of testosterone in males under the age of 55, which have been statistically associated to higher anti-S levels. Two circulating microRNAs that are highly expressed in females were found to positively correlate with anti-S/RBD levels after COVID-19 vaccination. Identification of sex-specific markers predictive of immune response to vaccination will contribute to customize health surveillance programs for healthcare professionals, accounting for sex disparities.
