Meet the Speakers

Chun-Chi Chang | Postdoc at Consiglio Lab

From Hormones to Host Defense: Immune Adaptation Following Androgen Deprivation Therapy 

Background: Androgen deprivation therapy (ADT), a cornerstone treatment for prostate cancer, has been associated with increased risk of community-acquired pneumonia (CAP). The immunological mechanisms underlying this association remain unclear, particularly how androgen suppression alters human antimicrobial immunity. We aimed to characterize early immune adaptations following ADT and assess their impact on host defense against major respiratory pathogens.

Methods: Peripheral blood immune cells were collected from prostate cancer patients prior to ADT initiation and four weeks after androgen suppression. Immune profiling was performed using high-dimensional spectral flow cytometry and single-cell RNA sequencing (scRNA-seq) on baseline samples and on cells cultured ex vivo under control conditions or stimulated with Streptococcus pneumoniae or influenza A virus (IAV), two leading causes of CAP. Primary outcomes included changes in immune cell composition, transcriptomic signatures, and functional responses relevant to antimicrobial immunity.

Preliminary results: ADT was associated with a trend toward increased conventional dendritic cells and reduced follicular helper T cells. B cells exhibited decreased CXCR5 and CCR6 expression following androgen suppression. Upon S. pneumoniae stimulation, B cells showed attenuated CD69 induction, while plasmacytoid dendritic cells demonstrated reduced CXCR3 and CCR10 expression. IAV stimulation revealed impaired activation and migratory marker expression across multiple subsets, including reduced CD69 and CXCR5 on B cells, CD69 on CD4+ T cells, CCR6 on pDCs, and CXCR3 on classical monocytes. scRNA-seq analyses confirmed reduced CD69 and CCR6 expression across immune subsets after ADT.

Conclusions: Short-term androgen suppression reshapes functional immune phenotypes without markedly altering overall cell composition, particularly impairing activation and homing capacities. These findings support androgen signaling as a key modulator of sex differences in antimicrobial immunity. As this study captures early immune adaptations, further studies are required to assess the changes over time.
Significance: Defining ADT-induced immune adaptation may inform strategies to mitigate infection risk while preserving oncologic efficacy.

Maria Markus | PhD Student at Trägårdh Lab

FDG-PET/CT-based treatment response predicts recurrence and overall survival in locally advanced cervical cancer 

Background: [18F]-fluorodeoxyglucose (FDG) positron emission tomography combined with computed tomography (PET-CT) is used for staging and treatment response evaluation in patients with locally advanced cervical cancer. While the maximum standardised uptake value (SUVmax) is a commonly used PET parameter, volume-based parameters (metabolic tumour volume (MTV) and total lesion glycolysis (TLG)) may offer superior predictive value. This study aimed to investigate the predictive value of different treatment response outcomes assessed with PET-CT, using both standard and volume-based PET parameters.

Methods: We retrospectively analyzed 133 patients with locally advanced cervical cancer treated with (chemo)radiotherapy who underwent PET-CT pre- and post-treatment. Treatment response was classified as complete metabolic response (CMR) (complete resolution of FDG uptake in all lesions), partial metabolic response (PMR) (> 25% reduction in the sum of SUVmax/MTV/TLG), progressive metabolic disease (PMD) (> 25% increase in the sum of SUVmax/MTV/TLG or appearance of new FDG-avid lesions) or stable disease (SD) (does not qualify for CMR, PMR or PMD). The three PET-parameters were analysed separately. Associations between treatment response and overall survival (OS) as well as recurrence (binary variable) were analysed through Cox and logistic regression models, respectively. 

Results: Compared to CMR, the odds ratio (OR) for recurrence was 1.2 for patients with PMR/SD for SUVmax, MTV and TLG, while in the PMD group, it was 1.7 for SUVmax and 1.8 for MTV and TLG. The hazard ratio (HR) for OS was 4.0 for patients with PMR/SD and 16.0 for those with PMD, based on SUVmax; for MTV and TLG, HRs were 2.9 for patients with PMR/SD and 16.0 for those with PMD. C-index from adjusted Cox models were comparable: 0.84 for SUVmax, and 0.86 for both MTV and TLG.

Conclusions: FDG-PET/CT-based assessment of treatment response is a strong predictor of recurrence and overall survival in advanced cervical cancer. SUVmax, MTV, and TLG demonstrated similar predictive performance.

Malavika Sreekumar Nair | Project Assistant at Pereira Lab

Epigenetic adjuvants for in vivo dendritic cell reprogramming 

Cancer immunotherapy has revolutionized oncology, but many patients do not benefit. Rational combinatorial strategies are needed to overcome resistance. We previously demonstrated that the transcription factors PU.1, IRF8, and BATF3 (PIB) reprogram cancer cells into Type 1 Conventional Dendritic Cell-like (cDC1-like) cells, remodeling the tumor microenvironment and synergizing with immune checkpoint blockade to elicit durable anti-tumor immunity. 

Here, we investigated whether epigenetic modulation could further enhance cDC1 reprogramming and immunogenicity. Twenty-seven human cancer cell lines were treated with PIB plus the histone deacetylase inhibitor valproic acid (VPA) or the DNA methyltransferase inhibitor 5-Azacytidine (AZA). Both enhanced reprogramming efficiency (CD45, HLA-DR expression), with VPA exhibiting the strongest effect (mean 4-fold vs. 2-fold for AZA), particularly in PIB-resistant lines. This effect was conserved in 15 murine cancer lines (~6-fold increase). 

Mechanistically, VPA facilitated early PU.1 chromatin binding enriched at IRF motifs, suggesting cooperative recruitment of cDC1 transcriptional machinery. Time-course transcriptomics on a high-responding line revealed VPA-driven early activation of antigen presentation pathways, co-stimulatory molecules, pro-inflammatory cytokines, and interferon-stimulated genes, showing accelerated immunogenicity. Functionally, PIB+VPA enhanced tumor antigen presentation and Pmel-specific CD8+ T-cell-mediated killing in vitro. In vivo, PIB+VPA induced tumor regressions in the YUMM1.7 melanoma model, achieving 9/10 complete responses versus 2/10 with PIB alone, when using low transduced cell numbers. All complete responders (9/9) rejected tumor rechallenge, demonstrating durable immunity. 

These findings establish VPA as a potent epigenetic dual-adjuvant that boosts both the efficiency and immunogenicity of cDC1 reprogramming, providing a compelling rationale for integration into next-generation cancer immunotherapy strategies.

Steffen Haupeltshofer | Postdoc at Paul-Visse Lab

Uncovering Pericyte-Macrophage Crosstalk in Ischemic Stroke 

Background: Pericytes (PC) line the microvasculature at the blood-brain interface and are early responders to hypoxia and acute ischemic stroke. PC are a highly dynamic cell type widely recognized as major contributors of the capillary constriction after stroke, but also as key effectors in the breakdown of the blood-brain-barrier that leads to influx of immune cells, ion imbalance and cerebral vascular oedema - all of which are associated with a poor stroke outcome. We hypothesize that their strategic position in the brain pinpoints PC as potential target cells to modulate downstream neuroinflammatory processes in acute ischemic stroke.

Methods: Ischemic stroke was induced in RGS5GFP/+ mice using permanent middle cerebral artery occlusion (pMCAO). To investigate the subtype variances in transcriptomic signatures within pericytes, we sacrificed mice timepoints 24 h after stroke induction and performed 10x single-RNA sequencing on non-neuronal cells. Using bioinformatic analysis we focused on the PC signalling, cellular interaction and communication capability with immune cells after stroke using CellChat.

Results: Stroke-related Atp13a5+ PC showed marked phenotypic plasticity and upregulation of immunomodulatory IL-6, CCL-2 and TGF-beta1 signalling. In particular, the pleiotropic cytokine IL-11, a member of the IL-6 superfamily, has been exclusively expressed by a specific pericyte population after stroke. Analysis of cellular communication revealed strong interactions between Atp13a5+ PC, PDGFR-beta+Acta2+ smooth muscle cells and CD45+ immune cells. Among these cell interactions, PC communication was strongly connected to macrophages. Interestingly, PC subtypes differed in their signalling preferences and communication strength with macrophage recruitment sites in the skull, meninges and periphery after stroke.

Conclusions: In acute ischemic stroke, PC acquire unique immunomodulatory phenotypes that constitutes a critical site for regulating cell-cell interactions, immune cell priming, and migration. Therefore, unravelling the role of PC in shaping neuroinflammatory responses may represent a promising therapeutic option in ischemic stroke treatment.

Léa Chicoisne | Postdoc at Consiglio Lab

Development of a systems-level immune profiling protocol for the menstrual blood  

The human endometrium is a hormonally responsive tissue lining the inner cavity of the uterus. It undergoes continuous remodeling throughout menstrual cycles, including degeneration (menstrual phase), regeneration (proliferative phase) and differentiation (secretory phase). It is composed of a dynamic population of leukocytes playing a critical role in reproductive processes, particularly for embryo implantation and the maintenance of pregnancy. Menstrual blood is a non-invasive specimen shown to closely resemble the uterine microenvironment. It is composed of endometrial and immune cells, blood and cervicovaginal mucus. Specific menstrual blood immune composition has been associated with recurrent pregnancy loss. Although menstrual blood has garnered increased attention in recent years, no standardized protocol for processing and analyzing samples have been established. Moreover, most studies analyzed individually the immune cell composition, transcriptome or proteome, lacking a holistic view.

We aim to develop and validate a standardized workflow to collect and process menstrual blood for systems-level immune analysis, by optimizing conditions (time of collection, viscosity, etc.) to profile simultaneously transcriptomics (bulk mRNA sequencing), plasma proteomics, immune cell population, and immune responses to viruses, fungi, and bacteria.
Our preliminary results show that time of collection (day vs. night) did not significantly affect volume, cell yield, or viability. In parallel, high viscosity negatively affects cell viability. Yet this viscosity is not related to the timing of collection but is more person- and cycle-dependent. Furthermore, cell yield decreases over menstrual cycle days, independently of the viscosity. 

Our findings suggest that, to obtain optimal cell yield and viability, menstrual blood must be collected at early menstruation, with adjustments for viscous samples. Menstrual blood presents a promising avenue for immune assessment, and this standardization will facilitate the obtention of reliable immune results between projects and support future biomarker discovery in reproductive disorders.

Wesley Sturgess | Postdoc at Wittrup Lab

Mapping the Proteomic Landscape of RNA-Induced Endosomal Damage

RNA-based therapies represent a promising yet challenging approach for the treatment of diseases such as cancer. Although RNA technology has been successfully applied in the treatment of liver diseases and in vaccines, including those developed against SARS-CoV-2, significant obstacles remain before these therapies can be fully realized in oncology.
One of the primary challenges is the inefficient escape of RNA cargo from endosomes following cellular uptake by endocytosis. Currently, only approximately 5% of internalized RNA successfully escapes into the cytosol, where it can exert its therapeutic effect. While several hypotheses attempt to explain this low efficiency, the precise mechanisms governing endosomal escape remain poorly understood.

Endosomal escape is generally thought to result from membrane damage induced by the RNA delivery vehicle. Such damage triggers the recruitment of proteins involved in the endosomal damage response machinery. Although some of these proteins have been well characterized, many others remain poorly defined, and their roles in endosomal disruption are still unclear.

To better understand the molecular machinery underlying endosomal damage, we are developing a method to isolate RNA-damaged endosomes. These isolated endosomes will then be analyzed using mass spectrometry and proteomic approaches to map the proteins recruited during this process. This strategy will enable us to identify the key pathways involved in endosomal damage and uncover potential targets that influence the efficiency of RNA release into the cytoplasm.

Anna Castellet | MSc at Consiglio Lab

Hormonal effects on the female immune system during IVF and menstrual cycle  

Infertility affects 15-25% of couples worldwide. In vitro fertilization (IVF) is the most common treatment, yet its success is limited, with average live birth rate of about 30% after the first attempt. Improving IVF outcomes requires understanding the mechanisms that shape reproductive success. One key determinant is the crosstalk between the endocrine and immune systems: sex hormones modulate immune cell composition, activation, and function, while the immune environment influences reproductive processes and success. Hormone treatments used in IVF alter immune cells proportions, potentially affecting implantation success.

Endocrine-immune crosstalk is crucial during the menstrual cycle and pregnancy. Hormonal fluctuations shape immune activity to support ovulation, implantation, and fetal tolerance. Inflammatory responses dominate the proliferative phase under rising estrogen levels, whereas progesterone promotes an immunosuppressive profile in the secretory phase. Despite evidence linking immunity to fertility, the specific hormonal effects on immune cells remain unaddressed, as most studies rely on single-hormone \textit{in vitro} experiments rather than holistic, system-level approaches.
Here we address these gaps through scRNA-seq analysis of endometrial tissue. From a publicly available dataset, 11 proliferative and 15 secretory phase endometrial samples were retained after quality filtering. Immune cell populations were identified and annotated, and differential expression analysis followed by functional enrichment analysis is performed between phases to further identify hormone-responsive immune gene programs.

Preliminary results show subtle immune shifts across menstrual phases. CD4+T, CD8+T and uNK cells are more abundant in the secretory phase, while other populations remain stable. The proliferative phase is enriched for inflammatory, cell cycle and cell death pathways, suggesting active tissue turnover. The secretory phase shows enrichment in immune tolerance, metabolism, and structural organization pathways, indicating tissue remodeling towards a receptive, implantation-ready environment.

Understanding how hormones regulate immune function during the menstrual cycle can reveal immune markers associated with reproductive outcomes, improving fertility treatments and pregnancy success.

Indra Putra Wendi | PhD Student at Wagner Lab

Utilizing polydopamine-functionalized electrospun  poly(ε-caprolactone) membrane to model upper airway epithelium at air-liquid interface

Identification and establishment of alternative insert materials to generate upper airway models is imperative to facilitate advancement in drug development for acute and chronic airway disease which are major public health concerns. An ideal model requires a scaffold with mechanical properties like those of human lungs and a poro-fibrous architecture that mimics the extracellular matrix (ECM) of lung tissue. Electrospun membranes generated using poly(ε-caprolactone) (PCL) was explored as an alternative to commercial inserts due to the fact that its mechanical properties are more similar to native airways. Due to its hydrophobic nature, different surface modification methods were explored to enhance cell seeding and proliferation, including surface etching, chemical coating, and matrix protein deposition. 

The efficiency of these methods was evaluated using water contact angle measurements for surface energy changes and Fourier transform infrared (FTIR) spectroscopy for evaluation of chemical changes on the surface of the membranes. Human airway cells were seeded onto the modified membranes to assess their impact on initial cell attachment, cell proliferation and monolayer formation prior to air lifting to reach air-liquid-interface culture. 

Our results indicated that polydopamine coating in conjunction with optimal seeding conditions was crucial for achieving a stable cell monolayer with barrier properties. This was confirmed through a combination of live-cell tracking dyes, FITC-dextran permeability assays, and transepithelial electrical resistance (TEER) measurements. Lastly, we evaluated morphological changes after lifting to air-liquid-interface culture using a custom-developed histological processing and paraffin embedding procedure to enable generation of transverse sections. When using human airway cells derived from a serous tumor, we observed morphological changes at ALI resembling those observed in vivo with these types of tumors. 
Overall, we established a method for using nanofibrous PCL scaffold with large-sized pores (± 5 um) to grow an airway ALI model with a barrier profile robust enough to be airlifted.

Zhimeng Fan | PhD Student at Swaminathan Lab

ECM remodeling overrides stiffness mediated dormancy checkpoint in breast cancer cells

Cancer recurrence is the leading cause of breast cancer-related mortality among women worldwide. During recurrence, disseminated tumor cells (DTCs) evade treatment by entering a dormant or quiescent state and are reactivated by changes in their surrounding environment at a later time. While several tumor niche factors contributing to DTC reactivation have been identified, the role of the physical properties of the extracellular matrix (ECM), including its stiffness and organization, remains poorly understood.

To address this, we investigated DTCs behavior using D2.0R and MCF7 cell lines and found that fibronectin (FN)-enriched but mechanically soft ECM keeps DTCs quiescent for extended periods while increasing the ECM stiffness triggered a switch to a proliferative phenotype. However, we discovered that enzymatic cleavage and remodeling of FN in a mechanically soft ECM was sufficient to override the stiffness-mediated dormancy checkpoint and induces DTCs reactivation, mimicking the effects of ECM stiffening. Mechanistically, through tumor surfaceome mapping (TSA) and quantitative high-resolution microscopy, we found that remodelling of FN in mechanically soft ECM results in downregulation of collagen expression which in turn leads to downregulation of the integrins α1 and α2. Furthermore, this reawakening is promoted by the activation of β1 integrin-FAK pathway suggesting a crosstalk between different integrin subtypes during this process. 

In summary, this study elucidates the mechanistic role of physical properties of the ECM and its remodeling in DTCs reactivation and highlights integrins as potential therapeutic targets to prevent breast cancer recurrence.

Milica Dimitrijevic | PhD Student at Leigh Lab

Transposable Elements as Regulators of Gene Expression During Embryonic Development in Giant Newt Genomes 

Salamanders are the primary tetrapod model for studying complex organ regeneration, a process that often reactivates developmental gene networks, highlighting the deep connection between regeneration and development. Another distinctive feature of salamanders is their gigantic genomes, largely expanded through the accumulation of repetitive elements, particularly long terminal repeat (LTR) transposons. While transposable elements (TEs) are recognized as important regulatory elements that can influence gene expression at multiple levels, their potential function in regulating gene expression during development and regeneration in salamanders is unknown. 

To address this, we generated CUT&RUN data for Iberian ribbed newt embryos (Pleurodeles waltl) and profiled the chromatin landscape of these animals at the middle hatching stage. We investigated histone modifications associated with active (H3K4me3 and H3K27ac) and repressive (H3K9me3) chromatin states. By integrating these data into the newly assembled, huge P. waltl genome (20.3 Gb), we identified putative cis-regulatory elements, including putative enhancers and promoters which originate from the annotated TEs. Our findings support the emerging view that transposable elements can serve as a reservoir of regulatory potential, and suggest that in salamanders, TEs may have been co-opted to facilitate the extensive gene regulation required for development and regeneration.

Our study lays a foundation for future in vivo studies to test the regulatory function of TE-derived elements, including CRISPR interference experiments to selectively silence candidate enhancers and promoters. Together, these insights shed new light on the functional significance of TE accumulation in regenerative species with giant genomes. 

Anindita Ray | MSc Student at Consiglio Lab

Sex Differences in Cardiovascular Disease: Single-cell RNA-seq analysis of Immune Responses Post-Myocardial Infarction 

Cardiovascular diseases (CVDs) remain the leading cause of mortality worldwide, with myocardial infarction (MI) being a common manifestation that triggers a tightly regulated innate immune response essential for cardiac repair but detrimental when dysregulated. Monocytes and neutrophils derived from the bone marrow are rapidly recruited to the injured myocardium, where they undergo functional reprogramming to coordinate inflammation and tissue repair. While excessive or prolonged innate immune activation amplifies adverse cardiac remodelling and heart failure, understanding the molecular drivers of this maladaptive response requires further investigation.

Emerging evidence indicates that biological sex significantly influences immune responses and clinical outcomes following MI, with males exhibiting stronger early inflammatory responses and females experiencing higher post-MI mortality. However, the cellular and molecular mechanisms underlying these sex-specific differences are poorly defined, limiting the development of precision therapies. The alarmin complex S100A8/A9 is a central amplifier of acute post-MI inflammation, and its pharmacological inhibition has shown therapeutic promise in preclinical models, yet its sex-dependent effects remain unexplored.

This study aims to define sex-specific immune cell composition and activation states following MI using a systems immunology approach. Single-cell CITE-seq data from male and female mice subjected to MI, with or without S100A8/A9 inhibition, were analysed across blood, heart, and bone-marrow compartments collected during the acute post-MI phase (day 3). Among the immune populations identified, males exhibited higher proportions of progenitor cells in bone-marrow, classical monocytes in heart and circulating neutrophils in blood compared to females, indicating sex-based differences in immune cell mobilisation following MI. These observations support the concept that acute inflammatory responses to MI are regulated in a sex-specific manner at single-cell level and may respond differentially to targeted anti-inflammatory intervention. 

Ongoing analyses are expected to further reveal distinct sex-dependent inflammatory gene signatures post-MI. This work highlights the importance of incorporating biological sex into immunomodulatory strategies to optimise post-MI therapies and advance precision cardiovascular medicine.

Konstantinos Kokkoros | Project Assistant at Pereira Lab

Modelling immunosenescence with direct reprogramming of fibroblasts to antigen presenting cells 
During aging the immune system ceases to effectively respond to pathogens and disease, a process known as immunosenescence. Antigen presenting cells (APCs), crucial for linking innate and adaptive immunity, are particularly susceptible, showing age-related declines in antigen presentation and thus T-cell activation. Beyond age, immune responses differ between sexes, with females typically exhibiting stronger immune responses but more side effects than males. These observations are indicative of cell-intrinsic programmes, which could illuminate differential disease susceptibility during aging.

Here, we model immune aging using direct reprogramming of human dermal fibroblasts (HDFs), derived from a library of >50 donors with equal distribution of biological sex,  consisting of 10 donors per life decade between 20-80 years old, into type-1 conventional dendritic cells (cDC1s). Preliminary reprogramming data reveals heterogeneous reprogramming efficiencies among individuals, with higher reprogramming efficiencies in younger donors. To dissect this variability, we will perform phenotypic, molecular, and functional characterization of reprogrammed cells focusing on their antigen-presenting capacity. Reprogramming efficiency will be quantified by flow cytometry using markers to assess phenotype (CD45, HLA-DR), fidelity (CD226, XCR1), and functionality (CD40, CD80-86). Transcriptional, epigenetic, and DNA methylation landscapes will be profiled before and after reprogramming using scRNA-seq, ATAC-seq, and CpG methylation analyses. Functional competence will be assessed by measuring cytokine secretion, antigen uptake, and processing capacity.

By integrating these datasets, we aim to link donor age and sex to reprogramming efficiency and APC function, thereby leveraging cell reprogramming as a human model to uncover intrinsic mechanisms driving age- and sex-related differences in immune competence.

Tianze Cao | PhD Student at Consiglio Lab

Sex hormones and metabolic features impact the immune serum proteome of females with polycystic ovary syndrome

Polycystic ovary syndrome (PCOS) is an endocrine disorder affecting 10% of women worldwide. PCOS patients display reproductive abnormalities including polycystic ovarian morphology and menstrual irregularities, and metabolic alterations such as hyperinsulinemia and an increased risk of type 2 diabetes. Chronic low-grade inflammation is increasingly acknowledged in PCOS, often characterized by elevated IL-6 and C-reactive protein. However, studies thus far have not distinguished whether this immune dysregulation is intrinsic to PCOS or a secondary effect of the syndrome’s associated clinical traits. Furthermore, standard analyses often overlook a holistic understanding of how cytokines function together as a coordinated immune event.

We profiled the immune landscape of the serum proteome of 106 PCOS patients and 58 female controls using the Olink Target 96 Inflammation panel. We utilized parallel mediation analysis to untangle immune alterations explained by dysregulated clinical traits, and employed Weighted Gene Co-expression Network Analysis (WGCNA) to identify coordinated cytokine modules in PCOS.

PCOS samples show significant increases in damage-associated molecular patterns (S100A12, IL-1A), inflammasome markers (IL-18, IL-18R1), and the immune exhaustion marker PD-L1, revealing immune activation and exhaustion in PCOS. Interestingly, for cytokines known to rise in PCOS, no difference in TNF was detectable in our cohort, while IL-6 showed the largest effect size (~30% level increase). Mediation analysis revealed that most cytokines’ elevation was found to be attributable to HOMA-IR (Homeostatic Model Assessment for Insulin Resistance), indicating the significant contribution of insulin resistance to immune cytokine dysregulation in PCOS. The elevations in S100A12, IL-1A and PD-L1 were independent of insulin resistance and testosterone, suggesting these markers represent a global consequence of the PCOS condition. 

Additionally, WGCNA identified a PCOS-unique co-expression module linking IL-10 with CXCL9-11 and IFNG, indicating aberrant regulation of type 1 immunity. Lastly, we observe two distinct PCOS immune subgroups, with TNFRSF9 being the most predictive biomarker for subgroup 2. TNFRSF9 (CD137) is a co-stimulatory receptor expressed on activated lymphocytes. High serum levels of TNFRSF9 in subgroup 2 indicate its binding events, which induce proliferation of activated T cells. We found a distinct metabolic profile between these two subgroups, where subgroup 2 was characterized by obesity and significant insulin resistance, while subgroup 1 exhibited a similar BMI level to controls and HOMA-IR level within the normal range, indicating TNFRSF9’s role in linking immune activation with metabolic dysfunction in PCOS.

Our findings reveal the immune activation and exhaustion as core immune dysregulation in PCOS, closely related to its insulin resistance. The PCOS immune subgroups exhibit distinct metabolic features, highlighting the heterogeneity of PCOS, emphasizing the potential of immunophenotyping to guide personalized treatment.

Elia Pennati | PhD Student at Bourgine Lab

A vascularized, mechanically active bone marrow-on-chip platform as a complex model of human bone marrow niches

The bone marrow (BM) is the primary site of hematopoiesis and a central component of the immune system. In its hematopoietically active regions, hematopoietic stem cells (HSCs) interact with stromal and vascular cells within a highly organized 3D microenvironment. These interactions are essential for regulating hematopoiesis and maintaining immune homeostasis. Growing interest in BM biology highlights the need for in-vitro models that reproduce its structural and functional complexity. Nevertheless, existing 3D BM models still oversimplify the native tissue, lacking full integration of endothelial, hematopoietic, mesenchymal, and bone compartments, as well as the mechanical cues present in-vivo.
To address these limitations, we developed a vascularized BM-on-chip (BMoC) platform coinceived to recreate the three main human BM niches: vascular, perivascular, and endosteal. To this aim, a five-channel microfluidic cell culture chamber was engineered for the spatially-controlled co-culture of mesenchymal stem cells (MSCs) differentiated toward osteogenic phenotype, in direct contact with a combination of MSCs, endothelial cells (HUVECs) and HSCs, all embedded in fibrin gel. To mimic the mechanical cues perceived in the BM, the culture chamber is coupled with an actuation compartment imposing a compressive mechanical stimulation.

Within the marrow compartment, a perfusable and interconnected microvascular network self-assembled within seven days. Vascular perfusion was validated by the dynamic intraluminal passage of fluorescent microbeads and FITC-Dextran, confirming the formation of functional vessels. Notably, a considerable number of CD34⁺ HSCs was also detected within the device, suggesting that the reconstructed in-vitro microenvironment supports preservation of stemness-associated phenotypes.

Our vascularized BMoC provides a biologically relevant model of the human BM microenvironment and represents a promising platform to study hematopoiesis, immune regulation, and disease mechanisms.
This project received support from the European Union’s Horizon Europe research and innovation programme under grant agreement No.101130604.