Kennedy Center

Abstract Extracellular particles (EP) released by cells are heterogenous, with extracellular vesicles (EV) believed to be the major component. To investigate different EV subpopulations, we conducted a panel screen of 15 fluorescent EV markers in 4 adherent cell lines. Nanoflow cytometry identifies TSPAN14 as the best EV marker in 3 of 4 cell lines. Surprisingly, less than 50% of EPs are labelled by the top 2 EV markers in half the cell lines. The EV marker negative EPs are detergent resistant, non-lipophilic, dense and have Thrombospondin shells by dSTORM; and were thus named Thrombospondin Encapsulated Dense particles (ThrEDs). ThrEDs are ubiquitous, contain extracellular matrix regulators and are released following cell stress. ThrEDs accelerate scratch-wound closure and downregulate the fibrotic phenotype in Dupuytren’s disease patient myofibroblasts. The TSP-1 C-terminus is exposed on the surface of ThrEDs, opening engineering applications.

Abstract Quantifying the structure and dynamics of species interactions in ecological communities is fundamental to studying ecology and evolution. While there are numerous approaches to analysing ecological networks, there is not yet an approach that can (1) quantify dissimilarity in the global structure of ecological networks that range from identical species and interaction composition to zero shared species or interactions and (2) map species between such networks while incorporating additional ecological information, such as species traits or abundances. To address these challenges, we introduce the use of optimal transport distances to quantify ecological network dissimilarity and functionally equivalent species between networks. Specifically, we describe the Gromov–Wasserstein (GW) and Fused Gromov–Wasserstein (FGW) distances. We apply these optimal transport methods to synthetic and empirical data, using mammal food webs throughout sub‐Saharan Africa for illustration. We showcase the application of GW and FGW distances to identify the most functionally similar species between food webs, incorporate additional trait information into network comparisons and quantify food web dissimilarity among geographic regions. Our results demonstrate that GW and FGW distances can effectively differentiate ecological networks based on their topological structure while identifying functionally equivalent species, even when networks have different species. The FGW distance further improves node mapping for basal species by incorporating node‐level traits. We show that these methods allow for a more nuanced understanding of the topological similarities in food web networks among geographic regions compared to an alternative measure of network dissimilarity based on species identities. Optimal transport distances offer a new approach for quantifying functional equivalence between networks and a measure of network dissimilarity suitable for a broader range of uses than existing approaches. OT methods can be harnessed to analyse ecological networks at large spatial scales and compare networks among ecosystems, realms or taxa. Optimal transport‐based distances, therefore, provide a powerful tool for analysing ecological networks with great potential to advance our understanding of ecological community structure and dynamics in a changing world.

Silicon Valley Clean Water (SVCW) provides wastewater treatment for over 220,000 users in the San Francisco mid-peninsula area. The treatment plant, established in the 1970s, faced significant challenges due to an aging piping system. Particularly, the plant’s sludge under their digester and secondary clarifier started showing signs of corrosion, exacerbated by their location near the saline San Francisco Bay. To address this issue, SVCW explored several rehabilitation options, ultimately identifying spray-in-place pipe (SIPP) lining and cured-in-place pipe (CIPP) as the most practical and cost-effective solutions. Despite severe corrosion, varying pipe diameters, difficult access, and tight schedules, the project team successfully implemented SIPP rehabilitation. This accomplishment highlights the importance of innovative thinking, skilled craftsmanship, and teamwork in resolving complex infrastructure challenges.

Abstract The autoimmune disease systemic lupus erythematosus (SLE) is associated with genetic variants in the X-linked gene CXORF21, which encodes the protein TASL. TASL acts as an adaptor in the IRF5 pathway and is necessary for the phosphorylation of IRF5 in response to TLR7 or TLR9 stimulation. Here, we investigate the role of TASL in the humoral immune response, and in the development of lupus in the B6.MRLlpr murine model of SLE. We find that while TASL is dispensable for their development, it is required for the full activation of B cells via endolysosomal TLR stimulation, and consequent interferon signalling and inflammatory cytokine expression. Additionally, TASL is crucial for the emergence of age-associated B cells (ABCs), a B cell population derived from the extrafollicular response that increases with age and is expanded in autoimmune disease, and the production of IgG2c antibodies. We also find that deletion of TASL prevents the onset of autoimmunity in the genetically-determined B6.MRLlpr model of lupus.

Targeting the JAK/STAT pathway has emerged as a key therapeutic strategy for managing Rheumatoid Arthritis (RA). JAK inhibitors suppress cytokine-mediated signaling, including the critical IL-6/STAT3 axis, thereby effectively targeting different aspects of the pathological process. However, despite their clinical efficacy, a subset of RA patients remains refractory to JAK inhibition, underscoring the need for alternative approaches. Here, we identify a novel JAK-independent mechanism of STAT3 activation, which is triggered by the formation of the immunological synapse (IS) in naive CD4+ T cells. Our data demonstrates that LCK mediates the TCR-dependent phosphorylation of STAT3 at the IS, highlighting this pathway as a previously unrecognized hallmark of early T cell activation. Furthermore, we show that the synaptic LCK/TCR-STAT3 pathway is compromised in RA. This discovery highlights a new therapeutic target for RA beyond JAK inhibitors, offering potential avenues for treating patients resistant to current therapies.

Ribosomally synthesized and post-translationally modified peptides (RiPPs) are a promising source of new pharmaceuticals, yet the therapeutic potential of fungal RiPPs remains largely underexplored. Here we report asperigimycins as a distinct class of fungal RiPPs, featuring a unique heptacyclic scaffold consisting of a benzofuranoindoline core and three additional macrocycles, primarily assembled by six distinct fungi-specific DUF3328 oxidases. Inspired by the enhancement of anticancer activity through the N-terminal pyroglutamate in naturally occurring asperigimycins C and D, we chemically modify the inactive asperigimycin B with a series of lipid substitutions at its N-terminus. A derivative with a C-11 linear fatty acid, 2-L6, achieves nanomolar anticancer potency comparable to that of clinically approved antileukemia drugs. High-throughput CRISPR screening identifies the SLC46A3 transporter as a critical factor mediating 2-L6 cellular uptake into human cells. Our findings highlight the promise of engineering asperigimycins as therapeutic leads for cancer treatment.

In Sjogren’s disease (SjD), the immune system targets the salivary and lacrimal glands, causing chronic inflammation, glandular dysfunction, and reduced tear and saliva production, leading to xerostomia and keratoconjunctivitis sicca1,2. The key immune mechanisms include B cell hyperactivity, marked by elevated circulating anti-SSA (Ro60 and Ro52), anti-SSB (La), and rheumatoid factors (RFs: IgM-RF, IgA-RF, and IgG-RF) antibodies, elevated expression of B cell activation and proliferation markers (e.g., CD86, CD38, HLA-DR, Ki67), increased secretion of IL6, BAFF, along with CD4+ T cell-driven inflammation and the formation of ectopic lymphoid structures, contributing to sustained autoimmunity

Abstract Background Leukocyte migration to the vasculature and the heart plays a critical role in hypertensive immune responses and is a carefully orchestrated process, regulated by the interactions of inflammatory chemokines with their receptors, notably CCR1, CCR2, CCR3, and CCR5 (collectively termed inflammatory chemokine receptors (iCCRs)). However, the inflammatory chemokine/receptor system is characterised by redundancy, ligand sharing and overlapping expression patterns. Consequently, our understanding of the specific and combinatory roles of chemokine receptors in cardiovascular inflammation remains incomplete, thus hindering the development of targeted therapies. To address this challenge, we have utilised two novel mouse models: iREP mice, carrying fluorescent reporters of inflammatory chemokine receptor expression, and TACKO mice, in which the entire iCcr locus containing Ccr1, Ccr2, Ccr3, and Ccr5 has been deleted. Purpose To evaluate the expression pattern of iCCRs in experimental hypertension and understand how the global deletion of iCCRs impacts disease progression. Methods We induced hypertension in mice by subcutaneously implanting osmotic minipumps administering angiotensin II (Ang II) for a duration of 14 days. Single-cell RNA sequencing was utilised for investigating iCCR expression in aortas during hypertension. Additionally, we employed flow cytometry and confocal microscopy to track iCCR expression and localization in aortas and hearts obtained from iREP mice. TACKO mice were utilised to assess the effect of global iCCr deficiency on hypertensive phenotypes. Blood pressure was measured via telemetry and vascular function was assessed using myography. Vascular and heart remodelling were evaluated through histology. Molecular mechanisms were analysed using Western Blotting and RT-qPCR, while immune cell changes in the aorta were evaluated through cytometry of time-of-flight (CyTOF). Results Ang II infusion induced higher iCCRs expression levels in both the aorta and the heart in comparison with sham treatment, particularly in fibrotic areas. After Ang II-treatment, global deletion of iCCRs in TACKO mice resulted in the prevention of endothelial dysfunction (n=5/group, P<0.05) and perivascular fibrosis (n=6-9, P<0.01), and a reduction in cardiomyocyte size (n=12-16, P<0.01) compared to wild type mice. However, no significant improvements were observed in other hypertensive phenotypes, including blood pressure, heart fibrosis, vascular NO synthase, and oxidative stress. CyTOF analysis revealed reductions in inflammatory monocytes and conventional type 2 dendritic cells in TACKO aorta, accompanied by a phenotypic switch in macrophages towards a less inflammatory phenotype. Conclusion Deficiency in iCCRs confers protective effects against vascular dysfunction in experimental hypertension, primarily by selectively reducing inflammatory myeloid cell homing to the aorta.

Background/Objectives: Iron (Fe) is a co-factor for enzymes of the developing brain necessitating sufficient supply. We investigated the effects of administering ferric derisomaltose/Fe isomaltoside (FDI) subcutaneously to Fe-deficient (ID) pregnant rats on cerebral and hepatic concentrations of essential metals and the expression of iron-relevant genes. Methods: Pregnant rats subjected to ID were injected with FDI on the day of mating (E0), 14 days into pregnancy (E14), or the day of birth (postnatal (P0)). The efficacy was evaluated by determination of cerebral and hepatic Fe, copper (Cu), and zinc (Zn) and gene expression of ferroportin, hepcidin, and ferritin H + L in pups on P0 and as adults on P70. Results: Females fed an ID diet (5.2 mg/kg Fe) had offspring with significantly lower cerebral and hepatic Fe compared to female controls fed a standard diet (158 mg/kg Fe). Cerebral Cu increased irrespective of supplying a standard diet or administering FDI combined with the standard diet. Hepatic hepcidin mRNA was significantly lower following ID. Cerebral hepcidin mRNA was hardly detectable irrespective of iron status. Conclusions: In conclusion, administering FDI subcutaneously to ID pregnant rats on E0 normalizes fetal cerebral and hepatic Fe. When applied at later gestational ages, supplementation with additional Fe to the offspring is needed to normalize cerebral and hepatic Fe.