Biology subjects

Ferrante, T. C.

Publications and source records attributed to Ferrante, T. C..

4 recordsLinked to original sources

bioRxiv2025

Cytokines control the physical state of immune tissue

Unlike most solid-like tissues, immune tissue is protean and reconfigurable--its component cells can patrol vast territories, find rare cellular partners, and dynamically self-organize into functional structures like germinal centers or tertiary lymphoid structures. The ability of immune cells to collectively behave as an active material that switches physical state is essential for adaptive immunity. Yet the principles governing the control of the physical properties of immune tissues, and how chemical and mechanical forms of control interplay remain obscure. Inspired by minimal immune cell components that recapitulate germinal center (GC) dynamics, we developed an agent-based model that predicted that a tissues ability to program its own physical properties governs the efficiency of antibody evolution. This motivated our search for biomolecular forms of control capable of tuning this material property. Using a tractable in vitro model, we found that CD40L-stimulated human B cells self-organize into active, liquid-like condensates that round, fuse, and internally mix. GC cytokines IL-4 and IL-21 tuned this effective temperature, driving phase transitions from cohesive, liquid-like states to dispersed, gas-like morphologies. These findings demonstrate that cytokines control tissue fluidity, suggesting a previously unrecognized biophysical form of immune regulation. Significance StatementThe bodys immune system is uniquely flexible, allowing cells to organize into temporary structures, like germinal centers or tertiary lymphoid structures, to fight intruders. How this physical flexibility is controlled is a major question. We discovered that the physical state of immune tissue is chemically programmable. We show that self-organized multicellular B cell structures behave like liquid droplets, and we used this framework to demonstrate that signals (cytokines) can modulate their physical properties. This finding identifies a new biophysical axis of immune control, suggesting the body actively tunes its tissue fluidity to optimize the immune response.

biophysics↗

bioRxiv2024

Organ Chips with integrated multifunctional sensors enable continuous metabolic monitoring at controlled oxygen levels

Despite remarkable advances in Organ-on-a-chip (Organ Chip) microfluidic culture technology, recreating tissue-relevant physiological conditions, such as the region-specific oxygen concentrations, remains a formidable technical challenge, and analysis of tissue functions is commonly carried out using one analytical technique at a time. Here, we describe two-channel Organ Chip microfluidic devices fabricated from polydimethylsiloxane and gas impermeable polycarbonate materials that are integrated with multiple sensors, mounted on a printed circuit board and operated using a commercially available Organ Chip culture instrument. The novelty of this system is that it enables the recreation of physiologically relevant tissue-tissue interfaces and oxygen tension as well as non-invasive continuous measurement of transepithelial electrical resistance, oxygen concentration and pH, combined with simultaneous analysis of cellular metabolic activity (ATP/ADP ratio), cell morphology, and tissue phenotype. We demonstrate the reliable and reproducible functionality of this system in living human Gut and Liver Chip cultures. Changes in tissue barrier function and oxygen tension along with their functional and metabolic responses to chemical stimuli (e.g., calcium chelation, oligomycin) were continuously and noninvasively monitored on-chip for up to 23 days. A physiologically relevant microaerobic microenvironment that supports co-culture of human intestinal cells with living Lactococcus lactis bacteria also was demonstrated in the Gut Chip. The integration of multi-functional sensors into Organ Chips provides a robust and scalable platform for the simultaneous, continuous, and non-invasive monitoring of multiple physiological functions that can significantly enhance the comprehensive and reliable evaluation of engineered tissues in Organ Chip models in basic research, preclinical modeling, and drug development.

bioengineering↗

bioRxiv2022

Optimizing CpG spatial distribution with DNA origami for Th1-polarized therapeutic vaccination

Multivalent presentation of ligands often enhances receptor activation and downstream signaling. DNA origami offers precise nanoscale spacing of ligands, a potentially useful feature for therapeutic nanoparticles. Here we introduce a "square block" DNA origami platform to explore the importance of spacing of CpG oligonucleotides, which engage Toll-like receptors and thereby act as danger signals for dendritic cells. Through in vitro cell-culture studies and in vivo tumor-treatment models, we demonstrate that square blocks induce Th1 immune polarization when CpG is spaced at 3.5 nm. We observe that this DNA origami vaccine enhances DC activation, antigen cross-presentation, CD8 T cell activation, Th1-polarized CD4 activation and NK cell activation. The vaccine also synergizes effectively with anti-PD-L1 for improved cancer immunotherapy in melanoma and lymphoma models and induces long-term T cell memory. Our results suggest that DNA origami may serve as an advanced vaccine platform for controlling adjuvant spacing and co-delivering antigens. One Sentence SummaryThis study developed a DNA origami-based cancer vaccine (DoriVac) that co-delivers antigen and CpG immune adjuvant with an optimal nanospacing for Th1 immune polarization.

synthetic biology↗

bioRxiv2019

Human colon-on-a-chip enables continuous in vitro analysis of colon mucus layer accumulation and physiology

Background & AimsThe mucus layer in the human colon protects against commensal bacteria and pathogens, and defects in its unique bilayered structure contribute to intestinal disorders, such as ulcerative colitis. However, our understanding of colon physiology is limited by the lack of in vitro models that replicate human colonic mucus layer structure and function. Here, we investigated if combining organ-on-a-chip and organoid technologies can be leveraged to develop a human-relevant in vitro model of colon mucus physiology. MethodsA human colon-on-a-chip (Colon Chip) microfluidic device lined by primary patient-derived colonic epithelial cells was used to recapitulate mucus bilayer formation, and to visualize mucus accumulation in living cultures non-invasively. ResultsThe Colon Chip supports spontaneous goblet cell differentiation and accumulation of a mucus bilayer with impenetrable and penetrable layers, and a thickness similar to that observed in human colon, while maintaining a subpopulation of proliferative epithelial cells. Live imaging of the mucus layer formation on-chip revealed that stimulation of the colonic epithelium with prostaglandin E2, which is elevated during inflammation, causes rapid mucus volume expansion via an NKCC1 ion channel-dependent increase in its hydration state, but no increase in de novo mucus secretion. ConclusionThis study is the first to demonstrate production of colonic mucus with a physiologically relevant bilayer structure in vitro, which can be analyzed in real-time non-invasively. The Colon Chip may offer a new preclinical tool to analyze the role of mucus in human intestinal homeostasis as well as diseases, such as ulcerative colitis and cancer.

cell biology↗