Biology subjects

Burnham, P.

Publications and source records attributed to Burnham, P..

9 recordsLinked to original sources

bioRxiv2026

Diversification of biofilm architecture among freshwater Pararheinheimera isolates

Air-liquid interfaces (ALIs) at the upper layer of oceans, lakes and rivers cover the majority of the earths surface. Microbes are known to accumulate at these resource-rich boundaries, but the mechanisms of ALI colonization are often assumed to mirror the formation of pellicle biofilms by non-aquatic organisms. Here, we analyzed ALI colonization by natural aquatic bacteria. We used samples from a freshwater lake to enrich for microbes that colonize the ALI in liquid growth medium. Mixed-species pellicles formed rapidly in these enrichments, were structurally stable for weeks and displayed a pronounced ecological succession. We isolated 31 members of the genus Pararheinheimera from early stages of mixed-species pellicle maturation. Five phylogenetically distinct Pararheinheimera clades were identified, each with a shared colony morphology. We used representative isolates to show that only one Pararheinheimera clade formed thin, adherent films at the ALI resembling classical pellicles. Isolates from the four remaining clades formed floating structures that could be categorized either as non-adhesive films or large viscous masses. Viscous mass (VM) pellicle formation was a polyphyletic trait that correlated with a highly mucoid appearance on agar plates, suggesting that the process is driven by copious secretion of extracellular matrix. Matrices from VM biofilms were largely non-adhesive, contained a mixture of acidic polysaccharides and proteins and formed thermally stable, shear-thinning hydrogels. Our results demonstrate that ALI colonization strategies vary widely even among closely related aquatic bacteria and identify VM pellicles as a distinct biofilm architecture with unique mechanical properties. ImportanceLakes, rivers and oceans contain a boundary between the air and the waters surface known as the air-liquid interface (ALI). Microbial communities that populate the ALI play crucial roles in nutrient cycling, but how aquatic microbes partition to these sites remains poorly characterized. Our study investigated how bacteria from a freshwater lake accumulate at the ALI. Lake water samples incubated in nutrient medium formed a layer of cells known as a pellicle biofilm at the ALI, and we isolated 31 different bacteria from a genus (Pararheinheimera) that was abundant during the early stages of pellicle formation. Only a subset of Pararheinheimera isolates formed traditional pellicle biofilms. Most formed either thin, non-adhesive films or large, gelatinous aggregates that appeared to persist at the ALI due to buoyancy. These findings expand our understanding of biofilm diversity in aquatic systems and suggest that the production of buoyant hydrogels may play an important role in structuring microbial communities at air-water boundaries.

microbiology↗

bioRxiv2025

Self-Organization Through Local Cell-Cell Communication Drives Intestinal Epithelial Zonation

The intestinal epithelium exhibits zonated gene expression along the crypt-villus axis, with distinct transcriptional programs in enterocytes at the villus top versus bottom. However, the mechanisms establishing these spatial patterns remain unclear. Three models could explain zonation: external gradients, cell-intrinsic temporal programs, or local self-organization. Using spatial transcriptomics and perturbations of two-dimensional intestinal organoids, we show that zonation emerges via spontaneous self-organization without mesenchymal, neural, or vascular inputs. Cell-intrinsic models were eliminated by transplanting cells into established monolayers; transplanted cells progressively adopted zonation profiles matching their new location, with strongly zonated genes showing the greatest adaptive responses. Pharmacological inhibition of EphA2 receptors disrupted zonation, revealing a previously unknown role for epithelial EphA-ephrin-A signaling in regulating enterocyte zonation. These findings demonstrate that self-organization through local epithelial cell-cell communication generates spatial patterns independently of external positional cues or cell-autonomous programs.

systems biology↗

bioRxiv2025

Multiscale Spatial Mapping of Microbial Communities for Biotherapeutic Development

Live biotherapeutic products (LBPs) are emerging as powerful tools to modulate the microbiome using well-defined microbial communities. Yet, designing, manufacturing, and delivering LBPs remains challenging, in part due to a lack of technologies capable of analyzing LBPs as complete, spatially organized consortia. Conventional sequencing-based methods lack sensitivity and specificity and do not provide critical spatial information. To address this, we present high-phylogenetic-resolution spatial mapping platform (HiPR-Map), a state-of-the-art spectral imaging technology that enables precise enumeration and spatial localization of microbial cells at species-level within complex communities. Through these advantages, HiPR-Map provides unique insights for LBP discovery and development. Applying HiPR-Map to an LBP designed to complement immune checkpoint therapy, we profiled over 1.8 million microbial cells engrafted in the murine gut. Our analysis revealed distinctive microbial spatial organization, underscoring the power of imaging-based microbiome profiling to optimize LBP design and characterization. This work highlights the transformative potential of spatial microbiome analysis for next-generation LBP development.

microbiology↗

bioRxiv2020

Cell-free DNA (cfDNA) and exosome profiling from a year-long humanspaceflight reveals circulating biomarkers

The health impact of prolonged space flight on the human body is not well understood. Liquid biopsies based on cell-free DNA (cfDNA) or exosome analysis provide a noninvasive approach to monitor the dynamics of genomic, epigenomic and proteomic biomarkers, and the occurrence of DNA damage, physiological stress, and immune responses. To study the molecular consequences of spaceflight we profiled cfDNA isolated from plasma of an astronaut (TW) during a year-long mission on the International Space Station (ISS), sampling before, during, and after spaceflight, and compared the results to cfDNA profiling of the subjects identical twin (HR) who remained on Earth, as well as healthy donors. We characterized cfDNA concentration and fragment size, and the positioning of nucleosomes on cfDNA, observing a significant increase in the proportion of cell-free mitochondrial DNA inflight, suggesting that cf-mtDNA is a potential biomarker for space flight-associated stress, and that this result was robust to ambient transit from the International Space Station (ISS). Analysis of exosomes isolated from post-flight plasma revealed a 30-fold increase in circulating exosomes and distinct exosomal protein cargo, including brain-derived peptides, in TW compared to HR and all known controls. This study provides the first longitudinal analysis of astronaut cfDNA during spaceflight, as well as the first exosome profiles, and highlights cf-mtDNA levels as a potential biomarker for physiological stress or immune system responses related to microgravity, radiation exposure, and other unique environmental conditions on the ISS.

cell biology↗

bioRxiv2020

Cell-free DNA Tissues-of-Origin Profiling to Predict Graft versus Host Disease and Detect Infection after Hematopoietic Cell Transplantation

Allogeneic hematopoietic cell transplantation (HCT) provides effective treatment for hematologic malignancies and immune disorders. Monitoring of post-transplant complications is critical, yet current diagnostic options are limited. Here, we show that cell-free DNA (cfDNA) in blood is a highly versatile analyte for monitoring of the most important complications that occur after HCT: graft-versus-host disease (GVHD), a frequent immune complication of HCT; infection; relapse of underlying disease; and graft failure. We demonstrate that these different therapeutic complications can be informed from a single assay, low-coverage bisulfite sequencing of cfDNA, followed by disease-specific bioinformatic analyses. To inform GVHD, we profile cfDNA methylation marks to trace the cfDNA tissues-of-origin and to quantify tissue-specific injury. To inform on infections, we implement metagenomic cfDNA profiling. To inform cancer relapse, we implement analyses of tumor-specific genomic aberrations. Finally, to detect graft failure we quantify the proportion of donor and recipient specific cfDNA. We applied this assay to 170 plasma samples collected from 27 HCT recipients at predetermined time points before and after allogeneic HCT. We found that the abundance of solid-organ derived cfDNA in the blood at one-month after HCT is an early predictor of acute graft-versus-host disease (area under the curve, 0.88). Metagenomic profiling of cfDNA revealed the frequent occurrence of viral reactivation in this patient population. The fraction of donor specific cfDNA was indicative of cell chimerism, relapse and remission, and the fraction of tumor specific cfDNA was informative of cancer relapse. This proof-of-principle study shows that cfDNA has the potential to improve the care of allogeneic HCT recipients by enabling earlier detection and better prediction of the complex array of complications that occur after HCT.

genomics↗

bioRxiv2019

Separating the signal from the noise in metagenomic cell-free DNA sequencing

Cell-free DNA (cfDNA) in blood, urine and other biofluids provides a unique window into human health. A proportion of cfDNA is derived from bacteria and viruses, creating opportunities for the diagnosis of infection via metagenomic sequencing. The total biomass of microbial-derived cfDNA in clinical isolates is low, which makes metagenomic cfDNA sequencing susceptible to contamination and alignment noise. Here, we report Low Biomass Background Correction (LBBC), a bioinformatics noise filtering tool informed by the uniformity of the coverage of microbial genomes and the batch variation in the absolute abundance of microbial cfDNA. We demonstrate that LBBC leads to a dramatic reduction in false positive rate while minimally affecting the true positive rate for a cfDNA test to screen for urinary tract infection. We next performed high throughput sequencing of cfDNA in amniotic fluid collected from term uncomplicated pregnancies or those complicated with clinical chorioamnionitis with and without intra-amniotic infection. The data provide unique insight into the properties of fetal and maternal cfDNA in amniotic fluid, demonstrate the utility of cfDNA to screen for intra-amniotic infection, support the view that the amniotic fluid is sterile during normal pregnancy, and reveal cases of intra-amniotic inflammation without infection at term.

genomics↗

bioRxiv2019

A cell-free DNA metagenomic sequencing assay that integrates the damage response to infection

High-throughput metagenomic sequencing offers an unbiased approach to identify pathogens in clinical samples. Conventional metagenomic sequencing however does not integrate information about the host, which is often critical to distinguish infection from infectious disease, and to assess the severity of disease. Here, we explore the utility of high-throughput sequencing of cell-free DNA after bisulfite conversion to map the tissue and cell types of origin of host-derived cell-free DNA, and to profile the bacterial and viral metagenome. We applied this assay to 51 urinary cfDNA isolates collected from a cohort of kidney transplant recipients with and without bacterial and viral infection of the urinary tract. We find that the cell and tissue types of origin of urinary cell-free DNA can be derived from its genome-wide profile of methylation marks, and strongly depend on infection status. We find evidence of kidney and bladder tissue damage due to viral and bacterial infection, respectively, and of the recruitment of neutrophils to the urinary tract during infection. Through direct comparison to conventional metagenomic sequencing as well as clinical tests of infection, we find this assay accurately captures the bacterial and viral composition of the sample. The assay presented here is straightforward to implement, offers a systems view into bacterial and viral infections of the urinary tract, and can find future use as a tool for the differential diagnosis of infections.

genomics↗

bioRxiv2018

Simultaneous multiplexed amplicon sequencing and transcriptome profiling in single cells

We describe Droplet Assisted RNA Targeting by single cell sequencing (DART-seq), a versatile technology that enables multiplexed amplicon sequencing and transcriptome profiling in single cells. We applied DART-seq to simultaneously characterize the non-A-tailed transcripts of a segmented dsRNA virus and the transcriptome of the infected cell. In addition, we used DART-seq to simultaneously determine the natively paired, variable region heavy and light chain amplicons and the transcriptome of B lymphocytes.

genomics↗

bioRxiv2017

Host-pathogen interactions in urinary tract infection from the structure and diversity of urinary cell-free DNA

Infections of the urinary tract are the most common form of infection in the human population. Here, we tested the utility of urinary cell-free DNA (cfDNA) to comprehensively monitor host and pathogen dynamics in the scope of bacterial and viral urinary tract infections. We assayed cfDNA isolated from 141 urine samples obtained from a cohort of 82 kidney transplant recipients by next-generation sequencing. We find that urinary cfDNA simultaneously informs about the composition of the bacterial and viral components of the microbiome, antimicrobial susceptibility, bacterial growth dynamics, kidney allograft injury, and the host response to infection. These different layers of information are accessible from a single assay and individually agree with corresponding clinical tests based on quantitative PCR, conventional bacterial culture, and urinalysis. In addition, cfDNA reveals the frequent occurrence of pathologies that remain undiagnosed in conventional diagnostic workups. Our work identifies urinary cfDNA as a highly versatile tool to monitor infections of the urinary tract.

genomics↗