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Felix, F.

Publications and source records attributed to Felix, F..

4 recordsLinked to original sources

SAMMBA is a high-throughput pipeline for isolating and phenotyping macroalgal strains

Anthropogenic climate change is causing the decline of seaweed forests in many parts of the world. Despite successful preservation efforts, their immense biodiversity is still severely underrepresented in germplasm biobanks throughout the world. These culture libraries can preserve genetic diversity and provide inoculum for marine forest restoration and mariculture ventures, and potentially accelerate the selection and breeding of climate-resilient and high-yielding strains. However, the complex life cycles and body plans of seaweeds pose a huge challenge for the development of standardized phenotyping and isolating protocols for microscopic stages, especially with the efficiency necessary to deal with the current pace of global climatic changes. Here, we present SAMMBA (Seaweed Automatable Microplate Microscopy for Breeding Approaches), an end-to-end pipeline for the high-throughput isolation, phenotyping and storage of macroalgal cells in 384-well plates (384WP). By optimizing fluorescence microscopy imaging and analysis, along with a novel fragmentation method and dilution-to-extinction isolation, different unialgal seaweed tissues could be regrown after thousand-fold dilutions. In a single plate, we successfully isolated 68 singlet gametophyte fragments of Laminaria ochroleuca (39 males, 29 females; 17.7% efficiency) and 60 spores of Phyllariopsis purpurascens (31.25% efficiency). Furthermore, the taxonomic versatility of SAMMBA was demonstrated through the successful isolation of 60 unialgal cultures of red algae (Halymenia sp., Hydrolithon sp., Erythrotrichia sp.) and 10 strains of the green alga Ulva sp, without cross-contamination. The viability and unialgal nature of the isolated strains were verified by distributing a single L. ochroleuca strain across an entire 384-well plate and imaging each well over 30 days. We found that the average specific daily growth rates (daily SGR) per well were 0.130 {+/-} 0.006 and 0.117 {+/-} 0.01 day-1 for males and females, respectively, showing a significant difference between sexes (n = 768; p = 1.27e-53), while edge effects significantly reduced daily SGR in males but not in females. This approach dramatically increases experimental reproducibility and statistical power compared to conventional methods. Due to its modular design and cost-effectiveness, SAMMBA is readily adaptable to macroalgal repositories globally. It supports high-throughput, selective recovery of unialgal strains without reliance on robotic platforms, while remaining fully compatible with automation. This system significantly expands the experimental and operational capacity in macroalgal hatcheries, providing a scalable foundation for phenomics, domestication programs, and standardized, verifiable biobanking of unialgal strains. Ultimately, SAMMBA could provide critical support for breeding strategies required to ensure the resilience of marine forests and aquaculture in a rapidly changing ocean.

plant biology↗

Integration of Protein Interactome Networks with Congenital Heart Disease Variants Reveals Candidate Disease Genes

Congenital heart disease (CHD) is present in 1% of live births, yet identification of causal mutations remains a challenge despite large-scale genomic sequencing efforts. We hypothesized that genetic determinants for CHDs may lie in protein interactomes of GATA4 and TBX5, two transcription factors that cause CHDs. Defining their interactomes in human cardiac progenitors via affinity purification-mass spectrometry and integrating results with genetic data from the Pediatric Cardiac Genomic Consortium revealed an enrichment of de novo variants among proteins that interact with GATA4 or TBX5. A consolidative score that prioritized interactome members based on variant, gene, and proband features identified likely CHD-causing genes, including the epigenetic reader GLYR1. GLYR1 and GATA4 widely co-occupied cardiac developmental genes, resulting in co-activation, and the GLYR1 missense variant associated with CHD disrupted interaction with GATA4. This integrative proteomic and genetic approach provides a framework for prioritizing and interrogating the contribution of genetic variants in disease.

genetics↗

A Transcriptional Switch Governing Fibroblast Plasticity Underlies Reversibility of Chronic Heart Disease

In diseased organs, stress-activated signaling cascades alter chromatin, triggering broad shifts in transcription and cell state that exacerbate pathology. Fibroblast activation is a common stress response that worsens lung, liver, kidney and heart disease, yet its mechanistic basis remains poorly understood1,2. Pharmacologic inhibition of the BET family of transcriptional coactivators alleviates cardiac dysfunction and associated fibrosis, providing a tool to mechanistically interrogate maladaptive fibroblast states and modulate their plasticity as a potential therapeutic approach3-8. Here, we leverage dynamic single cell transcriptomic and epigenomic interrogation of heart tissue with and without BET inhibition to reveal a reversible transcriptional switch underlying stress-induced fibroblast activation. Transcriptomes of resident cardiac fibroblasts demonstrated robust and rapid toggling between the quiescent fibroblast and activated myofibroblast state in a manner that directly correlated with BET inhibitor exposure and cardiac function. Correlation of single cell chromatin accessibility with cardiac function revealed a novel set of reversibly accessible DNA elements that correlated with disease severity. Among the most dynamic elements was an enhancer regulating the transcription factor MEOX1, which was specifically expressed in activated myofibroblasts, occupied putative regulatory elements of a broad fibrotic gene program, and was required for TGF{beta}-induced myofibroblast activation. CRISPR interference of the most dynamic cis-element within the enhancer, marked by nascent transcription, prevented TGF{beta}-induced activation of Meox1. These findings identify MEOX1 as a central regulator of stress-induced myofibroblast activation associated with cardiac dysfunction. The plasticity and specificity of the BET-dependent regulation of MEOX1 in endogenous tissue fibroblasts provides new trans- and cis- targets for treating fibrotic disease.

molecular biology↗

CCR3 antagonist impairs estradiol induced eosinophil migration to the uterus in ovariectomized mice

The eosinophils in the uterus was associated with the successful implantation of the embryo. In addition, they are prevalent in the female reproductive tract, where they may contribute to remodeling and maintenance of epithelial integrity. However, the mechanisms by which eosinophils migrate into the uterus are still not entirely clear, since there are a variety of chemotactic factors that can cause migration of these cells. Therefore, to evaluate the CCR3 participation on eosinophil migration, we using ovariectomized C57BL/6 mice treated with CCR3 antagonist SB328437 and 17-{beta}-estradiol (E2). Thus, it was possible to confirm the hypothesis that the CCR3 receptor has an important role in the migration of these cells to the uterus of mice, because it was clearly observed histologically that the antagonist reduces this migration and activity eosinophil peroxidase (EPO) corroborated this result. Concomitant to this, there was a 46% reduction in uterine edema, as demonstrated by the uterus weight and histology. Together, these data suggest that the CCR3 is an important receptor involved in the eosinophils regulation in uterus, including influencing the uterine edema.

physiology↗