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Biology subjects

Lester, J.

Publications and source records attributed to Lester, J..

4 recordsLinked to original sources

Genome-wide classification of tumor-derived reads from bulk long-read sequencing

DNA extracted from tissue samples typically derives from a complex mixture of cell types. Without single cell analysis, it has been generally impossible to determine the cell type of origin for most molecules. One clear example of this is in the complex milieu of a human neoplasm. Here, we develop ROCIT (https://github.com/tobybaker/rocit), a transformer-based model to classify the tumor or non-tumor origin of individual reads from bulk tumor samples sequenced with long-read whole genome sequencing. Using somatic mutations to derive training data, ROCIT uses read-level methylation patterns to accurately classify reads from any-where in the genome without requiring the adjacent normal tissue or the explicit identification of tumor differentially methylated regions. We apply ROCIT to a cohort of prostate and ovarian tumors and demonstrate high classification accuracy across the entire genome. We then demonstrate the potential of ROCIT predictions to improve somatic variant calling. ROCIT represents a major step forward in the analysis of bulk tumors with long-reads, enabling the accurate and sensitive identification of reads with specific cell types of origin genome-wide.

bioinformatics↗

Protective Effects of Butyrate on Retinal Neovascularization in Preclinical Retinopathy of Prematurity Models

Retinopathy of prematurity (ROP) remains a leading cause of childhood blindness worldwide, necessitating new therapeutic strategies. Current interventions targeting advanced disease stages often fail to prevent long-term visual impairment. This study investigates the potential of sodium butyrate (NaB), an orally administered short-chain fatty acid, in preclinical models of ROP. Using the oxygen-induced retinopathy (OIR) mouse model, we demonstrate that daily oral NaB supplementation significantly protects against pathological angiogenesis, impacting not only vascular but also neuronal and microglial pathology in the inner retina. Notably, NaB shows efficacy in early-phase ROP intervention, as evidenced by studies in postnatal day 9 (P9) OIR mice and a novel hyperglycemia-associated retinopathy (HAR) model that mimics the hyperglycemic conditions of many premature infants. These findings highlight NaB as a promising alternative or adjunct therapy to current anti-VEGF treatments, offering protection across multiple retinal cell types and stages of ROP development. The study underscores the need for further research to elucidate the specific mechanisms of NaBs action, paving the way for its potential clinical application in ROP management. This research marks the first exploration of butyrate as a preventative and therapeutic agent for ROP, setting the stage for additional preclinical evaluations and optimization.

pharmacology and toxicology↗

DNA methylation landscapes of matched primary and recurrent high grade serous ovarian cancers are preserved throughout disease progression and chemoresistance.

Little is known about the role of global DNA methylation in recurrence and chemoresistance of high grade serous ovarian cancer (HGSOC). We performed whole genome bisulfite sequencing (WGBS) and whole transcriptome sequencing (RNA-seq) to establish methylation and gene expression signatures in 62 primary and recurrent tumors from 28 patients diagnosed with stage III/IV HGSOC. Eleven of these patients carried pathogenic germline BRCA1/BRCA2 mutations. Genome-wide methylation and transcriptomic features identified in primary tumors were largely preserved in matched recurrent tumors from the same patient (P-value = 7.16 x 10-7 and 1.41 x 10-3 in BRCA1/2 and non-BRCA1/2 cases respectively). Tumors from BRCA1/2 carriers displayed high levels of heterogeneity, with significantly more shared methylation changes identified between primary and recurrent tumors from non-BRCA1/2 patients, which may be related to the poorer survival we observe in HGSOCs from non-BRCA1/2 carriers (P-value = 0.0056). Partially methylated domains (PMDs) dominated the epigenetic variation across all tumors, and were more hypomethylated in BRCA1/2 than non-BRCA1/2 cases. Differential gene expression analysis identified upregulation of genes from immune pathways including antigen processing and presentation in tumors from BRCA1/2 carriers, implicating increased immune response in the improved survival observed in these patients. In summary, this study shows a previously unreported conservation of methylation and gene expression in recurrent HGSOCs. These data have implications for the possible effectiveness of epigenetic based therapies to treat both primary and recurrent ovarian cancers.

genomics↗

Non-coding Somatic Mutations Converge on the PAX8 Pathway in Epithelial Ovarian Cancer

Transcriptional regulation is highly disease and cell-type specific. We performed H3K27ac chromatin immunoprecipitation and transcriptomic sequencing in primary tumors for the four different subtypes of invasive epithelial ovarian cancer (OC). Histotype-specific regulatory elements (REs) were enriched in enhancers (P<0.001). In silico prediction of putative target genes for histotype-specific REs identified genes (WFDC2, P=5.5x10-5) and pathways (PI3K-Akt signaling, P<0.002) known to be involved in OC development. Some genes (e.g. PAX8 and CA125) are associated with super-enhancers (SEs) in all OCs, while others are histotype-specific, including PPP1R3B which is associated with SEs specific to clear cell OC. Integrated analysis of active chromatin landscapes with somatic single nucleotide variants (SNVs) from whole genome sequencing (WGS) of 232 primary OCs identified frequently mutated REs, including the KLF6 promoter (P=8.2x10-8) and a putative enhancer at chromosome 6p22.1 (P<0.05). In high-grade serous OCs, somatic SNVs clustered in binding sites for the PAX8 binding partner TEAD4 (P=6x10-11), while the collection of cis regulatory elements associated with PAX8 was the most frequently mutated set of enhancers in OC (P=0.003). Functional analyses supported our findings: Knockdown of PPP1R3B in clear cell OC cells significantly reduced intracellular glycogen content, a signature feature of this histotype; and stable knockout of a 635 bp region in the 6p22.1 enhancer induced downregulation of two predicted target genes, ZSCAN16 and ZSCAN12 (P=6.6 x 10-4 and P=0.02). In summary, we have characterized histotype-specific epigenomic and transcriptomic landscapes in OC and defined likely functional REs based on somatic mutation analysis of ovarian tumors.

cancer biology↗