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Biology subjects

Xuan, J.

Publications and source records attributed to Xuan, J..

7 recordsLinked to original sources

MAE-UNETR++: Masked Autoencoder Pretraining for 3-D Lung Nodule Segmentation

Voxel-level annotation for volumetric medical imaging is expensive and difficult to scale, which makes training highcapacity 3-D segmentation models challenging in practice. Transfer learning (TL) from large public datasets is a common remedy, but it can under-perform when the source domain differs from the target anatomy and acquisition characteristics, as is often the case for pulmonary nodules. In this work, we propose a masked autoencoder (MAE) pretraining-based approach to break the data efficiency wall of domain difference and present a focused empirical study of domain-specific self-supervised learning (SSL) for 3-D lung nodule segmentation. We evaluate two experimental settings: first, Masked Autoencoder (MAE) pretraining versus random initialization across representative baselines; second, MAE versus Decathlon TL for UNETR++ while testing whether MAE-based pretraining also benefits a CNN baseline (V-Net). MAE pretraining on target-domain CT volumes achieves a Dice Similarity Coefficient (DSC) of 0.307, outperforming random initialization (0.136) and Decathlon weights (0.257). In addition, MAE improves the stability of V-Net in a "low-data" regime (i.e., with "insufficiently labeled" data), increasing DSC from 0.010 to0.071. Overall, these results suggest that MAE-based pretraining can provide a practical and robust initialization strategy for volumetric segmentation when labeled data are limited.

bioengineering↗

Comparative profiling of carnitine palmitoyltransferase 1 isoforms reveals vincamine as a selective carnitine palmitoyltransferase 1b inhibitor

Carnitine palmitoyltransferase 1 (CPT1) catalyzes the rate-limiting step of fatty acid oxidation and has emerged as a therapeutic target for metabolic diseases and cancer. CPT1 exists in three isoforms, CPT1a, CPT1b, and CPT1c, with distinct tissue distributions and enzymatic properties; however, limitations of previous platforms enabling parallel isoform comparison have undermined efforts to identify selective inhibitors that could minimize off-target effects. Here, we describe a DTNB-based enzyme activity assay adapted for high-throughput screening of CPT1b, the predominant isoform in cardiac and skeletal muscle. Mitochondrial extracts from Expi293F cells transfected with CPT1a or CPT1b expression plasmids served as sources of catalytically active enzymes. The assay was validated using three previously confirmed CPT1b inhibitors: (R)-(+)-etomoxir, perhexiline, and malonyl-CoA. We then generated side-by-side inhibitory profiles for both isoforms, identifying vincamine as a lead selective inhibitor of CPT1b. Furthermore, chlorpromazine, previously characterized only as a broad CPT1 inhibitor and subsequently shown to inhibit CPT1a, is demonstrated here to also inhibit CPT1b, expanding its known isoform profile. Together, these results establish a robust platform for comparative isoform profiling and demonstrate that selective modulation of CPT1b is achievable, with implications for targeted therapeutics in metabolic and oncological disease.

cell biology↗

Super-CUT&Tag: A Sensitive, Spatially Resolved Approach for Epigenomic Profiling in Tissue Sections

Spatial profiling of protein-DNA interactions is essential for understanding gene regulatory processes, but current methods suffer from low sensitivity, limited resolution, and complex workflows. Here we introduce Super-CUT&Tag (Solid-phase interface embedded with an Ultra-dense DNA barcode array to Profile Epigenomic landscapes for spatially Resolved CUT&Tag), a robust spatial epigenomic method that directly transfers in situ protein-DNA interactions from tissue sections onto a pre-fabricated spatially barcoded array. Incorporating 3D polyamidoamine (PAMAM) dendrimers markedly improves chromatin capture efficiency, achieving several-fold higher sensitivity compared to current spatial CUT&Tag approaches. Using Super-CUT&Tag, we profile H3K27ac spatial landscape and uncover previously hidden active-chromatin heterogeneity across mouse embryonic tissues and stages. Integration with spatial transcriptomics further reveals spatiotemporal enhancer regulation dynamics of Neurod2 during corticogenesis. By overcoming key bottlenecks in sensitivity, resolution and usability, Super-CUT&Tag establishes a powerful platform for spatial epigenomic profiling in complex tissues.

genetics↗

Local nucleolar surveillance mechanisms maintain rDNA stability

The nucleoli are subdomains of the nucleus that form around actively transcribed ribosomal RNA (rRNA) genes. The highly repetitive and transcribed nature of the rRNA genes (rDNA) by RNA polymerase I (Pol I) poses a challenge for DNA repair and replication machineries. Here, we profile the nucleolar proteome and the chromatin landscape of stalled replication sites upon rDNA damage to characterize the early steps of nucleolar DNA damage response (nDDR). We observed early dynamics in nucleolar-nucleoplasmic proteome localization and identified nucleolar replication stress signatures involving chromatin remodeling networks, transcription-replication conflicts and DNA repair. Our findings define localized surveillance mechanisms that activate the nDDR. Further, we identified that upon rDNA damage, nucleolar RNA Polymerase (Pol) II binds to intergenic rDNA sequences and generates R-loops (DNA:RNA hybrid structures) that are essential for recruiting nDDR factors. Using a boutique CRISPR-Cas9 synthetic lethal screen of DNA repair factors with inhibitors of RNA Pol I transcription, we identified an unexpected protective role for the DNA translocase RAD54L in nDDR. Loss of Rad54L increases nucleolar R-loops and rDNA damage leading to defects in nucleolar structure and enhanced sensitivity to PARP and RNA Pol I inhibitors. Altogether, our study uncovers localized surveillance networks within the nucleolus that respond to rDNA damage. These insights expand our understanding of the molecular mechanisms governing nDDR and opens new avenues for developing nDDR-targeting therapies.

molecular biology↗

Dual indexed design of in-Drop single-cell RNA-seq libraries improves sequencing quality and throughput

The increasing demand of single-cell RNA-sequencing (scRNA-seq) experiments, such as the number of experiments and cells queried per experiment, necessitates higher sequencing depth coupled to high data quality. New high-throughput sequencers, such as the Illumina NovaSeq 6000, enables this demand to be filled in a cost-effective manner. However, current scRNA-seq library designs present compatibility challenges with newer sequencing technologies, such as index-hopping, and their ability to generate high quality data has yet to be systematically evaluated. Here, we engineered a new dual-indexed library structure, called TruDrop, on top of the inDrop scRNA-seq platform to solve these compatibility challenges, such that TruDrop libraries and standard Illumina libraries can be sequenced alongside each other on the NovaSeq. We overcame the index-hopping issue, demonstrated significant improvements in base-calling accuracy, and provided an example of multiplexing twenty-four scRNA-seq libraries simultaneously. We showed favorable comparisons in transcriptional diversity of TruDrop compared with prior library structures. Our approach enables cost-effective, high throughput generation of sequencing data with high quality, which should enable more routine use of scRNA-seq technologies.

molecular biology↗

ChIP-BIT2: a software tool to detect weak binding events using a Bayesian integration approach

Transcription factor binding events play important functional roles in gene regulation. It is, however, a challenging task to detect weak binding events since the ambiguity in differentiation of weak binding signals from background signals. We present a software package, ChIP-BIT2, to identify weak binding events using a Bayesian integration approach. By integrating signals from sample and input ChIP-seq data, ChIP-BIT2 can detect both strong and weak binding events at gene promoter, enhancer or the whole genome effectively. The ChIP-BIT2 package has been extensively tested on ChIP-seq data, demonstrating its wide applicability in ChIP-seq data analysis.\n\nAvailability and ImplementationThe ChIP-BIT2 package is available at http://sourceforge.net/projects/chipbitc/.

bioinformatics↗

MSIGNET: a Metropolis sampling-based method for global optimal significant network identification

In this paper, we propose a novel approach namely MSIGNET to identify subnetworks with significantly expressed genes by integrating context specific gene expression and protein-protein interaction (PPI) data. Specifically, we integrate differential expression of each gene and mutual information of gene pairs in a Bayesian framework and use Metropolis sampling to identify functional interactions. During the sampling process, a conditional probability is calculated given a randomly selected gene to control the network state transition. Our method provides global statistics of all genes and their interactions, and finally achieves a global optimal sub-network. We apply MSIGNET to simulated data and have demonstrated its superior performance over comparable network identification tools. Using a validated Parkinson data set we show that the network identified using MSIGNET is consistent to previously reported results but provides more biology meaningful interpretation of Parkinsons disease. Finally, to study networks related to ovarian cancer recurrence, we investigate two patient data sets. Identified networks from independent data sets show functional consistence. And those common genes and interactions are well supported by current biological knowledge.

bioinformatics↗