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Shi, P.

Publications and source records attributed to Shi, P..

At least 19 recordsLinked to original sources

multiTEMPTED: Joint Dimensionality Reduction of Longitudinal Multi-omic Data with Modality-Specific Temporal Dynamics

Longitudinal multi-omic studies profile multiple molecular layers, such as microbiome composition, metabolomics, lipidomics, and proteomics, repeatedly over time. These layers reflect shared subject-level biological processes yet each may exhibit its own temporal dynamics. Most existing methods either integrate multiple omics modalities cross-sectionally or model a single modality longitudinally. The few methods that handle longitudinal multi-omic data assume a shared temporal trajectory across all modalities, limiting their ability to capture modality-specific dynamics, and can be computationally prohibitive at the scale of modern cohort studies. We introduce multiTEMPTED, an extension of the temporal tensor decomposition framework to M>=1 simultaneous omic modalities. The method jointly estimates subject components shared across modalities, modality-specific feature loadings identifying the contributing molecular features, and modality-specific temporal trajectories, while accommodating arbitrary, unaligned sampling schedules across subjects and modalities without imputation. In the MOMS-PI pregnancy cohort, joint analysis of the vaginal microbiome and cervicovaginal cytokines yields clearer separation between preterm and term birth outcomes than microbiome data alone, implicates Lactobacillus-dominant communities as protective, and identifies diverging cytokine trajectories in women who subsequently deliver preterm. In an exercise study profiling four plasma omics modalities, the leading unsupervised subject component from multiTEMPTED is strongly associated with sex, whereas the leading components from MEFISTO show no discernible association with known study phenotypes and appear numerically degenerate. In simulation experiments, multiTEMPTED accurately recovers the underlying component structure across a range of noise levels, outperforms MEFISTO in multiple settings, and runs several orders of magnitude faster. multiTEMPTED is implemented in the open-source R package multi.tempted (https://github.com/loulind/multi.tempted).

bioinformatics↗

Integrated metabolomics and proteomics from voxelated cortical hemispheres of adult rhesus monkeys

The spatial organization of molecular networks across cortex likely contributes to differences in local circuit vulnerability in aging and Alzheimers disease; yet many existing molecular datasets sacrifice spatial structure, sampling only a handful of regions per brain. Here, we present a framework for generating spatially registered, paired metabolomic and proteomic maps across an entire cortical hemisphere of an adult rhesus monkey, at millimeter resolution. One hemisphere each from two animals was harvested under controlled conditions, approximately flattened, and hand dissected at different sampling resolutions (roughly 2.5 and 4 mm/side) into tissue voxels. Each voxel was split after homogenization and extraction to provide matched aliquots for targeted metabolomics and deep untargeted proteomics. To handle these high dimensional data, we developed PChclust, a principal component guided feature clustering algorithm. For cross omic integration, we developed a spatially regularized sparse canonical correlation analysis (sr-sCCA), which incorporates spatial neighborhood structure via graph Laplacian smoothing. We recover meaningful biology: Molecular similarity between neighboring voxels decayed with distance in both modalities, confirming that voxelation captures spatially organized biological variance. The sr-sCCA identified joint proteome-metabolome components with coherent cortical gradients that were conserved across animals. Pathway enrichment analysis recovered brain relevant ontologies and reconstructed complete metabolic circuits from single voxels.

neuroscience↗

Decoding the MYC locus reveals a druggable ultraconserved RNA element

The human genome is dominated by noncoding sequences, most of which are poorly conserved across species. How genetic information is distributed between coding and noncoding regions remains a fundamental unresolved question. Using CRISPR saturation mutagenesis at base-pair resolution, we mapped the functional fitness landscape of the 10-kb human MYC locus with a near-PAMless, high-fidelity SpRY-Cas9. This unbiased interrogation revealed that the majority (67%) of functionally essential base-pairs in this locus are noncoding. Paradoxically, the phenotypic impact of noncoding sequences correlates inversely with evolutionary conservation, driven in part by rapidly diverging cis-regulatory DNA elements that remain functionally constrained in humans. Within this landscape, we identified an ultraconserved RNA element in the 3 untranslated region (UTR) that is indispensable for MYC-dependent cancer cells. Remarkably, steric-blocking antisense oligos targeting this RNA element selectively eliminate MYC-addicted cancer cells by suppressing MYC function without reducing MYC abundance. Mechanistically, this 3' UTR element promotes perinuclear localization of MYC mRNA and efficient nuclear import of the short-lived MYC protein, enabling its function as a nuclear transcription factor. Together, these findings highlight noncoding sequences as major carriers of functional genetic information, provide a comprehensive fitness map of the MYC locus, and uncover a therapeutically actionable RNA element that disables MYC-driven cancer.

genomics↗

Structural insights on the ligand selectivity of the mouse trace amine-associated receptor TAAR7e

Trace amine-associated receptors (TAARs), a subclass of G protein-coupled receptors involved in detecting volatile amines, are co-expressed with odorant receptors in the main olfactory epithelium. Although many behaviorally relevant amines have been identified for olfactory TAARs, the molecular mechanisms underlying their selectivity for amines with varying carbon chain lengths and degrees of amine substitution remain unclear. In this study, we used mTAAR7e as a model to investigate these mechanisms. Homology modeling revealed a conserved ligand-binding pocket, supported by sequence-structure covariance analysis across TAARs. Structure-activity relationship profiling revealed key chemical determinants integral to mTAAR7e-mediated odorant recognition and provided, for the first time, a structural explanation for its selective preference toward longer carbon chains and tertiary amines. Computationally predicted interactions between mTAAR7e and the representative ligand N,N-dimethylcyclohexylamine (DMCHA) were validated through site-directed mutagenesis. Furthermore, conformational dynamics of mTAAR7e during receptor activation were characterized, providing insights into activation-related structural rearrangements. Together, these findings offer novel insights into the molecular logic of TAAR ligand selectivity and may advance our understanding of how TAARs mediate both olfactory and systemic aminergic signaling.

molecular biology↗

A distinctive subset of microglia positioned at the paraventricular zone are dedicated to cleaning the cerebrospinal fluid

The interconnected ventricles in the central nervous system are filled with cerebrospinal fluid (CSF) which serves multifaceted roles including bring away metabolic wastes. However, whether there is an inherent "dislodging" mechanism, in addition to CSF drainage and reabsorption, at play for the removal of microparticles in the CSF is unclear. In this study, we identified a subset of microglia distributed in close proximity to the ependymal wall of ventricles. They differed from parenchymal microglia in morphology, behaviors and transcriptomes. In particular, the paraventricular microglia extended transependymal dendrites into the lumen of ventricles and were proficient in sequestering and phagocytosing intraventricular exogenous particles when present. A specific removal of the paraventricular microglia led to an acute ventriculomegaly due to an increased colloid osmotic pressure in the CSF resulting from protein accumulation. Thus, we identified the paraventricular microglia as specific guardians in monitoring and cleaning the CSF.

immunology↗

Infectome Analysis of Small Mammals in Southern China Reveals Ecological Associations and Emerging Threats from Diverse Pathogens

Small mammals harbor a diverse array of zoonotic pathogens. To date, however, metagenomic surveys of these species have primarily focused on viral diversity, with limited attention paid to bacteria and eukaryotic pathogens. Additionally, the ecological determinants of pathogen diversity within these mammals have not been systematically examined. Herein, we employed a metatranscriptomics approach to survey the pathogen infectome of 2,408 individual samples, representing lung, liver, and gut tissues from 858 animals collected throughout Guangdong province, China, with a study design that accounted for host species, tissue, season, and geographic location. We identified 76 pathogen species, including 29 RNA viruses, 12 DNA viruses, 5 bacteria, and 30 eukaryotic pathogens, 33 of which were newly discovered. Tissue distribution analysis revealed distinct organotropisms, suggesting varied transmission routes, while host distribution analysis showed that each animal carried an average of one pathogen, with 10 pathogens widely distributed among mammalian orders. Our characterization of the geographic and seasonal patterns revealed that pathogen richness was primarily influenced by region, host species, and season, while pathogen composition was largely shaped by host genetic distance. Collectively, these data provide the first comprehensive insight into the dynamics of the pathogen infectome in these key mammalian disease reservoirs, highlighting the major factors driving pathogen diversity and transmission.

microbiology↗

DTPSP: A Deep Learning Framework for Optimized Time Point Selection in Time-Series Single-Cell Studies

Time-series studies are critical for uncovering dynamic biological processes, but achieving comprehensive profiling and resolution across multiple time points and modalities (multi-omics) remains challenging due to cost and scalability constraints. Current methods for studying temporal dynamics, whether at the bulk or single-cell level, often require extensive sampling, making it impractical to deeply profile all time points and modalities. To overcome these limitations, we present DTPSP, a deep learning framework designed to identify the most informative time points in any time-series study, enabling resource-efficient and targeted analyses. DTPSP models temporal gene expression patterns using readily obtainable data, such as bulk RNA-seq, to select time points that capture key system dynamics. It also integrates a deep generative module to infer data for non-sampled time points based on the selected time points, reconstructing the full temporal trajectory. This dual capability enables DTPSP to prioritize key time points for in-depth profiling, such as single-cell sequencing or multi-omics analyses, while filling gaps in the temporal landscape with high fidelity. We apply DTPSP to developmental and disease-associated time courses, demonstrating its ability to optimize experimental designs across bulk and single-cell studies. By reducing costs, enabling strategic multi-omics profiling, and enhancing biological insights, DTPSP provides a scalable and generalized solution for investigating dynamic systems.

bioinformatics↗

Trajectory-guided dimensionality reduction for multi-sample single-cell RNA-seq data reveals biologically relevant sample-level heterogeneity

The analysis of single-cell RNA-sequencing (scRNA-seq) data with multiple biological samples remains a pressing challenge. We present MUSTARD, a trajectory-guided dimension reduction method for multi-sample multi-condition scRNA-seq data. This all-in-one decomposition reveals major gene expression variation patterns along the trajectory and across multiple samples simultaneously, providing opportunities to discover sample endotypes along with associated genes and gene modules. In data-driven simulation, MUSTARD achieves high accuracy in distinguishing sample-level group differences that existing methods fail to capture. MUSTARD also demonstrates a robust ability to capture gene markers and pathways associated with phenotypes of interest across multiple real-world case studies.

bioinformatics↗

Climate adaptation in P. trichocarpa: key adaptive loci identified for stomata and leaf traits

Identifying the genetic basis of traits underlying climate adaptation remains a key goal for predicting species responses to climate change, enabling the elucidation of gene targets for future climate-resilient crops. Here, we measured 14 leaf and stomatal traits under control (well-watered) and drought conditions, subsampling a diversity collection of over 1,300 Populus trichocarpa genotypes, a potential biofuel feedstock crop. Stomatal traits were correlated with the climate of origin for genotypes, such that those originating from environments subject to water deficit tended to have smaller stomata, but with higher density. Stomatal traits were also correlated with leaf morphology, with larger leaves having larger stomata and lower stomatal density mirrored in correlations to climate of origin. The direction of plastic responses - reduced stomatal size under drought - mirrors the correlations seen among genotypes with respect to the aridity of environmental origin. Genome-Wide Association Studies (GWAS) identified loci underlying trait diversity, including candidates contributing to stomatal size. We used climate of origin to predict stomatal size in genotypes with unknown trait values and found that these predicted phenotypes confirmed empirically measured allele effects. Finally, we found evidence that future climates may select for alleles contributing to decreased stomatal size, with the strength of selection depending on the availability of moisture. These findings reveal adaptive variation in stomatal and physiological traits along with underlying genetic loci, with implications for future selection and breeding - providing insights into the responses to future climate change. HighlightResearch on Populus trichocarpa reveals adaptation of physiological and stomatal traits linked to drought tolerance, with genotypes from arid regions exhibiting smaller stomata, offering insights for climate change adaptation and sustainable biofuel production.

plant biology↗

Singular Odorant Receptor Expression Orchestrated by Promoter Activation Specificity in Apis Mellifera Olfactory Sensory Neurons

Honeybee, positioned as an outgroup of flies and mosquitos, have exhibited intriguing duplications and expansions of OR genes in genomic studies. However, little is known about how the OR genes are expressed and regulated in honeybee olfactory sensory neurons (OSNs). In this study, we utilized a single-cell multi-omics approach to profile the transcriptome and chromatin accessibility in Apis mellifera antennal nuclei, aiming to elucidate OR gene expression and its underlying regulatory mechanisms. Our systematical analysis unveiled a similar singular expression pattern of ligand-specific receptors in Apis mellifera OSNs, parallel with those observed in Drosophila melanogaster. Mechanistically, we discovered that promoter activation of OR genes orchestrates receptor expression patterns. For instance, although multiple adjacent OR genes are co-expressed with a single active promoter through polycistronic transcription, only the first OR gene could produce functional receptor protein, as supported by transcriptome quantification in OSNs. Additionally, we found that co-expression of receptor proteins might occur only when co-expressed OR genes possess multiple accessible promoters. This scenario is less common than expected, considering the number and evolutionary age of OR genes in Apis mellifera, suggesting a selection favoring the specialization of rapidly expanded OR genes in Apis mellifera. Overall, our study provides significant insights into the insect olfaction system and the regulatory mechanisms of OR genes expression.

neuroscience↗

Aurora Kinase A Inhibition plus Tumor Treating Fields Suppress Glioma Cell Proliferation in a Cilium-Independent Manner

Tumor Treating Fields (TTFields) have been shown to extend the survival of glioblastoma (GBM) patients. TTFields interfere with a broad range of cellular processes which may contribute to their efficacy. Among these, TTFields disrupt primary cilia stability on GBM cells. Here we asked if concomitant treatment of TTFields with other agents that interfere with GBM ciliogenesis can further suppress GBM cell proliferation in vitro. Aurora Kinase A (AURKA) promotes both cilia disassembly and GBM growth in vitro and in xenograft models. Inhibitors of AURKA such as Alisertib have been previously demonstrated to inhibit cilia disassembly and increase the frequency of cilia in various cell types. However, here we show that physiological concentrations of Alisertib treatment significantly reduced GBM cilia frequency in gliomaspheres across multiple patient derived cell lines, and in patient biopsies treated ex vivo with Alisertib. This activity of Alisertib seems to be glioma cell specific as it did not reduce neuronal or glial cilia frequencies in mixed primary cell cultures from mouse forebrain. Furthermore, Alisertib depletion of glioma cilia appears specific to AURKA inhibition, as a potent AURKB inhibitor, AZD1152, had no effect on GBM ciliary frequency. Treatment of two different GBM patient-derived cell lines with TTFields and Alisertib resulted in a significant reduction in cell proliferation compared to either treatment alone. However, this effect was not cilia-dependent as the combined treatment reduced proliferation in cilia-depleted cell lines lacking, ARL13b, or U87MG cells which are naturally devoid of ARL13B+ cilia. This result is not surprising given the wide range of pathways regulated by AURKA in addition to cilia. Nonetheless, Alisertib-mediated effects on glioma cilia may be a useful biomarker of drug efficacy within tumor tissue. Considering Alisertib has been shown to cross the blood brain barrier and inhibit intracranial growth of xenografted tumor models, our data warrant future studies to explore whether concomitant Alisertib and TTFields exposure prolongs survival of brain tumor-bearing animals in vivo.

cancer biology↗

Evolutionary prediction for new echolocators

It was suggested over 60 years ago that echolocation is widespread in animals. Although echolocation has been found to evolve independently in several mammalian lineages, this hypothesis remains largely untested due to the difficulty of determining the presence of echolocation. To address this issue, we combined molecular convergence analyses from 190 high-quality mammalian genomes with machine learning to predict potential new mammalian echolocators. Our model predicted three promising lineages of echolocating mammals. Behavioral experiments confirmed that the gracile shrew mole (Uropsilus gracilis), the highest- ranking species among predicted echolocators, as well as almost all shrew moles (Uropsilinae), are capable of echolocation through the use of ultrasonic pulses. In contrast to most bats that use laryngeal echolocation, the characteristics of calls, the morphology of the stylohyal bone, and the results of tongue ligation experiments all suggest that shrew moles produce ultrasonic clicks with their tongues for echolocation. Finally, we estimated at least 20% of all living mammalian orders with echolocation ability, thus empirically supporting Griffins hypothesis that echolocation is widespread among animals. Our findings not only provide evidence that three novel lineages of echolocating mammals, but also demonstrate that phylogenetically replicated phenotypes can be predicted through genetic convergence. One sentence summaryShrew moles are capable of echolocation.

evolutionary biology↗

PVN microglia via P2Y12 transmit hemodynamic signal to promote sympathetic excitation in hypertension

Hypertension is usually accompanied with an elevated sympathetic tonicity, but how sympathetic hyperactivity is triggered is not fully understood. Recent advances reveal that microglia-centered neuroinflammation contributes to sympathetic excitation in hypertension. In this study, we performed a temporospatial analysis of microglia at both morphological and transcriptomic levels, and found that microglia in the hypothalamic paraventricular nucleus (PVN) were early responders to hypertensive challenges. PVN is the central hub for maintaining cardiovascular function via regulation of fluid balance and sympathetic outflow. Comprehensive vasculature analyses unveiled that PVN was characterized by high capillary density, thin vessel diameter, and complex vascular topology among brain regions. As such, PVN is susceptible to the penetration of ATP released from the vasculature in response to hemodynamic disturbance after blood pressure increase. ATP ligation to microglial P2Y12 receptor is responsible for the microglial accumulation and activation in the PVN. Furthermore, either pharmacological blockade or genetic ablation of microglial P2Y12 could substantially restrain blood pressure increase under hypertensive challenge. Together, these findings disclose that a unique vasculature pattern results in the vulnerability of PVN pre-sympathetic neurons to hypertension-associated insults, which is mediated by microglia.

neuroscience↗

Time-Informed Dimensionality Reduction for Longitudinal Microbiome Studies

Longitudinal studies are crucial for understanding complex microbiome dynamics and their link to health. We introduce TEMPoral TEnsor Decomposition (TEMPTED), a time-informed dimensionality reduction method for high-dimensional longitudinal data that treats time as a continuous variable, effectively characterizing temporal information and handling varying temporal sampling. TEMPTED captures key microbial dynamics, facilitates beta-diversity analysis, and enhances reproducibility by transferring learned representations to new data. In simulations, it achieves 90% accuracy in phenotype classification, significantly outperforming existing methods. In real data, TEMPTED identifies vaginal microbial markers linked to term and preterm births, demonstrating robust performance across datasets and sequencing platforms.

bioinformatics↗

MitoSort: robust demultiplexing of pooled single-cell genomics data using endogenous germline mitochondrial variants

Multiplexing across donors has emerged as a popular strategy to increase throughput, reduce costs, overcome technical batch effects, and improve doublet detection in single-cell genomic studies. Using endogenous genetic barcodes eliminates the need for additional experimental processing steps. Among the available choices for endogenous barcodes, the unique features of mtDNA variants render them a more computationally efficient and robust option compared to genome variants. Here we present MitoSort, a method that uses mtDNA germline variants to assign cells to their donor of origin and identify cross-genotype doublets. We evaluated the performance of MitoSort by in silico pooled mtscATAC-seq libraries and experimentally multiplexed data using cell hashing method. MitoSort achieve both high accuracy and efficiency on genotype clustering and doublet detection for mtscATAC-seq data, which fills a void left by the inadequacies of current computational techniques tailored for scRNA-seq data. Moreover, MitoSort exhibits versatility and can be applied to various single-cell sequencing approaches beyond mtscATAC-seq, as long as the mtDNA variants can be reliably detected. Furthermore, through a case study, we demonstrated that demultiplexing 8 individuals assayed at the same time with MitoSort, enables the comparison of cell composition without batch effects.

bioinformatics↗

High-level production of nervonic acid in the oleaginous yeast Yarrowia lipolytica by systematic metabolic engineering

Brain and neurological diseases are influencing more than one billion worlds people. Nervonic acid (cis-15-tetracosenoic acid, C24:1 {Delta}15) benefits the treatment of neurological diseases and the health of brain. Currently, the sources of nervonic acid are limited to the seeds of a couple of plants. In this study, we employed the oleaginous yeast Yarrowia lipolytica to overproduce nervonic acid oil by systematic metabolic engineering. First, engineering the fatty acid elongation (FAE) pathway by expressing a heterologous {beta}-ketoacyl-CoA synthase gene CgKCS enabled the production of nervonic acid in Y. lipolytica. Second, modulation of endogenous pathways by expressing a C16:0-acyl-CoA preferred fatty acid elongase gELOVL6 together with a C18:0-acyl-CoA preferred fatty acid desaturase MaOLE2 increased the content of nervonic acid in total fatty acids (TFA). Third, iterative expression of CgKCS, gELOVL6 and MaOLE2 at the genomic loci of rDNA, FAD2, TGL4, GSY1 and SNF1 dramatically improved the production of nervonic acid. Fourth, the biosynthesis of both nervonic acid and lipids were further enhanced by expression of the MaOLE2-CgKCS fusion protein and glycerol-3-phosphate acyltransferases (GPAT) and diacylglycerol acyltransferases (DGAT) from Malania oleifera in the endoplasmic reticulum (ER) membrane. Fifth, an ER structure regulator YlINO2 was identified in Y. lipolytica and the overexpression of YlINO2 led to a 39.3% increase in lipid production. Next, pilot-scale fermentation in 50-L reactor using the strain YLNA9 exhibited a lipid titer of 96.7 g/L and a nervonic acid titer of 17.3 g/L, the highest reported titer to date for de novo nervonic acid production. We also found that disruption of the AMP-activated S/T protein kinase SNF1 increased the ratio of nervonic acid (C24:1) to lignoceric acid (C24:0) by 61.6% and a ratio of 3.5:1 (nervonic acid to lignoceric acid) was achieved in the strain YLNA10. Finally, a proof-of-concept purification and separation of nervonic acid were performed and the purity of it reached 98.7%. This study suggested that oleaginous yeasts are attractive hosts for the cost-efficient production of nervonic acid and possibly other very long-chain fatty acids (VLCFAs).

bioengineering↗

Sequencing-free Tissue-wide Spatial Profiling of Post-transcriptional Regulations

The importance of genetic or epi-genetic heterogeneity has been increasingly recognized, but it has been challenging to profile intracellular post-transcriptional targets with sufficient throughput and resolution at across large-scale tissue samples. This study describes a technique, Spectrum-FISH, for high-throughput, sequencing-free, and tissue-wide spatial profiling of various post-transcriptional targets in acute tissue sections with subcellular resolution. The platform uses a biochip with an array of vertically aligned nanoprobes to effectively extract intracellular molecules for downstream analysis in the coordinates of the large-scale of cells within a tissue slice. As a proof-of-concept, the Spectrum-FISH is used to profile the spatial dynamics of 24 miRNAs and 9 m6A-modified messenger RNAs (m6A-mRNA) in acute olfactory bulb (OB) slices of millimeter scale. The results showed potentially multiomics spatial heterogeneity for the examined post-transcriptional regulations in rodent OB, especially in the outer plexiform layer and granule layer, where highly correlated miRNAs and m6A-mRNAs groups were identified, indicating a potential cooperative involvement of different post-transcriptional regulations at these OB regions.

genomics↗

Increasing Ciliary ARL13B Expression Drives Active and Inhibitor-Resistant SMO and GLI into Glioma Primary Cilia

ADP-ribosylation factor-like protein 13B (ARL13B), a regulatory GTPase and guanine exchange factor (GEF) enriches in primary cilia and promotes tumorigenesis in part by regulating Smoothened (SMO), GLI, and Sonic hedgehog (SHH) signaling. Gliomas with increased ARL13B, SMO and GLI2 expression are more aggressive but the relationship to cilia is unclear. Previous studies showed increasing ARL13B in glioblastoma cells promoted ciliary SMO accumulation, independent of exogenous SHH addition. Here we show SMO accumulation is due to increased ciliary, but not extraciliary ARL13B. Increasing ARL13B expression promotes the accumulation of both activated SMO and GLI2 in glioma cilia, but not in NIH3T3 fibroblast cilia. ARL13B-driven increases in ciliary SMO and GLI2 are resistant to SMO inhibitors, GDC-0449 and cyclopamine. Finally, temozolomide chemotherapy which increases ARL13B expression in glioma, stimulates SMO and GLI2 into glioma cilia, but not fibroblast cilia. Collectively, our data suggest factors that elevate ARL13B may drive drug-resistant SMO and GLI into cilia. This suggests the ARL13B-associated mechanism that leads to ciliary SMO/GLI recruitment may promote treatment resistance in glioma.

cancer biology↗