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Cheng, Y.

Publications and source records attributed to Cheng, Y..

At least 55 records · Page 3Linked to original sources

Changing Pacific salmon nursery lake ecosystem dynamics over centuries to millennia: insights from sedimentary DNA metabarcoding

Pacific salmon are keystone species to North Pacific freshwater, coastal, and oceanic ecosystems, but many populations have declined or become more variable in recent decades due to anthropogenic impacts and climate change. Long-term records are needed to understand past changes, identify ecosystem stressors, and guide restoration. We used sedimentary DNA (sedDNA), an emerging paleoecological approach offering broader taxonomic information than traditional methods, to reconstruct ecosystem changes across five Pacific salmon nursery lakes in British Columbia (Canada). DNA metabarcoding targeting the 18S ribosomal RNA gene V7 region was used to track shifts in eukaryotic communities including algae and invertebrates over centuries to millennia. Most lakes showed notable algal community shifts over the past two centuries, with declining green algae and rising diatom relative abundances. Chrysophytes and dinoflagellates also increased over the past century in most lakes, likely driven by stronger thermal stratification, which favored these motile and mixotrophic algae that are capable of vertical migration and flexible nutrient acquisition. We contextualized the trajectories of each core through an ordination analysis based on 98 lakes distributed across British Columbia, which identified land-use changes and longer growing seasons as potential drivers. Network analyses of the sedDNA time series revealed decreasing modularity and increasing connection across lakes, suggesting a shift in resilience mechanisms from between-module buffering by compartmentalized specialists to within-guild insurance via functional overlap among generalists. Our findings demonstrate that sedDNA provides taxonomically rich, long-term insights into aquatic ecological dynamics, which are foundational for understanding and protecting Pacific salmon nursery habitats.

ecology↗

A PSMA-Targeted Up-conversion Nanoplatform for Deep-Tissue dual activation Photodynamic and Sonodynamic Therapy of Castration Resistant Prostate Cancer.

Photodynamic therapy (PDT) for prostate cancer is limited by the shallow penetration of visible light into deep-seated tumors. In contrast, sonodynamic therapy (SDT) enables noninvasive deep-tissue activation using ultrasound, while near-infrared photodynamic therapy (NIR-PDT) enhances tissue penetration through up-conversion-mediated activation. Based on these advantages, we developed a prostate-specific membrane antigen (PSMA)-targeted nanotheranostic platform (UCNPs@mSiO2/HPPH@TCS) to compare NIR-PDT and SDT in vitro for penetration-enhanced and targeted prostate cancer therapy. The nanoformulation consists of NaYF4:Yb3+, Er3+ up-conversion nanoparticles coated with mesoporous silica for high loading of HPPH (Photochlor), which functions as both photosensitizer and sonosensitizer, and a PSMA-targeted chitosan shell for selective tumor targeting. Physicochemical characterization confirms a uniform core-shell structure ([~]63 nm). Tissue-mimicking depth studies demonstrate that SDT and NIR-PDT achieve greater penetration than conventional 665 nm PDT in thicker tissue layers. Intracellular reactive oxygen species (ROS) analysis shows that SDT generates higher total ROS levels, whereas NIR-PDT produces greater singlet oxygen generation. Three-dimensional spheroid models further validate efficacy, demonstrating rapid spheroid collapse via apoptosis, with SDT inducing earlier apoptosis and more uniform penetration than PDT. Collectively, the results suggest SDT provides superior overall therapeutic efficacy compared with NIR-PDT, highlighting the potential of this nanoplatform for precision prostate cancer therapy.

cancer biology↗

Ubiquitination-mediated mitochondrial protein degradation ensures seedling emergence by regulating ER-mitochondrial interaction and mitophagy

Seedling emergence is a pivotal step of plant survival, requiring rapid hypocotyl elongation for soil penetration. This energy-demanding process necessitates active mitochondrial respiration, which inevitably induces oxidative damage. Therefore, plants evolved a quality control mechanism that selectively removes dysfunctional mitochondria through the mitophagy pathway. Here, we identified SPL2, a mitochondrial E3 ligase which is essential for hypocotyl elongation and seedling emergence through degrading mitochondrial outer membrane proteins, such as TRB1 and FIS1A. Intriguingly, these proteins also interact with an ER protein, VAP27-1, forming a complex at the ER-mitochondria contact sites, which is essential for mitophagy initiation. The spl2 mutant exhibits enhanced ER-mitochondrial tethering and mitophagy activation, whereas its overexpression has the opposite effects. The expression of SPL2 increases after light perception, in agreement with the reduced mitophagy. Collectively, our findings reveal novel mechanistic insights into seedling emergence, which are coordinated through protein ubiquitination, ER-mitochondrial interaction, and mitophagy.

plant biology↗

Cell DiffErential Expression by Pooling (CellDEEP) highlights issues in differential gene expression in scRNA-seq

Accurate identification of differentially expressed genes (DEGs) in single-cell RNA sequencing (scRNA-seq) data remains challenging. Single-cell-specific statistical models often report large numbers of candidate genes but can exhibit inflated false positive rates, whereas pseudobulk approaches improve false discovery control at the cost of reduced sensitivity. To overcome the noise and bias that other tools have, and allow the user to have more control of the DEG process, we present CellDEEP, which uses a cell aggregation (metacell) approach. This tool provides a framework for flexible selection of pooling strategies and parameterisation for differential expression analysis (DE). Benchmarking on simulated and real datasets, including COVID-19 and rheumatoid arthritis, shows that CellDEEP often outperforms other methods, consistently reduces false positives compared to single-cell methods and recovers more true positives than pseudobulk methods. Our work shifts the focus from selecting a single "best" method to an approach that reduces cell-level noise while preserving biological signal, together with transparent validation framework, advancing more reliable differential-expression analysis in single-cell transcriptomics. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=189 HEIGHT=200 SRC="FIGDIR/small/710522v1_ufig1.gif" ALT="Figure 1"> View larger version (35K): org.highwire.dtl.DTLVardef@14692f9org.highwire.dtl.DTLVardef@5b37d6org.highwire.dtl.DTLVardef@aece11org.highwire.dtl.DTLVardef@5ade3d_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioinformatics↗

Effects of the Coenzyme Q10 Analog 6-Bromo-ubiquinone (6-Br-Q0C10) on Mammalian Cell Growth

Synthetic 6-Br-Q0C10 has been shown to exhibit a partial electron transfer activity of native coenzyme Q in the isolated mitochondria. It reduces energy coupling efficiency by approximately 30%, suggesting that it may be useful in modulating cell growth in tissue culture. Whether or not it behaves in the same way in the whole cells, or animal, however, has not yet been fully examined. Recently we have investigated the effect of 6-Br-Q0C10 across multiple cell lines using three detection methods. Treatment with 6-Br-Q0C10 reduces cell proliferation in all cell lines tested, with different effectiveness. Obesity-related cell lines were the most susceptible, and a pronounced inhibitory effect was also observed in cancer cell lines. These results strengthen the idea of using 6-Br-Q0C10 to manage obesity or to retard the growth of rate cancer cells and thus prolonging life.

biochemistry↗

Beyond Student's t: A Systematic Exploration of Heavy-Tailed Residual Densities for Outlier Handling in Population PK Modeling

BackgroundReliable population pharmacokinetic (PopPK) parameter estimation can be compromised by outliers under Gaussian residual error models. A common mitigation strategy is post hoc filtering based on conditional weighted residuals (CWRES); however, this approach can be insensitive due to model "masking" driven by variance inflation. Practical barriers to implementing robust likelihoods in standard software have motivated interest in computationally simpler exponential-tail alternatives such as the Laplace and exponential power distribution (EPD). MethodsWe implemented a one-compartment PopPK model using a custom likelihood workaround in Monolix to benchmark four residual error distributions: Normal, Laplace, Generalized Error Distribution (GED), and Students t. We assessed CWRES sensitivity under extreme contamination and compared estimation performance using theoretical tail-behavior analysis, controlled simulation studies spanning multiple contamination severities, and a real-world caffeine PK case study with influential terminal-phase deviations. ResultsSimulations showed that CWRES-based diagnostics can be unreliable: extreme outliers frequently produced |CWRES| < 6 because the Normal model inflated residual variance, thereby masking contamination. Exponential-tail models (Laplace, GED) improved robustness for mild to moderate outliers but failed under extreme deviations due to insufficiently heavy tails compared to power-law decay. In contrast, the Students t model, via power-law tail behavior, maintained stable and minimally biased structural parameter estimates across contamination scenarios. Consistent patterns were observed in the caffeine case study, where the Students t model provided improved fit and physiologically plausible parameter estimates. ConclusionsCWRES-driven outlier handling is methodologically fragile because influential contamination can be masked by variance inflation and induce biased inference. Among robust residual error models, exponential-tail distributions may be insufficient for extreme outliers, whereas the Students t distribution provides more stable inference across contamination severities. These findings support adopting Students t residual modeling as a default robust option in routine PopPK workflows when outlier contamination is plausible.

pharmacology and toxicology↗

Decoupling CAR-T Expansion, Conversion, and Decay Timing: Physiologically Aligned Semi-Mechanistic Modeling with Smooth Gating and a Cauchy Likelihood Residual Model

Chimeric antigen receptor (CAR) T-cell therapies exhibit complex cellular kinetics characterized by rapid expansion, contraction, and long-term persistence, often with substantial inter-individual variability, frequent below-quantification (BLQ) observations, and occasional influential data points. These features can destabilize inference under Gaussian residual assumptions and motivate robust, likelihood-based approaches that can jointly accommodate outliers and censoring. In prior work, we showed that combining Students t residuals with likelihood-based BLQ handling (M3 censoring) improves robustness in CAR-T cellular kinetics modeling; however, implementing Students t censoring likelihoods is not straightforward in all modeling platforms because the Students t cumulative distribution function (CDF) lacks a simple elementary closed-form. Here, we evaluated the Cauchy residual likelihood as a practical, heavy-tailed alternative to Students t that provides closed-form expressions for both the probability density function (PDF) and CDF. In a two-compartment IV pharmacokinetic simulation with terminal-phase outlier contamination, Cauchy residuals preserved stable parameter recovery comparable to Students t while avoiding the bias and instability observed under Normal residuals. In a real-data integrated CAR-T cellular kinetics application fitted with full Bayesian inference and likelihood-based BLQ handling, replacing Students t with Cauchy yielded highly concordant posterior inference and similar subject-level predictions. We further extended a semi-mechanistic CAR-T cellular kinetics framework by replacing piecewise switching with smooth S-shaped time-varying rate functions and allowing process-specific transition times. Full Bayesian posterior summaries supported asynchronous transition timing, including earlier conversion relative to expansion and later decay-related transitions. Collectively, these results support Cauchy likelihoods as an implementation-friendly robust option and demonstrate that smooth, process-specific transition modeling can enhance physiological plausibility for CAR-T kinetics.

pharmacology and toxicology↗

Delay Differential Equation (DDE) Modeling of CAR-T Cellular Kinetics: Application to BCMA-Targeted (Ide-cel, Orva-cel) and CD19-Targeted (Liso-cel) Therapies

Chimeric antigen receptor (CAR) T-cell therapies undergo rapid in vivo expansion followed by contraction and variable long-term persistence after a single infusion, yielding cellular kinetic (CK) profiles that differ fundamentally from conventional small-molecule and biologic pharmacokinetics. Piecewise, phase-based CK models are widely used, but discontinuous switching and constant expansion assumptions can create numerical instability around the transition window and bias the characterization of early expansion and near-peak behavior. Building on our prior saturable expansion framework (Vmax/Km), we advanced CAR-T CK modeling by introducing (i) smooth S-shaped gating to replace discontinuous phase switching and enable continuous time-varying expansion dynamics, and (ii) delay differential equation (DDE) components to evaluate whether longitudinal data support explicit lags in downstream biological processes. Data were pooled from three CAR-T trials (TRANSCEND, KarMMa-3, and EVOLVE) spanning two BCMA-targeted products (ide-cel, orva-cel) and one CD19-targeted product (liso-cel). Models were estimated in Monolix using SAEM with importance sampling for final likelihood evaluation. Model selection relied on likelihood-based criteria (AIC, BIC, and Monolix BICc) and diagnostic assessments. Relative to a constant-expansion baseline, saturable expansion improved fit and reduced systematic model misspecification at high transgene levels (e.g., qPCR transgene copies/{micro}g). Among multiple DDE placements evaluated, the data most strongly supported a delay on effector-to-memory conversion; delays in effector-like and/or memory-like decay were not favored. Simulations indicated that the conversion delay primarily modulated the timing and magnitude of the memory-like trajectory, with minimal impact on total-cell trajectories during the expansion phase at the evaluated scale. In a shared covariate framework with product as a categorical effect, BCMA-targeted products exhibited higher baseline levels and expansion capacity than liso-cel, with stronger evidence for slower effector-like decay for orva-cel than ide-cel. Overall, smooth-gated saturable expansion with DDE-based delayed conversion provides a parsimonious, biologically motivated framework for CAR-T CK and supports cross-product comparisons under harmonized structural assumptions.

pharmacology and toxicology↗

A Global Genomic Resource for Outcrossing Arabidopsis lyrata and Arabidopsis arenosa

Genetic studies leveraging natural variation in Arabidopsis species have improved our understanding of evolutionary genetic processes underlying ecologically important and adaptive traits. Integrating the thorough functional knowledge accumulated in A. thaliana with the extensive natural variation in outcrossing Arabidopsis species is a powerful approach to study the basis of adaptation in natural evolutionary and ecological contexts. Here we present an integrated genomics database of sequenced genomes from several studies in A. lyrata (1018 genomes in total) and A. arenosa (736 genomes in total), spanning the geographic ranges of these two ploidy-variable, outcrossing taxa. We provide a searchable genome browser with population data mapped to respective reference genomes, an interactive geographic map of population structure clusters, and an efficient way to subsample the full dataset of genetic variation, available at arabidopsislyrata.org. To demonstrate its utility, we perform a genome-wide association study on a latitudinal cline of A. lyrata and find strong associations of several loci with latitude, including variants in key regulators of photoperiodic growth. This resource provides access to genetic diversity data in a single repository, enabling further studies of comparative genetics and local adaptation, as well as of individual genes of interest.

evolutionary biology↗

Arborist: Prioritizing Bulk DNA Inferred Tumor Phylogenies via Low-pass Single-cell DNA Sequencing Data

Cancer arises from an evolutionary process that can be reconstructed from DNA sequencing and modeled by tumor phylogenies. High coverage bulk DNA sequencing (bulk DNA-seq) is widely available, but tumor phylogeny inference requires deconvolution, often resulting in non-uniqueness in the solution space. Single-cell DNA sequencing (scDNA-seq) holds potential to yield higher resolution tumor phylogenies, but the sparsity of emerging low-pass sequencing technologies poses challenges for the study of single-nucleotide variants. Increasing availability of data sequenced with both modalities provides an opportunity to capitalize on the advantages of these technologies. While inference methods exist for bulk DNA-seq and for low-pass scDNA-seq, no joint inference methods currently exist. As a first step, we propose a method named ARBORIST that prioritizes tumor phylogenies inferred via bulk DNA-seq using low-pass scDNA-seq data. ARBORIST takes as input a candidate set of trees with corresponding SNV clustering, along with variant and total read count data from scDNA-seq and uses variational inference to approximate a lower bound on the marginal likelihood of each tree in the candidate set. On simulated data, matching characteristics of current scDNA-seq data, ARBORIST outperforms both bulk and low-pass single-cell reconstruction methods. On a biological dataset, ARBORIST conclusively resolves the evolutionary relationship between different SNV clusters on a malignant peripheral nerve sheath tumor, which is supported by orthogonal validation via a proxy for copy number. ARBORIST provides a principled framework for integrating bulk DNA-seq and low-pass scDNA-seq data, improving confidence in tumor phylogeny reconstruction. Availabilityhttps://github.com/VanLoo-lab/Arborist

bioinformatics↗

An injectable soft implant for long-acting, reversible, ultra-stable release of therapeutics

Providing long-term (>6 months) zero-order drug release from easily administered formulations is a key challenge in improving patient adherence and facilitating access. Herein, we report the design and development of an injectable, biodegradable, long-acting polymeric microparticle-embedded hydrogel platform for prolonged, zero-order release of therapeutics. This "soft implant" is injectable for ease of administration and can be retrieved via a small incision, allowing for discontinuation of therapy if desired. Central to the platform are surface-eroding poly(orthoester) (POE) microparticles, which were molecularly tailored to tune zero-order drug release across a wide range of timeframes. We demonstrate the clinical potential of the "soft implant" using levonorgestrel, a contraceptive agent requiring sustained dosing. In vitro, we observed zero-order release for 300 days, projected for >12 months, with behavior consistent with surface erosion further supported through Raman chemical mapping. In vivo studies confirmed zero-order release for six months, projected to 12 months, from a subcutaneous injection in rats. We envision that our platform could transform therapies that require long-term, regular drug dosing, significantly improving compliance and therapy outcomes.

bioengineering↗

A Single-Cell Temporal Atlas of Mouse Nasal Embryonic Development

The mammals breathe and perceive odors through anatomically and functionally distinct respiratory and olfactory regions of the nose; however, how the early embryo endows this system remains largely unknown. Here, we present a temporal high-resolution single-cell RNA sequencing (scRNA-seq) atlas of mouse nasal development, comprising 183,247 cells collected from 20 embryos spanning embryonic day 10.5 (E10.5) to E18.5. We identified 7 major cell clusters and 52 molecularly defined subtypes, including previously unrecognized rare populations and niche-derived signaling components involved in progenitor cell maintenance. Lineage reconstruction of epithelial and mesenchymal compartments revealed dynamic cell fate transitions and highlighted candidate regulators of lineage specification, including Foxa1 as a regulator of nasal respiratory ciliated epithelium differentiation. Integration with adult nasal epithelium datasets further delineated transcriptional dynamics of olfactory stem cells. This study provides a molecular roadmap of nasal development and offers a valuable resource for future research into nasal biology and regenerative strategies.

developmental biology↗

Tirzepatide improves pancreatic β-cell function in mice and patients with type 2 diabetes.

The dual incretin receptor agonist tirzepatide improves {beta}-cell function in T2D patients, but the underlying mechanism remains unclear. This study aimed to elucidate the molecular pathway through which tirzepatide restores {beta}-cell functional improvement. High-fat diet (HFD)-fed C57BL/6J mice were treated with vehicle, a GIP analogue, semaglutide or tirzepatide. Tirzepatide significantly reduced body weight and improved glucose tolerance in HFD-fed mice without altering {beta}-cell mass, proliferation, or apoptosis. Instead, tirzepatide reversed {beta}-cell dedifferentiation, as indicated by reduced ALDH1A3 expression and restored levels of the identity transcription factors PDX1 and MAFA. Single-cell RNA sequencing (scRNA seq) and in vitro studies revealed that tirzepatide up-regulated FOXO1, reactivating the FOXO1-PDX1/MAFA axis. In T2D patients, tirzepatide improved glycemic control, reduced insulin demand, increased HOMA-{beta}, and decreased HOMA-IR. Improvement in HOMA-{beta} correlated positively with baseline insulin resistance. Hence, our study suggested that tirzepatide restores {beta}-cell function in T2D by reprogramming stressed {beta} cells and re-establishing {beta}-cell identity through FOXO1-dependent transcriptional reactivation. These findings provide a mechanistic basis for the superior efficacy of dual incretin receptor agonism in T2D management. ARTICLE HIGHLIGHTSO_LITirzepatide restores {beta} cell identity and function without altering {beta} cell mass in HFD induced diabetic mice. C_LIO_LITirzepatide reverses {beta}-cell dedifferentiation and restores key {beta}-cell transcription factors (PDX1, MAFA) through reactivation of the AKT-FOXO1 signaling pathway. C_LIO_LITirzepatide increases HOMA-{beta} and decreases HOMA-IR in T2D patients, and improvements in HOMA-{beta} positively correlate with baseline insulin resistance. C_LIO_LIThese results demonstrate that tirzepatides therapeutic benefits are not only metabolic but also involve direct restoration of {beta}-cell identity and function. This highlights {beta}-cell reprogramming as a novel therapeutic avenue, thus supporting the broader clinical adoption of dual incretin receptor agonists. C_LI

physiology↗

Structure-guided design of a targeted autoantibody degrader for neurologic disease

Despite rapid progress in the diagnosis of autoantibody-mediated neurologic diseases, standard-of-care therapeutic options remain limited to nonspecific immunosuppression. Here, we report an alternative therapeutic strategy using targeted protein degradation to eliminate pathogenic autoantibodies while leaving the rest of the immune system intact. We previously discovered autoimmune vitamin B12 central deficiency (ABCD), a neurologic condition in which autoantibodies targeting the transcobalamin receptor (CD320) impair the transport of cobalamin (B12) from the blood into the central nervous system (CNS). Combining scanning alanine mutagenesis by phage display, cryo-electron microscopy, and computational modeling, we elucidated a highly conserved anti-CD320 epitope and defined the structural determinants of antigen-autoantibody binding. Next, we synthesized a lysosome-targeting chimera (LYTAC) comprising the lysosome targeting glycan, triGalNAc, fused to the antigenic epitope of CD320 as autoantibody bait. In vitro, this LYTAC promoted the specific lysosomal internalization and extracellular clearance of anti-CD320, restoring homeostatic cellular uptake of B12. In a passive transfer mouse model of ABCD, LYTAC treatment rapidly cleared anti-CD320 from circulation and prevented penetration of anti-CD320 into the CNS. These findings uncover the mechanism of autoantibody-antigen binding in ABCD and demonstrate targeted autoantibody degradation as a therapeutic strategy that may be generalizable to other autoimmune neurologic diseases.

neuroscience↗

Differential adenosine to inosine RNA editing of SINE B2 non-coding RNAs in mouse unveils a novel type of epi-transcriptome response to amyloid beta neuro-toxicity

Alzheimers disease (AD) is characterized by early molecular responses to amyloid beta neuro-toxicity that remain poorly defined. RNA editing by adenosine-to-inosine (A-to-I) conversion is a major epitranscriptomic mechanism, yet its contribution to non-coding RNA regulation during neurodegeneration is largely unknown. Short Interspersed Nuclear Element (SINE)-derived RNAs, including mouse B2 RNAs, represent the dominant substrates of A-to-I editing and have recently been shown to regulate gene expression through ribozyme-mediated processing. Here, we introduce and validate a repeat-aware bioinformatics framework that enables position-specific quantification of A-to-I editing within highly repetitive SINE RNAs--an analysis that has been challenging using standard genome-based pipelines. Applying this approach, we uncover discrete editing hotspots in B2 RNAs whose modification is selectively increased during the earliest stages of amyloid beta pathology. Elevated B2 RNA editing is consistently observed across hippocampal or neocortical tissue from two independent mouse models of amyloid beta accumulation and in hippocampal neurons exposed to acute amyloid beta toxicity. Functional perturbation of ADAR activity alters both B2 RNA editing levels and the expression of B2 RNA regulated stress-response genes, directly linking RNA editing to SINE-mediated transcriptional control. Independent validation using Nanopore direct RNA sequencing indicates increased RNA modification signals at the same B2 RNA regions identified by short-read Illumina sequencing. Together, our findings establish a previously unrecognized epitranscriptomic response to amyloid beta neurotoxicity mediated by site-specific A-to-I editing of SINE RNAs. This work defines a new analytical paradigm for studying RNA editing in RNAs from repetitive elements and reveals a regulatory axis connecting amyloid beta toxicity, ADAR activity, SINE RNA editing, and stress-responsive gene expression, with implications for conserved mechanisms underlying Alzheimers disease and related neurodegenerative disorders.

bioinformatics↗

Nanoscale Protonation Limits and Charge Density in Polymer Films Govern the Activity of Immobilized LacZ under Acid Stress

Under acidic conditions, polycationic polymer coatings function as protective immobilization supports through protonation-mediated local pH buffering. However, it remains unclear how polymer support design parameters, such as film thickness and charge density, govern that vital protonation process. Leveraging the precise control of film thickness and copolymer composition enabled by initiated chemical vapor deposition (iCVD), we systematically investigated how these parameters govern the protonation behavior of poly[glycidyl methacrylate-co-2-(dimethylamino)ethyl methacrylate] (pGD) thin films and, in turn, the activity of immobilized {beta}-galactosidase (LacZ). Infrared spectroscopy suggests that proton penetration was capped at a depth of [~]250 nm in pGD with 65% DMAEMA, limiting the polycationic thickness in pGD films thicker than this value. Consistent with this limit, immobilized LacZ activity under acidic stress (pH 4) increased with protonated thickness up to [~]250 nm and then plateaued. Raising the polycationic monomer content from 25 to 65 mol% increased LacZ activity at pH 4 by up to 83%, consistent with a higher positive charge density providing stronger local pH buffering. To test whether this behavior depends on immobilization sites, we evaluated two approaches: random immobilization (via amine-epoxy ring-opening reactions) and site-directed immobilization (via SpyCatcher/SpyTag binding). Directed immobilization preserved higher LacZ activity than random immobilization, but the protonation-dependent protection trend remained consistent for both strategies. These findings establish protonation depth and charge density as tunable design parameters for polycationic immobilization supports that stabilize enzymes under acidic conditions.

bioengineering↗

Leveraging Human Pangenome for Improved Somatic Variant Detection

Somatic variant detection is technically challenging due to low variant allele fractions, the confounding presence of germline variation, and reference bias. Linear references such as GRCh38 miss sample-specific variation, causing misalignments and incorrect variant calls. Although telomere-to-telomere donor-specific assemblies (DSAs) accurately represent individual genomes, their application is limited by cost and technical barriers. Alternatively, the graph-based human pangenome provides a scalable framework to improve read alignment and perform genome inference. Here, we benchmarked somatic variant detection using GRCh38, graph-based pangenomes, and pangenome-inferred DSAs with a HapMap mixture dataset and the COLO829 melanoma cell line. Pangenome-guided alignment improves read mapping and somatic variant calling accuracy. Furthermore, personalized pangenomes partially reconstruct donor-specific genomic content, improving accuracy, reducing germline contamination, and enabling detection of events in loci absent or poorly represented in GRCh38. These findings demonstrate that graph-based and personalized pangenomes are effective strategies for enhancing somatic variant detection compared with GRCh38.

genomics↗

Infection of 5xFAD mice with a mouse-adapted SARS-CoV-2 does not alter Alzheimer's disease neuropathology yet induces wide-spread changes in gene expression across diverse cell types

Alzheimers disease (AD) is a progressive neurodegenerative disorder and the leading cause of dementia. It is characterized by cognitive decline and accumulation of amyloid beta (A{beta}) plaques and neurofibrillary tangles. Accumulating evidence indicates that viral infection may worsen and/or increase development of established AD pathology. The COVID-19 pandemic has brought attention to the link between SARS-CoV-2 infection and neurologic conditions that vary in severity and duration, as well as the worsening of clinical symptoms in elderly people with dementia. To better understand potential mechanisms by which SARS-CoV-2 infection impacts AD neuropathology, aged 5xFAD and wildtype (WT) mice were intranasally infected with mouse-adapted SARS-CoV-2 (MA10). Intranasal infection of aged-matched (10-14 month) 5xFAD or wild type (WT) C57BL/6 mice with MA10 resulted in viral infection of the lungs that correlated with acute viral pneumonia characterized by lymphocyte inflammation and antiviral immune responses. Viral RNA was not detected within the central nervous system (CNS) of either WT or 5xFAD mice at days 7 or 21 post-infection (p.i.), nor were there signs of overt glial activation or neuroinflammation. There were no differences in either A{beta} plaque volume or number within the brains of MA10-infected 5xFAD mice compared to uninfected 5xFAD mice. However, bulk RNA sequencing and spatial transcriptomics revealed evidence of altered expression of genes associated with neuronal and glial dysfunction, as well as reduced expression of genes encoding adhesion molecules in vascular endothelial cells. Collectively, these findings demonstrate that MA10 infection did not affect A{beta} plaque size or numbers in 5xFAD mice, yet in both WT and 5xFAD mice, there were numerous down-stream effects on gene expression associated with resident CNS cell function.

pathology↗