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Wilson, K. A.

Publications and source records attributed to Wilson, K. A..

15 recordsLinked to original sources

DNA damage drives a unique, Alzheimer's disease-relevant senescent state in neurons

Alzheimers disease (AD) shares molecular hallmarks with the canonical drivers of cellular senescence. Senescent cells have also been shown to accumulate in the brain with age, yet the mechanisms linking AD pathology to the accumulation of senescent cells in the brain remain unclear. Here, we demonstrate that DNA damage in patient-derived directly induced neurons (iNs) drives a senescent-like cell state with relevance to AD. DNA damage-induced senescent iNs show significant transcriptional concordance with human AD neurons and a weighted gene co-expression network analysis (WGCNA) uncovers candidate regulators associated with the senescent-like state in neurons. Direct comparison of iNs to the original patient fibroblasts reveals striking cell-type specific senescence signatures following DNA damage. iNs adopt a p21-associated senescent-like state characterized by a senescence-associated secretory phenotype (SASP) and predicted activation of NF-{kappa}1. In contrast, fibroblasts develop a p16-associated senescent state lacking a SASP phenotype and show a predicted repression of NF-{kappa}1. Early responses to DNA damage further reveal divergent DNA damage response (DDR), with neurons exhibiting higher accumulation of damage lesions relative to fibroblasts. Together, these findings demonstrate that DNA damage drives a unique senescent-like neuronal state that models molecular features of AD, while also revealing fundamental cell-type specific differences in senescent-like phenotypes and DDR.

cell biology↗

Alzheimer's disease risk factor APOE4 exerts dimorphic effects on female bone

Individuals diagnosed with Alzheimers disease (AD) are at an increased risk of bone fractures. Conversely, a diagnosis of osteoporosis in women is the earliest known predictor for AD. However, mechanisms responsible for the coupled decline in cognitive and skeletal health remain unclear. Proteomic analysis of cortical bone from aged mice revealed neurological disease-associated proteins that are highly enriched in aged mouse bones, including apolipoprotein E (Apoe) and amyloid precursor protein. Further, Apoe localized specifically to bone-embedded osteocytes with expression twice as high in aged female bone as in young or male counterparts. In humans, APOE allele variants carry differing AD risk with age. To investigate APOE allelic roles in bone, we utilized a humanized APOE knock-in mouse model that expresses either the protective APOE2, the neutral APOE3, or the AD risk factor APOE4, and analyzed bone and hippocampus from the same mice. APOE4 exerted strong sex-specific effects on the bone transcriptome and proteome, relative to APOE2 or APOE3. Interestingly, the APOE4-associated perturbation in the female bone proteome was more pronounced than the corresponding alterations observed in the hippocampus. APOE4 protein causes bone fragility in females, but not males, even without changes in cortical bone structure. These bone quality deficits arose from suppression of osteocyte perilacunocanalicular remodeling. We find that APOE4 is a new molecular culprit capable of disrupting osteocyte maintenance of bone quality as early as midlife in a manner that disproportionately affects females. These findings highlight osteocytes as potential targets for early diagnosis of age-related cognitive impairment, and treatment for bone fragility, in females.

molecular biology↗

Intervertebral disc impairments in a mouse model of Alzheimer's Disease

Chronic low back pain, frequently associated with intervertebral disc (IVD) degeneration, is highly prevalent in individuals with Alzheimers disease (AD), and the pain intensity is highly correlated with the degree of cognitive impairment. While the incidences of both afflictions increase dramatically in the elderly population, it is unknown whether AD exacerbates the health of the IVD. Utilizing one-year-old male and female 5xFAD mice that constitutively express human APP and PSEN1 transgenes with five AD-linked mutations, we measured the lumbar IVDs extracellular matrix composition, the three-dimensional structure, histopathological degeneration, and mechanical behavior. The collagen, glycosaminoglycans, and advanced glycation end-products content of the IVD were not appreciably different between the 5xFAD animals and their wild-type littermates. Likewise, the 5xFAD IVDs were not histopathologically degenerated. However, the IVD volume, measured by contrast-enhanced microCT, was larger in the 5xFAD animals. Furthermore, dynamic microcompression revealed that 5xFAD IVDs exhibited higher loss tangent, indicating altered tissue damping and fluid-flow dynamics within the disc. These results suggest that although the IVDs of mice with AD are not more degenerated, they may be more susceptible to damage accumulation due to the elevated absorption of energy. Elderly individuals with AD may thus be more prone to IVD injuries that lead to eventual degeneration and spinal pain. Future work will focus on defining the molecular mechanisms and the consequences of these mechanical and structural changes in the IVD and their consequences to low back pain in individuals with AD.

bioengineering↗

Examining the Biophysical Properties of the Inner Membrane of Gram-Negative ESKAPE Pathogens

The World Health Organization has identified multidrug-resistant bacteria as a serious global health threat. Gram-negative bacteria are particularly prone to antibiotic resistance, and their high rate of antibiotic resistance has been suggested to be related to the complex structure of their cell membrane. The outer membrane of Gram-negative bacteria contains lipopolysaccharides that protect the bacteria against threats such as antibiotics, while the inner membrane houses 20-30% of the bacterial cellular proteins. Given the cell membranes critical role in bacterial survival, antibiotics targeting the cell membrane have been proposed to combat bacterial infections. However, a deeper understanding of the biophysical properties of the bacterial cell membrane is crucial for developing effective and specific antibiotics. In this study, Martini coarse-grain molecular dynamics simulations were used to investigate the interplay between membrane composition and biophysical properties of the inner membrane across four pathogenic bacterial species: Klebsiella pneumoniae, Pseudomonas aeruginosa, Enterobacter cloacae, and Escherichia coli. The simulations indicate the impact of species-specific membrane composition on dictating the overall membrane properties. Specifically, cardiolipin concentration in the inner membrane is a key factor influencing the membrane features. Model membranes with varying concentrations of bacterial lipids (phosphatidylglycerol, phosphatidylethanolamine, and cardiolipin) further support the significant role of cardiolipin in determining the membrane biophysical properties. The bacterial inner membrane models developed in this work pave the way for future simulations of bacterial membrane proteins and for simulations investigating novel strategies aimed at disrupting the bacterial membrane to treat antibiotic-resistant infections.

biophysics↗

An RNA interference approach for functional studies in the sea urchin ands its use in analysis of Nodal signaling gradients

Dicer substrate interfering RNAs (DsiRNAs) destroy targeted transcripts using the RNA-Induced Silencing Complex (RISC) through a process called RNA interference (RNAi). This process is ubiquitous among eukaryotes. Here we report the utility of DsiRNA in embryos of the sea urchin Lytechinus variagatus (Lv). Specific knockdowns phenocopy known morpholino and inhibitor knockdowns, and DsiRNA offers a useful alternative to morpholinos. Methods for designing and obtaining specific DsiRNAs that lead to destruction of targeted mRNA are described. DsiRNAs directed against pks1, an enzyme necessary for pigment production, show how successful DsiRNA perturbations are monitored by RNA in situ analysis and by qPCR to determine relative destruction of targeted mRNA. DsiRNA-based knockdowns phenocopy morpholino- and drug-based inhibition of nodal and lefty. Other knockdowns demonstrate that the RISC operates early in development as well as on genes that are first transcribed hours after gastrulation is completed. Thus, DsiRNAs effectively mediate destruction of targeted mRNA in the sea urchin embryo. The approach offers significant advantages over other widely used methods in the urchin in terms of cost, and ease of procurement, and offers sizeable experimental advantages in terms of ease of handling, injection, and knockdown validation. HighlightsDsiRNA provides an RNAi approach for perturbation of sea urchin embryos. A dilution series of DsiRNA oligos reveals properties of the Nodal gradient in establishing the Dorsal-Ventral axis.

developmental biology↗

Single-cell transcriptomics reveals colonic immune perturbations during amyloid-β driven Alzheimer's disease in mice

The "gut-brain axis" is emerging as an important target in Alzheimers disease (AD). However, immunological mechanisms underlying this axis remain poorly understood. Using single-cell RNA sequencing of the colon immune compartment in the 5XFAD amyloid-{beta} (A{beta}) mouse model, we uncovered AD-associated changes in ribosomal activity, oxidative stress, and BCR/plasma cell activity. Strikingly, levels of colon CXCR4+ antibody secreting cells (ASCs) were significantly reduced. This corresponded with accumulating CXCR4+ B cells and gut-specific IgA+ cells in the brain and dura mater, respectively. Consistently, a chemokine ligand for CXCR4, CXCL12, was expressed at higher levels in 5XFAD glial cells and in in silico analyzed human brain studies, supporting altered neuroimmune trafficking. An inulin prebiotic fiber diet attenuated AD markers including A{beta} plaques and overall frailty. These changes corresponded to an expansion of gut IgA+ cells and rescued peripheral Tregs levels. Our study points to a key glia-gut axis and potential targets against AD. Study HighlightsO_LIAD is associated with altered immune parameters in the gut of 5XFAD mice. C_LIO_LI5XFAD colon has reduced ASCs, including CXCR4+ cells with a migratory gene signature. C_LIO_LI5XFAD brain gliosis includes increased CXCL12 expression. C_LIO_LICXCR4+ B cells and gut-specific IgA+ ASCs accumulate in the 5XFAD brain and/or dura mater. C_LIO_LIInulin diet attenuates AD disease parameters while boosting IgA+ cell and Treg levels. C_LI

immunology↗

Machine Learning identifies conserved traits that influence lifespan and healthspan responses to dietary restriction

Dietary restriction (DR) is a potent method to enhance lifespan and healthspan, but individual responses are influenced by genetic variations. Understanding how metabolism-related genetic differences impact longevity and healthspan are unclear. To investigate this, we used metabolites as markers to reveal how different genotypes respond to diet to influence longevity and healthspan traits. We analyzed data from Drosophila Genetic Reference Panel strains raised under AL and DR conditions, combining metabolomic, phenotypic, and genome-wide information. Employing two computational methods across species--random forest modeling within the DGRP and Mendelian randomization in the UK Biobank--we pinpointed key traits with cross-species relevance that influence lifespan and healthspan. Notably, orotate was linked to parental age at death in humans and counteracted DR effects in flies, while threonine extended lifespan, in a strain- and sex-specific manner. Thus, utilizing natural genetic variation data from flies and humans, we employed a systems biology approach to elucidate potential therapeutic pathways and metabolomic targets for diet-dependent changes in lifespan and healthspan.

systems biology↗

Membrane cholesterol regulates inhibition and substrate transport by the glycine transporter, GlyT2

Membrane cholesterol binds to and modulates the function of the specific SLC6 transporters. Here we investigate how cholesterol binds to and modulates the rate of glycine transport by the SLC6 glycine transporter GlyT2, and how this impacts lipid inhibition of GlyT2. Bioactive lipid inhibitors of GlyT2 are analgesics that bind to the lipid allosteric site of the outward facing GlyT2 conformation that is accessible from the extracellular solution. Using molecular dynamics simulations, mutagenesis and cholesterol depletion experiments, we show that bioactive lipid inhibition of glycine transport is modulated by the recruitment of membrane cholesterol to a cholesterol binding site formed by transmembrane helices 1, 5 and 7. Recruitment involves cholesterol flipping from its membrane orientation, and insertion of the 3 hydroxyl group into the cholesterol binding cavity to interact with the base of the lipid allosteric site and the bound inhibitor. The recruitment of membrane cholesterol by allosteric GlyT2 inhibitors is a potential avenue for the development of high-potency, specific pain analgesics and could provide alternative therapeutics that target GlyT2 and other SLC6 neurotransmitter transporters.

neuroscience↗

Longitudinal fundus imaging and its genome-wide association analysis provide evidence for a human retinal aging clock

Biological age, distinct from an individuals chronological age, has been studied extensively through predictive aging clocks. However, these clocks have limited accuracy in short time-scales. Deep learning approaches on imaging datasets of the eye have proven powerful for a variety of quantitative phenotype inference tasks and provide an opportunity to explore organismal aging and tissue health. Here we trained deep learning models on fundus images from the EyePACS dataset to predict individuals chronological age. These predictions led to the concept of a retinal aging clock, "eyeAge", which we employed for a series of downstream longitudinal analyses. eyeAge was used to predict chronological age on timescales under a year using longitudinal fundus imaging data from a subset of patients. To further validate the model, it was applied to a separate cohort from the UK Biobank. The difference between individuals eyeAge and their chronological age, hereafter "eyeAgeAccel", was computed and used for genome-wide association analysis (GWAS). EyeAge predicted chronological age more accurately than other aging clocks (mean absolute error of 2.86 and 3.30 years on quality-filtered data from EyePACS and UKBiobank, respectively). Additionally, eyeAgeAccel was highly independent of blood marker-based measures of biological age (e.g. "phenotypic age"), maintaining an all-cause mortality hazard ratio of 1.026 even in the presence of phenotypic age. Longitudinal studies showed that the resulting models were able to predict individuals aging, in time-scales less than a year, with 71% accuracy. The individual-specific component to this prediction was confirmed with the identification of multiple GWAS hits in the independent UK Biobank cohort. The knockdown of the fly homolog to the top hit, ALKAL2, which was previously shown to extend lifespan in flies, also slowed age-related decline in vision in flies. In conclusion, predicted age from retinal images can be used as a biomarker of biological aging that is independent from assessment based on blood markers. This study demonstrates the potential utility of a retinal aging clock for studying aging and age-related diseases and quantitatively measuring aging on very short time-scales, opening avenues for quick and actionable evaluation of gero-protective therapeutics.

bioinformatics↗

Lipid fingerprints are similar between SLC6 transporters in the neuronal membrane

We use molecular dynamics simulations to characterise the local lipid annulus, or "fingerprint", of three SLC6 transporters (dDAT, hSERT, and GlyT2) embedded into a complex neuronal membrane. New membrane analysis tools were created to improve leaflet detection and leaflet-dependent properties. Overall, lipid fingerprints are comprised of similar lipids when grouped by headgroup or tail saturation. The enrichment and depletion of specific lipids, including sites of cholesterol contacts, varies between transporters. The subtle differences in lipid fingerprints results in varying membrane biophysical properties near the transporter. Through comparisons to previous literature, we highlight that the lipid-fingerprint in complex membranes is highly dependent on membrane composition. Furthermore, through embedding these transporters in a simplified model membrane, we show that the simplified membrane is not able to capture the biophysical properties of the complex membrane. Our results further characterise how the presence and identity of membrane proteins affects the complex interplay of lipid-protein interactions, including the local lipid environment and membrane biophysical properties. HIGHLIGHTSO_LILipid fingerprints are comprised of similar lipid classes C_LIO_LISites of specific lipid contacts, including CHOL, varies between transporters C_LIO_LIChanges in lipid annulus result in variable local membrane biophysical properties C_LIO_LIMembrane composition, including that of complex membranes, affects lipid annulus C_LI GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=91 SRC="FIGDIR/small/427530v1_ufig1.gif" ALT="Figure 1"> View larger version (25K): org.highwire.dtl.DTLVardef@ae6ee9org.highwire.dtl.DTLVardef@1f39af0org.highwire.dtl.DTLVardef@412256org.highwire.dtl.DTLVardef@355f1c_HPS_FORMAT_FIGEXP M_FIG C_FIG

biophysics↗

The inhibitory activity of GlyT2 targeting bioactive lipid analgesics are influenced by formation of a deep lipid cavity

The human glycine transporter 2 (GlyT2 or SLC6A5) has emerged as a promising drug target for the development of new analgesics to manage chronic pain. N-acyl amino acids inhibit GlyT2 through binding to an allosteric binding site to produce analgesia in vivo with minimal overt side effects. In this paper we use a combination of medicinal chemistry, electrophysiology, and computational modelling to explore the molecular basis of GlyT2 inhibition at the allosteric site. We show how N-acyl amino acid head group stereochemistry, tail length and double bond position promote enhanced inhibition by deep penetration into the binding pocket. This work provides new insights into the interaction of lipids with transport proteins and will aid in future rational design of novel GlyT2 inhibitors.

biochemistry↗

The power of mixed survey methodologies for detecting decline of the Bornean orangutan

For many threatened species, it is difficult to assess precisely for large areas the change in their abundances over time and the relative impacts of climate and anthropogenic land use. This is because surveys of such species are typically restricted to small geographic areas, are conducted during short time periods, and use different survey protocols. We assessed the change in the abundance of Bornean orangutan Pongo pygmaeus morio in Sabah, Malaysia, and to identify environmental drivers affecting the change by integrating different types of survey data. We used nest count data obtained from aerial and ground transect surveys and occurrence data obtained from reconnaissance walks and interview survey over the past decade. We built a spatially-explicit dynamic population model within the Bayesian framework allowing these varying survey data to be analyzed jointly by explicitly accounting for each surveys sampling rate. We found that sampling rates vary across survey types, reflecting each surveys associated effort. Orangutan survival rates were strongly determined by natural forest extent and moderately by temperature. Orangutan migration rates across more than 1 km distance between forest patches were low, which underlines the importance of maintaining ecological connectivity. The paucity of species abundance data collected in a consistent manner over many years across broad extents often hinders the assessment of species population trend and their persistence across regional scales. We demonstrate that this can be addressed by integrating multiple survey data across different localities, provided that sampling rate inherent to each survey is accounted for.

ecology↗

The effects of data adequacy and calibration size on the accuracy of presence-only species distribution models

O_LIPresence-only data used to develop species distribution models are often biased towards areas that are frequently surveyed. Furthermore, the size of calibration area with respect to the area covered by the species occurrences has been shown to affect model accuracy. However, existing assessments of the effect of data inadequacy and calibration size on model accuracy have predominately been conducted using empirical studies. These studies can give ambiguous results, since the data used to train and test the model can both be biased.\nC_LIO_LIThese limitations were addressed by applying simulated data to assess how inadequate data coverage and the size of calibration area affect the accuracy of species distribution models generated by MaxEnt and BIOCLIM. The validity of four presence-only performance measures, Contrast Validation Index (CVI), Boyce index, AUC and AUCratio, was also assessed.\nC_LIO_LICVI, AUC and AUCratio ranked the accuracy of univariate models correctly according to the true importance of their defining environmental variable, a desirable property of an accuracy measure. Contrastingly, Boyce index failed to rank the accuracy of univariate models correctly and a high percentage of irrelevant variables produced models with a high Boyce index.\nC_LIO_LIInadequate data coverage and increased calibration area reduced model accuracy by reducing the correct identification of the dominant environmental determinant. BIOCLIM outperformed MaxEnt models in predicting the true distribution of simulated species with a symmetric dominant response. However, MaxEnt outperformed BIOCLIM in predicting the true distribution of simulated species with skew and linear dominant responses. Despite this, the standard performance measures consistently overestimated the performance of MaxEnt models and showed them as always having higher model accuracy than the BIOCLIM models.\nC_LIO_LIIt has been acknowledged that research should be directed towards testing and improving species distribution modelling tools, particularly how to handle the inevitable bias and scarcity of species occurrence data. Simulated data, as demonstrated here, provides a powerful approach to comprehensively test the performance of modelling tools and to disentangle the effects of data properties and modelling options on model accuracy. This may be impossible to achieve using real-world data.\nC_LI

ecology↗

Musashi expression in intestinal stem cells attenuates radiation-induced decline in intestinal homeostasis and survival in Drosophila

Exposure to genotoxic stress by environmental agents or treatments, such as radiation therapy, can diminish health span and accelerate aging. We have developed a Drosophila melanogaster model to study the molecular effects of radiation-induced damage and repair. Utilizing a quantitative intestinal permeability assay, we performed an unbiased GWAS screen (using [~]150 strains from the DGRP reference panel) to search for natural genetic variants that regulate radiation-induced gut permeability. From this screen, we identified the RNA binding protein, Musashi (msi).Msi overexpression promoted intestinal stem cell proliferation, which increased survival after irradiation and rescued radiation-induced gut permeability. We identified that a novel role of Msi in ISC proliferation a novel role for Msi in enhancing ISC function following radiation-induced gut damage, which identifies Msi as a potential therapeutic target.

molecular biology↗

Genome-wide analysis reveals distinct genetic mechanisms of diet-dependent lifespan and healthspan in D. melanogaster

Dietary restriction (DR) robustly extends lifespan and delays age-related diseases across species. An underlying assumption in aging research has been that DR mimetics extend both lifespan and healthspan jointly, though this has not been rigorously tested in different genetic backgrounds. Furthermore, nutrient response genes important for lifespan or healthspan extension remain underexplored, especially in natural populations. To address these gaps, we utilized over 150 DGRP strains to measure nutrient-dependent changes in lifespan and age-related climbing ability to measure healthspan. DR extended lifespan and delayed decline in climbing ability on average, but there was no evidence of correlation between these traits across individual strains. Through GWAS, we then identified and validated jughead and Ferredoxin as determinants of diet-dependent lifespan, and Daedalus for diet-dependent physical activity. Modulating these genes produced independent effects on lifespan and climbing ability, further suggesting that these age-related traits are likely to be regulated through distinct genetic mechanisms.

genetics↗