Biology subjects

Thompson, P. R.

Publications and source records attributed to Thompson, P. R..

8 recordsLinked to original sources

bioRxiv2026

PAD2 knockout reduces myelin protein aggregates, modulates neuroinflammation and protects motor neurons, axons and neuromuscular junction in a SOD1-ALS mouse model

BackgroundDysregulated peptidyl deiminase 2 (PAD2) and aberrant protein citrullination (PC), a posttranslational modification (PTM), are involved in various inflammatory and neurodegenerative diseases. We previously showed in transgenic mice and postmortem human tissues that PC and PAD2 are altered in amyotrophic lateral sclerosis (ALS), a neurodegenerative disease characterized by motor neurons loss, paralysis, and death. Herein, we investigated the role of PAD2 in ALS by PAD2 knockout in a SOD1-ALS mouse model. MethodsTo investigate the role of PAD2-induced citrullination in ALS pathogenesis, we generated PAD2 knockout (PAD2KO) in SOD1G93A ALS mouse model and investigated the consequent modulation on the neuropathology and clinical symptoms, using molecular biology techniques such as qPCR, Western blotting, confocal microscopy, and electron microscopy. Additionally, we identified C3 as being citrullinated in human ALS using ionFinder. ResultsOur results show that PAD2KO blocked the increased PC and reduced myelin basic protein (MBP) aggregates in the ALS model. PAD2KO also improved motor neuron survival and the integrity of myelin, axons, and neuromuscular junctions, and reduced microgliosis in the white matter and C3 protein levels in astrocytes. Clinically, data from monitoring the body weight changes suggests that PAD2KO modulates the course of the disease in the ALS mouse model, accelerating the onset while slowing the progression after the onset, and modestly extending the survival of male mice. ConclusionThese results show that PAD2 is responsible for the increased PC in ALS and PC contributes to neuroinflammation and degeneration of motor neurons and myelinated axons. The modest modulation of the disease phenotype suggests that the role of PC in ALS is complex, involving altered PC in numerous proteins and in multiple cell types. Future studies are needed to investigate how PC modulates individual protein functions in various cell types to understand the contribution of PC to ALS pathogenesis.

neuroscience↗

bioRxiv2025

SARM1, the executioner of axon degeneration, is an ADP-ribosyl transferase and autoMARylation negatively regulates its activation

Axon degeneration is a hallmark of nearly all neurodegenerative diseases. SARM1 plays a central role in this process by degrading NAD+ into nicotinamide and ADPR or cADPR. SARM1 also catalyzes a base exchange reaction between NAD+-phosphate (NADP+) and nicotinic acid (NA) to generate NAADP. These second messengers (i.e., ADPR, cADPR, and NAADP), and NAD+ consumption, are thought to drive axon degeneration. Herein, we identify a fourth reaction catalyzed by SARM1: mono-ADP-ribosylation (MARylation). Specifically, we show that SARM1 MARylates itself and other proteins with a catalytic efficiency (kcat/Km) higher than its NAD+ hydrolase activity. We further show that auto-MARylation promotes a phase transition and renders SARM1 responsive to regulation by NMN. Notably, endogenous SARM1 is MARylated at mitochondria, suggesting that MARylation may regulate SARM1 localization. Together, these findings uncover new regulatory mechanisms and expand the known signaling functions of SARM1. SignificanceSARM1 is an NAD+ hydrolase that executes axon degeneration in myriad neurodegenerative diseases. In addition to NAD+ hydrolysis, SARM1 catalyzes NAD+ cyclization and a base exchange reaction with NADP+ and nicotinic acid. These studies show that SARM1 also catalyzes the transfer of single ADPR moieties to proteins, including itself. Notably, we show that this auto-modification regulates SARM1 activity, allowing the protein to respond to NMN and to permit the phase transition. We also show that endogenous SARM1 is modified at the mitochondria, suggesting that this post-translational modification regulates SARM1 subcellular localization. These findings offer valuable mechanistic insights into SARM1 regulation that will ultimately inform the development of inhibitors targeting SARM1 for neurodegenerative diseases.

biochemistry↗

bioRxiv2024

High-throughput screening of more than 30,000 compounds for anthelmintics against gastrointestinal nematode parasites

Gastrointestinal nematodes (GINs) are amongst the most common parasites of humans, livestock, and companion animals. GIN parasites infect 1-2 billion people worldwide, significantly impacting hundreds of millions of children, pregnant women, and adult workers, thereby perpetuating poverty. Two benzimidazoles with suboptimal efficacy are currently used to treat GINs in humans as part of mass drug administrations, with many instances of lower-than-expected or poor efficacy and possible resistance. Thus, new anthelmintics are urgently needed. However, screening methods for new anthelmintics using human GINs typically have low throughput. Here, using our novel screening pipeline that starts with human hookworms, we screened 30,238 unique small molecules from a wide range of compound libraries, including ones with generic diversity, repurposed drugs, natural derivatives, known mechanisms of action, as well as multiple target-focused libraries (e.g., targeting kinases, GPCRs, and neuronal proteins). We identified 55 compounds with broad-spectrum activity against adult stages of two evolutionary divergent GINs, hookworms (Ancylostoma ceylanicum) and whipworms (Trichuris muris). Based on known databases, the targets of these 55 compounds were predicted in nematode parasites. One novel scaffold from the diversity set library, F0317-0202, showed good activity (high motility inhibition) against both GINs. To better understand this novel scaffolds structure-activity relationships (SAR), we screened 28 analogs and created SAR models highlighting chemical and functional groups required for broad-spectrum activity. These studies validate our new and efficient screening pipeline at the level of tens of thousands of compounds and provide an important set of new GIN-active compounds for developing novel and broadly-active anthelmintics.

pharmacology and toxicology↗

bioRxiv2024

Muscle-Specific Pyruvate Kinase Isoforms, Pkm1 and Pkm2, Regulate Mammalian SWI/SNF Proteins and Histone 3 Phosphorylation During Myoblast Differentiation

Pyruvate kinase is a glycolytic enzyme that converts phosphoenolpyruvate and ADP into pyruvate and ATP. There are two genes that encode pyruvate kinase in vertebrates; Pkm and Pkl encode muscle- and liver/erythrocyte-specific forms, respectively. Each gene encodes two isoenzymes due to alternative splicing. Both muscle-specific enzymes, Pkm1 and Pkm2, function in glycolysis, but Pkm2 also has been implicated in gene regulation due to its ability to phosphorylate histone 3 threonine 11 (H3T11) in cancer cells. Here, we examined the roles of Pkm1 and Pkm2 during myoblast differentiation. RNA-seq analysis revealed that Pkm2 promotes the expression of Dpf2/Baf45d and Baf250a/Arid1A. Dpf2 and Baf250a are subunits that identify a specific sub-family of the mammalian SWI/SNF (mSWI/SNF) of chromatin remodeling enzymes that is required for activation of myogenic gene expression during differentiation. Pkm2 also mediated the incorporation of Dpf2 and Baf250a into the regulatory sequences controlling myogenic gene expression. Pkm1 did not affect expression but was required for nuclear localization of Dpf2. Additionally, Pkm2 was required not only for the incorporation of phosphorylated H3T11 in myogenic promoters, but also for the incorporation of phosphorylated H3T6 and H3T45 at myogenic promoters via regulation of AKT and protein kinase C isoforms that phosphorylate those amino acids. Our results identify multiple unique roles for Pkm2 and a novel function for Pkm1 in gene expression and chromatin regulation during myoblast differentiation.

molecular biology↗

bioRxiv2023

Inhibiting the GPI Transamidase Subunit GPAA1 Abolishes CD24 Surface Localization and Enhances Macrophage-Mediated Phagocytosis of Ovarian Cancer Cells

The CD24-Siglec10 signaling axis is an immune checkpoint pathway that shields ovarian cancer cells from phagocytosis by tumor-associated macrophages (TAMs), making it an appealing immunotherapeutic target. Here, we investigate factors influencing CD24 cell surface expression and assess their suitability as drug targets. Using a CRISPR-based knockout screen, we identify GPAA1 (glycosylphosphatidylinositol anchor attachment-1) as a positive regulator of CD24 cell surface expression. GPAA1 is a crucial component of the multi-subunit GPI transamidase complex, which facilitates the attachment of GPI lipid anchor to the C-terminus of CD24, enabling its surface localization. Reducing the activity of GPAA1 in ovarian cancer cells, either by genetic ablation or targeting with an aminopeptidase inhibitor bestatin, disrupts GPI attachment to CD24. This disruption impairs CD24 cell surface localization, enhances phagocytosis by TAMs, and suppresses tumor growth in mice. Our study highlights the potential of GPAA1 targeting as a therapeutic approach for CD24-positive ovarian cancers.

cancer biology↗

bioRxiv2023

A parametric model for estimating the timing and intensity of animal migration

Animals of many different species, trophic levels, and life history strategies migrate, and the improvement of animal tracking technology allows ecologists to collect increasing amounts of detailed data on these movements. Understanding when animals migrate is important for managing their populations, but is still difficult despite modelling advancements. We designed a model that parametrically estimates the timing of migration from animal tracking data. Our model identifies the beginning and end of migratory movements as signaled by changes in step length and turning angle distributions. To this end, we can also use the model to estimate how an animals movement changes when it begins migrating. We tested our model on three datasets: migratory ferruginous hawks (Buteo regalis) in the Great Plains and barren-ground caribou (Rangifer tarandus groenlandicus) in northern Canada, and non-migratory brown bears (Ursus arctos) from the Canadian Arctic. We estimated the beginning and end of migration in caribou and hawks to the nearest day, while confirming a lack of migratory behaviour in the brown bear population. The flexibility of our modelling framework allowed us to assess intricacies associated with each dataset: long-term stopover behaviour in ferruginous hawks and a priori knowledge of caribou calving areas and behaviour. In addition to estimating when caribou and ferruginous hawks migrated, our model also identified differences in how the two populations migrated; ferruginous hawks achieved efficient migrations by increasing their movement rates while caribou migration was achieved through significant increases in directional persistence. Our approach is broadly applicable to many animal movement studies. We anticipate that rigorous assessment of migration metrics will aid understanding of both how and why animals move.

ecology↗

bioRxiv2021

Time-dependent memory and individual variation in Arctic brown bears (Ursus arctos)

1Animal movement modelling provides unique insight about how animals perceive their landscape and how this perception may influence space use. When coupled with data describing an animals environment, ecologists can fit statistical models to location data to describe how spatial memory informs movement. We performed such an analysis on a population of brown bears (Ursus arctos) in the Canadian Arctic using a model incorporating time-dependent spatial memory patterns. Brown bear populations in the Arctic lie on the periphery of the species range, and as a result endure harsh environmental conditions. In this kind of environment, effective use of memory to inform movement strategies could spell the difference between survival and mortality. The model we fit tests four alternate hypotheses (some incorporating memory; some not) against each other, and we found a high degree of individual variation in how brown bears used memory. We found that 52% (11 of 21) of the bears used complex, time-dependent spatial memory to inform their movement decisions. These results, coupled with existing knowledge on individual variation in the population, highlight the diversity of foraging strategies for Arctic brown bears while also displaying the inference that can be drawn from this innovative movement model.

ecology↗

bioRxiv2019

Predictor species: Improving assessments of rare species occurrence by modelling environmental co-responses

Designing an effective conservation strategy requires understanding where rare species are located. Although species distribution models are primarily used to identify patterns at large spatial scales, their general methodology is relevant for predicting the occurrence of individual species at specific locations. Here we present a new approach that uses Bayesian networks to improve predictions by modelling environmental co-responses among species. For species from a European peat bog community, our approach consistently performs better than single-species models, and better than conventional multi-species models for rare species when calibration data are limited. Furthermore, we identify a group of "predictor species" that are relatively common, insensitive to the presence of other species, and can be used to improve occurrence predictions of rare species. Predictor species are distinct from other categories of conservation surrogates such as umbrella or indicator species, which motivates focused data collection of predictor species to enhance conservation practices.

ecology↗