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Hulsman, M.

Publications and source records attributed to Hulsman, M..

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snpXplorer: an interactive platform for haplotype-aware exploration and integrated annotation of GWAS data

BackgroundGenome-wide association studies (GWAS) have identified thousands of loci associated with complex traits and diseases, yet translating these signals into biological insight remains challenging. Most associated variants are non-coding and reside in linkage disequilibrium (LD) blocks, where multiple correlated variants jointly contribute to association signals. These clusters, or haplotypes, may capture shared regulatory and functional contexts. Interpreting GWAS signals thus requires approaches that integrate regulatory, functional, and cross-trait evidence, while preserving the broader haplotypic context of disease-associated loci. At the same time, the rapid growth of publicly available GWAS summary statistics has enabled large-scale cross-trait analyses, but also introduced redundancy across closely related phenotypes. Efficient interpretation of GWAS data therefore requires tools that integrate heterogeneous data sources while preserving genomic and biological contexts. ResultsWe present snpXplorer, an interactive web platform for haplotype-aware exploration and annotation of GWAS data. The platform incorporates >10,000 GWAS datasets from OpenGWAS and enables multi-scale analysis across variants, haplotypes, genes, and traits. Key features include (i) a haplotype-based representation of association signals derived from LD structure, (ii) a unified variant annotation framework integrating clinical annotations (ClinVar), allele frequencies (gnomAD), functional predictions (CADD, AlphaGenome), quantitative trait loci (GTEx), structural variation, and GWAS associations, and (iii) cross-trait exploration using semantic similarity-based clustering of phenotypes. Use cases centered on Alzheimers disease illustrate this utility: for example, at the TMEM106B locus, snpXplorer identified a haplotype linked to eleven distinct traits, revealing synergistic pleiotropy across neurological and behavioral phenotypes alongside antagonistic pleiotropy with height. ConclusionssnpXplorer allows users to browse, filter, and inspect variant-, haplotype-, gene- and trait-level evidence, lowering the barrier to biological interpretation of GWAS results. Compared with existing tools that focus on specific aspects of GWAS interpretation, the strength of snpXplorer is that it reduces the need for fragmented queries across databases.

genetics↗

Soluble SORL1 in cerebrospinal fluid as a marker for functional impact of rare SORL1 variants.

BackgroundAs part of the retromer, sortilin-related receptor (SORL1) sorts cargo proteins away from the endosome to the trans-Golgi network or to the cell surface, where SORL1 is cleaved into sSORL1 and shed into the interstitial fluid and CSF. SORL1s cargo includes APP and A{beta}, which may explain why protein-truncating genetic variants (PTVs) in SORL1 are observed almost exclusively in Alzheimers Disease (AD) patients, and that rare, predicted pathogenic missense variants have been associated with a 10-fold increased risk of AD. However, functional evidence supporting variant pathogenicity is warranted. Here we investigated whether soluble SORL1 (sSORL1) concentrations in cerebrospinal fluid (CSF) offer a potential in vivo biomarker to support impaired SORL1 function in variant carriers. MethodsUsing an ELISA assay for SORL1 (ABCAM), we determined sSORL1 concentrations in CSF from 218 participants of the Alzheimer Dementia Cohort (ADC) (54% females). We compared sSORL1 in CSF derived from 90 carriers of diverse SORL1 variants with concentrations observed in 78 SORL1-WT AD patients, and 50 SORL1-WT controls for whom CSF-pTau-181, CSF-tTau, CSF-A{beta}42 concentrations were available. In a subset of 36 individuals, we used Western blotting (WB) to validate sSORL1 concentrations as determined by ELISA. ResultsCSF-sSORL1 concentrations did not differ between SORL1-WT AD patients and controls. While CSF-sSORL1 did not correlate with CSF-A{beta}42 concentrations in SORL1-WT AD patients (p=0.62), it correlated with sCSF-ptau-181 (p=9.7x10-6). The mean CSF-sSORL1 concentration in the SORL1 WT AD cases and controls was 466 pg/ml, with wide variance (SD = 133). Comparatively, concentrations were significantly lower in PTV carriers (260 pg/ml, p=3.2x10-7) and in carriers of predicted damaging SORL1 missense variants (323 pg/ml, p=2.4x10-7). CSF-sSORL1 concentrations measured by ELISA correlated strongly with concentrations estimated by WB ({rho}=0.552; p=5.0x10-4). ConclusionCSF-sSORL1 increases with CSF-ptau, suggesting that increased SORL1-retromer activity may serve to rescue cellular stress associated with AD-related processes. However, impairing SORL1 genetic variants may preclude SORL1 trafficking to the cell surface, as supported by lower CSF-sSORL1 protein concentrations in carriers. While further refinements are necessary, we present first evidence for the applicability of ELISA-based quantification of sSORL1 in CSF to evaluate the functional impact of rare SORL1 variants.

neuroscience↗

Increased Genetic Protection Against Alzheimer's Disease in Centenarians

We constructed a polygenic protective score specific to Alzheimers disease (AD PPS) based on the current literature among the participants enrolled in five studies of healthy aging and extreme longevity in the US, Europe, and Asia. This AD PPS did not include variants on Apolipoprotein E (APOE) gene. Comparisons of AD PPS in different data sets of healthy agers and centenarians showed that centenarians have stronger genetic protection against AD compared to individuals without familial longevity. The current study also shows evidence that this genetic protection increases with increasingly older ages in centenarians (centenarians who died before reaching age 105 years, semi-supercentenarians who reached age 105 to 109 years, and supercentenarians who reached age 110 years and older). However, the genetic protection was of modest size: the average increase in AD PPS was approximately one additional protective allele per 5 years of gained lifetime. Additionally, we show that the higher AD PPS was associated with better cognitive function and decreased mortality. Taken together, this analysis suggests that individuals who achieve the most extreme ages, on average, have the greatest protection against AD. This finding is robust to different genetic backgrounds with important implications for universal applicability of therapeutics that target this AD PPS.

genetics↗

SMAC mimetics induce human macrophages to phagocytose live cancer cells

Macrophages engulf apoptotic bodies and cellular debris as part of homeostasis, but they can also phagocytose live cells such as aged red blood cells. Pharmacologic reprogramming with the SMAC mimetic LCL161 in combination with T cell-derived cytokines can induce macrophages to phagocytose live cancer cells in mouse models. Here we extend these findings to encompass a wide range of monovalent and bivalent SMAC mimetic compounds, demonstrating that live cell phagocytosis is a class effect of these agents. We demonstrate robust phagocytosis of live pancreatic and breast cancer cells by primary human macrophages across a range of healthy donors. Unlike mouse macrophages where combination of SMAC mimetics with lymphotoxin enhanced phagocytosis, human macrophages were more efficiently polarized to phagocytose live cells by the combination of SMAC mimetics and IFN{psi}. We profiled phagocytic macrophages by transcriptional and proteomic methodologies, uncovering a positive feedback loop of autocrine TNF production.

immunology↗

Neuronal downregulation of PLCG2 impairs synaptic function and elicits Alzheimer disease hallmarks

We developed a high content screening to investigate how Alzheimer disease (AD) genetic risk factors may affect synaptic mechanisms in rat primary neuronal cultures. Out of the target genes identified, we found that Plcg2 downregulation in mouse dentate gyrus neurons consistently disrupted dendritic morphology and synaptic function. In human neuronal cultures (hNCs), PLCG2 downregulation also impaired synaptic function and increased A{beta} levels and Tau phosphorylation. Very rare PLCG2 loss-of-function (LoF) variants were associated with a 10-fold increased AD risk. PLCG2 LoF carriers exhibit low mRNA/protein PLCG2/PLC{gamma}2 levels and the R953* LoF mutation compromised synaptic function and increased AD hallmarks in hNCs. Single nuclei RNAseq analyses confirmed that the downregulation of PLCG2 impacted pathways related to synaptic and neuronal functions, potentially through neurexin in neurons. In conclusion, PLC{gamma}2 downregulation could increase AD risk by impairing synaptic functions and increasing the A{beta} levels and Tau phosphorylation in neurons.

neuroscience↗

Characterising tandem repeat complexities across long-read sequencing platforms with TREAT

Tandem repeats (TR) play important roles in genomic variation and disease risk in humans. Long-read sequencing allows for the accurate characterisation of TRs, however, the underlying bioinformatics perspectives remain challenging. We present otter and TREAT: otter is a fast targeted local assembler, cross-compatible across different sequencing platforms. It is integrated in TREAT, an end-to-end workflow for TR characterisation, visualisation and analysis across multiple genomes. In a comparison with existing tools based on long-read sequencing data from both Oxford Nanopore Technology (ONT, Simplex and Duplex) and PacBio (Sequel 2 and Revio), otter and TREAT achieved state-of-the-art genotyping and motif characterisation accuracy. Applied to clinically relevant TRs, TREAT/otter significantly identified individuals with pathogenic TR expansions. When applied to a case-control setting, we significantly replicated previously reported associations of TRs with Alzheimers Disease, including those near or within APOC1 (p=2.63x10-9), SPI1 (p=6.5x10-3) and ABCA7 (p=0.04) genes. We finally used TREAT/otter to systematically evaluate potential biases when genotyping TRs using diverse ONT and PacBio long-read sequencing datasets. We showed that, in rare cases (0.06%), long-read sequencing suffers from coverage drops in TRs, including the disease-associated TRs in ABCA7 and RFC1 genes. Such coverage drops can lead to TR mis-genotyping, hampering the accurate characterisation of TR alleles. Taken together, our tools can accurately genotype TR across different sequencing technologies and with minimal requirements, allowing end-to-end analysis and comparisons of TR in human genomes, with broad applications in research and clinical fields.

bioinformatics↗

MotifScope: a multi-sample motif discovery and visualization tool for tandem repeats

Tandem repeats (TRs) constitute a significant portion of the human genome, exhibiting high levels of polymorphism due to variations in size and motif composition. These variations have been associated with various neuropathological disorders, underscoring the clinical importance of TRs. Furthermore, the motif structure of these repeats can offer valuable insights into evolutionary dynamics and population structure. However, analysis of TRs has been hampered by the limitations of short-read sequencing technology, which lacks the ability to fully capture the complexity of these sequences. With long-read data becoming more accessible, there is now also a need for tools to explore and characterize these TRs. In this study, we introduce MotifScope, a novel algorithm for visualization of TRs in their population context based on a de novo k-mer approach for motif discovery. Comparative analysis against three established tools, uTR, TRF, and vamos, reveals that MotifScope can identify a greater number of motifs and more accurately represent the actual repeat sequence. Additionally, MotifScope enables comparison of sequencing reads within an individual and assemblies across different individuals, showing its applicability in diverse genomic contexts. We demonstrate potential applications of MotifScope in diverse fields, including population genetics, clinical settings, and forensic analyses.

genomics↗

Relying on the relationship with known disease-causing variants in homologous proteins to predict pathogenicity of SORL1 variants in Alzheimer's disease

SORL1 encodes the retromer-associated receptor SORLA that functions in endosomal recycling. Rare variants in SORL1 have been associated with Alzheimers disease (AD) and rare pathogenic variants are estimated to occur in up to 2.75% of early onset AD patients and in 1.5% of unrelated late onset AD patients. While truncation mutations are observed almost exclusively in AD patients, it is currently unknown which among the hundreds of rare missense variants identified in SORL1, are pathogenic. Here we address this question by relying on SORLAs distinct molecular architecture. First, we completed a structure-guided sequence alignment for all the protein domains. Next, we identified proteins that contain domains homologous to those of SORLA, which include pathogenic variants for monogenic diseases. We identified the analogous domain positions of these variants in the SORLA protein sequence and showed that variants in these positions similarly impair SORL1, and lead to AD. Together, our findings represent a comprehensive compendium on SORLA protein variation and functional effects, which allowed us to prioritize SORL1 genetic variants into high or moderate priority mutations. We envision that this compendium will be used by clinical geneticists for assessing variants they identify in patients, allowing further development of diagnostic procedures and patient counseling strategies. Utimately, this compendium will inform investigations into the molecular mechanisms of endosomal recycling which will support the development of therapeutic treatment strategies for SORL1 variant-carrying patients.

biochemistry↗

snpXplorer: a web application to explore SNP-associations and annotate SNP-sets

Genetic association studies are frequently used to study the genetic basis of numerous human phenotypes. However, the rapid interrogation of how well a certain genomic region associates across traits as well as the interpretation of genetic associations is often complex and requires the integration of multiple sources of annotation, which involves advanced bioinformatic skills. We developed snpXplorer, an easy-to-use web-server application for exploring Single Nucleotide Polymorphisms (SNP) association statistics and to functionally annotate sets of SNPs. snpXplorer can superimpose association statistics from multiple studies, and displays regional information including SNP associations, structural variations, recombination rates, eQTL, linkage disequilibrium patterns, genes and gene-expressions per tissue. By overlaying multiple GWAS studies, snpXplorer can be used to compare levels of association across different traits, which may help the interpretation of variant consequences. Given a list of SNPs, snpXplorer can also be used to perform variant-to-gene mapping and gene-set enrichment analysis to identify molecular pathways that are overrepresented in the list of input SNPs. snpXplorer is freely available at https://snpxplorer.net. Source code, documentation, example files and tutorial videos are available within the Help section of snpXplorer and at https://github.com/TesiNicco/snpXplorer. Key points: O_LIsnpXplorer shows GWAS summary statistics, regional information and helps deciphering GWAS outcomes C_LIO_LIsnpXplorer interactively compares association levels of a genomic region across phenotypes C_LIO_LIsnpXplorer performs variant-to-gene mapping and gene-set enrichment analysis C_LI O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=151 SRC="FIGDIR/small/377879v2_ufig1.gif" ALT="Figure 1"> View larger version (47K): org.highwire.dtl.DTLVardef@13a9ba5org.highwire.dtl.DTLVardef@c07b9aorg.highwire.dtl.DTLVardef@f2ce4forg.highwire.dtl.DTLVardef@c6d228_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioinformatics↗

A NINE-YEAR LONGITUDINAL STUDY IN THE BLOOD OF A SUPERCENTENARIAN REVEALS AN EXTENSIVE SUBCLONAL ARCHITECTURE AND ONGOING SUBCLONAL DYNAMICS

The aged hematopoietic system is characterized by decreased immuno-competence and by a reduced number of hematopoietic stem cells (HSCs) that actively generates new blood cell (age-related clonal hematopoiesis, ARCH). While both aspects are commonly associated with an increased risk of aging-related diseases, it is currently unknown to what extent these aspects co-occur during exceptional longevity. Here, we investigated these aspects in blood cells of an immuno-hematopoietically normal female who reached 111 years. Blood samples were collected across a 9-year period at ages 103, 110 and 111 years. We applied several genetic sequencing approaches to investigate clonality in peripheral blood samples and sorted cell subsets. Immuno-competence was characterized using flow cytometry, T-cell receptor excision circle (TREC) assays, and in vitro proliferation assays. We identified a single DNMT3A-mutated HSC clone with a complex subclonal architecture and observed ongoing subclonal dynamics within the 9-year timeframe of our sampling. The mutated HSC generated 78-87% myeloid cells, 6-7% of the B-cells, 6% of CD8+ T-cells, and notably 22% of the CD4+ T-cells. Intriguingly, we found that T-cells were capable of robust proliferation when challenged in vitro. Moreover, we observed a surprisingly high TREC content, indicative of recent generation of naive T-cells. Concluding, we observed long-term stability of extreme ARCH with ongoing clonal dynamics combined with functional T-cell immunity. Our results indicate that extreme ARCH does not compromise immuno-competence and that a clonally expanded CD4+ T-cell subset may serve as a potential hallmark of the supercentenarian immune system. Key pointsO_LILongitudinal blood sampling from a female aged 103-111 revealed a dynamic clonal hematopoiesis contributing to myeloid and lymphoid subsets C_LIO_LIDespite the highly advanced age and extreme clonal hematopoiesis we observed functional T-cell immunity C_LI

immunology↗

Neuropsychological Test Performance of Cognitively Healthy Centenarians: Normative data from the Dutch 100-plus Study

BackgroundThe population who reaches the extreme age of 100 years is growing. At this age, dementia incidence is high and cognitive functioning is variable and influenced by sensory impairments. Appropriate cognitive testing requires normative data generated specifically for this group. Currently, these are lacking. We set out to generate norms for neuropsychological tests in cognitively healthy centenarians while taking sensory impairments into account.\n\nMethodsWe included 235 centenarians (71.5% female) from the 100-plus Study, who self-reported to be cognitively healthy, which was confirmed by an informant and a trained researcher. Normative data were generated for 15 tests that evaluate global cognition, pre-morbid intelligence, attention, language, memory, executive and visuo-spatial functions by multiple linear regressions and/or percentiles. Centenarians with vision and/or hearing impairments were excluded for tests that required these faculties.\n\nResultsSubjects scored on average 25.6{+/-}3.1 (range 17-30, interquartile-range 24-28) points on the MMSE. Vision problems and fatigue often complicated the ability to complete tests, and these problems explained 41% and 22% of the missing test scores respectively, whereas hearing problems (4%) and task incomprehension (6%) only rarely did. Sex and age showed a limited association with test performance, whereas educational level was associated with performance on the majority of the tests.\n\nConclusionsNormative data for the centenarian population is provided, while taking age-related sensory impairments into consideration. Results indicate that, next to vision impairments, fatigue and education level should be taken into account when assessing cognitive functioning in centenarians.

epidemiology↗

Centenarian Controls Increase Variant Effect-sizes by an average two-fold in an Extreme Case-Extreme Control Analysis of Alzheimer’s Disease

The detection of genetic loci associated with Alzheimers disease (AD) requires large numbers of cases and controls because variant effect-sizes are mostly small. We hypothesized that variant effect-sizes should increase when individuals who represent the extreme ends of a disease spectrum are considered, as their genomes are assumed to be maximally enriched or depleted with disease-associated genetic variants.\n\nWe used 1,073 extensively phenotyped AD cases with relatively young age at onset as extreme cases (66.3{+/-}7.9 years), 1,664 age-matched controls (66.0{+/-}6.5 years) and 255 cognitively healthy centenarians as extreme controls (101.4{+/-}1.3 years). We estimated the effect-size of 29 variants that were previously associated with AD in genome-wide association studies.\n\nComparing extreme AD-cases with centenarian-controls increased the variant effect-size relative to published effect-sizes by on average 1.90-fold (SE=0.29, p=9.0x10-4). The effect-size increase was largest for the rare high-impact TREM2 (R74H) variant (6.5-fold), and significant for variants in/near ECHDC3 (4.6-fold), SLC24A4-RIN3 (4.5-fold), NME8 (3.8-fold), PLCG2 (3.3-fold), APOE-{varepsilon}2 (2.2-fold) and APOE-{varepsilon}4 (2.0-fold). Comparing extreme phenotypes enabled us to replicate the AD association for 10 variants (p<0.05) in relatively small samples. The increase in effect-sizes depended mainly on using centenarians as extreme controls: the average variant effect-size was not increased in a comparison of extreme AD cases and age-matched controls (0.94-fold, p=6.8x10-1), suggesting that on average the tested genetic variants did not explain the extremity of the AD-cases. Concluding, using centenarians as extreme controls in AD case-controls studies boosts the variant effect-size by on average two-fold, allowing the replication of disease-association in relatively small samples.

genetics↗

Neuropathology and cognitive performance in centenarians

AbstractWith aging, the incidence of neuropathological hallmarks of neurodegenerative diseases increases in the brains of cognitively healthy individuals. It is currently unclear to what extent these hallmarks associate with symptoms of disease at extreme ages. Forty centenarians from the 100-plus Study cohort agreed to post-mortem brain donation. Centenarians self-reported to be cognitive healthy at baseline, which was confirmed by a proxy. Objective ante-mortem measurements of cognitive performance were associated with the prevalence, distribution and quantity of age- and AD-related neuropathological hallmarks. Despite self-reported cognitive health, objective neuropsychological testing suggested varying levels of ante-mortem cognitive functioning. Post-mortem, we found that most neuropathological hallmarks related to age and neurodegenerative diseases such as A{beta} and Tau pathology, as well as atherosclerosis, were abundantly present in most or all centenarians, whereas Lewy body and pTDP-43 pathology were scarce. We observed that increased pathology loads correlated across pathology subtypes, and an overall trend of higher pathology loads to associate with a lower cognitive test performance. This trend was carried especially by the presence of neurofibrillary tangles (NFTs) and granulovacuolar degeneration (GVD) and to a lesser extent by A{beta}-associated pathologies. Cerebral Amyloid Angiopathy (CAA) specifically associated with lower executive functioning in the centenarians. In conclusion, we find that while the centenarians in this cohort escaped or delayed cognitive impairment until extreme ages, their brains reveal varying levels of disease-associated neuropathological hallmarks, some of which associate with cognitive performance.

neuroscience↗

The 100-plus Study of Dutch cognitively healthy centenarians: rationale, design and cohort description

RATIONALEAlthough the incidence of dementia increases exponentially with age, some individuals reach >100 years with fully retained cognitive abilities. To identify the characteristics associated with the escape or delay of cognitive decline, we initiated the 100-plus Study (www.100plus.nl).\n\nDESIGNThe 100-plus Study is an on-going prospective cohort study of Dutch centenarians who self-reported to be cognitively healthy, their first-degree family members and their respective partners. We collect demographics, life history, medical history, genealogy, neuropsychological data and blood samples. Centenarians are followed annually until death. PET-MRI scans and feces donation are optional. Almost 30% of the centenarians agreed to post-mortem brain donation.\n\nCOHORT DESCRIPTIONTo date (September 2018), 332 centenarians were included in the study. We analyzed demographic statistics of the first 300 centenarians (25% males) included in the cohort. Centenarians came from higher socio-economic classes and had higher levels of education compared to their birth cohort; alcohol consumption of centenarians was similar, and most males smoked during their lifetime. At baseline, the centenarians had a median MMSE score of 25 points (IQR: 22.0-27.5); the large majority lived independently, retained hearing and vision abilities and was independently mobile. Mortality was associated with cognitive functioning: centenarians with a baseline MMSE score [&ge;]26 and <26 points had a mortality percentage of respectively 17% and 42% per annual year in the second year after baseline (p=0.003). The cohort was 2.1-fold enriched with the neuroprotective APOE-{varepsilon}2 allele relative to 60-80 year-old population controls (p=4.8x10-7), APOE-{varepsilon}3 was unchanged and the APOE-{varepsilon}4 allele was 2.3-fold depleted (p=6.3x10-7).\n\nCONCLUSIONSComprehensive characterization of the 100-plus cohort of cognitively healthy centenarians might reveal protective factors that explain the physiology of long-term preserved cognitive health.

neuroscience↗