Molecular Loci Research Articles

A compendium of genetic variations associated with promoter usage across 49 human tissues

Promoters play a crucial role in regulating gene transcription. However, our understanding of how genetic variants influence alternative promoter selection is still incomplete. In this study, we implement a framework to identify genetic variants that affect the relative usage of alternative promoters, known as promoter usage quantitative trait loci (puQTLs). By constructing an atlas of human puQTLs across 49 different tissues from 838 individuals, we have identified approximately 76,856 independent loci associated with promoter usage, encompassing 602,009 genetic variants. Our study demonstrates that puQTLs represent a distinct type of molecular quantitative trait loci, effectively uncovering regulatory targets and patterns. Furthermore, puQTLs are regulating in a tissue-specific manner and are enriched with binding sites of epigenetic marks and transcription factors, especially those involved in chromatin architecture formation. Notably, we have also found that puQTLs colocalize with complex traits or diseases and contribute to their heritability. Collectively, our findings underscore the significant role of puQTLs in elucidating the molecular mechanisms underlying tissue development and complex diseases.

Multi-omics analysis of the molecular response to glucocorticoids - insights into shared genetic risk from psychiatric to medical disorders

BackgroundAlterations in the effects of glucocorticoids have been implicated in mediating some of the negative health effects associated with chronic stress, including increased risk for psychiatric disorders as well as cardiovascular and metabolic diseases. This study investigates how genetic variants influence gene expression and DNA methylation (DNAm) in response to glucocorticoid receptor (GR)-activation, and their association with disease risk. MethodsWe measured DNAm (n=199) and gene expression (n=297) in peripheral blood before and after GR-activation with dexamethasone, with matched genotype data available for all samples. A comprehensive molecular quantitative trait locus (QTL) analysis was conducted, mapping GR-response methylation (me)QTLs, GR-response expression (e)QTLs, and GR-response expression quantitative trait methylation (eQTM). A multi-level network analysis was employed to map the complex relationships between the transcriptome, epigenome, and genetic variation. ResultsWe identified 3,772 GR-response meCpGs corresponding to 104,828 local GR-response meQTLs that did not strongly overlap with baseline meQTLs. eQTM and eQTL analyses revealed distinct genetic influences on gene expression and DNAm. Multi-level network analysis uncovered GR-response network trio QTLs, characterized by SNP-CpG-transcript combinations where meQTLs act as both eQTLs and eQTMs. GR-response trio variants were enriched in GWAS for psychiatric, respiratory, autoimmune and cardiovascular diseases and conferred a higher relative heritability per SNP than GR-response meQTL and baseline QTL SNP. ConclusionsGenetic variants modulating the molecular effects of glucocorticoids are associated with psychiatric as well as medical diseases and not uncovered in baseline QTL analyses.

XQTLatlas: a comprehensive resource for human cellular-resolution multi-omics genetic regulatory landscape.

Understanding how genetic variants influence molecular phenotypes in different cellular contexts is crucial for elucidating the molecular and cellular mechanisms behind complex traits, which in turn has spurred significant advances in research into molecular quantitative trait locus (xQTL) at the cellular level. With the rapid proliferation of data, there is a critical need for a comprehensive and accessible platform to integrate this information. To meet this need, we developed xQTLatlas (http://www.hitxqtl.org.cn/), a database that provides a multi-omics genetic regulatory landscape at cellular resolution. xQTLatlas compiles xQTL summary statistics from 151 cell types and 339 cell states across 55 human tissues. It organizes these data into 20 xQTL types, based on four distinct discovery strategies, and spans 13 molecular phenotypes. Each entry in xQTLatlas is meticulously annotated with comprehensive metadata, including the origin of the tissue, cell type, cell state and the QTL discovery strategies utilized. Additionally, xQTLatlas features multiscale data exploration tools and a suite of interactive visualizations, facilitating in-depth analysis of cell-level xQTL. xQTLatlas provides a valuable resource for deepening our understanding of the impact of functional variants on molecular phenotypes in different cellular environments, thereby facilitating extensive research efforts.

Selection of marker systems for determining the allelic composition of high molecular glutenin loci in winter wheat

Aim. Selection of marker systems and conducting multiplex polymerase chain reactions to determine the allelic composition of the Glu-A1, Glu-B1 and Glu-D1 loci, which determine the high quality of flour in common wheat samples. Methods. Polymerase chain reaction (PCR) using specific primers for the Glu-A1, Glu-B1 and Glu-D1 loci of wheat. Results. PCR analysis was performed to detect the insertion at 43 b. p. in the MAR region, in varieties with the Glu-B1al allele, the presence of an amplicon 563 b. p. long was observed, for varieties that carry other alleles of the locus – an amplicon 520 b. p. long. The results on the Glu-D1 locus showed that for № 2, № 3, № 4, № 7, № 8, № 10, № 11, № 19, № 20, № 21 subunits (5 + 10) were identified. Conclusions. With the help of allele-specific primers, the allelic composition of loci of high molecular weight glutenins was determined in 22 lines wheat. Three alleles Glu-A1a, Glu-A1b, Glu-A1c were found in the Glu-A1 locus; in the Glu-B1 locus, only the Glu-B1al allele was detected, there are two alleles at the Glu-D1 locus: Glu-D1a and Glu-D1d (the Glu-D1a allele predominated). As a result of our research, samples of soft wheat (lines № 2, № 7, № 21) were selected, which can be attributed to genotypes with high baking quality of the flour.

Addition to the genus Harmajaea (Agaricales, Pseudoclitocybaceae): a new and a known species from China

A new species, Harmajaea xizangense, discovered in Tibet, China, is illustrated and described based on morphological and molecular data. Morphologically, H. xizangense is mainly characterized by its small pileus, relatively larger and longer spores, and unique geographical distribution. Phylogenetic analyses were performed based on nucleotide sequences of four molecular loci: the internal transcribed spacer (ITS), the nuclear ribosomal large subunit (nrLSU), the DNA-directed RNA polymerase II subunit 2 (RPB2), and the translation elongation factor (TEF1-α). The results indicated that the specimens belonged to the genus Harmajaea, occupying an isolated position in the phylogenetic trees. Furthermore, three collections of H. harperi are introduced for the first time from Liaoning Province of China. This study provides full descriptions, color illustrations, and phylogenetic trees of the two Harmajaea species.

Penicillium species associated with postharvest blue mold rots of garlic in Serbia.

Garlic (Allium sativum L.) is an important plant because of its medicinal and gastronomical benefits. The host can be affected by different plant pathogens, among which Penicillium species are one of the most important rot agents, of stored garlic and other stored food products. This research resolves the etiology of blue mold rots associated with garlic in Serbia. Penicillium-like isolates were collected (2016-2022) from diseased stored garlic bulbs in Serbia and identified using a polyphasic approach. Detailed characterization of the obtained cultures was performed using three solid media, three incubation temperatures, molecular and phylogenetic analyses of four molecular loci [internal transcribed spacer (ITS), beta-tubulin (BenA), calmodulin (CaM) and DNA-dependent RNA polymerase II second largest subunit (RPB2)] and the pathogenicity assay. Five Penicillium species, i.e. P. allii, P. glabrum, P. italicum, P. polonicum, and P. psychrotrophicum were identified and confirmed as postharvest pathogens of garlic. Herein, we report for the first time the pathogenicity of P. psychrotrophicum and P. italicum on garlic. Findings from this study provide insights into the previously unknown diversity of Penicillium fungi responsible for garlic bulb decay in Serbia, and will help in the assessment of the phytosanitary status of this crop, as well as the creation and application of effective disease management strategies.

DNA-binding factor footprints and enhancer RNAs identify functional non-coding genetic variants

BackgroundGenome-wide association studies (GWAS) have revealed a multitude of candidate genetic variants affecting the risk of developing complex traits and diseases. However, the highlighted regions are typically in the non-coding genome, and uncovering the functional causative single nucleotide variants (SNVs) is challenging. Prioritization of variants is commonly based on genomic annotation with markers of active regulatory elements, but current approaches still poorly predict functional variants. To address this, we systematically analyze six markers of active regulatory elements for their ability to identify functional variants.ResultsWe benchmark against molecular quantitative trait loci (molQTL) from assays of regulatory element activity that identify allelic effects on DNA-binding factor occupancy, reporter assay expression, and chromatin accessibility. We identify the combination of DNase footprints and divergent enhancer RNA (eRNA) as markers for functional variants. This signature provides high precision, but with a trade-off of low recall, thus substantially reducing candidate variant sets to prioritize variants for functional validation. We present this as a framework called FINDER—Functional SNV IdeNtification using DNase footprints and eRNA.ConclusionsWe demonstrate the utility to prioritize variants using leukocyte count trait and analyze variants in linkage disequilibrium with a lead variant to predict a functional variant in asthma. Our findings have implications for prioritizing variants from GWAS, in development of predictive scoring algorithms, and for functionally informed fine mapping approaches.

Fine mapping of candidate effector genes for heart rate.

An elevated resting heart rate (RHR) is associated with increased cardiovascular mortality. Genome-wide association studies (GWAS) have identified > 350 loci. Uniquely, in this study we applied genetic fine-mapping leveraging tissue specific chromatin segmentation and colocalization analyses to identify causal variants and candidate effector genes for RHR. We used RHR GWAS summary statistics from 388,237 individuals of European ancestry from UK Biobank and performed fine mapping using publicly available genomic annotation datasets. High-confidence causal variants (accounting for > 75% posterior probability) were identified, and we collated candidate effector genes using a multi-omics approach that combined evidence from colocalisation with molecular quantitative trait loci (QTLs), and long-range chromatin interaction analyses. Finally, we performed druggability analyses to investigate drug repurposing opportunities. The fine mapping pipeline indicated 442 distinct RHR signals. For 90 signals, a single variant was identified as a high-confidence causal variant, of which 22 were annotated as missense. In trait-relevant tissues, 39 signals colocalised with cis-expression QTLs (eQTLs), 3 with cis-protein QTLs (pQTLs), and 75 had promoter interactions via Hi-C. In total, 262 candidate genes were highlighted (79% had promoter interactions, 15% had a colocalised eQTL, 8% had a missense variant and 1% had a colocalised pQTL), and, for the first time, enrichment in nervous system pathways. Druggability analyses highlighted ACHE, CALCRL, MYT1 and TDP1 as potential targets. Our genetic fine-mapping pipeline prioritised 262 candidate genes for RHR that warrant further investigation in functional studies, and we provide potential therapeutic targets to reduce RHR and cardiovascular mortality.

Effects of heat exposure from late sowing on agronomic traits and the technological quality of hexaploid wheat

Heat stress reduces wheat grain yield and affects quality traits relevant to trading and end use. In this study, 98 landraces and 152 elite hexaploid wheat varieties and breeding lines were trialled at two irrigated field sites in southern NSW to better understand heat stress responses and identify promising genotypes for heat tolerance breeding. Potential heat tolerance was defined using performance in heat-exposed late-sown (LS) plots relative to normal-sown (NS) plots, using a regression approach to calculate response indices uncorrelated with awn emergence time and performance per se in NS. The best individual responders for grain yield included genotypes from both Australia and Mexico. On average, lines selected in CIMMYT's High Temperature Wheat Yield Trials (HTWYT) performed well for yield per se in NS as well as for yield response to LS. CIMMYT lines previously selected for large grains under heat stress conditions performed well for grain size in NS and for LS responses of grain size and test weight. Dough peak resistance and time to peak resistance in a mixograph were increased by LS and these traits were positively correlated with grain protein and the occurrence of the Glu-D1d high molecular weight glutenin locus allele, respectively.

Polymorphisms of ITGA9 Gene and Their Correlation with Milk Quality Traits in Yak (Bos grunniens).

A single-nucleotide polymorphism (SNP) is a genome-level trait that arises from a variation in a single nucleotide, leading to diversity in DNA sequences. SNP screening is commonly used to provide candidate genes for yak breeding efforts. Integrin Subunit Alpha 9 (ITGA9) is an integrin protein. It plays an important role in cell adhesion, signalling, and other processes. The aim of this study was to discuss the association between genetic polymorphisms in the ITGA9 gene and milk quality traits and to identify potential molecular marker loci for yak breeding quality. We genotyped 162 yaks using an Illumina Yak cGPS 7K liquid chip and identified the presence of polymorphisms at nine SNP loci in the ITGA9 gene of yaks. The results showed that the mutant genotypes in the loci g.285,808T>A, g.306,600T>C, and g.315,413C>T were positively correlated with the contents of casein, protein, total solids (TS), and solid nonfat (SNF) in yak milk. In other loci, heterozygous genotypes had a positive correlation with nutrient content in yak milk. Then, two ITGA9 haplotype blocks were constructed based on linkage disequilibrium, which facilitated a more accurate screening of ITGA9 as a candidate gene for yak milk quality improvement. In conclusion, we identified SNPs and haplotype blocks related to yak milk quality traits and provided genetic resources for marker-assisted selection in yak breeding.

Two invalid genera in the family Bythinellidae Locard, 1893 (Caenogastropoda: Truncatelloidea)

Within the genus Bythinella Moquin-Tandon, 1856, with about 250 nominal species, the separation of several genera would be helpful in understanding the phylogenetic relationships between these snails, but neither morphology nor molecular loci studied so far can be used to distinguish any natural taxon over the range of species within this genus. Apart from Bythinella there are two other genera listed in the Bythinellidae by WoRMS: Terrestribythinella Sitnikova, Starobogatov et Anistratenko, 1992, and Strandzhia Georgiev et Glöer, 2013. Considering the cytochrome c oxidase subunit I (COI) sequences, as well as poor morphological data, Terrestribythinella should be considered a junior subjective synonym of Bythinella. In the case of Strandzhia, both COI and 18S sequences studied by us, as well as morphology re-examined (misinterpretation of the penis morphology in the original description) clearly show the assignment of Strandzhia to the genus Grossuana Radoman, 1983, thus Strandzhia becomes a junior objective synonym of Grossuana.

A new look at genetic variability in pomegranate: Landscape genetics and structural equation modeling (SEM)

Pomegranate (Punica granatum L.) is an important horticultural plant in Iran and has been a food source since ancient times. Despite several genetic diversity studies conducted on these taxa, there are no reports on the landscape genetics of these important plants in Iran, nor is there any information on the effects of environmental, spatial, and climatic variables on the genetic structure of pomegranate. Therefore, this study used genetics, structural equation modeling (SEM), and species distribution modeling (SDM) approaches to explore the above-mentioned objectives. We used both cytogenetic and molecular analyses (SSR and SCoT markers) of several pomegranate samples along with environmental, spatial, and climatic variables. The redundancy analysis (RDA) analysis showed a significant positive association between terminal chiasmata with the longitudinal distribution of the pomegranate accession, while total chiasma number and interstitial chiasmata have an association with the altitude. The spatial PCA analysis indicated the role played by both global and local spatial variables in the cytogenetic variability like the univalent and terminal chiasmata formation. The Mantel test indicates that the cytogenetic difference in pomegranate plants is not related to their geographical distance, while the Moran I test result shows that geographical regions with similar spatial conditions affect the cytogenetic structuring of pomegranate plants in a similar way. SEM methods showed a strong negative association between bivalent formation and the environmental variables, while they showed both positive and negative associations between some of the environmental variables and different kinds of chiasmata. The sPCA analysis of molecular data showed a significant global, and local, spatial structuring of the molecular data and the genetic cline formation within the pomegranate accessions studied. Similarly, the spatial PCA analysis of climate variables, precipitation, and temperature revealed a significant structuring of genetic diversity and the formation of genetic clines by these factors. RDA analysis identified some of the loci that are significantly associated with temperature and precipitation. Therefore, both geographical/ or spatial variables and climate factors contribute to the genetic structure of the pomegranate accessions studied. SEM models based on genetic data revealed a positive association between environmental and climate variables with genetic diversity parameters and SSR-SCoT molecular loci studied. Climate change is unlikely to hurt pomegranate growing areas due to the high genetic diversity of pomegranate plants and the diverse genetic lineages present in the country.

Establishment of Start Codon Targeted Polymorphism Molecular Markers System and Genetic Diversity Analysis of Fritillaria thunbergii

After years of selective breeding, numerous cultivars of Fritillaria thunbergii have been cultivated in various regions of Zhejiang Province, showing significant genomic and epigenetic differences. Molecular marker loci developed so far are mainly based on non-coding regions of the genome, and their identification efficiency is limited when applied to these cultivars. Therefore, this study used start codon targeted polymorphism molecular marker to detect PCR amplification of 100 Fritillaria thunbergii samples from various sources. The SCoT-PCR reaction system was optimized using a single-factor and orthogonal testing to determine the optimal conditions: DNA concentration of 4.2 ng • μL−1, primer concentration of 0.88 μmol • L−1, dNTP concentration of 0.35 mmol • L−1, Mg2+ concentration of 2.5 mmol • L−1, Supercharm Fidelity enzyme amount of 1.2 U, and total reaction volume of 25 μL. Moreover, 16 effective primers were identified, including SCoT1, SCoT2, SCoT3, SCoT7, SCoT11, SCoT12, SCoT14, SCoT17, SCoT19, SCoT23, SCoT25, SCoT26, SCoT29, SCoT35, and SCoT37. The annealing temperature for each primer was determined. Among them, SCoT14, SCoT25, SCoT29, SCoT35, and SCoT41 were specific for identifying six main cultivars: “Zhebei 1”, “Zhebei 2”, “Zhebei 3”, “Xuansanzi”, “Sanzi”, and “Duozi”. SCoT7, SCoT14, and SCoT35 were useful in tracking mutations in progeny of Zhebei 3 after radiation treatment, while SCoT29 could distinguish Zhebei 1 from Chunan. In addition, cluster analysis was conducted to simulate the genetic relationship between different cultivars, demonstrating that genetic similarity analysis is practically significant for predicting genetic correlation among different cultivars. The results not only improve the identification ability of Fritillaria thunbergii cultivars, but also help objectively evaluate their genetic stability and environmental adaptability. These achievements provide important scientific evidence for the ecological protection and utilization of germplasm resources of Fritillaria thunbergii and serve as a reference for exploring genetic diversity and evolution of traditional Chinese medicine.

Phylogeny of the genus Loxospora s.l. (Sarrameanales, Lecanoromycetes, Ascomycota), with Chicitaea gen. nov. and five new combinations in Chicitaea and Loxospora.

Loxospora is a genus of crustose lichens containing 13 accepted species that can be separated into two groups, based on differences in secondary chemistry that correlate with differences in characters of the sexual reproductive structures (asci and ascospores). Molecular phylogenetic analyses recovered these groups as monophyletic and support their recognition as distinct genera that differ in phenotypic characters. Species containing 2'-O-methylperlatolic acid are transferred to the new genus, Chicitaea Guzow-Krzem., Kukwa & Lendemer and four new combinations are proposed: C.assateaguensis (Lendemer) Guzow-Krzem., Kukwa & Lendemer, C.confusa (Lendemer) Guzow-Krzem., Kukwa & Lendemer, C.cristinae (Guzow-Krzem., Łubek, Kubiak & Kukwa) Guzow-Krzem., Kukwa & Lendemer and C.lecanoriformis (Lumbsch, A.W. Archer & Elix) Guzow-Krzem., Kukwa & Lendemer. The remaining species produce thamnolic acid and represent Loxospora s.str. Haplotype analyses recovered sequences of L.elatina in two distinct groups, one corresponding to L.elatina s.str. and one to Pertusariachloropolia, the latter being resurrected from synonymy of L.elatina and, thus, requiring the combination, L.chloropolia (Erichsen) Ptach-Styn, Guzow-Krzem., Tønsberg & Kukwa. Sequences of L.ochrophaea were found to be intermixed within the otherwise monophyletic L.elatina s.str. These two taxa, which differ in contrasting reproductive mode and overall geographic distributions, are maintained as distinct, pending further studies with additional molecular loci. Lectotypes are selected for Lecanoraelatina, Pertusariachloropolia and P.chloropoliaf.cana. The latter is a synonym of Loxosporachloropolia. New primers for the amplification of mtSSU are also presented.

Accounting for isoform expression increases power to identify genetic regulation of gene expression.

A core problem in genetics is molecular quantitative trait locus (QTL) mapping, in which genetic variants associated with changes in the molecular phenotypes are identified. One of the most-studied molecular QTL mapping problems is expression QTL (eQTL) mapping, in which the molecular phenotype is gene expression. It is common in eQTL mapping to compute gene expression by aggregating the expression levels of individual isoforms from the same gene and then performing linear regression between SNPs and this aggregated gene expression level. However, SNPs may regulate isoforms from the same gene in different directions due to alternative splicing, or only regulate the expression level of one isoform, causing this approach to lose power. Here, we examine a broader question: which genes have at least one isoform whose expression level is regulated by genetic variants? In this study, we propose and evaluate several approaches to answering this question, demonstrating that "isoform-aware" methods-those that account for the expression levels of individual isoforms-have substantially greater power to answer this question than standard "gene-level" eQTL mapping methods. We identify settings in which different approaches yield an inflated number of false discoveries or lose power. In particular, we show that calling an eGene if there is a significant association between a SNP and any isoform fails to control False Discovery Rate, even when applying standard False Discovery Rate correction. We show that similar trends are observed in real data from the GEUVADIS and GTEx studies, suggesting the possibility that similar effects are present in these consortia.

Liver regulatory mechanisms of noncoding variants at lipid and metabolic trait loci

Genome-wide association studies (GWASs) have identified hundreds of risk loci for liver disease and lipid-related metabolic traits, although identifying their target genes and molecular mechanisms remains challenging. We predicted target genes at GWAS signals by integrating them with molecular quantitative trait loci for liver gene expression (eQTL) and liver chromatin accessibility QTL (caQTL). We predicted specific regulatory caQTL variants at four GWAS signals located near EFHD1, LITAF, ZNF329, and GPR180. Using transcriptional reporter assays, we determined that caQTL variants rs13395911, rs11644920, rs34003091, and rs9556404 exhibit allelic differences in regulatory activity. We also performed a protein binding assay for rs13395911 and found that FOXA2 differentially interacts with the alleles of rs13395911. For variants rs13395911 and rs11644920 in putative enhancer regulatory elements, we used CRISPRi to demonstrate that repression of the enhancers altered the expression of the predicted target and/or nearby genes. Repression of the element at rs13395911 reduced the expression of EFHD1, and repression of the element at rs11644920 reduced the expression of LITAF, SNN, and TXNDC11. Finally, we showed that EFHD1 is a metabolically active gene in HepG2 cells. Together, these results provide key steps to connect genetic variants with cellular mechanisms and help elucidate the causes of liver disease.

Molecular quantitative trait loci in reproductive tissues impact male fertility in cattle.

Breeding bulls are well suited to investigate inherited variation in male fertility because they are genotyped and their reproductive success is monitored through semen analyses and thousands of artificial inseminations. However, functional data from relevant tissues are lackingin cattle, which prevents fine-mapping fertility-associated genomic regions. Here, we characterize gene expression and splicing variation in testis, epididymis, and vas deferens transcriptomes of 118 mature bulls and conduct association tests between 414,667 molecular phenotypes and 21,501,032 genome-wide variants to identify 41,156 regulatory loci. We show broad consensus in tissue-specific and tissue-enriched gene expression between the three bovine tissues and their human and murine counterparts. Expression- and splicing-mediating variants are more than three times as frequent in testis than epididymis and vas deferens, highlighting the transcriptional complexity of testis. Finally, we identify genes (WDR19, SPATA16, KCTD19, ZDHHC1) and molecular phenotypes that are associated with quantitative variation in male fertility through transcriptome-wide association and colocalization analyses.

Interaction molecular QTL mapping discovers cellular and environmental modifiers of genetic regulatory effects

Bulk-tissue molecular quantitative trait loci (QTLs) have been the starting point for interpreting disease-associated variants, and context-specific QTLs show particular relevance for disease. Here, we present the results of mapping interaction QTLs (iQTLs) for cell type, age, and other phenotypic variables in multi-omic, longitudinal data from the blood of individuals of diverse ancestries. By modeling the interaction between genotype and estimated cell-type proportions, we demonstrate that cell-type iQTLs could be considered as proxies for cell-type-specific QTL effects, particularly for the most abundant cell type in the tissue. The interpretation of age iQTLs, however, warrants caution because the moderation effect of age on the genotype and molecular phenotype association could be mediated by changes in cell-type composition. Finally, we show that cell-type iQTLs contribute to cell-type-specific enrichment of diseases that, in combination with additional functional data, could guide future functional studies. Overall, this study highlights the use of iQTLs to gain insights into the context specificity of regulatory effects.

Integration of Alzheimer’s Disease GWAS with Molecular QTLs Reveals Prioritized Risk Genes and Disease‐associated Molecular Alterations

AbstractBackgroundDespite the success of genome‐wide association studies (GWAS) to identify genetic variants associated with complex diseases, as the majority of these variants reside in non‐coding genome, one important task is to link these variants to disease‐associated molecular alterations in risk genes. To this end, we developed a systematic gene prioritization methodology that integrated our recent Alzheimer’s disease (AD) GWAS meta‐analysis with molecular quantitative trait loci (QTL) in disease‐relevant tissues and identified highly likely candidate risk genes and molecular alterations within the majority of 42 novel loci identified. In this subsequent study, we updated our pipeline with new molecular QTL catalogues and analyzed the complete landscape of AD risk.MethodWe performed GWAS‐QTL integration in AD‐relevant tissues and cells by querying the lead variants in each locus in the molecular QTL catalogues, (ii) investigating genetic colocalization between GWAS and molecular QTL signals using coloc, and (iii) conducting transcriptome, methylome, and proteome‐wide association studies (TWAS/MWAS/PWAS) using the EADB stage I meta‐analysis GWAS on 85,934 cases and 401,577 controls. The updated gene prioritization pipeline included new analyses based on new brain protein expression QTL (pQTL), cell‐type‐specific expression QTL (ct‐eQTL), expression QTL (eQTL), histone acetylation QTL (haQTL), and methylation QTL (mQTL) catalogues. Moreover, we used Nanopore RNA sequencing to further investigate risk‐associated splicing events revealed by splicing QTL (sQTL) integration.ResultOur brain ct‐eQTL coloc results showed that while the most risk‐associated gene expression alterations are specific to microglia, the proportion of astrocyte‐specific eQTL colocalizations were doubled compared to the previous studies. These included EGFR, a previously prioritized gene due to bulk brain eQTL coloc hits, whose downregulation was regulated by the protective signal specifically in astrocytes. Importantly, in a previously unresolved locus we could prioritize HS3ST5 through astrocyte‐specific eQTLs as well. Furthermore, pQTL coloc and PWAS prioritized SNX1, a previously prioritized gene at a lower confidence together with neighboring FAM96A gene, by implicating its genetic downregulation with increased AD risk.ConclusionOur updated pipeline systematically prioritized AD risk genes and disease‐associated molecular alterations in the majority of risk loci, and contributed to better interpretation of AD GWAS and understanding of genetically‐associated molecular mechanisms underlying AD risk.

Polymorphisms of CCSER1 Gene and Their Correlation with Milk Quality Traits in Gannan Yak (Bos grunniens).

Coiled-coil serine-rich protein 1 (CCSER 1) gene is a regulatory protein gene. This gene has been reported to be associated with various economic traits in large mammals in recent years. The aim of this study was to investigate the association between CCSER1 gene single nucleotide polymorphisms (SNPs) and Gannan yaks and to identify potential molecular marker loci for breeding milk quality in Gannan yaks. We genotyped 172 Gannan yaks using Illumina Yak cGPS 7K liquid microarrays and analyzed the correlation between the three SNPs loci of the CCSER1 gene and the milk qualities of Gannan yaks, including milk fat, protein and casein. It was found that mutations at the g.183,843A>G, g.222,717C>G and g.388,723G>T loci all affected the fat, protein, casein and lactose traits of Gannan yak milk to varying extents, and that the milk quality of individuals with mutant phenotypes was significantly improved. Among them, the milk fat content of AG heterozygous genotype population at g.183,843A>G locus was significantly higher than that of AA and GG genotype populations (p < 0.05); the casein and protein content of mutant GG and CG genotype populations at g.222,717C>G locus was significantly higher than that of wild-type CC genotype population (p < 0.05); and the g.388,723G>T locus of the casein and protein contents of the mutant TT genotype population were significantly higher (p < 0.05) than those of the wild-type GG genotype population. These results provide potential molecular marker sites for Gannan yak breeding.